WO2000059909A1 - Heterocyclic compounds having affinities for serotonin receptors - Google Patents

Abstract

Compounds represented the general formula (I) exhibit affinities for various serotonin receptors (such as 5HT-1A, 2, 6, 7, and so on) and are therefore useful as remedies for central nervous system diseases or the like wherein R?1 and R2¿ are each independently hydrogen, lower alkyl, or the like; R?3 and R4¿ are each independently hydrogen, lower alkyl, or the like; or alternatively R?1 and R3¿ may be united to form a single bond; n is 0 or 1; R5 is hydrogen; R6 is hydrogen, hydroxyl, or the like; R7 is hydrogen; R?8 and R9¿ are each independently hydrogen, lower alkylthio, or the like; or alternatively R?7 and R9¿ may be united to form a single bond; R10 is hydrogen, lower alkyl, or the like; R11 is hydrogen, lower alkyl, or the like; and R12 is hydrogen or lower alkyl

Description

 TECHNICAL FIELD Heterocyclic compound having affinity for cepatonin receptor

 The present invention relates to novel heterocyclic compounds. Since the present compound has an affinity for serotonin receptor, it is useful as a medicament such as a therapeutic drug for central nervous system diseases. It is also useful as a synthetic intermediate thereof.

Background art

 Serotonin (5-hydroxytryptamine) is one of the biological amines and has various physiological activities. For example, it is present in the basal granule cells of the intestine and promotes intestinal motility. In addition, it is released into the blood from platelets during bleeding and is involved in hemostasis by contracting capillaries. Separately, serotonin also acts as a neurotransmitter in the brain and is involved in the regulation of mental activity, pain threshold, body temperature, sleep-wake cycle, etc. via serotonin receptors [Physiol. Rev. 72 (1992) 165-229].

 Serotonin receptors are roughly classified into seven families, and at least 14 species have been discovered including their subtypes [Pia / ^ aco Rev. 46 (1994) 157-203]. It has been reported that each receptor is involved in various physiological functions and disease states, and serotonin receptor binding agents exhibit agonist or antagonist action to prevent or prevent various diseases. Harmacol. Eev. 43 (1991) 509-525, which is expected to be a therapeutic drug.

Among them, the 5-HT _{5 A} , 5-HT _{5 B} , 5-HT ₆ and 5-HT ₇ receptors are the most recently discovered and cloned serotonin receptors [i ^? Lett. 355 (1994) 242-6, FEB S Lett. 333 (1993) 25-31, J. Neurochem. 66 (1996) 47-56, Neuron, 11 (1993) 449-458], its selective operation. Drugs (agonist) and antagonists (anthus gonist) are rarely reported. All of these receptors are mainly distributed in the central nervous system. Has obviously summer, for example, _{5-HT 5 A, 5- HT} 5 B receptors to a depth not the hippocampus and cerebral cortex involved in learning and memory ^{[i 1 ^? Et. 355} (1994) 242-6, FEBS Lett. 333 (1993) 25-3 1], 5-HT ₆ receptor is involved in the striatum involved in motor function [J. Neurochem. 66 (1996) 47-56], 5-HT ₇ receptor Is reported to be present in the suprachiasmatic nucleus, a mammalian biological clock [^ uro / i, 11 (1993) 449-458]. Therefore, selective agonist or antagonist of these receptors may be an anti-dementia drug, an anti-parkinson's disease drug, an antipsychotic, a drug for treating circadian rhythm disorder, and the like. _{Furthermore, 5-HT 5 Α, 5} -ΗΤ 5 Β, already developed selective Agonisu preparative Ya antagonists DOO for the treatment of each ¾ diseases against _{5-ΗΤ 6, 5-ΗΤ} 7 receptor other serotonin receptors However, by showing affinity for multiple serotonin receptor subtypes and other receptors, they may be more effective or have fewer side effects.

As the tetracyclic fused heterocyclic compound, for example, the following are known. The W096 / 32944, Erugorin (ergoline) compounds having antagonistic preparative effect on 5-HT ₇ receptors have been described. W095 / 28403 describes aveo-ergoline derivatives acting on 5-HT receptors. JP55-124784 (A) describes an indole derivative having a hypotensive action. GB1482871 (A) describes an indole derivative having an adrenergic antagonistic activity and the like. All of these compounds contain N atoms as ring-constituting heteroatoms, but do not contain 0 atoms. Therefore, it cannot suggest the compound of the present invention.

Under the circumstances, serotonin receptors, particularly 5-HT ₆ and 5 - has been desired to develop a medicine containing novel compounds and it has an affinity against HT ₇ receptor.

Disclosure of the invention

 As a result of intensive studies, the present inventors have found that the novel condensed tetracyclic heterocyclic compound and its derivative have affinity for various serotonin receptors, and completed the present invention described below. did.

1. Equation (1):

 (Where

1 ¹ Oyobi 11 ^2, it it independently hydrogen, lower alkyl or halogen; R ³ and R ⁴ are each independently hydrogen, lower alkyl, lower alkenyl, lower alkylcarbonyl, § optionally substituted Reel carbonyl or cyano; R ¹ and R ³ may together form a single bond;

n is 0 or 1, provided that when n = 1, neither R ³ nor R ⁴ is hydrogen; R ⁵ is hydrogen;

R ⁶ is hydrogen, hydroxy, lower alkoxy or lower alkyl carboxy;

R ⁷ is hydrogen;

R ⁸ and are each independently hydrogen, lower alkylthio or halogen;

R ⁷ and R ⁹ may together form a single bond;

R ^{1 Q} is hydrogen, lower alkyl, lower alkylcarbonyl, lower alkyloxyl carbonyl, optionally substituted arylcarbonyl, optionally substituted heteroaryl.

—Lucarponyl, lower alkylsulfonyl, optionally substituted arylsulfonyl or optionally substituted heteroarylsulfonyl;

R ¹¹ is hydrogen, lower alkyl or halogen;

R ¹² represents hydrogen or lower alkyl)

Or a pharmaceutically acceptable salt thereof, a prodrug thereof, or a hydrate thereof. 2. The compound according to 1 above, wherein R ¹ and R ³ together form a single bond.

3. The compound according to the above 1, wherein R ² is hydrogen or lower alkyl.

4. The compound according to 1 above, wherein R ⁴ is lower alkyl.

5. The compound according to above 1, wherein R ⁶ is hydrogen.

6. The compound according to 1 above, wherein R ⁷ and R ⁹ together form a single bond.

7. The compound according to 1 above, wherein R ⁸ is hydrogen.

8. The compound according to 1 above, wherein R ^{1 Q} is hydrogen.

9. The compound according to 1 above, wherein R ¹¹ and R ¹² are hydrogen.

10. R ¹ and R ³ together form a single bond; R ² is hydrogen or lower alkyl; R ⁴ is lower alkyl; R ⁶ is hydrogen; R ⁷ and R ⁹ are together A bond is formed; R ⁸ is hydrogen; R ¹ . Is hydrogen; R ¹¹ and R ¹² are hydrogen; n is 0.

R ¹ and R ³ together form a single bond; R ² is hydrogen or lower alkyl; R ⁴ is lower alkyl; R ⁶ is hydrogen; R ⁷ and R ⁹ are together Forming a bond; R ⁸ is hydrogen, lower alkylthio or halogen; R ¹ . Wherein R ¹¹ and R ¹² are each independently hydrogen or lower alkyl; and n is ◦, wherein R is hydrogen, lower alkyloxycarbonyl, or optionally substituted arylcarbonyl. Compound.

1 2. R ¹ and R ³ together form a single bond; R ² is hydrogen or lower alkyl; R ⁴ is lower alkyl; R ⁶ is hydrogen; R ⁷ and R ⁹ are together R ⁸ is hydrogen; R ¹ . Is a substituted or unsubstituted arylsulfonyl; R ¹¹ and R ¹² are hydrogen; n is 0.

1 3. R ¹ and R ³ together form a single bond; R ² is hydrogen or lower alkyl; R ⁴ is lower alkyl; R ⁶ is hydrogen; R ⁷ , R ⁸ and R ⁹ are both hydrogen R ⁸ is hydrogen; R ¹ ° is optionally substituted carbonyl or optionally substituted arylsulfonyl; R ¹¹ and R ¹² are hydrogen; n is 0; 2. The compound according to the above 1. 1 4. R ¹ and R ³ together form a single bond; R ² is hydrogen or lower alkyl; R ⁴ is lower alkyl; R ⁶ is hydrogen; R ⁷ and R ^{9 are —}緖to form a single bond; R ⁸ is halogen; R ^{1 G} is hydrogen; R ^{1 1} is hydrogen or halogen; R ^{1 2} is hydrogen; is n is 0, compounds of claim 1, wherein the.

15. R ¹ and R ³ together form a single bond; R ² is hydrogen or lower alkyl; R ⁴ is lower alkyl; R ⁶ is hydrogen; R ⁷ and R ⁹ are both hydrogen or R ⁸ is hydrogen; ϋ ^{1 G} is hydrogen; R ¹¹ and R ¹² are hydrogen; n is 1.

 16. A pharmaceutical composition comprising the compound according to any one of the above 1 to 15.

 1 7. An agent for preventing or treating a disease mediated by a mouth tonine receptor, comprising the compound according to any one of the above 1 to 15.

 18. An agent for preventing or treating central nervous system diseases, comprising the compound according to any one of the above items 1 to 15.

 19. The preventive or remedy according to the above 18, wherein the central nervous system disease is anxiety, depression, schizophrenia, circadian rhythm disorder, stroke, dementia, pain or Parkinson's disease.

 20. A microorganism belonging to the genus Streptomyces, which is capable of producing the compound according to any one of 1 to 15 above.

 21. The microorganism according to 20 above, which is capable of producing the compound according to 15 above.

22. The microorganism according to 20 above, which is Streptomyces sp. PA-48561.

 2 3. The method according to 1 above, comprising culturing the microorganism according to any one of 20 to 22 above, separating and purifying a compound produced from the obtained culture solution, and then chemically modifying if necessary. A method for producing a compound according to any one of claims 1 to 15.

 2 4. A method for preventing or treating a disease mediated by an oral tonin receptor, which comprises administering the compound according to any one of the above items 1 to 15.

2 5. To manufacture a prophylactic or therapeutic agent for a disease mediated by serotonin receptors, Use of the compound according to any one of the above 1 to 15.

BEST MODE FOR CARRYING OUT THE INVENTION

 Each group of the compound (I) will be described below. Unless otherwise noted, each term has a common meaning either alone or in combination with other terms. Halogen includes fluorine, chlorine, bromine and iodine.

 Lower alkyl includes straight-chain or branched C 1 -C 6 alkyl, such as methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, sec-butyl, tert-butyl, n-butyl. —Pentyl, i-pentyl, neo-pentyl, tert-pentyl, n-hexyl and the like. Preferably, it is C 1 -C 4 alkyl, especially methyl, n-propyl, tert-butyl and the like.

 Lower alkenyl includes straight-chain or branched C2-C6 alkenyl, and includes vinyl, aryl, 1-brodinyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-hexenyl and the like. Is exemplified. Preferably, it is 2-propenyl or the like.

 The lower alkoxy includes the oxy bonded to the lower alkyl, and examples thereof include methoxy, ethoxy, i-propoxy, t. Ert-butoxy, pentyloxy, and hexyloxy. Preferably, it is C1-C4 alkoxy, especially methoxy, ethoxy and the like.

 Aryl means a mono- or condensable aromatic hydrocarbon group, for example, phenyl, hy-naphthyl, naphthyl, anthryl, indenyl, phenanthryl and the like. Preferably it is phenyl.

 Heteroaryl means an aromatic monocyclic or polycyclic group containing the same or different heteroatoms selected from 0, S and N.

The monocyclic group includes a 5- to 7-membered group containing 1 to 4 heteroatoms, and includes furyl, thienyl, tetrazolyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, thiadiazolyl, pyridinyl, oxazinyl, and toxinyl. Liazinyl and the like are exemplified. The polycyclic group includes a bicyclic or tricyclic heterocyclic group containing 1 to 5 heteroatoms. And benzofurael, isobenzofuranyl, benzochenyl, indolyl, isoindolyl, ingzolyl, benzimidazolyl, benzoxazolyl, benzothiazolyl, benzotriazolyl and the like.

 Substituents on the aryl or heteroaryl include halogen (eg, Br), hydroxy, amino, carboxy, cyano, nitro, carbamoyl, sulfamoyl, lower alkyl (eg, methyl) , Lower alkyl halides (eg, trifluoromethyl), lower alkyl rubamoyl (eg, methylcarbamoyl), lower alkylsulfamoyl (eg, methylsulfamoyl), lower alkoxy (eg, methoxy), lower alkoxycarbonyl (eg, Examples: ethoxycarbonyl) and the like, but preferably, nitrogen, nitro, lower alkyl and lower alkoxy.

R ¹ is preferably form connexion single bond such together with R ^3.

R ² is preferably hydrogen, lower alkyl (eg, methyl), or halogen (eg, B r), and particularly preferably hydrogen.

R ⁴ is preferably hydrogen, lower alkyl (eg, methyl, n-propyl), lower alkenyl (eg, 2-propenyl), or cyano, and particularly preferably methyl.

R ⁶ is preferably hydrogen, hydroxy, lower alkoxy (eg, methoxy, ethoxy, etc.), or lower alkyl carbonyloxy (eg, acetyloxy, etc.), and particularly preferably hydrogen.

R ⁷ is preferably form connexion single bond such with R ^9.

R ⁸ is preferably hydrogen, lower alkylthio (eg, methylthio), or halogen (eg, Br, C 1, etc.), and particularly preferably hydrogen.

R ^{1 (1} is preferably hydrogen, lower alkyl (eg, methyl, etc.), lower alkyl carbonyl (eg, acetyl, etc.), arylcarbonyl (eg, benzoyl, Ρ-Β-benzoyl, ρ—Ν) 0 ₂ - Benzoiru etc.), lower alkyl O alkoxycarbonyl (e.g. tert - butoxide deer Lupo sulfonyl, etc.), or § Li one Rusuruhoniru (e.g. toluenesulfonyl Honyl, p-Br-benzenesulfonyl, p-N〇2-benzenesulfonyl), and particularly preferably hydrogen.

R ¹¹ is preferably hydrogen, lower alkyl (eg, tert-butyl, etc.), or halogen (eg, Br, C 1 etc.), preferably hydrogen.

R ^{1 2} is preferably hydrogen or lower alkyl (eg: tert - heptyl etc.) Ru Der, preferably hydrogen.

 n is preferably 0.

Examples of the pharmaceutically acceptable salt of compound (I) include salts formed with inorganic bases, ammonia, organic bases, inorganic acids, organic acids, basic amino acids, halogen ions and the like, or internal salts. . Examples of the inorganic base include alkali metals (Na, K, etc.), alkaline earth metals (Ca, Mg, etc.), and examples of the organic base include trimethylamine, triethylamine, choline, proforce, ethanolanol, and the like. Is exemplified. Examples of the inorganic acid include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like. Examples of the organic acid include p-toluenesulfonic acid, methanesulfonic acid, formic acid, trifluoroacetic acid, and maleic acid. Examples of the basic amino acid include lysine, arginine, ornithine, histidine and the like. Hydrates, solvates, racemates, optically active forms, and prodrugs of the compound (I) are all included in the present invention. The prodrug is a derivative of the compound (I) having a group that can be chemically or metabolically degraded, and is a compound that becomes a pharmaceutically active compound of the present invention by solvolysis or under physiological conditions in vivo. is there. Methods for selecting and producing suitable prodrug derivatives are described, for example, in Design of Prodrugs, Elsevier, Amsterdam 1985. When the compound has a carboxyl group, an ester derivative produced by reacting the base acidic compound with a suitable alcohol, or a base derivative prepared by reacting the base acidic compound with a suitable amine Prodrugs such as amide derivatives are exemplified. When the present compound has a hydroxyl group, for example, a compound having a hydroxyl group and a suitable acyl halide or Illustrative are open-drugs, such as the acyloxy derivatives prepared by reacting with appropriate acid anhydrides. When the compound has an amino group, prodrugs such as amide derivatives produced by reacting a compound having an amino group with a suitable acid halide or a suitable mixed acid anhydride are exemplified. You.

 Compound (I) is obtained by culturing a Streptomyces strain capable of producing any of the compounds represented by the above formula (I) in a suitable medium, collecting and isolating from the medium, purifying, and optionally, It can be produced by chemical modification.

1.Culture conditions

 Cultivate a strain belonging to the genus Streptomyces, in a solid or liquid medium with an appropriate medium composition and medium conditions. The medium may contain a carbon source, a nitrogen source, sterile salts, vitamins and the like. Examples of the carbon source include glucose, sucrose, soluble starch, dextrin, glycerin, fatty acids, organic acids and the like. Nitrogen sources include, for example, soy flour, pinna, corn starch, beef extract, yeast extract, cottonseed powder, polypeptone, wheat malt, sodium nitrate, ammonium nitrate, and the like. Examples of the inorganic salts include calcium carbonate, sodium chloride, potassium chloride, magnesium sulfate, copper sulfate, manganese chloride, zinc sulfate, cobalt chloride, and various phosphates. The culturing temperature is usually about 12 to 44 ° C, preferably about 20 to 40 ° C. The pH of the medium is usually about 6.0 to 8.0, preferably about 6.5 to 7.2. The culture time is usually about 4 to 12 days.

 The above-mentioned strain is preferably an actinomycete, particularly preferably / • epio ^ ces sp, PA-48561 strain (this strain is the Institute of Biotechnology, Institute of Industrial Science and Technology, Ministry of International Trade and Industry (location: Tsukuba East, Ibaraki Prefecture) 1-chome 1-3), deposited on January 11, 1998 under accession number FERM P-17052. On February 7, 2000, changed to an international deposit under accession number FERM BP-7023. ).

2. Form

This strain PA-48561 is a Gram stain-positive, acid-fast stain-negative bacterium. Branches, 0.5 ~ 1.0〃 πι in width, does not break. The spores form a spiral spore chain at the tip of the spore stem branched from the aerial hypha. The spore chains are relatively short, most spores are cylindrical to sausage, 1.0-2.0〃111 long, 0.5-1.0 0.5πι wide, and their surfaces are smooth. Neither sporangia nor sclerotia are found.

 3.Culture properties (28 ° C, 14 days)

 Aerial hyphae grew well on starch inorganic salt agar medium and glycerin-asparagine agar medium, but no aerial hyphae were formed on yeast malt agar medium and oatmeal agar medium. The colonies on the starch inorganic salt agar medium are white and light yellow brown with sporulation. Soluble dyes do not recognize this.

 4.Physiological properties

 (1) Growth temperature range: It grows at 12-44 ° C on the inorganic salt agar medium (ISP-4), and shows the best growth especially at 28-41 ° C.

 (2) Liquefaction of gelatin: negative

 (3) Coagulation of milk: negative

 (4) Milk peptone: negative

 (5) Tyrosinase reaction: negative

 (6) Production of melanoid pigment: negative

 (7) Starch hydrolysis: positive

 (8) Salt tolerance: In Bennett's medium, it grows well up to 7% salt concentration.

(9) Utilization of carbon source

(table 1 )

5. Cell components

 LL-diaminopimelic acid and glycine were detected from the cell wall, suggesting that the cell wall was type I. For the major saccharide components of all cells, glucose and arabinose were detected, but no galactose and xylose were found, and the sugar pattern was type C. For menaquinone, MK-9 (H6 and 8) was detected. As for phospholipids, phosphatidylethanolamine and phosphatidylinositol were detected, but phosphatidylcholine and unknown glucosamine-containing phospholipids were not detected. II.

As shown by these mycological properties, PA-48561 strain belongs to the genus Streptomyces. Regarding the taxonomic position of this bacterium, see “Bergey's Manual of Systematic Bacteriology Volume 4”, Shiling, EB and Gottl ieb, D. Corporate's' Discipline of Ob 'type'Cultures' of Ob 'Streptomyces II. Speech's' Disc Ription' from 'First' Susday (Cooperative description of type cultures of Streptomyces.il. Species description from first study.), "Inuichi Nishinare, Giananole, Saibu, Systematic, Pacteriology," Vol. 18 (1968) (Int. J. Syst. Bacteriol., 18, 69-189, 1968) and other published literatures. As a result, Streptomyces shioyaensis was listed as the most closely related strain to this strain. This strain and the standard strain IF012820 of Streptomyces shioyaensis were cultured under the same conditions, and the two were compared. As a result, the test items shown in Table 2 were used. Significant differences were observed between the two strains. Therefore, this bacterium was regarded as a new species of the genus Streptomyces and named Streptomyses sp. (Table 2)

本発明者らは、

PA- 48561を培養し、 その発酵液から常法によ り抽出、 クロマ ト分離'精製等を行うことにより、 以下に示す天然物 （主成分 ht- 13- A、 マイナ一成分 ht- 13-B) を得た。

We have:

PA-48561 is cultured, and the following natural products (main component ht-13-A, minor component ht-13-) are extracted from the fermented liquor by a conventional method, followed by chromatographic separation and purification. B) was obtained.

The chemical structures of both were deduced from physicochemical data such as IR, MS, and NMR, and finally, 3-Brht-13-A and 5,5a-Dihydro-4, derivatives of M-13-A -The absolute stereochemical structure of p-bromobenzoyl ht-13-A was determined by X-ray crystallography. Similarly, the chemical structure of ht-13-Β was determined by comparison with ht-13-Α. As a result, it was found that each was a new tetracyclic alkaloid. In addition, the present inventors have appropriately modified these natural products by well-known reactions (eg, oxidation, reduction, protection, deprotection, C-C cleavage reaction, halogenation, hydroxylation, alkylation, alkylthiolation). , Demethylation, 0-alkylation, 0- Various derivatives were synthesized by acylation, N-alkylation, N-alkenylation, N-acylation, N-cyanoation, N-sulfonylation and the like.

Since compound (I) has affinity for various serotonin receptors, it functions as an agonist or antagonist. Therefore, serotonin receptors, especially to 5 - Various central nervous diseases (examples HT ₆ or 5 -HT ₇ is interposed: sleep-wake disorders, Gaibiri rhythm disorders, anxiety, depression, schizophrenia, stroke , Dementia, pain, Parkinson's disease, etc.) and can be orally or parenterally administered to animals including humans. Examples of the dosage form include granules, tablets, capsules, injections, suppositories and the like. Upon formulation, various additives may be used as required, for example, excipients (lactose, mannitol, crystalline cellulose, starch, etc.), disintegrants (potassium melomerose, hydroxypropyl methylcellulose, polyvinylpolypyrrolidone, etc.). , A binder (methylcellulose, hydroxypropyl cellulose, polyvinyl alcohol, etc.), a lubricant (magnesium stearate, talc, etc.), a stabilizer, a coloring agent, and a coating agent. The dosage may vary depending on the subject's age, body weight, symptoms, administration method, etc., but is usually about 0.001 mg to lg / day per adult, orally or parenterally, and the frequency of administration is 1 ~ Several times a day.

 EXAMPLES Examples of the present invention will be shown below, but these do not limit the present invention in any way.

 (Abbreviation)

TLC: Thin layer chromatography

HPLC: High performance liquid chromatography

Example 1 (Isolation of ht-13-A)

 (Method 1)

 [Culture]

 Glucose 2.0%, potato starch 2.0%, defatted soy flour (GP-SL, manufactured by Yoshiwara Oil Co., Ltd.) 2.0%, yeast extract (Difco) 0.5%, sodium chloride 0.25%, copper sulfate pentahydrate 0.0 005%, manganese chloride.0.4 hydrate 0.005%, zinc sulphate / 7 hydrate 0.0 0.005%, carbonic acid Dispense a medium with a composition of 0.35% water, tap water (adjusted to pH 7.0, diluted caustic soda) into a 500 mL wide-necked Erlenmeyer flask at 121 ° C for 20 minutes. Sterilized. Agar plate cultures of SrepioHycessp. PA-48561 were punched into this medium with stainless steel pipes of 6 mm diameter, inoculated with 2 pieces of agar, 28 mm in amplitude 70 mm, 180 rpm per minute at 28 ° C. Shaking culture was performed for 4 days.

 [Extract]

 The culture-finished solution of 96 flasks was separated into about 1.5 Kg of wet cells and 8 L of the supernatant using a centrifuge (Beckman J-6M, 4200 rpm, 15 minutes). The cells were leached with 1.5 L of ethanol for 1 day and filtered by suction. The ethanol filtrate containing water was subjected to solvent distillation under reduced pressure, and the remaining aqueous layer was extracted with 1 L of ethyl acetate, and then washed with saturated saline. The ethyl acetate layer was concentrated to dryness to obtain 0.5 g of a crude extract. 0.5 g of the crude extract was used as a 50 mL solution of ethyl acetate, and extracted with 50 mL of 0.1 N hydrochloric acid. The hydrochloric acid layer was adjusted to pH 8 with saturated aqueous sodium hydrogen carbonate, extracted with 100 mL of ethyl acetate, washed with saturated saline, and concentrated to dryness to obtain 25 mg of a base fraction.

 [Separation and purification]

The above base fraction 25 ^ 1 was added to C (Merck Precoated TLC plate silica gel 60 F254, thickness 0.5 mm, developing solvent: porcine form / medium = 9/1) ), Followed by HPLC [column: Devosyl ODS—HG5, 20 x 250 mm (Nomura Chemical Co., Ltd.), mobile phase: 0.1% trif-acetate (20% acetate) Ril-water), flow rate: 6 mL / min] to obtain a fraction containing ht-13-A. After evaporating the solvent, the residual aqueous layer was adjusted to pH 8 with saturated aqueous sodium hydrogen carbonate and extracted with 50 mL of ethyl ethyl dichloride. Then, it was washed with saturated saline, and the ethyl acetate layer was concentrated to dryness to obtain ht-13-A (4 mg).

 1) Name: ht-13-Α: (6aR, 9aS) -7-methyl-6a, 8, 9, 9a-tetrahydro-4H, 6H, 7H-benzo [i, j] pyrrolo [2, 3-f] [10, 4] oxazazulene

2) Physicochemical properties:

Shape: Pale yellow needles

Solubility: Soluble in ethyl acetate, acetone, black form, and methanol; insoluble in water Melting point: 234 ~ 237 ° C

[a] _D -199.0 ± 4.7 ° (C = 0.504, methanol, 29 ° C)

として Elemental analysis:

As

Calculated values: C: 73.66%, H: 7.06%, N: 12.27%

Measured value: C: 73.48%, H.-7.06%, N: 12.09%

LSIMS: m / z 229 [M + H] ⁺

IR (KBr) cm— ¹ : 3144, 3098, 3060, 3033, 1621, 1510, 1456, 1344, 1293, 1242,

1065, 777, 741

 UV (methanol) ma ma (£): 221 (28,500), 275 (6,200), 287 (6,000), 297 (5,600)

) 11 NMR (double-mouthed form, 600 MHz) δ: 2.09 (IH, dddd, J = 13.7, 7.9, 2.9, 1.2 Hz), 2.43 (3H, s), 2.50 (IH, m), 2.57 (IH, m), 2.60 (1H, m), 2.65 (1H, br.ddd, J = 13.6,13.6,1.4Hz), 3.12 (IH, dd, J = 8.4,8.4 Hz), 3.42 (1H, ddd, J = 14.7, 3.6, 0.7 Hz), 4.47 (IH, ddd, J = 9.7, 6.2, 2.9 Hz), 6.63 (1H, dd, J = 7.8, 0.9 Hz), 6.98 (IH, dd, J = 8.0, 0.9 Hz) ), 7.00 (1H, br.s), 7.07 (IH, dd, J = 7.9,7.9 Hz), 8.10 (1H, br.s)

¹³ C NMR (double-mouthed form, 150 MHz) δ: 29.19, 30.81, 40.52, 55.60, 72.93, 86.65, 104.00, 106.15, 111.03, 117.12, 120.06, 122.82, 138.71, 151.82 Color reaction: Positive for ラ, 'Raken'-Dondorf reagent,, and sulfuric acid

 TLC: Rf value 0.36 (Merck Pre-Trade TLC pre-silicone gel 60F254, mouth mouth Holm: methanol = 9: 1)

HPLC: Retention time 9.0 minutes (Devosyl 0DS-T-5 4.6 φ x 150 mm (Nomura Chemical Co., Ltd.), mobile phase 18% acetonitrile-reluate water (0,1¾ trifluoroacetic acid), flow rate 1 mL / min, (Detection UV 220 nm)

 (Method 2)

 [Culture]

 0.5% soluble starch, 0.5% glucose, 0.5% polypeptone (fine granules, manufactured by Nippon Pharmaceutical Co., Ltd.), 0.5% beef extract (manufactured by Difco), 0.5% yeast extract (Difco) 0.25% sodium chloride, 0.25% sodium chloride, and tap water (adjusted to pH 7.0, diluted caustic soda) into a 2 L triangular flask with a branch. 800 mL was dispensed and sterilized at 121 ° C for 20 minutes. This medium was inoculated with 4 mL of a 20% glycerin-storing vial (storage at 180 ° C) of Streptomyces sp. PA—485851 strain, and the amplitude was 7 O mm, and the rotation was 180 rpm. Shaking culture was performed for 28 and 24 hours.

 Separately, soluble starch 1.5%, glucose 1.0%, dried yeast (beast, manufactured by Iwaki Seiyaku Co., Ltd.) 0.5%, defatted soy flour (GP-SL, manufactured by Yoshiwara Oil Co., Ltd.) 2.0 %, Sodium chloride 0.25%, Adekinol LG-121 (manufactured by Asahi Denka Kogyo Co., Ltd.) 0.03%, medium 30 L of tap water (without pH adjustment) Was injected into a 50 L capacity fermenter, sterilized at 121 ° C for 25 minutes, inoculated with the entire amount of the above culture, and aerated and stirred at 23 ° C for 6 days (aerated). : 150 L / min, stirring: 150 rotations / min).

 [Extract]

53.6 L of culture termination solution is KS type ultra high speed centrifuge (Kansai Centrifuge) The mixture was subjected to continuous centrifugation at 18, 212 revolutions per minute (19, 292) at a flow rate of 5 L / min to obtain 6.7 kg of wet bacterial cells. Acetone (18 L) was added to the wet cells, and the mixture was stirred for about 1 hour (at this time, the pH was adjusted to 2.0 with a dilute hydrochloric acid aqueous solution). A filter cloth centrifugal filter (Kansai Centrifuge Co., Ltd. Company, KAN15) was used to obtain 24 L of filtrate.

 The filtrate was adjusted to pH 7.0 using sodium bicarbonate, and concentrated under reduced pressure to obtain 6 L of an aqueous phase. To this aqueous phase, 5 L of water was added, and extracted twice with ethyl acetate (7 L, 3 L). This ethyl acetate solution was extracted twice with 0.1 N aqueous hydrochloric acid (5 L, 5 L) to obtain 10 L of 0.1 N aqueous hydrochloric acid. This 0.1 N hydrochloric acid aqueous solution was adjusted to pH 7.0 using sodium bicarbonate, and extracted twice with ethyl acetate (3 L, 2 L). This ethyl acetate solution was washed with 800 mL of water and concentrated under reduced pressure to obtain 0.37 g of an oily substance.

[Separation and purification]

 The crude extract was fractionated by preparative HPLC (0 DSS—15 / 305 cm 0 × 5 O cm (YMC), 15% acetonitrile (0.1% TFA)). An effective fraction of 450 mL was obtained. This fraction was adjusted to pH 7.3 using a dilute sodium hydroxide solution, and concentrated under reduced pressure to obtain an aqueous phase of about 370 mL. The aqueous phase was extracted twice with 80 mL of ethyl acetate, the ethyl acetate phase was concentrated under reduced pressure, and crystallized from a small amount of ethyl acetate and n-hexane to obtain the compound (ht-13-A) Was obtained as colorless needle crystals (62 mg in total). Example 2 (Isolation of ht-13-A and ht-13-B)

 [Fermentation]

 0.5% soluble starch, 0.5% glucose, 0.5% polypeptone (fine granules, manufactured by Nippon Pharmaceutical Co., Ltd.), 0.5% beef extract (manufactured by Difco), 0.5% yeast extract (Difco) 0.25% sodium chloride, 0.25% sodium chloride, and tap water (adjusted to pH 7.0, diluted caustic soda) into a 2 L triangular flask with a branch. 800 mL was dispensed and sterilized at 121 ° C for 20 minutes. In this medium

Streptomyces sp. PA— Inject 4 mL of 20% glycerin storage vial (stored at 180 ° C) of 4 strains, and shake at 28 ° C at an amplitude of 70 mm at 180 rpm. C, 24:00 Shaking culture was performed.

 Separately, soluble starch 0.5%, glucose 0.5%, polypeptone (fine granules, manufactured by Nippon Pharmaceutical Co., Ltd.) 0,5%, beef extract (Difco) 0,5%, water extract extract (Manufactured by Difco) 0.25%, sodium chloride 0.25%, Adecanol LG—121 (manufactured by Asahi Denka Kogyo Co., Ltd.) 0.03%, tap water (pH 7. Adjust to 0, inject 20 L of medium with the composition of dilute caustic soda) into a 30 L jar fermenter, sterilize at 121 ° C for 25 minutes, inoculate the entire amount of the above culture, Aeration and agitation culture was performed at 28 ° C for 24 hours (aeration: 10 L / min, agitation: 150 rotations / min). Separately, soluble starch 4.0%, glucose 2.0%, dried yeast (Bist, Iwaki Seiyaku Co., Ltd.) 1.0%, defatted soy flour (GP-SL, Yoshihara Oil Co., Ltd.) 4 0%, sodium chloride 0.25%, Adecanol LG-121 (manufactured by Asahi Denka Kogyo Co., Ltd.) 0.03%, medium with tap water (without pH adjustment) 150 L was poured into a 250 L tank, sterilized at 121 C for 25 minutes, inoculated with 6 L of the above culture, and aerated and stirred at 23 ° C for 1 day (aeration). : 75 LZ min, stirring: 120 revolutions / min).

 [Extract]

 30 L of water was added to 138 L of the culture termination solution, and the pH was adjusted to 1.5 using a diluted sulfuric acid solution. 8 kg of celite (filter aid 'Hyflo') was added, and the mixture was filtered with a Fil Yuichi Press (Noritake Seisakusho, size: 500 SQ) to obtain about 130 L of filtrate. The filtrate was adjusted to ρΗ80 using a diluted sodium hydroxide solution, and extracted with 54 L of ethyl acetate. This ethyl acetate solution was extracted twice with 0.1 L hydrochloric acid solution (10 L, 8 L) to obtain 18 L of 0.1 N hydrochloric acid solution. This 0.1 N hydrochloric acid aqueous solution was adjusted to pH 7.5 using a dilute sodium hydroxide solution, and extracted twice with ethyl acetate (8 L, 5 L). The ethyl acetate solution was washed with 1.0 L of water and concentrated under reduced pressure to obtain 1.06 g of an oily substance.

[Separation and purification]

The crude extract was divided into two portions and subjected to preparative HPLC (0 DSS—15 / 305 c Fractionation was performed using møx50 cm (YMC), 15% acetonitril (0.1% TFA), flow rate: 6 OmL / min, to obtain 2.0 L of an effective fraction. This fraction was adjusted to pH 7.6 using a dilute caustic soda solution, and concentrated under reduced pressure to obtain about 1.7 L of an aqueous phase. This aqueous phase was extracted twice with 200 mL of ethyl acetate, and the ethyl acetate phase was concentrated under reduced pressure to dryness, crystallized from a small amount of ethyl acetate and n-hexane, and ht-13-A was colorless. Acicular crystals (a total of 420 mg) were obtained.

 On the other hand, in this preparative HPLC process, 1.5 L of a UV absorption fraction eluted after the ht-13-A fraction was obtained. This fraction was adjusted to pH 7.6 with dilute sodium hydroxide solution and concentrated under reduced pressure to obtain about 1.3 L of an aqueous phase. This aqueous phase was extracted twice with 150 mL of ethyl acetate, and the ethyl acetate phase was concentrated under reduced pressure, washed with a small amount of petroleum ether, and ht-13-B (38.7 mg, light brown powder) ) Got.

 1) Name: ht-13-B: (6aR, 8R, 9aS) -7,8-dimethyl-6a, 8,9,9a-tetrahydro-4H, 6H, 7H-benzo [i, j] pyrrolo [2, 3-f] [10, 4] oxazazulene

 2) Physicochemical properties:

Shape: Pale yellow needles

Solubility: Ethyl acetate, Acetone, Black form, Soluble in methanol, Insoluble in water Melting point: 182 ~ 184 ° C

[a] _D -177.9 ± 4.3 ° (C = 0.502, methanol, 29 ° C)

Elemental analysis: as C.SHHNJO O. IHJO

Calculated values: C: 73.80%, H: 7.51%, N: 11.48%

Found: C: 73.58%, H: 7.41%, N: 11.58%

ESI-MS: m / z 243 [M + H] ⁺

IR (Or) cm- ¹ : 3144, 3101, 3054, 3033, 1621, 1509, 1454, 1342, 1295, 1245, 1065, 774, 739

UV (methanol) λ max nm (£): 221 (29,600), 275 (5,700), 287 (5,500), 297 (5,100) MR (double-mouthed form, 600 MHz) δ 1.16 (3Η, d, J = 6.6 Hz), 1.84 (1H, ddd, J = 13.8, 3.6, 2.4 Hz), 2.42 (3H, s), 2.58 (1H, br.td, J = 13.2, 1.2 Hz), 2.71 (1H, ddd, J = 13.8, 9.6, 7.2 Hz), 3.12 (1H, ddd, J = 12.0, 6.0, 3.6 Hz), 3.38 (1H, ddd, J = 14.4, 3.6, 0.6 Hz), 3.40 (1H, m), 4.43 (1H, ddd, J = 9.6, 6.0, 3.6 Hz), 6.64 (1H, dd, J = 7.8, 0.9 Hz), 6.98 (1H , dd, J = 7.8, 0.9 Hz), 7.00 (1H, br.s), 7.07 (1H, dd, J = 7.8,7.8 Hz), 8.10 (1H, br.s)

¹³ C NMR (double-mouthed form, 150 MHz) δ: 14.22, 29.52, 35.11, 38.84, 57.96, 67.18, 86.52, 103.90, 106.07, 111.22, 117.12, 120.17, 122.78, 138.79, 152.15 Color reaction: Drägendorff Positive for reagent, chloride and sulfuric acid.

 TLC: Rf value 0.31 (Merck Pleated TLC plate silica gel 60F254, ku mouth form: methanol = 9: 1)

HPLC: retention time 12.5 min (Deberoshiru ODS- T-5 4.6 φ x 150 mm ( Nomura Chemical Co., Ltd.), mobile phase 18% § Se Tonito Lil monohydrate (0.1 ¾ preparative Rifuroro acetate), flow rate 1 _m L / min, Hereinafter, derivatives of the above compounds (ht-13-A and ht-13-B) were synthesized.

(Note: Each number indicated by "Ex" indicates 列 M column number.)

Example 3 (Synthesis of 4-Meht-13-A)

 (6aR, 9aS) -4,7-dimethyl-6a, 8,9,9a-tetrahydro-4H, 6H, 7H-benzo [i, j] pyrrol o [2,3-f] [10,4] oxazazulene

 ht-13-Α (10 mg) was dissolved in THF (2 mL), and NaH (about 60% oil suspension, 4 mg) was added. After stirring at room temperature under a nitrogen stream for 5 minutes, methyl iodide (20 μL) was added, and the mixture was stirred at room temperature under a nitrogen stream for 2 hours. The reaction solution was diluted with a saturated aqueous sodium hydrogen carbonate solution and partitioned with ethyl acetate. After washing with a saturated saline solution and drying, the solvent was distilled off, and the residue was separated by silica gel TLC (form: methanol = 95: 5) to obtain the title compound 5.8.

[Physicochemical properties]

Solubility: Ethyl acetate, Acetone, Cloth form, Soluble in methanol, Insoluble in water ESI-MS: m / z 243 [] *

 'Η NMR (hologram, 300 MHz) δ: 2.07 (1H, m), 2.42 (3H, s), 2.45-2.69 (4H, m), 3.10 (lH, dd, J = 8.3, 8.3 Hz) , 3.39 (1H, dd, J = 14.1, 2.7 Hz), 3.73 (3H, s), 4.46 (1H, ddd, J = 9.3, 5.7, 3.0 Hz), 6.63 (1H, dd, J = 8.0, 0.8 Hz) ), 6.85 (1H, br.s), 6.90 (lH, dd, J = 8.0, 0.8 Hz), 7.10 (1H, dd, J = 8.0, 8.0Hz) Color reaction: Dragendorff reagent, Example 4 (Synthesis of 4-Benzoyl ht-13-Α)

(6ai?, 9a5 -4-benzoyl-7-methyl-6a, 8,9,9a-tetrahydro-4H, 6H, 7H-benzo [i,] pyrrolo [2,3-] [10,4] oxazazulene ht- 13-Α (lmg) was dissolved in THF (200 μL), NaH (oil suspension containing about 60% oil, lmg) was added, and the mixture was stirred at room temperature under a nitrogen stream for 5 minutes. The reaction mixture was diluted with a saturated aqueous solution of sodium bicarbonate and partitioned with acetic acid;! Tyl. After washing with saturated saline and drying, the solvent was removed. After evaporation, the residue was fractionated by silica gel / TLC (cut-mouth form: metasol = 95: 5) to give 0.3 mg of the title compound. [Physicochemical properties]

Solubility: Ethyl acetate, Acetone, Soluble in black form, insoluble in water

ESI-MS: m / z 333 [M + H] ⁺

iH NMR (hologram form, 300 MHz) δ 2.07 (1H, m), 2.39 (3H, s), 2.44- 2.65 (4H, n), 3.10 (lH, ^ /, J = 7.5,7.5 Hz), 3.39 (1H, dd, J = 13.7,1.5 Hz), 4.41 (1H, m), 6.87 (1H, dd, J = 8.1,0.9 Hz), 7.08 (1H, br.s), 7.28 (lH, n) , 7.49-7.64 (3H, m), 7.70- 7.73 (2H, m), 8.10 (1H, dd, J = 8.1, 0.9Hz)

Color reaction: Positive for G, Rake, Ndolph's reagent, K-, and sulfuric acid Example 5 (Synthesis of 4-B0C ht-13-Α)

(6aJ? '9a <S) -4-i-butoxycarbony) -7-inetliy 6a, 8'9'9a-tetrahydro-4H'6H'7H-benzo [j'] pyrrolo [2,3-] [10 , 4] oxazazulene ht-13-Α (50 mg) was dissolved in acetate; tolyl (10 raL), and di-tert-butyldicarbonate (300 mg) was added to 4-('methyl) reaminohi. Resin (60 _mg ) was added and the mixture was stirred at room temperature for 15 hours. The solvent was distilled off, and the residue was fractionated by silica gel TLC (cloth form: methanol = 95: 5). 49 mg of the title compound were obtained.

[Physicochemical properties]

Solubility: Soluble in ethyl acetate, acetate, and black form, insoluble in water

LSIMS: m / z 329 [Μ + Η] +, 657 [2Μ + Η] +

ΐΗ NMR (double-mouthed form, 300 MHz) (5: 1.65 (9H, s), 2.07 (1H,), 2.43 (3H, s), 2.40-2.70 (4H, _m), 3.13 (lH,), 3.36 (1H, dd, J = 15.3, 2.7Hz), 4.40 (1H, m), 6.78 (1H, dd, J = 8.1, 0.9Hz), 7.19 (lH, dd, J = 8.1, 8.1Hz), 7.39 ( 1H, s), 7.83 (1H, d,

(J = 8.lHz)

Color reaction: Positive for ref, Re, Re, and Sulfuric acid

Example 6 (Synthesis of 3-Br ht-13-Α)

(ea ^ Da ^ -S-romo-T-methyl-ea ^^. ea-tetraliydro ^ HeHTH-benzo [i ', y] pyrrolo [2,3-i] [10,4] oxazazulene ht-13-A (10 mg) was dissolved in chloroform (10 mL), 10 mg (1.25 equivalents) was gradually added to N-Fu, moss cincinimi and stirred for 14 hours. After evaporating the solvent, separate by HPLC (column: Devosyl 0DS-T-5, 10 ø X 250 mm, mobile phase: 25% acetonitrile-water (0.1% trifluoro!) Acetic acid), concentrate and concentrate in aqueous ammonia The extract was combined and extracted with 10 mL of ethyl acetate. The solvent layer was washed with saturated saline and then distilled off to obtain 6 mg of the title compound.

[Physicochemical properties]

Solubility: Insoluble in water soluble in ethyl acetate, acetone, black form, and methanol ESI-MS: m / z 307 ([M + H] +: Br), 309 ([M + H] ⁺ : siBr)

iH NMR (double-mouthed form, 300 MHz) δ: 2.07 (1Η, m), 2.42 (3H, s), 2.44- 2.70 (4H, n) _; 3.11 (lH, n), 3.40 (1H, dd, J = 14.7, 3.3 Hz), 4.43 (1H, m), 6.54 (1H, d, J = 8.1 Hz), 7.06 (1H, br.s), 7.19 (lH, rf, J = 8.1 Hz), 8.24 (1H , br.s)

Color reaction: Positive for,, raque, 'N'-Dolph's reagent, lye', sulfuric acid

 HPLC: Retention time 9.2 min (Develosil ODS-T-5 4.6 φ X 150 mm (Nomura Chemical Co., Ltd.), mobile phase 25% acetonitrile monohydrate (0.1% trifluoroacetic acid), flow rate 1 mL / min, (Detection UV 210 nm)

Example 7 (Synthesis of 5-SMe ht-13-Α)

 5-SMe ht-13-Α: (6aR, 9aS) -5-methylthio-7-methyl-6a, 8, 9, 9a-tetrahydro-4H, 6H, 7H-benzo [i, j] pyrrolo [2,3 -f] [10,4] oxazazulene

 110 mg of sulfuryl chloride dissolved in 2.5 mL of dichloromethane was added dropwise at −20 ° C. to 40 mg of dimethyl disulfide dissolved in 2.5 mL of methane, and stirred at room temperature for 1 hour. 2.5 mL of the above reaction solution was added dropwise to a solution of ht-13-Α in dichloromethane (50 mg / 10 mL), and the mixture was stirred at -35 ° C for 1 hour. Thereafter, the temperature was gradually returned to room temperature, partitioned with 0.1 M aqueous ammonia, and the dichloromethane layer was washed with saturated saline, dried, and the solvent was distilled off. The residue was separated by silica gel TLC (chloroform: methanol: = 95: 5) to give 20 mg of the title compound.

[Physicochemical properties] 9Z

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TP 'HI 9

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璲 ikr) 氺, ¾ £ k2Mf—,, ^ α α, Be 4, ^^ 翘 digging: Z fZZlO / OOdf / IDJ 6066S / 00 O 8.10 (1H, br.s)

 TLC: Rf value 0.45 (Merck Pleated TLC plate silica gel 60F254, chloroform: methanol = 9: 1)

 HPLC: retention time 3.4 min (Devosyl 0DS-5 4.6 φ x 150 mm (Nomura Chemical Co., Ltd.), mobile phase 30% acetonitrile monohydrate (0.1% trifluoroacetic acid), flow rate 1 mL / min , Detection UV 210 nm)

 [Physicochemical properties of 3-C1 ht-13-A]

Solubility: Ethyl acetate, Acetone, Soluble in black form, insoluble in water

Positive ion ESI-MS: m / z 263 [M + H] ⁺

Negative Vion ESI-MS: m / z 261 [M-H]-

NMR (polychrome form, 300 MHz) δ: 2.07 (1H, m), 2.44 (3H, s), 2.40-

2.76 (4Η, Λ2), 3 ・ 13 (ΐΗ, ί¾ J = 7.8,7.8Hz), 3.40 (1H, dd = 14.4,3.0 Hz), 4.44 (1H, m),

6.57 (1H, d, J = 8A Hz), 7.04 (1H, d, J = 8.4Hz), 7.06 (1H,, 8.27 (1H, br.s)

¹³ CNMR (double-mouthed form, 75 MHz) δ 29.56, 31.13, 40.82, 55.86, 72.97, 87.12, 107.19, 108.68, 112.65, 118.40, 120.84, 122.03, 135.35, 150.94

 Τ L C: Rf value 0.35 (Merck Pleated TLC Plate Silica Gel 60F254, 口 mouth form: methanol = 9: 1)

 HPLC: retention time 4.4 min (Devosyl 0DS-5 4.6 φ x 150 mm (Nomura Chemical Co., Ltd.), mobile phase 30% acetonitrile monohydrate (0.1 trifluoroacetic acid), flow rate 1 mL / min, detection UV 210 nm)

Example 9 (Synthesis of 1 Butvl-5-SMe ht-13-Α and 1,3-di Butyl-5-SMe ht-13-Α)

(1) li-Butyl-5-SMe ht-13-Α: (6ai2,9a) -li-Butyl-5-metliylthio-7-metliyl- 6a, 8,9,9a-tetrahydro-4H, 6H7H-benzo [ iy] pyrrolo [2,3-i] [10 ₎ 4] oxazazulene

(2) l, 3-di-i-Butyl-5-SMe ht-13-Α: (Sa ^ Sa ^ -l'S-di-i-Butyle 5-methylthio-7-methyle Sa'S'S'Sa-tetrahydro ^ H'SHTH-benzo ^ ipyrrolo 'S- ^ l ^ lO' ^ oxazazulene 5-SMeht-13-A (20 mg) of Example 7 was dissolved in THF (1 mL), and -Butyl acetate (100 μ¾) was added thereto, followed by stirring at 50 ° C for 6 hours. The ethyl acetate layer was washed with saturated saline, dried over anhydrous sodium sulfate, and the solvent was distilled off.The residue was subjected to HPLC (column: "D-Sil ODS-5 10 x 150 mm, mobile phase: 0.1% trifluoroacetic acid-50% acetonitrile (4 minutes), 0.1% trifluoroacetic acid [50% → (4 minutes) 90%] acetonitrile monohydrate, lash, 1-Butyl-5-SMe, 0.1% triflate!] 95% acetonitrile acetate (2 min), flow rate: 5 mL / min, detection: UV 220 nm) 7.7 mg of ht-13-Α and 7.8 mg of l, 3-di-i-Butyl-5-SMe ht-13-Α were obtained.

[Physicochemical properties of l-i-Butyl-5-SMe ht-13-Α]

Solubility: Ethyl acetate, acetate, black form, soluble in methanol, insoluble in water Positive ion ESI-MS: m / z 331 [M + H] ⁺

Negative Vion ESI-MS: m / z 329 [M-H]-, 283 [M-SCHJ "

iH NMR (double-mouthed form, 300ΜΗζ) δ 1.43 (9H, s), 2.17 (1H, m), 2.36 (3H, s), 2.480 (3H, s), 2.35-2.67 (4H,), 3.18 (1H ,), 3.57 (1H, d, J = 10.8Hz), 4.43 (1H, m), 6.86 (1H, d, J = 8.6 Hz), 7.19 (1H, d, J = 8.6 Hz), 7.96 (1H, br.s)

Color reaction: Positive for tracing, 'Ndorff reagent, chloride, sulfuric acid'

 TLC: Merck Pre-Trade TLC plate Silica gel 60F254, Rf value was 0.46 when developing solvent: Cloth form: methanol = 95: 5.

HPLC: 0.1% trifluorocoacetic acid [30% → (7 minutes) 95%] acetonitrile monohydrate was used as the mobile phase using a silica gel ODS-54.6 ø X 150 mm (Nomura Chemical Co., Ltd.) as the column. When the flow rate was set to 1 mL / min and the detection was performed at UV 210 nm, the retention time was 7.5 minutes.

 [Physicochemical properties of l, 3-di-t-Butyl-5-SMe]

Solubility: soluble in ethyl acetate, acetate, black form, methanol, insoluble in water ESI-MS: m / z 387 [M + H] ⁺

iH NMR (double-mouthed form, 300 MHz) δ 1.42 (9H, s), 1.46 (9H, s), 2.18QH, m), 2.37 (3H, s), 2.54 (3H, br.s), 2.4-2.8 (4H, m), 3.25 (1H, m), 3.60 (1H, d, J = 10.8Hz),

4.46 (1H, m), 7.14 (lH, s), 8.07 (1H.br.s)

 Color reaction: Positive for "Raken" Ndorff reagent, h-, sulfuric acid

 TLC: Merck Pre-Ted TLC plate Silica gel 60F254 was used, and Rf value was 0.52 when developing solvent: Cloth form: methanol = 95: 5. HPLC: Using 'II Sil ODS-54.60 x 150 mm (Nomura Chemical Co., Ltd.) in the column, • 0.1% trifluoroacetic acid mono [30% → (7 min) 95%] acetonitrile monohydrate slurry as the mobile phase Example 10 (Synthesis of 1-Butyl ht-13-Α) when detection was performed at UV 210 nm with a flow rate of 1 mL / min using a reagent and a flow rate of 1 mL / min.

(6a ^, 9a6) -li-butyl-7-methyl-6a, 8,9 _; 9a-tetrahydro-4H, 6H, 7H-benzo [j,] pyrrolo [2,3-/] [10 ₎ 4] oxazazulene

 L-i-Butyl-5-SMeht-13-A (4 mg) of Example 9 was dissolved in ethanol (1 mL), and Raney nickel (50 mg) was added. After heating under reflux for 1 hour, Raney nickel was removed by mouth and the solvent was distilled off under reduced pressure. The residue was fractionated by TLC (Merck Pre-Trade TLC Blade Silica Gel 60F254, thickness 0.25 mm, form: methanol: 95: 5) to obtain 3.5 mg of the title compound.

 [Physicochemical properties]

Solubility: soluble in ethyl acetate, acetone, black form, methanol, insoluble in water LSIMS: m / z 284 [Μ] ⁺ , 285 [Μ + Η] ⁺

HR-LSIMS, m / z: Ci ₈ H ₂₅ N ₂₀

 Calculated value: 285.1966

 Actual value: 285.1970

 iH NMR (double-mouthed form, 600 MHz) δ: 1.44 (9H, s), 2.19 (1H, m), 2.48 (3H, s), 2.5-2.8 (4H, m), 3.20 (1H, m), 3.44 (1H, dd, J = 14.4, 3.6 Hz), 4.45 (1H, m), 6.92 (1H, d, J = 8.4 Hz), 6.98 (1H, br.s), 7.18 (1H, d, J = 8.4 Hz), 7.98 (1H, br.s)

i ³ C NMR (double-mouthed form, 150 MHz) δ: 29.11, 30.33, 30.52, 34.64, 40.35, 55.53, 72.74, 85.89, 103.15, 110.77, 118.21, 120.56, 121.37, 127.07, 137.61, 149.86

Color reaction: Positive for "Rakes", Ndolph's reagent, anode, sulfuric acid

 TLC: Merck Pleitted TLC plate Silica gel 60F254, and the Rf value was 0.21 when using a developing solvent of black form: methanol = 95: 5.

HPLC: Using a column of TE'V'D Sill ODS-54.6 ø X 150 mm (Nomura Chemical Co., Ltd.), and using a mobile bed as a moving bed, 0.1% trif DD acetate 30% → 95% acetonitrile / water 7 minutes, 0.1% trif

Using D1 acetic acid (95% acetonitrile / water for 3 minutes) and a flow rate of 1 mL / min, the retention time when detecting with UV 210 mn was 7.1 minutes.

Example 11 (Synthesis of 1,3-di-^-Butyl ht-13-Α)

[2,3-/1 [10, 4]oxazazulene (6ai?, 9a5 -l, 3-di-i-Butyl-7-methyl-6a, 8,9,9a-tetrahydro-4H, 6H, 7H-

[2,3- / 1 [10, 4] oxazazulene

 The l, 3-di-i-Butyl-5-SMe ht-13-Α of Example 9 (4 mg) was dissolved in ethanol (1 mL),

Raney nickel (50 mg) was added. After heating under reflux for 1 hour, Raney nickel was removed by mouth and the solvent was distilled off under reduced pressure. The residue was separated by TLC (Merck Pre-Trade TLC Plate Silica Gel 60F254, thickness 0.25 mm, chloroform: methanol = 95: 5) to give 3.0 mg of the title compound.

 [Physicochemical properties]

Solubility: Soluble in ethyl acetate, acetone, black form, methanol, insoluble in water LSIMS: m / z 340 [M] +, 341 [Μ + Η] ⁺ , 681 [2Μ + Η] +

HR-LSIMS, m / z: C22H ₃₃ N ₂₀

Calculated value: 341.2592

Actual value: 341.2597

iH NMR (double-mouthed form, 600 MHz) 6: 1.44 (9H, s), 1.48 (9H, s), 2.18 (1H, m), 2.48 (3H, br.s), 2.50-2.80 (4H, m ), 3.21 (1H, w), 3.45 (1H, dd, J = 14.5, 3.9 Hz), 4.44 (1H, m), 7.01 (1H, br.s), 7.12 (1H, s), 8.07 (1H, br.s) i3C NMR (deuterochlorohonorem, 150 MHz) δ: 29.35, 30.19, 30.55, 30.59, 34.29, 34.89, 40.31, 55.47, 72.54, 85.64, 110.26, 118.55, 118.81, 120.31, 124.61, 126.41, 134.85, 148.09

Color reaction: G, 'Raken' Ndorff reagent, G-, positive for sulfuric acid

TLC: Merck Pleitted TLC plate using silica gel 60F254. Mouth form: Rf value was 0.25 when developing solvent was methanol = 95: 5. HPLC: Using a column, "Herosil ODS-54.6 ø X 150 mm (Nomura Chemical Co., Ltd.)" as the mobile phase,,, lash, ent (0.1% trif acetic acid 30 ° / ₀ → 95% acetonitrile) Using water / water 7 min, 0.1% trif D mouth acetic acid 95% acetonitrile / water 7 min), and the flow rate was lm L / min, the retention time was 8.6 min when detected at UV 210 nm.

Solubility: Soluble in ethyl acetate, acetone, black form, methanol, insoluble in water. Implementation _Example 12 (Synthesis of 5,5a-Dihydro ht-13-A)

 5,5a-Dihydro ht-13-Α: (5aS, 6aR, 9aS) -7-methyl -5,5a, 6a, 8, 9, 9a-hexahydro-4H, 6H, 7H-benzo [i, j] pyrrolo [2,3-f] [10,4] oxazazulene

 ht-13-Α (10 mg) was dissolved in 1 mL of acetic acid at triflou mouth, 5 nig of sodium borohydride was added, and the mixture was stirred at room temperature for 1 hour. Then, the mixture was diluted with 2 mL of water under ice cooling, neutralized with ammonia water, and extracted with 10 mL of ethyl acetate. After the ethyl acetate layer was washed with a saturated saline solution, the solvent was distilled off to obtain 10.4 iug of a residue. After separation by HPLC (column: Devosyl 0DS-T-5, 10 φ x 250 mm, mobile phase: 5% acetonitrile-water containing 0.1% trifluoroacetic acid), the mixture was concentrated, neutralized with aqueous ammonia and neutralized with acetic acid. Extracted with 10 mL of ethyl acetate. The ethyl acetate layer was washed with a saturated saline solution and then distilled off to obtain 6 mg of the title compound.

[Physicochemical properties]

Solubility: Ethyl acetate, Acetone, Black form, Soluble in methanol, Insoluble in water ESI-MS: m / z 231 [Μ + Η] ÷

'Η NMR (double-mouthed form, 300 MHz) 6: 1.28 (1H, m), 1.95 (1H, m), 2.17 (1H, a), 2.32 (1H,) 2.38 (3H, s), 2.35-2.50 (2H, m), 3.11 (1H, JB), 3.20-3.38 (2H, Jn), 3.74 (1H, dd, J = 7.8, 7.8 Hz), 3.89 (ΙΗ, ΤΗ), 6.36 (1H, d, J = 7.8 Hz),

6.39 (1H, d, J = 7.8 Hz), 6.95 (1H, dd, J = 7.8, 7.8 Hz),

Color reaction: Positive for Dragen-Dolph reagent, chloride and sulfuric acid

Example 13 (Formation of 5,5a-Dihydro-4-p-bromob_enzyl j? T- 丄 3-)

(5aS, 6aR, 9aS) -4-p-bromobenzoyl-7-methyl-5, 5a, 6a, 8, 9, 9a-hexahydro-

4H, 6H, 7H-benzo [i,] pyrrol o [2, 3-f] [10,4] oxazazulene

 5,5a-Dihydro ht-13-Α (5.5 mg) from Example 12 was dissolved in 1 mL of tetrahydrofuran, and 6 mg of sodium hydride (oil suspension containing about 60%) was added under ice-cooling. At room temperature

Stir for 30 minutes. After that, 10 mg of brombenzoyl alk mouth light was added, and the mixture was further stirred at room temperature for 1 hour. The reaction solution was diluted with a saturated aqueous solution of sodium hydrogen carbonate and partitioned with ethyl acetate. The ethyl acetate layer was separated. After washing with saturated saline and drying, the solvent is distilled off, and the residue is subjected to silica gel TLC (Merck Pre-Trade TLC plate silica gel 60-254, chloroform: methanol = 9: 1, 1 ^ = 0.52-0.59) ) To give 6 mg of the title compound.

〖Physical and chemical properties]

Solubility: Soluble in ethyl acetate, acetone, black form, methanol, insoluble in water

ESI-MS: m / z 413 [M + H] ÷: "Br, 415 [M + H] ^÷ : ⁸¹ Br

Color reaction: Positive for Dragendorf reagent, chloride and sulfuric acid

 HPLC: retention time 11.7 min (Devosyl 0DS-T-5 4.6 φ x 150 hall (Nomura Chemical Co., Ltd.), mobile phase 30% acetonitrile monohydrate (0.1% trifluoroacetic acid), flow rate 1 mL / min , Detection UV 210 nm)

Example 14 (Synthesis of 6-0H ht-13-Α)

 (6R, 6aR, 9aS) -6-hydroxy-7-methyl-6a, 8, 9, 9a-tetrahydro-4H, 6H, 7H-benzo [i, j] pyrrolo [2, 3-f] [10, 4 ] oxazazulene

ht-13--13 (lOmg) was dissolved in 1 mL of THF-water (9/1), and 20 mg of 2,3-dichloro-5,6-dicyano-P-benzoquinone was added under ice-cooling in a nitrogen stream. In addition, the mixture was stirred at room temperature for 3 hours. After evaporating the solvent, the residue was dissolved in 90% ethyl acetate-methanol, and 4 mg of a crude product of the title compound was obtained by alumina column chromatography (2 g; ethyl acetate-methyl alcohol). This was purified by silica gel TLC (Merck Pre-Coated TLC plate silica gel 60F254, black form: methanol = 85:15, Ri = 0.45 to 0.36) to obtain 3 mg of the title compound.

[Physicochemical properties]

Solubility: Ethyl acetate, Acetone, Black form, Soluble in methanol, Insoluble in water ESI-MS: m / z 245 [M + H] ⁺

 NMR (polychrome form, 300 MHz) δ: 2.11 (IH, m), 2.39 (IH, m), 2.45 (3H, s), 2.77 (1H, g-like, ^ 9.0 Hz), 2.85 (IH, dd, J = 6.0, 2.4 Hz), 3.21 (IH, td, J = 8.7, 2.1 Hz), 5.15 (IH, m), 5.26 (IH, d, J = 2A Hz), 6.62 (1H, dd, J = 8.1, 0.9 Hz), 6.97 (1H, dd, J = 8A, 0.9 Hz), 7.06 (IH, dd, J = 8.1,8.1 Hz), 7,34 (1H, d, J = 2.7 Hz), 8.29 (IH, br.s)

l ³ C NMR (double-mouthed form, 150 MHz) δ: 30.21, 39.79, 55.56, 62.38, 76.00, 79.33, 104.28, 106.56, 113.85, 116.03, 123.19, 124.14, 138.41, 151.98 Color reaction: Dragen-Gendorf reagent Positive for sulfuric acid

 HPLC: Retention time 5.2 min (Devosyl 0DS-T-5 4.6 φ x 150 mm (Nomura Chemical Co., Ltd.), mobile phase 18% acetonitrile monohydrate (0.1% trifluoroacetic acid), flow rate 1 mL / min , Detection UV 210 nm)

Real 15 (synthesis of 6-0Me ht-13-Α)

(6, 6a5, 9a5) -6 -methoxy-7-methyl-6a, 8 3 9, 9a-tetrahydro-4, 6, ΊΗ- benzo [j] pyrrolo [2,3- /] [10,4] oxazazulene

 ht-13-Α (20 mg) was dissolved in methanol (4 mL), DDQ (44 mg) was added at room temperature in a nitrogen stream, and the mixture was stirred for 1 hour. 36 mL of I-yl acetate was added to the reaction mixture, and the mixture was made □-into an alumina column (4 g of aluminum metal oxide). 90% ethyl acetate-methanol 50 m

Eluted with L. The residue obtained by evaporating the solvent from the eluate was crystallized in acetonitrile. This gave 18.8 mg of the title compound.

[Physicochemical properties]

Solubility: Soluble in ethyl acetate, acetone, black form, and methanol. Insoluble in water. LSIMS: m / z 227 [M-0CHJ +, 259 [M + H] 517 [2M + H] +

'HNMR (double-mouthed form, 300 MHz) (5 (ppm): 2.06 (1H, m), 2.40 (1Η, m), 2.47 (3Η, s), 2.60 (1Η, Μ), 2.8K1H, dd _} J = 6.3, 2.1 Hz), 3.20 (1Η, dd, J = 8.1, 8.1 Hz), 3.39 (3H, s), 4.73 (1H, d, J = 2.1 Hz), 5.23 (1H, ddd, J = 9.3 , 5.7, 2.4Hz), 6.64 (1H, dd, J = 8.0, 1.2Hz), 6.97 (1H, dd, 8.0, 1.2Hz), 7.08 (1H, dd, J = 8.0, 8.0Hz), 7.16 (1H , D, J = ZA Hz), 8.65 (1H, br.s)

1 ³ CR (heavy black port Holm, 150 MHz) (5: 31.09 , 40.14, 56.03, 56.05, 71.85, 77.24, 79.58, 103.96, 106.77, 111.45, 116.03, 123.30, 123.45, 138.87, 152.16 Color reaction: Dora one Gendorf reagent, chloride, positive for sulfuric acid

 TLC: Merck Pre-Ted TLC plate Silica gel 60F254 was used, and Rf value was 0.23 when developing solvent: chloroform: methanol = 9: 1.

HPLC: Use ^ S "0DS-5 4.6 X 150 mm (Nomura Chemical Co., Ltd.) as the mobile column, and use a mobile phase as a mobile phase, such as ash, lash, ent (0.1% trif acetic acid, 30% → 95% acetonitrile / water). 7 min, 0.1 ° /. Trif D mouth acetic acid, 95 acetonitrile / water 3 min), the retention time was 3.0 min when the flow rate was 1 mL / min and the detection was performed at UV 220 nm.

Example 16 (Synthesis of 6-OEt ht-13-A)

(6R, 6a.R, 9a5) -6-ethoxy-7-methyl-6a, 8, 9, 9a-tetrahydro-4 ^, also H, ΊΗ-benzo [i, J] pyrrolo [2, 3- /] [10,4] oxazazulene

ht-13-Α (20 mg) was dissolved in ethanol (4 mL), DDQ (44 mg) was added at room temperature in a nitrogen stream, and the mixture was stirred for 30 minutes. 36 mL of ethyl acetate was added to the reaction solution, and the mixture was subjected to D-application to an alumina column (4 g of Merck aluminum oxide), and eluted with 50 mL of a solvent (ethyl acetate: methanol = 9: 1). The solvent was distilled off from the eluate, and 31 mg of the residue obtained was crystallized in acetate nitrile to give 10 mg of the title compound. [Physicochemical properties 性

Solubility: Soluble in ethyl acetate, acetone, black form, methanol, insoluble in water. Molecular formula: CnHjoNiOj

Positive ion ESI-MS: m / z 227 [M-0CH _! CH ₃ ] ⁺ , 273 [M + H] +

Negative Vion ESI-MS: m / z 271 [M-H]-

NMR (polychrome form, 300 MHz) δ (ppm): 1.16 (3H, t, J = 6.9 Hz), 2.06 (1H, m), 2.38 (1H, m), 2.45 (3H, s), 2.57 ( 1H, m), 2.80 (1H, dd, J = 5.7, 1.8 Hz), 3.18 (1H, dd, J = 8.1, 8.1 Hz), 3.66 (2H, m), 4.89 (1H, d, J = 1.8 Hz) ), 5.23 (1H, ddd, J = 9.0, 5.7, 2.1 Hz), 6.63 (1H, dd, J = 8.0, 0.9 Hz), 6.96 (1H, dd, = 8.0, 0.9 Hz), 7.07 (1H, dd , J = 8.0, 8.0 Hz), 7.15 (1H, d, J = 2A Hz), 8.42 (1H, br.s) Color reaction: Positive for Dragendorf reagent, chloride and sulfuric acid

 TLC: Merck Pre-Ted TLC plate Silica gel 60F254, Rf value was 0.24 when developing solvent: chloroform: methanol = 9: 1.

HPLC: Using a column of 、, D, D-sil 0DS-5 4.6 φ χ 150 mm (Nomura Chemical Co., Ltd.), and using 0.1% triflo-D acetic acid-30% acetate :: tolyl / water as the moving bed and a flow rate of 1%. The retention time in the case of detection at UV 220 nm as mL / min was 3.8 minutes.

Example 17 (Synthesis of 4-Acetyl-6-acetoxy ht-13-A)

 (65,6ai ?, 9a5) -4-acetyl-6-acetoxy-7-methyl-6a, 8,9,9a-tetrahydro-4 ^, 6ff, ΊΗ-benzo [j] pyrrolo [2,3-] [ 10,4] oxazazulene

The 6-OH ht-13-A (8 mg) of Example 14 was chick. Dissolved in water (2 mL), acetic anhydride (200 μL) was added, and the mixture was stirred at room temperature for 15 hours. Methanol (5 mL) was added to the reaction solution, and the mixture was stirred for 5 minutes, and then the solvent was distilled off. Aqueous sodium bicarbonate was added, the mixture was partitioned with ethyl acetate, and the ethyl acetate layer was washed with saturated brine, dried with polish, and the solvent was distilled off. The residue was dissolved in methanol, and 5.1 mg of a crude product of the title compound was obtained by TLC (Merck Pre-coated TLC Plate silica gel 60F254, chloroform: methanol = 9: 1). This was developed. The solvent was changed and the product was purified again by TLC (ethyl acetate: methanol = 9: 1, Rf = 0.4). OAV §Re d

White

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+ [H + w] ovz ^z ro: sH-isa ^-^ ^

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69) ν-εΐ- —. . Κ3- λ ¾ ^ LVO- 88'0 =, L2) V-CI- ^ NO- A ¾ ^ ^ 9O ~ L ^ O = JH 'Π iC-i- ^ 翘 翘 Drilling: T1: Okina ¾ϋ 暂'

^^ ίι 4 —: Ye, ^ —Yeong ^) 0 Ye ¾ ³ ∞. ¾ nZlOmd £ / lDd 6066S / 00OAV 7.01 (1H, br.s), 7.02 (1H, dd, J = 7.8, 1.2 Hz), 7.08 (1H, dd, J = 7.8.7.8 Hz), 8.15 (1H, br.s)

i3C NMR (Heavy mouth form, 150 MHz) δ (ppm): 28.94, 29.40, 35.12, 39.04, 64.87, 80.74, 105.32, 106.96, 108.61, 117.19 .. 117.95, 120.62, 123.19, 138.00, 150.75 Color reaction : ョ-, positive for sulfuric acid

 TLC: Merck Pleitted TLC plate Silica gel 60F254, Rf value is 0.69 when developing solvent is used as solvent: methanol: methanol = 9: 1, toluene: ethyl acetate = 2: 1 as developing solvent In this case, the Rf value was 0.34.

HPLC: Develosyl ODS-T-54.6 φ x 150 mm (Nomura Chemical Co., Ltd.) for the column, 0.1% trifluoroacetic acid [40% acetonitrile-water] as the mobile phase, and the flow rate was 1 mL / min. As a result, the retention time when detected at UV 210 nm was 12.6 minutes. Example 19 (Synthesis of 7-Demethyl ht-13-A)

 7-CNht-13-A (27 mg) of Example 18 was dissolved in 18 mL of 33% acetic acid, zinc (Mallinckrodt) (300 mg) was added, and the mixture was stirred at 70 ° C for 3 hours. After removing zinc from the reaction mixture, it was made alkaline with aqueous ammonia and extracted twice with 50 mL of ethyl acetate. Then, after washing with saturated saline, ethyl acetate was distilled off to obtain 28 mg of a residue. 28 mg of the residue was purified by TLC (Merc Pre-coated TLC Plate silica gel 60F254, developing solvent; methanol: methanol = 4: 1, Rf = 0.11 to 0.32) to obtain 17 mg of the title compound.

[Physicochemical properties]

Solubility: Soluble in ethyl acetate, acetone, black form, methanol, insoluble in water ESI-MS: m / z 215 [M + H] +

iH NMR (double-mouthed form, 300 MHz) δ (ppm): 2.22 (1H, m), 2.50 (1H, m), 2.74 (1H, br.t), 3.21 (2H, m), 3.42 (2H, m), 4.37 (1H, m), 6.65 (1H, dd, J = 7.8, 0.9 Hz), 6.98 (1H, br.s), 6.99 (1H, dd, J = 7.8, 0.9 Hz), 7.08 (1H , dd, J = 7.8,7.8 Hz), 8.11 (1H, br.s)

Color reaction: Positive for ト, Raquen's "Ndorff Reagent, Hot", sulfuric acid TLC: Merck Pre-Trade TLC Plate Silica gel 60F254, Rf value was 0.11 when developing solution was used with a form solvent: methanol = 4: 1.

HPLC: using Deverosil ODS-T-54.6 ø X 150 mm (Nomura Chemical Co., Ltd.) for the column, and 0.1% trifluoroacetic acid mono [20% → (15 minutes) 50%] acetonitrile reluuwater gradient as the mobile phase The retention time was 6.4 minutes when the flow rate was ImL / min and the detection was performed at UV 210 nm.

Perform ^ 20 (Synthesis of 7-Propenyl ht-13-A)

 7-Demethyl ht-13-Α (7 mg) of Example 19 was dissolved in dimethylformamide (0.7 mL), and sodium hydrogen carbonate (14 mg) and arylpromide (14 mg) were added. Room temperature Stir for 1 hour. After distilling off dimethylformamide under reduced pressure, 5 mL of water was added, and the mixture was extracted twice with 10 mL of ethyl acetate. Then, after washing with saturated saline, ethyl acetate was distilled off to obtain 7 mg of a residue. The residue (7 mg) was purified by TLC (Merck Pre-Trade TLC plate silica gel 60F254, developing solvent; chloroform: methyl = 9: 11 ^ = 0.49 to 0.58) to obtain 6 mg of the title compound.

[Physicochemical properties]

Solubility: Soluble in ethyl acetate, acetone, black form, methanol, insoluble in water Positive ion ESI-MS: 255 [M + H] ⁺

Negative ion ESI-MS: 253 [MH] -iH NMR (double-mouthed form, 300 MHz) δ (ppm): 2.09 (1H, m), 2.49 (2H,). 2.69 (2H, m), 2.88 (1H , Dd, J = 13.2, 7.8 Hz), 3.16 (1H, m), 3.46 (1H, d, J-12.0 Hz), 3.58 (1H, m), 4.47 (1H, m), 5.16 (1H, d, J = 10.2 Hz), 5.25 (1H, d, J = 17.1 Hz), 5.95 (1H, m), 6.63 (1H, dd, J = 7.8, 0.9 Hz), 6.98 (1H, dd, J = 0.78, 0.9 Hz), 6.99 (1H, br.s), 7.07 (1H, dd, J = 7.8,7.8 Hz), 8.08 (1H, br.s)

13C NMR (double-mouthed form, 75 MHz) δ (ppm): 29.40, 30.61, 52.25, 56.84, 70.99, 86.56, 104.03, 106.16, 111.12, 117.16, 117.78, 120.15, 122.89, 135.12, 138.76, 151.90 Color reaction: Positive for sulfate and sulfuric acid

 TLC: Merck Pre-Ted TLC plate Using silica gel 60F254, the Rf value was 0.54 when the developing solvent was 9: 1 chloroform-form: methanol.

HPLC: Develosil ODS-T-54.6 φ x 150 mm (Nomura Chemical Co., Ltd.) on the column, 0.1% trifluoroacetic acid mono [20% → (15 minutes) 50%] acetonitrile monohydrate gradient as the mobile phase The retention time was 9.2 minutes when the flow rate was lm L / min and the detection was performed at UV 210 nm.

Example 21 (Synthesis of 7-n-Propyl ht-13-A)

 7-Demethyl ht-13-Α (8 mg) of Example 19 was dissolved in dimethylformamide (0.8 mL), and sodium hydrogencarbonate (16 mg) and then n-propyl bromide (16 mg) were added. Stir at ° C for 5 hours. After distilling off dimethylformamide under reduced pressure, add 5 mL of water and extract twice with 10 mL of ethyl acetate. Then, after washing with saturated saline, ethyl acetate was distilled off to obtain 7.5 mg of a residue. 7.5 mg of the residue was purified by TLC (Merck Pre-Trade TLC Plate Silica Gel 60F254, developing solvent; mouth form: methanol = 9: 1, Rf = 0.44 to 0.54) to obtain 7 mg of the title compound. Was.

[Physicochemical properties]

Solubility: Ethyl acetate, Acetone, Black form, Soluble in methanol, Insoluble in water Positive ion ESI-MS: 257 [M + H] ⁺

Negative ion ESI-MS: 255 [MH]-iH NMR (form chromatography, 300 MHz) 5 (ppm): 0.95 (3H, t, .7 = 7.5 Hz), 1.57 (2H, m), 2.14 (2H , m), 2.46 (2H, m), 2.65 (2H, m), 2.83 (1H, m), 3.23 (1H, m), 3.44 (1H, m), 4.46 (1H, m), 6.63 (1H, dd, J = 7.8, 0.9 Hz), 6.98 (1H, dd, J = 7.8, 0.9 Hz), 6.99 (1H, br.s.), 7.07 (1H, dd, J = 7.9,7.9 Hz), 8.07 ( 1H, br.s)

13C NMR (double-mouthed form, 75 MHz) δ (ppm): 12.16, 21.29, 29.52, 30.75, 52.14, 56.22, 71.79, 86.51, 103.98, 106.13, 111.31, 117.20, 120.09, 122.87, 138.78, 151.95 Color reaction: G, 'Raken'Ndorff's reagent, K-, positive for sulfuric acid

TLC: Merck Pleitted TLC plate Silica gel 60F254, and the Rf value was 0.44 in the case of using a developing solvent of black form: methanol = 9: 1. HPLC: Derosil ODS-T-54.6 φ x 150 mm (Nomura Chemical Co., Ltd.) on the column, 0.1% trifluoroacetic acid monoacetic acid [20 ° /. → (15 min) 50%] Using acetonitrile Lilue water gradient, the retention time was 9.5 min when the flow rate was I mL / min and detected at UV 210 nm.

The reaction scheme of Examples 22 to 25 is shown below,

h-,, _λ Toluenesulfonyl chloride / NaH / THF

 or v?

 p-Bromobenzenenesulfonyl chloride / NaH / THF

 or (4-p-Methoxybenzenesulfonyl ht-13-Α) Ex 24 p-Methoxybenzenesulfonyl chloride / NaH / THF

 or

 p-Nitrobenzenesulfonyl chloride / NaH / THF

-N0 ₂

 6 '

 (4-p-Nitrobenzenesulfonyl ht-i 3-A) Ex 25 Example 22 (Synthesis of 4-Toluenesulfonyl ht-13-A)

 (6aE, 9aS) -4-Toluenesulfonyl-7-methyl-6a, 8, 9, 9a-tetrahydro-4H, 6H, 7H-benzo [i, j] yrrol o [2,3-f] [10,4] oxazazulene

ht-13-A (10 mg) was dissolved in tetrahydrofuran (2 mL), and sodium hydride (oil suspension containing about 60% oil, 6 mg) was added under ice-cooling. After stirring at room temperature under a nitrogen stream for 20 minutes, toluenesulfonyl chloride (17 _mg ) was added, and the mixture was stirred at room temperature under a nitrogen stream for 2 hours. The reaction was diluted with saturated aqueous sodium bicarbonate and partitioned with ethyl acetate. After washing with saturated saline and drying, the solvent was distilled off, and the residue was fractionated by thin layer gel thin layer chromatography (chloroform: methanol = 9: 1) to obtain 15 mg of the title compound. Was.

[Physicochemical properties]

Solubility: Ethyl acetate, Acetone, Black form, Soluble in methanol, Insoluble in water ESI-MS: m / z 383P] +

Ή NMR (double-mouthed form, 300 MHz) δ: 2.01 (1H, m), 2.35 (3H, s), 2.40 (3H, s), 2.44 to 2 · 63 (4Η, m), 3.09 (1H, t -like), 3.33 (1H, dd, J = 3.3 _; 15.3 Hz), 4.33 (1H, m), 6.75 (11-1, dd, J = 0.9, 8.1 Hz), 7.17 (lH, dd, J = 8.4 , 8.4 Hz), 7.23 (2H, d, J = 8.4 Hz), 7.36 (1H, br.s), 7.61 (1H, dd, J = 0.9, 8.1 Hz), 7.77 (2H, d, J = 8.4 Hz)

Color reaction: Positive for "Raken", Ndorff Reagent, Hot, sulfuric acid

 TLC: Rf value 0.62 (Merck Pleated TLC plate silica gel 60F254, black form: methanol = 9: 1)

HP LC: Retention time 6.91 min (Devosyl ODS-T-54.6 ø X 150 mm (Nomura Chemical Co., Ltd.), mobile phase 40% acetonitrile monohydrate (0.1% trifluoroacetic acid), flow rate lmL / min, detection (UV 210 nm)

Example 23 (Synthesis of 4-p-Bromobenzenesulfonyl ht-13-A)

 (6aR, 9aS) -4-p-Bromobenzenesulfonyl-7-methyl-6a, 8, 9, 9a-tetrahydro-4H, 6H, 7H-benzo [i, j] pyrrolo [2,3-f] [10, 4] oxazazulene

 ht-13-II (10 mg) was dissolved in tetrahydrofuran (2 mL), and sodium hydride (oil suspension containing about 60% oil, 10 mg) was added under ice-cooling. After stirring at room temperature under a nitrogen stream for 20 minutes, parabromobenzenesulfonyl chloride (23 mg) was added, and the mixture was stirred at room temperature under a nitrogen stream for 1 hour. The reaction solution was diluted with a saturated aqueous sodium hydrogen carbonate solution and partitioned with ethyl acetate. After washing with saturated saline and drying, the solvent was distilled off, and the residue was fractionated by thin layer gel thin layer chromatography (chloroform: methanol = 9: 1) to obtain 13 mg of the title compound. Was.

[Physicochemical properties]

Solubility: Soluble in ethyl acetate, acetone, black form, methanol, insoluble in water ESI-MS: m / z 447 ([M + H] ⁺ ; "Br), 449 ([Μ + Η] ⁺ : ⁸¹ ΒΓ)

ΐΗ NMR (double-mouthed form, 300 MHz) δ: 2.01 (1Η, m), 2.40 (3Η, s), 2.42 to 2.63 (4Η, m), 3.09 (1Η, t-like), 3.33 (1Η, dd, J = 2.7, 15.3 Hz), 4.33 (1H, m), 6.77 (1H, d, J = 8.4 Hz), 7.19 (1H, dd, J = 8.4, 8.4 Hz), 7.32 (1H, br) s), 7.58 (2H, d, J = 8.7 Hz), 7.68 (1H, d, J = 8.4 Hz), 7.73 (2H, d, J = 8.7 Hz)

Color reaction: Positive for G, Randolph's reagent, chloride, sulfuric acid

TLC: Rf value 0.59 (Merck Precoated TLC Plate silica gel 60F254, Black mouth form: methanol = 9: 1)

HPLC: Retention time 17.07 minutes (Devosyl ODS-T-5 4.6 φ X 150 mm (Nomura Chemical Co., Ltd.), mobile phase 35% acetate nitrile water (0.1% trifluoroacetic acid), flow rate 1 m

(L / min, detection UV 210 nm)

Example 24 (Synthesis of 4-p-MethoxybenzenesulfonyI ht-13-A)

 (6aR, 9aS) -4-p-Methoxybenzenesulionyl-7-methyl-6a, 8,9,9a-tetrahydro-

4H, 6H, 7H-benzo [i, j] pyrrolo [2,3-f] [10,4] oxazazulene

 ht-13-Α (10 mg) was dissolved in tetrahydrofuran (2 mL), and sodium hydride (about 60% oil suspension, 10 mg) was added under ice-cooling. After stirring at room temperature under a nitrogen stream for 20 minutes, paramethoxybenzenesulfonyl chloride (19 mg) was added, and the mixture was stirred at room temperature under a nitrogen stream for 1 hour. The reaction solution was diluted with a saturated aqueous sodium hydrogen carbonate solution and partitioned with ethyl acetate. After washing and drying with a saturated saline solution, the solvent is distilled off, and the residue is subjected to thin gel gel thin layer chromatography (black form: methanol).

= 9: 1) to give 15 mg of the title compound.

[Physicochemical properties]

Solubility: Soluble in ethyl acetate, acetone, black form, methanol, insoluble in water

ESI-MS: m / z 399 [M + H] ⁺

iH NMR (double-mouthed form, 300 MHz) δ: 2.01 (1H, m), 2.40 (3H, s), 2.42 to 2.64 (4H, m), 3.09 (lH, t-like), 3.33 (1H, dd , J = 3.3, 15.3 Hz), 3.80 (3H, s), 4.33 (1H, m), 6.75 (1H, dd, J = 0.9, 8.4 Hz), 6.89 (2H, d, J = 9.0 Hz), 7.12 (1H, dd, J = 8.4, 8.4 Hz), 7.50 (1H, br.s), 7.61 (lH, dd, J = 0.9, 8.1 Hz), 7.82 (2H, d, J = 9.0 Hz) : Positive for G, 'Raque', Ndolph's reagent, G-, and sulfuric acid

TLC: Rf value 0.54 (Merck Pleated TLC plate silica gel 60F254, black form: methanol = 9: 1) HPLC: Retention time 10.08 minutes (Develosyl ODS-T-54 · 6 ø x l50 mm (Nomura Chemical Co., Ltd. Company), mobile phase 35% acetonitrile Lilue water (0.1% trifluoroacetic acid), flow rate 1 m (L / min, detection UV 210 nm)

Example 25 (Synthesis of 4-p-Nitrobenzenesulfonyl ht-13-A)

 (6aR, 9aS) -4-p-Nitrobenzenesulfonyl-7-meth l-6a, 8, 9, 9a-tetrahydro-

4H, 6H, 7H-benzo [i, j] pyrrolo [2,3-f] [10,4] oxazazulene

 ht-13-Α (10 mg) was dissolved in tetrahydrofuran (2 mL), and sodium hydride (oil suspension containing about 60% oil, 10 mg) was added under ice-cooling. After stirring at room temperature under a nitrogen stream for 20 minutes, paranitrobenzenesulfonyl chloride (20 mg) was added, and the mixture was stirred at room temperature under a nitrogen stream for 1 hour. The reaction solution was diluted with a saturated aqueous sodium hydrogen carbonate solution and partitioned with ethyl acetate. After washing with a saturated saline solution and drying, the solvent was distilled off, and the residue was fractionated by thin layer gel thin layer chromatography (chloroform: methanol = 9: 1) to obtain 5 mg of the title compound. Was.

[Physicochemical properties]

Solubility: Ethyl acetate, Acetone, Black form, Soluble in methanol, Insoluble in water ESI-MS: m / z 414 [Μ + Η] ⁺

iH NMR (double-mouthed form, 300 MHz) δ: 2.02 (1H, m), 2.40 (3H, s), 2.42 to 2.63 (4H, m), 3.10 (1H, t-like), 3.33 (1H, dd , J = 3.3, 15.6 Hz), 4.33 (lH, m), 6.79 (1H, dd, J = 0.9, 8.1 Hz), 7.22 (1H, dd, J = 8.1, 8.1 Hz), 7.33 (1H, br. s), 7.61 (1H, dd, J = 0.9, 8.1 Hz), 8.05 (2H, d, J = 9.0 Hz) .. 8.25 (2H, d, J = 9.0 Hz),

Color reaction: positive for G, Randolph's reagent, G, and sulfuric acid

 TLC: Rf value 0.57 (Merck Pleated TLC plate silica gel 60F254, black-mouthed form: medium = 9: 1)

HPLC: Retention time 10.68 min (Develosil ODS-T-54.6 φ X 150 mm (Nomura Chemical Co., Ltd.), mobile phase 35% acetonitrile-water (0.1% trifluoroacetic acid), flow rate 1 mL / min, (Detection UV 210 nm) The reaction schemes of Examples 26 to 28 are shown below.

 (4-Toluenesulfonyl-5,5a-dihydro ht-13-Α) g Example 26 (Synthesis of 4-Benzoyl-5,5a-dihydro ht-13-Α)

 (5aS, 6aR, 9aS) -4-Benzoyl-7-methyl-5, 5a, 6a, 8, 9, 9a-hexahydro-4H, 6H, 7H-benzo [i, j] pyrrolo [2,3-f] [10,4] oxazazulene

 5,5a-Dihydroht-13-A (10 mg) obtained in Example 12 was dissolved in tetrahydrofuran (2 mL), and sodium hydride (oil suspension containing about 60% oil, mg) and stirred at room temperature for 20 minutes. Thereafter, Benzoiruku mouth light (13 mg) was added, and the mixture was further stirred at room temperature for 30 minutes. The reaction solution was diluted with a saturated aqueous solution of sodium hydrogen carbonate and partitioned with ethyl acetate. After the ethyl acetate layer was washed with saturated saline and dried, the solvent was distilled off, and the residue was subjected to silica gel thin layer chromatography (Merck Precoated TLC plate, silica gel 60F254, gel form: methanol = 92: 8). , Rf = 0.42-0.56) to give 15 mg of the title compound.

[Physicochemical properties]

Solubility: Ethyl acetate, Acetone, Black form, Soluble in methanol, Insoluble in water ESI-MS: m / z 335 [M + H] ⁺

iHNMR (double-mouthed form, 300 MHz) δ: 1.25 (1H, m), 1.96 (1H, m), 2.17 (1H, m), 2 · 27 to 2 · 36 (2Η, m), 2.34 (3H, s), 2.39-2.50 (2H, m), 3.11 (1H, m), 3.44 (1H, m), 3.74 (1H, t-like, J = 10.8 Hz), 3.84 (1H, m), 4.26 (1H , br.s), 6.72 (1H, d, J = 8.1 Hz), 726 to 756 (7Η, m)

Color reaction: Positive for --ke, 'Ndorff Reagent, Hot ", sulfuric acid

TLC: Rf value 0.60 (Merck Pleated TLC plate silica gel 60F254, 917

¾ ^ q> ^ ii) UIUI OST x ø d-9-i-s o a ^) ^ go'i m ^: 〇 dt dt H

 (1: 6 = / ^ —: Ma ^ ψ a a ^ ζΖ ^ 32 ^ 09 ^ ^ c ^ 4 ー ΟΊΙ i {一 匚 4 ^^) IS'O door · 0 Ί 1

 ^ ^ mm \ -H-E ^ ΜΉ ^ Λ ^^ Α: ¾¾GO

( ^Z H i'8 = f '') V2'S '( ^sj q ΉΤ) Τ0-8' ( ^Ζ Η Δ "8 = Γ 'Ρ') 2 L '(s-iq' HT) 8T i '(V ^ 'Ηΐ) 8n9'(s' ^ q 'm) ZV' Hl) 98-8 '(^ H 90T = f' ^ Π- ^ 'm) ^' (ra (HI) S '(ra' ΗΙ) Η · ε '' (ra 'HZ) 9fZ'(s' H8) S8'S '(∞' liZ) l Z OZ ~ 6TS '(ra' HI) i6'I '(^' Hl) l8'T: P (zHK 008 'Ma Ψ aa heavy) Hi

+ [H + H] 08S z / m: SW-IS3 氺^v ¾ k 1 (-r, Ma ^ ψα α, ^ & 翘 翘

 [¥ kSi¾ ^ S 呦]

. ¾ S m 8 坳 aq 齄 辇, 迪 ^^ (69 · 0 ~ ε 0 = ίΗΊ: 6 = SI (— r ^: Ma ^ a a ^ gsi09 Λ (^ — 0ΊΧ il, ^ — 匚: ΰ

、 っ辛畕 ¾翁¾ 、¾翁 ¾Λ ¾ST 氺 ¾葶¾ ェ翘 ¾ 。 つ ェ翊 ¾ 、っ^娑 -c

畕, 畕 畕 ¾ ¾, ¾ ¾Λ ¾ ¾ ST ¾ 翘 翘 翘 ¾.翊 翊 翊 ¾

¾ 氺 ma · n

(3 m 9ΐ) ίΐ yy. ^ A Futari®. Φί¾ · & Ο Ζ 遝 § ¥ ΟΪ (s ui οι '{^ rnm ^^ ° 09 ^) Ma> Mine 氺 ^^ ^ m ^ (^ Ζ) ^ ^ a' ^ (S ui oi) ν- εΐ-ΐ ^ oa qi (i-¾g ^c g ¾ # ϋ Z \ W

Ή^¾9 'S^BS) -oa iCq¾x8 -¾6 '6' 8 '¾9 ^e ¾S

(Ή ^¾ 9 'S ^B S)

 (¾tf a, ® v-SI- ^ q oa ^ qi -Bg'g-iiozua oj ^ iN-ci- ^) S (ran 0TS ΛΠ W i ^ / Ί ^ ΐ, [4be ^ '(P- ^

ΓΗ-^ ^)% 09 (^ si)% 9]-n a d% ro Ψ¾Ι Ψ¾Ι ゝ (¾ ^^

r ^ara osi x ø e ^ gi-sao a ^) S¾¾ ^: o τ d H

(Γ · 6 = ^ —, ： Ma ^ a mouth fZZlO / OOdf / IDd 6066S / 00OAV Mobile phase 30% acetonitrile-water (0.1% trifluoroacetic acid), flow rate lmL / min, detection UV 210 nm)

Example 28 (Synthesis of 4-Toluenesulfonyl-5,5a-dihydro ht-13-A)

(5aS, 6aR, 9aS) -4-Toluenesulfonyl-7-meth l-5,5a, 6a, 8, 9, 9a-hexahydro-4H.6H, 7H-benzo [i, j] pyrrolo [2,3-f ] [10,4] oxazazulene

 5,5a-Dihydro ht-13-Α (10 mg) obtained in Example 12 was dissolved in tetrahydrofuran (1 mL), and sodium hydride (oil suspension containing about 60% oil, 10 mL) was added under ice cooling. mg) and stirred at room temperature for 20 minutes. Thereafter, toluenesulfonyl chloride (17 mg) was added, and the mixture was further stirred at room temperature for 20 hours. The reaction solution was diluted with a saturated aqueous sodium hydrogen carbonate solution and partitioned with ethyl acetate. After the ethyl acetate layer was washed with saturated saline and dried, the solvent was distilled off, and the residue was subjected to silica gel thin-layer chromatography (Melk Plecote TLC plate, silica gel 60F254, gel form: methanol = 9: l) , Rf = 0.43-0.59) to give 6 mg of the title compound.

[Physicochemical properties]

Solubility: Ethyl acetate, Acetone, Black form, Soluble in methanol, Insoluble in water ESI-MS: m / z 385 [M + H] ⁺

iHNMR (double-mouthed form, 300ΜΗζ) δ: 1.25 (1H _; m), 1.88 (1H, m), 2.07 (1H, m), 2.22 (1H, m), 2.34 (3H, s), 2.38 (3H, s), 2.30-2.45 (2H, m), 3.11 (2H, m), 3.39 (1H, t-like, J = 10.2 Hz), 3.68 (1H, m), 4.29 (1H, t-like, J = 9.0 Hz), 6.65 (1H, dd, J = 0.9, 8.1 Hz), 7.12 (1H, dd, J = 8.1, 8.1 Hz), 7.23 (2H, d, J = 8.4 Hz), 7.38 (1H, dd, J = 0.9, 8.1 Hz), 7.67 (2H, d, J = 8.4 Hz)

Color reaction: Positive for sulfate and reagent

 TLC: Rf value 0.57 (Merck Pleated TLC plate silica gel 60F254, black form: methanol = 9: 1)

HP LC: Retention time 8.21 min (Develosyl 003-1 ¹ -54.60 150 ^^ 1 (Nomura Chemical Co., Ltd.), mobile phase 35% acetonitrile-water (0.1% trifluoroacetate), flow rate 1 mL / Min, detection UV 210 nm)

The reaction scheme of Examples 29 to 32 is shown below,

-Bromo ht-13-Α 3,5-Dibromo ht-13-Α

 ht-13-A-7-oxide (Ex 30)

 ht-13-B-7-oxide (Ex 31)

 5,5a-Dihydro ht-13-A-7-oxide (Ex 32)

Example 29 (Synthesis of 5-Bromo ht-13-A and 3,5-Dibromo ht-13-A)

 (1) 5-Bromo ht-13-A: (6aR, 9aS) -5-Bromo-7-methyl-6a, 8, 9, 9a-tetrahydro-4H, 6H, 7H-benzo [i, j] pyrrolo [ 2, 3-f] [10, 4] oxazazulene

(2) 3,5-Dibromo ht-13-Α: (6aR, 9aS) -3, 5-Dibromo-7-methyl-6a, 8, 9, 9a-tetrahydro-4H, 6H, 7H-benzo [i, j] pyrrolo [2, 3-f] [10,4] oxazazulene ht-13-Α (30 mg) was dissolved in black-mouthed form (7 mL), N-promosuccinic imide (28 mg) was added, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated and separated by thin-layer chromatography (Merck Precoated TLC Plate, silica gel 60F254, developing solvent: black-mouthed form: methanol = 93: 7), and 3,5-Dibromo ht-13-A (7 mg) and crude 5-Bromo ht-13-A. Crude 5-Bromo ht-13-A is for high performance liquid chromatography (column: Develosil ODS-T-510-5 x 250 mm (Nomura Chemical Co., Ltd.), mobile phase: 0.1 ° /. Trifluoroacetic acid [20 ° /.→ (30 min) → 25% (acetonitrile monohydrate) gradient], flow rate: 3 mL / min, retention time: 19 to 24 min), and the eluate is sodium hydrogen carbonate After neutralization with an aqueous solution, the mixture was extracted with ethyl acetate. The ethyl acetate layer was treated in a conventional manner to give 5-Bromo ht-13-A (6 mg).

[Physicochemical properties of 5-Bromo ht-13-Α]

Solubility: Soluble in ethyl acetate, acetone, black form, methanol, insoluble in water ESI-MS: m / z 307 ([M + H] +: ⁷⁹ Br), 309 ([M + H] +: "Br)

iH NMR (deuterium chloroform, 300 MHz) δ: 2.06 (1H, m), 2.45 (3H, s), 2.42 to 2.64 (4H, m), 3.12 (1H, t-like), 3.26 (lH, dd, J = 2.1, 13.8 Hz), 4.44 (1H, m), 6.63 (1H, dd, J = 0.9, 8.1 Hz), 6.90 (1H, dd, J = 0.9, 8.1 Hz), 7.05 (1H, dd, J = 8.1, 8.1 Hz), 8.14 (1H, br.s)

Color reaction: Positive for G, 'Raque', 'Ndorff Reagent, Gamma, and sulfuric acid'

 TLC: Rf value 0.32 (Merck Pleated TLC plate silica gel 60F254, black form: methanol = 95: 5)

 HP LC: Retention time 9.43 min (Develosil ODS-T-54.6 ø X 150 mm (Nomura Chemical Co., Ltd.), mobile phase 0.1% trifluoroacetic acid [20% → (15 minutes) → 50% (acetonitrile water) ) Gradient], flow rate l mL / min, detection UV210nm)

 [Physicochemical properties of 3,5-Dibromo ht-13-A]

Solubility: Soluble in ethyl acetate, acetone, black form, methanol, insoluble in water

389([M+H]+: siBr X 2) ESI-MS: m / z 385 ([M + H] ⁺ : "BrX 2), 387 ([M + H] ⁺ :

389 ([M + H] +: siBr X 2)

iH NMR (double-mouthed form, 300 MHz) δ: 2.04 (1Η, m), 2.43 (3Η, s), 2.41 to 2.61 (4H, m), 3.10 (1H, t-like), 3.24 (lH , Dd, J = 1.8, 13.2 Hz), 4.40 (lH, m), 6.54 (1H, d, J = 8.4 Hz), 7.16 (1H, (1, J = 8.4 Hz) .. 8.22 (1H, br. s)

Color reaction: Positive for ラ, Laundrdorf reagent,, and sulfuric acid

 TLC: Rf value 0.43 (Merck pre-coated TLC plate silica gel 60F254, porch mouth: medium = 95: 5)

 HP LC: Retention time 12.79 min (Develosil ODS-T-54.6 ø X 150 mm (Nomura Chemical Co., Ltd.), mobile phase 0.1 ° /. Trifluoroacetic acid— [20% → (15 min) —50% (acetonit Ril-water) gradient], flow rate l mL / min, detection UV210nm)

Implementation_Example 30 (Synthesis of ht-13-A-7-oxide)

 (6aR, 9aS) -7-methyl-6a, 8, 9, 9a-tetrahydro-4H, 6H, 7H-benzo [i, j] pyrrolo [2,3-f] [10,4] oxazazulene 7-oxide

 ht-13-A (10 mg) was dissolved in dichloromethane (1.Ί mL), and methacrylic acid perbenzoic acid (9 mg) was added, followed by stirring at room temperature for 30 minutes. The solvent was distilled off from the reaction mixture, and the residue was subjected to thin-layer chromatography (Merck Pre-Trade TLC Plate Siri Kiki 60F254, developing solvent: black form: methanol: concentrated aqueous ammonia = 40: 10: 1, Rf = 0.38 to 0.29) to give 4.5 mg of the title compound.

[Physicochemical properties]

Solubility: Soluble in methanol, ethanol and dimethyl sulfoxide

ESI-MS: m / z 245 [M + H] ⁺

iH NMR (Heavy Metal, 300 MHz) δ: 2.28 (1H, m), 3.00 (1H, m), 3.28 (3H, s), 3.34 to 3.48 (2H, m), 3.63 (1H, m ), 3.88 (1H, m) .4.02 (1H, m), 4.71 (1H, m), 6.53 (1H, dd, J = 2.1, 8.1 Hz), 6.96 (1H, dd, J = 8.1, 8.1 Hz) , 7.00 (1H, dd, J = 2.1, 8.1 Hz), 7.14 (1H, s)

Color reaction: Positive for ト, LaΓndorf reagent,-, sulfuric acid TLC: Rf value 0.24 (Merck Pleated TLC plate silica gel 60F254, mouth form: methanol: concentrated aqueous ammonia = 40: 10: 1)

 HP LC: Retention time 9.82 min (Develosil ODS-T-54.6 ø X 150 mm (Nomura Chemical Co., Ltd.), mobile phase 20% acetonituril water (0.1% trifluoroacetate), flow rate 1 mL / min, (Detection UV 210 nm)

Example 31 (Synthesis of ht-13-B-7-oxide)

 (6aR, 8R, 9aS) -7,8-diiiiethyl-6a, 8,9,9a-tetrahydro-4H, 6H, 7H-benzo [i, j] yrrol o [2,3-f] [10,4] oxazazulene 7-oxide

 ht-13-B (2 mg) was dissolved in 0.4 mL of dichloromethane, to which was added peroxybenzoic acid (2.5 mg), followed by stirring at room temperature for 1 hour. The solvent was distilled off from the reaction mixture, and the residue was subjected to thin-layer mouth chromatography (Merck Pleated TLC plate silica gel 60F254, developing solvent: mouth mouth form: methanol: concentrated ammonia water = 40: 10: 1, Rf = 0.41 to 0.34) to give 1.5 mg of the title compound.

[Physicochemical properties 性

Solubility: Soluble in methanol, ethanol and dimethyl sulfoxide

ESI-MS: m / z 259 [M + H] ⁺

iH NMR (deuterated methanol, 300 MHz) δ 1.57 (3Η, d, J = 7.2 Hz), 2.02 (1H, dd, J = 2.1, 14.1 Hz), 3.19 (3H, s), 3.25 (1H, m), 3.39 ~ 3.43 (2H, m), 3.90 (1H, dd, J = 1.5, 7.5 Hz), 4.06 (1H, dd, J = 6.6, 9.0 Hz), 4.70 (1H, m), 6.53 (1H, dd, J = 1.4, 8.4 Hz), 6.96 (1H, dd, J = 8.4, 8.4 Hz), 7.00 (1H, dd, J = 1.5, 8.4 Hz), 7.14 (1H, s)

Color reaction: Positive for "Traake," "Ant", "Ruff's reagent, Ford", sulfuric acid

 TLC: Rf value 0.30 (Merck Pleated TLC plate silica gel 60F254, mouth form: methanol: concentrated aqueous ammonia = 40: 10: 1)

HPLC: Retention time 11.21 min (Devosyl ODS-T-54.6 ø X 150 mm (Nomura Chemical Co., Ltd.), mobile phase 22% acetonitrile-water (0.1% trifluoroacetic acid), flow rate 1 mL / min, detection (UV 210 nm) Example 32 (Synthesis of 5,5a-Dihydro ht-13-A-7-oxide)

 (5aS, 6aR, 9aS) -7-methyl -5,5a, 6a, 8, 9, 9a-hexahydro-4H, 6H, 7H-benzo [i, j] pyrrolo [2,3-f] [10,4 ] oxazazulene 7-oxide

5,5a-Dihydroht-13-A (10 mg) obtained in Example 12 was dissolved in 1.5 mL of dichloromethane, and metabenzo-perbenzoic acid (9 _mg ) was added thereto. Stir for 2 hours. After distilling off the solvent, high-performance liquid chromatography {Column: Devosyl ODS-T-510 $ 5 x 250 mm (Nomura Chemical Co., Ltd.), Moving bed: 0.1% Trifluoroacetic acid— [5% → (25 minutes) 50% ( [Acetonitrile-water) gradient], flow rate: 3 mL / min, retention time: 11 to 13 minutes}, neutralized with aqueous ammonia, and concentrated. The salt was desalted with MC IGELCHP 20 P (75 to 150) (Mitsubishi Chemical Corporation) to obtain 7 mg of the title compound.

[Physicochemical properties]

Solubility: Soluble in methanol, ethanol, dimethyl sulfoxide, water

ESI-MS: m / z 247 [M + H] ⁺

iH NMR (deuterated methanol, 300 MHz) δ: 1.83 (1Η, m), 2.20 (1Η, m), 2.46 (1Η, m),

2.86 (1Η, m), 3.16 to 32828 (2Η, m), 3 · 25 (3Η, s), 3.51 to 3.64 (2Η, m), 3.74 (1Η, t-like,

J = 7.8 Hz), 3.98 (1H, m), 4.33 (1H, m), 6.34 (1H, d, J = 8.1 Hz), 6.40 (lH, d, J = 8.1

Hz), 6.92 (lH, dd, J = 8.1, 8.1 Hz)

Color reaction: Positive for,,,,,, and sulfuric acid

TLC: Rf value 0.27 (Merck Pleated TLC plate silica gel 60F254, mouth form: methanol: concentrated ammonia water = 40: 10: 1)

 HP LC: Retention time 8.33 min (Develosil ODS-T-54.6 ø X 150 mm (Nomura Chemical Co., Ltd.), mobile phase 0.1% trifluoroacetic acid [5% → (15 minutes) → 50% (acetonitrile Water) gradient], flow rate lmL / min, detection UV210nm)

Test example 1

Cell membrane preparation obtained from rat brain or HEK293 cells expressing the receptor After incubating the mixture of the radioligand and the test compound at several concentrations under the respective conditions, the mixture is suction-filtered over a Whatman GF / C filter. Liquid scintillation <Measure the radioactivity on the filter with a counter and calculate the 50¾ inhibitory concentration (1 „value) of the test compound for each specific binding, and calculate the Cheng-Prusofi [Biochem. Pharmacol. 22 (1973) 3099-3108], the Ki value was determined from the equation Ki = IC _s ./(l+[L]/Kd).

[L] indicates the radioligand concentration used, and Kd indicates the dissociation constant. Table 3 shows the experimental conditions for each receptor. Table 4 shows the results.

 (Table 3)

 Radioactive ligand from the receptor

Hippocampus 1 nM [ ³ H] 8-OH-DPAT 25 ° C 30 min

5-HT ₂ rat cerebral cortex 1 nM [ ³ H] Ketanserin 37 ° C 30 min Rat 5-HT 6

5-HT ₆ 8 nM [ ³ H] 5HT 25 ° C 120min

 (Thigh 293)

5-HT ₇ Hit 5-HT ₇ (HEK293) 0.5 nM [ ³ H] 5CT 25 ° C 120min

(Table 4-1)

 Ki (nM) SE

Example Compound number

5-HT! _A 5-HT ₂ 5-HT ₆ 5-HT ₇

1 ht-13-A 1.3 ± 0.4 180 ± 46 0.38 ± 0.12 2.5 ± 0.3

2 ht-13- B 15.0 ± 2.8 130 2.7 ± 2.1 370 ± 97

3 4- Me ht-13-Α 9.9 79> 8100 150

4 4-Benzoyl ht-13-Α 560 2400 2.4 ± 0.71 400

5 4-Boc ht-13-Α 250 ± 39 1300 ± 280 94 ± 23 1400 ± 210

6 3- Br ht- 13-A 4.5 190 270 24

7 5-SMe ht- 13- A 130 2000 2.3 ± 2.1 130

8 (1) 5-CI ht-13- A 34 ± 9.3 1400 ± 180 0.19 ± 0.06 270 ± 43

8 (2) 3-C1 ht-13-Α 3.6 ± 1.3 86 ± 14 1.4 ± 0.56 35 ± 2.8

_Q , lt-Butyl-5-SMe ht-

710 1900 6.6 ± 3.1 440 id— A

 1 0 1-t-Butyl ht-13-Α 0.27 ± 0.03 260 1.5 ± 1.4 2.8

_{1 1} 1,3-di-t-Butyl

 140 2600 590 ± 14 1200

XI 丄 d-Λ

 1 2 5,5a-dihydro ht-13-Α 4.3 3000 34 36

1 ₀ 5,5a-dihydro-p-Br-

6600 30000 53 ± 17 14000

DGIlZO 1 III »丄 0 Λ

 1 4 6- OH ht- 13-A 2000 29000 540 ± 170 470

18 (1) 7-CN ht-13-Α 4200〉 72000 38 ± 19 88000

18 (2) 7-CN seco ht-13-Α 35 10000 5.0 ± 1.5 2600

1 9 7-demethyl ht-13-A 1.9 ± 0.21 3200 ± 570 0.28 ± 0.08 72 ± 1.4

2 0 7-propenyl ht- 13- A 1 240 7.0 ± 0.1 7.8

2 1 7-n-propyl ht-13-Α 0.19 96 7.6 ± 2.3 1.5 (Table 4-2)

Formulation Example 1 The compound of the present invention showed strong affinity for various serotonin receptors.

 An appropriate amount of ht-13-A, microcrystalline cellulose, and magnesium stearate of Example 1 is mixed and tableted to give a tablet.

Formulation Example 2

 An appropriate amount of ht-13-A of Example 1, lactose, and magnesium stearate are mixed, granulated, and sized to obtain granules.

Formulation Example 3

A capsule is obtained by filling the granules obtained as in Formulation Example 2 into capsules. Industrial applicability

 This compound has an affinity for various serotonin receptors and is therefore useful as a preventive or therapeutic agent for various central diseases.

Claims

The scope of the claims 1. Equation (1):

 (Where R ¹ and R ² are each independently hydrogen, lower alkyl or halogen; R ³ and R ^4, which it independently hydrogen, lower alkyl, lower alkenyl, lower alkylcarbonyl, optionally substituted § reel carbonyl or Shiano; 11 ¹ Oyobi 1 ^3, together such connexion single bond May be formed; n is 0 or 1, provided that when n = l, wherein neither hydrogen R ³ and R ^4; 11 ⁵ is hydrogen; R ⁶ is hydrogen, hydroxy, lower alkoxy or lower alkyl carboxy; R ⁷ is hydrogen; R ⁸ and R ⁹ are each independently hydrogen, lower alkylthio or halogen; R ⁷ and R ⁹ may be taken together to form a single bond; R ^{1 D} is hydrogen, lower alkyl, lower alkylcarbonyl, lower alkyloxycarbonyl, optionally substituted arylcarbonyl, optionally substituted heteroarylcarbonyl, lower alkylsulfonyl, optionally substituted arylsulfonyl. Or optionally substituted heteroarylsulfonyl; R ¹¹ is hydrogen, lower alkyl or halogen; R ¹² represents hydrogen or lower alkyl) Or a pharmaceutically acceptable salt thereof, a prodrug thereof, or a hydrate thereof. 2. The compound of claim 1, wherein R ¹ and R ³ together form a single bond. 3. The compound according to claim 1, wherein R ² is hydrogen or lower alkyl. 4. R ⁴ is lower alkyl, a compound according to claim 1. 5. R ⁶ is hydrogen The compound of claim 1, wherein. 6. R ⁷ and R ⁹ together form a connexion single bond, a compound according to claim 1. 7:. R ⁸ is hydrogen, a compound according to claim 1. 8. The compound of claim 1, wherein R ^{1 Q} is hydrogen. 9. R ^{1 1} and R ^{1 2} is hydrogen The compound of claim 1, wherein. 10. R ¹ and R ³ together form a single bond; R ² is hydrogen or lower alkyl; R ⁴ is lower alkyl; R ⁶ is hydrogen; R ⁷ and R ⁹ are together The compound according to claim 1, wherein R ⁸ is hydrogen; R ¹ Q is hydrogen; R ¹¹ and R ¹² are hydrogen; n is 0; 1 1. R ¹ and R ³ combine to form a single bond; R ² is hydrogen or lower alkyl; R ⁴ is lower alkyl; R ⁶ is hydrogen; R ⁷ and R ⁹ are combined R ⁸ is hydrogen, lower alkylthio or halogen; R ¹⁰ is hydrogen, lower alkyloxycarbonyl, or optionally substituted arylcarbonyl; R ¹¹ and R ¹² are The compound of claim 1, wherein each independently is hydrogen or lower alkyl; 1 2. R ¹ and R ³ together form a single bond; R ² is hydrogen or lower alkyl; R ⁴ is lower alkyl; R ⁶ is hydrogen; R ⁷ and R ⁹ are together R ⁸ is hydrogen; R ¹ . Is a substituted arylsulfonyl; R ¹¹ and R ¹² are hydrogen; and n is 0. 1 3 .: R ¹ and R ³ together form a single bond; R ² is hydrogen or lower alkyl; R ⁴ is lower alkyl; R ⁶ is hydrogen; R ⁷ , R ⁸ and R ⁹ are Hydrogen; R ⁸ Hydrogen; R ¹ . There optionally substituted § Li one also ylcarbonyl or unsubstituted or good I § Li - Rusuruhoniru; R ^{1 1} and R ^{1 2} is hydrogen; is n is 0, a compound according to claim 1. 14. R ¹ and R ³ together form a single bond; R ² is hydrogen or lower alkyl; R ⁴ is lower alkyl; R ⁶ is hydrogen; R ⁷ and R ⁹ are together bond to form; R ⁸ is halogen; R ¹ Q is hydrogen; R ^{1 1} is hydrogen or halogen; R ^{1 2} is hydrogen; is n is 0, a compound according to claim 1. 15. R ¹ and R ³ together form a single bond; R ² is hydrogen or lower alkyl; R ⁴ is lower alkyl; R ⁶ is hydrogen; R ⁷ and R ⁹ are both hydrogen or together R ⁸ is hydrogen; R ¹ . Is hydrogen; R ¹¹ and R ¹² are hydrogen; n is 1.  16. A pharmaceutical composition comprising the compound according to any one of claims 1 to 15. 17. A preventive or therapeutic agent for a disease mediated by a mouth tonine receptor, comprising the compound according to any one of claims 1 to 15. 18. An agent for preventing or treating central nervous system diseases, comprising the compound according to any one of claims 1 to 15.  19. The preventive or therapeutic drug according to claim 18, wherein the central nervous system disease is anxiety, depression, schizophrenia, circadian rhythm disorder, stroke, dementia, pain or Parkinson's disease. 20. A microorganism belonging to the genus Streptomyces, which is capable of producing the compound according to any one of claims 1 to 15.  21. The microorganism according to claim 20, capable of producing the compound according to claim 15. 22. The microorganism according to claim 20, which is Streptomyces sp. PA-48561. 23. A method comprising culturing the microorganism according to any one of claims 20 to 22 and isolating and purifying a compound produced from the obtained culture solution, followed by optionally chemical modification. Item 16. The method for producing a compound according to any one of Items 1 to 15. 24. A method for preventing or treating a serotonin receptor-mediated disease, comprising administering the compound according to any one of claims 1 to 15.  25. Use of the compound according to any one of claims 1 to 15 for the manufacture of a prophylactic or therapeutic agent for a disease mediated by a serotonin receptor.