JPH06199892A - Tan-1666 related comopund - Google Patents

Abstract

PURPOSE:To obtain a new subject compound having tachykinin antagonism and effective in treatment and prevention for tachykinin-intercalated diseases such as asthma, atopic dermatitis and nettle rash, by culturing a specific microorganism belonging to the genus Aspergillus. CONSTITUTION:A microorganism belonging to the genus Aspergillus and having ability capable of producing a compound (TAN-1666A) expressed by formula I or a compound (TAN-1666B) expressed by formula II [e.g. Aspergillus terreus FL-29694 (FERM P-13126), etc.] is inoculated into a culture medium and subjected to shake culture at 28 deg.C for 48hr to afford a seed culture. The seed culture is transplanted to a culture medium in a stainless-steel tank and subjected to spinner culture under aeration at 24 deg.C and then, pH of culture medium is adjusted to pH 2-4 and the cultured mixture is extracted with dichloromethane, etc., and the extracted liquid is extracted with a dilute alkali water. Thereby, TAN-1666B is dissolved in a water layer and TAN-1666A is left in an organic solvent layer and these layers are purified to provide the objective TAN-1666 related compound expressed by formula III [R1 is H; R2 is (substituted)alkoxy, etc., or R2 forms direct bond together with R1].

Description

Detailed Description of the Invention

[0001]

The present invention relates to a novel peptide compound TA
N-1666 [Hereinafter, it may be abbreviated as TAN-1666. ] Related compounds, more specifically, tachykinin antagonism, particularly substance P antagonism, substance K
Novel peptide compound TAN having pharmacological activity such as (neurokinin A) antagonism and neurokinin B antagonism
-1666 related compound and its manufacturing method.

[0002]

2. Description of the Related Art Substance P is a peptide belonging to the tachykinin group together with substance K (neurokinin A) and neurokinin B, and is known to exhibit various physiological activities [M. Otuka and Kay.・ K. Yoshioka, Physiological Previews, Volume 1, Page 1 (1991)
Year)〕. In particular, since it plays an important role as a neurotransmitter involved in the pain sensation of the unmyelinated sensory nerve projected to the dorsal root of the spinal cord and as a mediator of inflammation, its antagonist is applied as a novel analgesic and anti-inflammatory agent. There is expected. The compounds having substance P receptor antagonism so far include actinomycete metabolite WS-9326 and its derivatives (JP-A-3-148227) and related peptides (JP-A-3-27399), FR113680-related. Compounds (International Publication WO91 / 12266) and the like are known. In addition, cyclic peptides (Japanese Patent Laid-Open No. 3-2
197), peptide compounds (JP-A-4-2109).
No. 96) and the like are said to have the above effect.

[0003]

Examples of tachykinin-mediated diseases in humans or animals include respiratory diseases (eg, asthma, bronchitis, rhinitis, cough, sputum, etc.), eye diseases (eg, conjunctivitis, spring catarrh, etc.), Skin diseases (eg contact dermatitis, atopic dermatitis, urticaria, other eczema-like dermatitis), inflammatory diseases (eg rheumatoid arthritis, osteoarthritis etc.), pain (eg migraine, headache, Toothache, cancer pain, back pain, etc., cardiovascular disease (eg, hypertension, angina, heart failure, thrombosis, etc.), brain disease (eg, dementia, schizophrenia, etc.) and the like. Therefore, there is a need for compounds with tachykinin antagonism that are useful in the treatment or prevention of these diseases.

[0004]

In view of the above situation, the present inventors have found that if a tachykinin receptor antagonist having a novel skeleton can be found, a drug useful for treating or preventing a tachykinin-mediated disease can be obtained. I was convinced that it could be obtained, and I sought to find its origin in microbial metabolites, and conducted extensive research. As a result, among a large number of microorganisms isolated from soil, a certain microorganism produces a new substance, that the microorganism belongs to the genus Aspergillus, and by culturing the microorganism in an appropriate medium, a tachykinin receptor is obtained. Knowing that active compounds that strongly antagonize could accumulate in the medium, two active compounds were isolated and named TAN-1666A and B. The present inventors completed the present invention by examining the physicochemical and biological properties of these compounds in detail and confirming that they are novel substances. That is, the present invention provides (1) the general formula

[Chemical 4] [Wherein R ₁ represents hydrogen, R ₂ represents a bond together with a hydroxyl group, an optionally substituted hydrocarbon oxy group, an optionally substituted amino group or R ₁ ] A compound or a salt thereof, (2) the compound according to item ₁ , wherein R ₁ is hydrogen and R ₂ is an alkylamino group, (3) the compound according to item 2, wherein the alkylamino group is an ethylamino group,
(4) Belonging to the genus Aspergillus, the formula

[Chemical 5] Compound TAN-1666A represented by or a formula

[Chemical 6] The compound TAN-1666A or B or a salt thereof, which comprises culturing a microorganism having the ability to produce the compound TAN-1666B represented by the formula (1) in a medium, allowing the compound to be produced and accumulated in the culture, and collecting the compound. And a tachykinin antagonist comprising the compound according to item (5).

In the general formula [I], the hydrocarbon group of the optionally substituted hydrocarbon oxy group represented by R ₂ is
Preferably, it is a hydrocarbon group having 1 to 20 carbon atoms, and examples thereof include an alkyl group, a cycloalkyl group, an alkenyl group, an alkynyl group, an aryl group and an aralkyl group.
Of these, an alkyl group and a cycloalkyl group are preferred. More preferably, it is an alkyl group. As the above-mentioned alkyl group, an alkyl group having 1 to 10 carbon atoms is preferable, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, tert-pentyl, 1- Examples include ethylpropyl, hexyl, isohexyl, 1,1-dimethylbutyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 2-ethylbutyl and the like. More preferably, it is an alkyl group having 1 to 6 carbon atoms, and examples thereof include methyl, ethyl, propyl, isopropyl, butyl, isobutyl and pentyl. The cycloalkyl group is preferably a cycloalkyl group having a carbon number of 3 to 8, and examples thereof include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and the like. More preferably, it has 4 to 6 carbon atoms.
Cycloalkyl groups such as cyclobutyl. Examples include cyclopentyl and cyclohexyl. The alkenyl group is preferably an alkenyl group having 2 to 10 carbon atoms, and examples thereof include vinyl, allyl, 2-butenyl, 3-butenyl, 2
-Pentenyl, 5-hexenyl, 4-decenyl and the like can be mentioned. The alkynyl group has 2 to 10 carbon atoms.
The alkynyl group of is preferably ethynyl, 2-propynyl (propargyl), 2-butyn-1-yl, 3-
Examples include butyn-2-yl, 1-pentyn-3-yl, 3-pentyn-1-yl, 4-pentyn-2-yl, 3-hexyn-1-yl and the like. The aryl group is preferably an aryl group having a carbon number of 6 to 14, and examples thereof include phenyl, naphthyl, anthryl, biphenylyl and the like. The aralkyl group has 7 to 20 carbon atoms.
Is preferred, and examples thereof include benzyl, phenethyl, benzhydryl, trityl, naphthylethyl, and anthrylethyl.

In the general formula [I], the substituent in the optionally substituted hydrocarbon oxy group represented by R ₂ is, for example, hydroxy, carboxyl, sulfo, cyano, nitro, halogen, mercapto or substituted. Amino group (eg, amino, mono- or di-alkylamino, etc.), optionally substituted carbamoyl group (eg, carbamoyl, mono- or di-alkylcarbamoyl, etc.), optionally substituted sulfamoyl group (eg, , Sulfamoyl, mono- or di-alkylsulfamoyl, etc.), alkoxy (alkyloxy) groups,
Examples thereof include a cycloalkyloxy group, an alkenyloxy group, an aryloxy group, an aralkyloxy group, an alkylthio group, a cycloalkylthio group, an arylthio group, an aralkylthio group, an acyl group and a heterocyclic group. Preferably, for example, hydroxy, carboxyl, sulfo, halogen, optionally substituted amino group, alkoxy (alkyloxy) group, alkylthio group, heterocyclic group and the like can be mentioned. 1 to 3 of these substituents may be substituted at appropriate positions on the hydrocarbon group.

Here, examples of the halogen include fluorine, chlorine, bromine and iodine. The alkyl group in the mono- or di-alkylamino group, the mono- or di-alkylcarbamoyl group, the mono- or di-alkylsulfamoyl group, the alkoxy (alkyloxy) group and the alkylthio group is preferably an alkyl group having 1 to 6 carbon atoms. A group (eg, methyl, ethyl, propyl, etc.). The cycloalkyl group in the cycloalkyloxy group and the cycloalkylthio group preferably has 3 to 8 carbon atoms.
Is a cycloalkyl group (eg, cyclopentyl, cyclohexyl, etc.). The alkenyl group in the alkenyloxy group is preferably an alkenyl group having 2 to 10 carbon atoms (allyl, pentenyl, etc.). The aralkyl group in the aralkyloxy group and the aralkylthio group is preferably an aralkyl group having 7 to 20 carbon atoms (benzyl, phenethyl and the like). The aryl group in the aryloxy group and the arylthio group is preferably an aryl group having 6 to 14 carbon atoms (phenyl, naphthyl, etc.).

The acyl group is preferably an acyl group derived from an organic carboxylic acid, and examples thereof include a formyl group, an alkanoyl (alkylcarbonyl) group, an arylcarbonyl group, an aralkylcarbonyl group, an alkylcarbonyloxy group and an arylcarbonyloxy group. , Aralkylcarbonyloxy group, heterocyclic carbonyl group and the like. The alkyl group in the alkanoyl (alkylcarbonyl) group and the alkylcarbonyloxy group, the aryl group in the arylcarbonyl group and the arylcarbonyloxy group, the aralkylcarbonyl group and the aralkyl group in the aralkylcarbonyloxy group are the same as those defined in the above hydrocarbon group. Things are used. As the heterocyclic group in the heterocyclic carbonyl group, the same heterocyclic group as the substituent of the hydrocarbonoxy group defined below is used. Specific preferred examples of the acyl group include formyl, acetyl, propionyl, butyryl, pionyl, pivaloyl, benzoyl, formyloxy, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, tert-butoxycarbonyl, iso Butoxycarbonyl and the like. Examples of the heterocyclic group include 5- to 6-membered heterocyclic groups containing 1 to 4 nitrogen atoms, oxygen atoms and / or sulfur atoms, and specific examples thereof include pyrrolidino, 2-oxopyrrolidino, pyrrolidinyl, pyrrolyl,
Pyrazolyl, imidazolyl, furyl, thienyl, oxazolyl, isoxazolyl, isothiazolyl, thiazolyl, piperidino, piperidinyl, pyridyl, pyridazinyl, pyrazinyl, piperazinyl, pyrimidinyl, pyranyl, tetrahydropyranyl, tetrahydrofuryl, indolyl, 1,3,4-oxadiazolyl, 1 , 2,3-
Thiadiazolyl, 1,3,4-thiadiazolyl, 1,
2,3-triazolyl, 1,2,4-triazolyl,
1,3,4-triazolyl, tetrazolyl, 1,3-dioxolanyl, morpholino, morpholinyl and the like can be mentioned.

In the general formula [I], the substituent in the optionally substituted amino group represented by R ₂ is, for example, sulfo, amidino, carbamoyl, mono- or di-alkylcarbamoyl, alkyl group, cycloalkyl group,
Examples thereof include an aryl group, an aralkyl group, an acyl group and a heterocyclic group. An alkyl group, an alkyl group, a cycloalkyl group in a mono- or di-alkylcarbamoyl group,
As the aryl group, aralkyl group and acyl group, the same substituents as the above-mentioned hydrocarbon oxy group represented by R ₂ can be used, but further suitable substituents (eg, substituted with C _1-6 alkyl) Good amino group, hydroxy group,
1 or 2 may be substituted with a carboxyl group or the like). Examples of the heterocyclic group include 5- to 6-membered heterocyclic groups containing 1 to 4 nitrogen atoms, oxygen atoms and / or sulfur atoms having a bond on a carbon atom. The heterocyclic group may be further substituted with one or two suitable substituents (eg, amino group _optionally substituted with C _1-6 alkyl, hydroxy group, carboxyl group, etc.). The substituent in the optionally substituted amino group represented by R ₂ is
Preferred are, for example, an alkyl group having 1 to 4 carbon atoms which may be substituted with an amino group which may be substituted with C _1-6 alkyl, an aralkyl group having 7 to 20 carbon atoms and the like.

In the general formula [I], the optionally substituted amino group represented by R ₂ may form a nitrogen-containing heterocyclic group having a bond on the nitrogen atom. The nitrogen-containing heterocyclic group having a bond on the nitrogen atom is preferably a 5- to 7-membered heterocyclic group which may further contain an oxygen atom as a ring-constituting atom. Specific preferred examples thereof include 1-pyrrolidinyl, piperidino, piperazinyl, 4-morpholinyl, 1-azacycloheptoyl and the like. These are substituted with 1 to 3 substituents such as an alkyl group, a cycloalkyl group, an alkenyl group, an alkynyl group, an aryl group or an aralkyl group, which are defined as the substituents of the hydrocarbon group in the above-mentioned hydrocarbon oxy group. May be. R ₂ is preferably alkoxy, aralkyloxy, alkylamino optionally substituted with a mono- or di-substituted amino group, aralkylamino optionally substituted with a mono- or di-substituted amino group, alkyl It is a 6-membered heterocyclic group containing 1-2 nitrogen atoms which may be substituted.

The compounds TAN-1666A and B are produced by filamentous fungi belonging to the genus Aspergillus. For example, one strain FL-2 newly isolated from Indian soil.
It can be produced by 9694. TAN-16
The 66-producing strain has the following properties. a) Morphological characteristics This strain FL-29694 grows well on malt extract agar medium and potato / glucose agar medium, but conidial formation is poor. Growth and conidia formation are poor on Czapek agar. Moderate growth on oatmeal agar, but rich conidia formation. The hyphae are transparent and have septa. Conidia stalks arise from aerial hyphae,
Slightly curved, with septum visible. Its surface is smooth and the tip is enlarged to become a capsular bag. The capsule is hemispherical, 15-20
The size of the diameter in μm is shown. It produces more metula than the top of the capsule. The metre is 3.5 to 4.5 × 1.
It has a cylindrical shape of 5 to 2.0 μm, and 2 to 4 bottle-shaped phialides are bundled on the tip thereof. The surface of phialide is smooth and has a size of 5 to 6 × 1.0 to 2.0 μm, and spherical or subspherical conidia are formed at the tip thereof. The surface of conidia is smooth and the size is 2.0-2.5x
Long chains are formed at 1.0 to 2.0 μm.

B) Properties on agar medium Malt extract agar medium Medium growth, the colony diameter becomes 47 mm after 2 weeks when kept at 24 ° C. The surface is flat and consists of a velvety mycelium with a raised central part, with regular outer edges. Aerobic hyphae are well developed but conidia are poorly formed. The color tone of the surface of the colony is pale yellowish white from the central portion to the peripheral portion, and dark yellow to pale yellowish brown from the central portion of the back surface to the peripheral portion. No formation of soluble pigment is observed. Potato-glucose agar medium Growth is moderate, and the colony diameter becomes 54 mm after 2 weeks when kept at 24 ° C. The surface is flat, consisting of a myrtle in the shape of a pillow with a slightly raised central portion, with a slightly irregular outer edge. Although the development of aerial hyphae is good,
Conidia are poorly formed. The color tone of the surface of the colony is light gray from the center to the middle, and white at the periphery. On the back side, the middle part is dark reddish brown or reddish brown, the middle part is reddish brown, and the peripheral part is light yellowish brown. No formation of soluble pigment is observed. Czapek agar medium Growth is poor, and the colony diameter becomes 22 mm after 2 weeks when kept at 24 ° C. The surface is velvety with a slightly raised central part, rarely composed of wool-like mycelium, and radial wrinkles are observed from the middle part to the peripheral part. The outer edge is slightly irregular. Aerobic hyphae are moderately developed but conidia are poorly formed. The color of the colony surface is
The central part has a pale yellowish gray color, the middle part has a cream color, and the peripheral part has a pale grayish white color. Also, many water droplets are observed from the central part to the middle part. The center of the backside of the village is pale yellowish white, and it is pale reddish brown from the center to the periphery. No formation of soluble pigment is observed. Oatmeal agar medium Medium growth, the colony diameter becomes 57 mm after 2 weeks when kept at 24 ° C. The surface is slightly raised, consisting of powdery to velvety mycelium, with some depressions in the center and thinning in the periphery. The outer edge is regularly bordered. Development of aerial hyphae and conidia formation are good. The central part of the surface of the village has a yellowish brown color with a slight grayish white mixture, but the middle part has an annular light yellowish brown color, and the middle part to the peripheral part has a cream color or a light grayish white color. The central part of the back surface is light yellow-brown or cream-colored, and the middle part to the peripheral part is light yellow-brown or ivory colored. No soluble dye formation is observed.

C) Physiological properties The growth conditions of this strain were examined on a potato-glucose agar medium. It grows in any of pH 3 to pH 12, the growth temperature range is 13 ° C to 45 ° C, and the optimum temperature is 25 ° C to 35 ° C. It grows even at 45 ° C. When the above properties are compared with the identification search table described in "Mold Separation, Culture and Identification" by D. Malloch, translated by Shunichi Udagawa (Medical and Dental Publishing Co., Ltd., 1983), page 51. , This spore consists of 1 cell, colony, conidia and other organs are colorless or light-colored, conidia stalk has apical sac, and forms a bottle-shaped fearide. Therefore, Aspergillus spp.
It is clear that it belongs to s). In addition, KB
"THE GENUS Aspergillus" by KB Raper and others (1965,
When compared with the characteristics of the Aspergillus strain described in The Williams and Wilkins Company,
This strain forms a metre on the top of the apical sac, a phialide occurs on it, and the conidia head is cylindrical and yellowish brown, but the conidia stalk is colorless, so Aspergillus terreus (Aspergillus terreus) ) Group, and Aspergillus terreu
s) It was identified as No. FL-29694. This strain was designated by the Fermentation Research Institute (IFO) under the accession number IFO 32451 on June 17, 1992, and also on August 26, 1992.
Contract number F to the Ministry of International Trade and Industry Industrial Technology Research Institute (FRI)
Deposited as ERMP-13126 respectively.

TAN-166 belonging to the genus Aspergillus
Like the other filamentous fungi, the 6-producing strain can be mutated by irradiation with ultraviolet rays, X-rays, radiation, etc., monospore separation, various mutation treatments, and other means. Even if it is a mutant strain obtained, or a mutant strain obtained by using genetic engineering or other general genetic techniques, it is substantially different species in comparison with the taxonomic characteristics described above. Any material having the property of producing the compound, which is not sufficient, can be used in the present invention.

The medium used for culturing the compound-producing bacterium may be liquid or solid as long as it contains a nutrient source that can be utilized by the bacterium, but when treating a large amount, it is more suitable to use a liquid medium. is there. The medium is appropriately mixed with a carbon source, a nitrogen source, an inorganic substance, and a trace nutrient source that can be assimilated by the compound-producing bacterium. Examples of carbon sources include glucose, lactose, sucrose, maltose, dextrin, starch, glycerin, mannitol, sorbitol, oils and fats (eg, soybean oil, lard oil, chicken oil, etc.), n-paraffin, etc. Is, for example, meat extract, yeast extract,
Dry yeast, soybean flour, corn steep liquor, peptone, cottonseed flour, molasses, urea, ammonium salts (eg,
Ammonium sulfate, ammonium chloride, ammonium nitrate, ammonium acetate, etc.) are used. Further, salts containing sodium, potassium, calcium, magnesium, etc., metal salts such as iron, manganese, zinc, cobalt, nickel etc., salts such as phosphoric acid, boric acid etc. and salts of organic acids such as acetic acid, propionic acid etc. may be used appropriately. Good. Other amino acids (eg, glutamic acid, aspartic acid,
Alanine, lysine, methionine, proline, etc.), peptides (eg, dipeptides, tripeptides, etc.), vitamins (eg, B ₁ , B ₂ , nicotinic acid, B _{1 2} , C, etc.),
Nucleic acids (eg, purines, pyrimidines, derivatives thereof, etc.) may be contained. Of course, inorganic or organic acids or alkalis, buffers and the like may be added for the purpose of adjusting the pH of the medium, or appropriate amounts of oils and fats, surfactants and the like may be added for the purpose of defoaming. In liquid culture, the pH of the medium is preferably near neutral, and particularly preferably pH 5.5 to 7.
The culturing temperature is about 20 ° C to 30 ° C, and the culturing time is about 48 hours
168 hours is preferred.

TAN produced with the progress of culture
-1666 has ¹²⁵ I-BHSP (Bolton Hunter (Bol)
tonand Hunter) Mark [Lys ³ ] Substance P, 8
It is quantified using a binding inhibition assay with 1.4 TBq / mmol, NEN Research Products (USA) as a radio ligand.
Usually, the maximum production of TAN-1666 is reached in 4 to 6 days of culture. Target compound TAN-16 from culture
Examples of the method of collecting 66A and 66B include the following. These are fat-soluble substances, TAN
Since -1666A shows neutrality and TAN-1666B shows acidity, a general means utilizing this property may be adopted. Further, since both of these components are contained in both the bacterial cells and the filtrate, the following purification method is adopted. First, the culture solution is adjusted to pH 1.5 to 6, preferably pH 2 to 4, and then an organic solvent immiscible with water, for example, dichloromethane, ethyl acetate, methyl isobutyl ketone, butanol, etc. is added and filtered to obtain TAN-1666A or TAN-1.
Extract 666B. When the organic solvent layer was extracted with dilute alkaline water, TAN-1666B was transferred to the aqueous layer,
-1666A remains in the organic solvent layer. The TAN-1666B is extracted into the organic solvent layer by adjusting the phase-dissolved water layer to be acidic again and extracting with the above-mentioned water-immiscible organic solvent.
If these two organic solvent layers are washed with water and then concentrated, T
Crude materials containing AN-1666A or B respectively are obtained. The crude material was further purified to give pure TAN-16
Various chromatographic methods are advantageously used to obtain 66. For example, when ordinary column chromatography is used, silica gel, crystalline cellulose, adsorptive resin (for example, Diaion HP-20 (manufactured by Mitsubishi Kasei), Amberlite XAD-I or II) is used as a carrier.
(Rohm and Haas, USA), Sephadex LH-20 (Pharmacia, Sweden), etc. are used. To elute the active substance from the column, depending on the type of carrier, a suitable organic solvent such as dichloroethane, toluene, ethyl acetate, acetone,
A single solvent or a mixed solvent such as methanol, or a mixed solvent of an organic solvent miscible with water and an aqueous solution such as water, dilute alkaline water, dilute acidic water or a buffer solution is used.

TAN-1666A or T can also be measured by preparative high performance liquid chromatography (HPLC).
AN-1666B can be purified. As the carrier, octadecylsilane (ODS) type and silica gel type carriers can be advantageously used. For example, in the case of ODS, a mixed solution of methanol or acetonitrile and a salt-containing aqueous solution is advantageously used. The organic solvent eluate containing the active substance is concentrated, or, if it contains an aqueous solution, it is extracted with a suitable organic solvent immiscible with water and concentrated, and the residue is pulverized or the concentrated residue is converted into a suitable crystallization solvent such as diethyl ether. Ether, chloroform, ethyl acetate, acetone,
Dissolve it in methanol or a mixture thereof and leave it in a cool place to obtain crystals. Since TAN-1666B is an acidic substance, it can be treated with an appropriate base to produce TA.
A salt of N-1666B is obtained. These are prepared by a method known per se. The type of salt is, for example, an alkali metal salt that is a salt with an inorganic salt (eg, sodium salt,
Examples thereof include potassium salts), alkaline earth metal salts (eg, calcium salts), ammonium salts and the like.
These salts are also within the scope of the present invention.

Next, the TAN-obtained in the examples described later
1 shows the physicochemical properties of 1666A and B. [TAN-1666A] (1) Appearance: White powder (2) Optical rotation: -100 ° (c 0.503, dimethylformamide (DMF), 26 ° C) (3) Molecular weight: m / z 496 (M + H) ⁺ , (SI (From mass spectrum) (4) Elemental analysis value: (%) (water content: 0.25 mol) Actual value: C, 72.22; H, 6.10; N, 8.33 Calculated value: C, 72.05; H, 5.95; N, 8.40; O, 13.60 (5) Molecular formula: C ₃₀ H ₂₉ N ₃ O ₄ (495) (6) Ultraviolet absorption (UV) spectrum: in methanol Maximum: 274 nm (ε 25,000) (7) Infrared absorption (IR ) Spectrum: major absorption in KBr tablets (wave number, cm ^-1 ) 3380, 3330, 1690, 1620, 1450, 1210, 1190, 700 (8) ¹³ C nuclear magnetic resonance (NMR) spectrum: 75 MHz, DMS
Δppm in Od ₆ [Fig. 1] 175.0 (s), 169.7 (s), 166.8 (s), 164.5 (s), 139.2 (d),
135.7 (s), 135.1 (s), 134.7 (s), 129.9 (d), 129.9 (d), 1
29.4 (d), 129.4 (d), 129.4 (d), 128.8 (d), 128.8 (d), 12
8.3 (d), 128.3 (d), 128.1 (d), 128.1 (d), 127.4 (d), 12
7.4 (d), 127.2 (d), 126.4 (d), 121.3 (d), 57.3 (d), 54.
9 (d), 49.7 (d), 37.3 (t), 36.3 (t), 17.1 (q) [where s is a quaternary carbon, d is CH, t is CH ₂ and q is CH ₃ ] ( 9) Color reaction: positive; phosphomolybdic acid, Greig-Lieback reaction negative; Sakaguchi, Dragendorff, ninhydrin, sulfuric acid,
Potassium permanganate reaction (10) High Performance Liquid Chromatography (HPLC): Carrier; YMC Pack A312 S-5 120A ODS 6 × 150 Mobile phase; 60% Acetonitrile (CH ₃ CN) /0.02M Phosphate buffer pH 6.3 Flow rate: 2.0 ml / min Detection method: 250 nm (UV) Elution time: 4.9 min (11) Thin layer chromatography (TLC): Carrier: Kieselgel 60 F ₂₅₄ 0.25 mm (E. Merck A
G.) Developing solvent; toluene: acetone = 1: 1 Rf value; 0.47 (12) Property: neutral lipophilicity (13) Amino acid analysis: L-alanine (1 mol), phenylalanine (D-, L-each) 1 mol)

[TAN-1666B] (1) Appearance: colorless crystals (2) Melting point: 202-203.5 ° C. (3) Optical rotation: + 34.5 ° (c 0.413, DMF, 26
(° C) (4) Molecular weight: m / z 513 (M ⁺ ), (from EI mass spectrum) (5) Elemental analysis value: (%) (water content: 0.25 mol) Actual value: C, 69.65; H, 6.14; N, 8.26 Calculated value: C, 69.55; H, 6.13; N, 8.11; O, 16.21 (6) Molecular formula: C ₃₀ H ₃₁ N ₃ O ₅ (513) (7) UV spectrum: maximum value in methanol: 275 nm (ε 29,000) (8) IR spectrum: major absorption in KBr tablets (wavenumber, cm ⁻¹ ) 3290, 1730, 1650, 1540, 1220, 700 (9) ¹³ C NMR spectrum: 75 MHz, DMSO- d ₆ δp
pm [Fig.2] 172.7 (s), 171.8 (s), 170.6 (s), 164.4 (s), 138.9 (d),
137.6 (s), 137.4 (s), 134.8 (s), 129.3 (d), 129.1 (d), 1
29.1 (d), 129.1 (d), 129.1 (d), 128.8 (d), 128.8 (d), 12
8.0 (d), 128.0 (d), 127.7 (d), 127.7 (d), 127.4 (d), 12
7.4 (d), 126.3 (d), 126.0 (d), 121.9 (d), 53.4 (d), 53.
4 (d), 48.2 (d), 37.7 (t), 37.0 (t), 18.2 (q) (10) Color reaction: Positive; Phosphomolybdic acid, Greig-Reeback reaction Negative; Sakaguchi, Dragendorf, Ninhydrin, sulfuric acid,
Potassium permanganate reaction (11) HPLC: Carrier; YMC Pack A312 S-5 120A ODS 6 × 150 Mobile phase; 35% CH ₃ CN / 0.02M phosphate buffer pH 6.3 Flow rate; 2.0 ml / min Detection method 250 nm (UV) elution time; 5.2 min (12) TLC: carrier; Kieselgel 60 F ₂₅₄ 0.25 mm (E. Merck AG.) Developing solvent; toluene: acetone: formic acid = 50: 50: 2 Rf value; 0 .52 (13) Properties: acidic fat-soluble (14) Amino acid analysis: L-alanine (1 mol), phenylalanine (D-, L-each 1 mol)

From the above data and detailed examination of the NMR spectrum, TAN-1666A (Compound 1) and B (Compound 2) are presumed to have the following structural formulas. These are the corresponding dihydro TAN-1666A by catalytic reduction.
(Compound 3) and dihydroTAN-1666B (Compound 4) are provided. TAN-1666A

[Chemical 7] TAN-1666B

[Chemical 8]

Next, a method for producing a derivative using TAN-1666B (Compound 2) will be described. By subjecting the compound to, for example, an esterification or amidation reaction,
A compound having a more potent tachykinin antagonistic action can be obtained. Specific examples of the method for producing the derivative are shown below. [Esterification of Carboxylic Acid] General formula by subjecting TAN-1666B or a salt thereof to an esterification reaction

[Chemical 9] [Wherein R ₁ ′ represents hydrogen and R ₂ ′ represents an optionally substituted hydrocarbon oxy group] or a salt thereof will be described below. The esterification reaction of TAN-1666B (Compound 2) or a salt thereof is performed by a method known per se, for example, the following method. 1) Mixing the compound 2 with a diazoalkane (eg, diazomethane, phenyldiazomethane, diphenyldiazomethane, etc.) and a solvent that does not inhibit the reaction (eg, ethers such as tetrahydrofuran, dioxane, esters such as ethyl acetate, nitriles such as acetonitrile) React in liquid. The reaction temperature is about 0 ° C to the reflux temperature. The reaction time is about 2 minutes to 2 hours. 2) Alkali metal salt of compound 2 (eg sodium salt,
Activated alkyl halides such as potassium salt and lithium salt (eg methyl iodide, benzyl bromide, p-
Nitro-benzyl bromide, m-phenoxybenzyl bromide, pt-butylbenzyl bromide, pivaloyloxymethyl chloride, etc.). This reaction is
A solvent that does not interfere with the reaction (eg, dimethylformamide,
Amides such as dimethylacetamide and hexamethylphosphoramide). The reaction temperature is about 0 ° C to 60 ° C. The reaction time is about 2 minutes to 4 hours. Coexistence of triethylamine or the like in this reaction solution does not affect the progress of the reaction. 3) reacting compound 2 with an alcohol, such as methanol, ethanol, benzyl alcohol, etc. This reaction is carried out in the presence of a carbodiimide condensing agent (eg dicyclohexylcarbodiimide). The reaction temperature is about 0 ° C to the reflux temperature. The reaction time is about 15
Minutes to 18 hours. As the solvent, solvents that do not inhibit the reaction, for example, halogenated hydrocarbons such as chloroform, dichloromethane and dichloroethane are used. 4) The acid anhydride of compound 2 obtained by reacting compound 2 with an acid halide (eg, ethyl chloroformate, benzyl chloroformate, etc.) is reacted with alcohol (eg, methanol, ethanol, benzyl alcohol, etc.) The reaction is carried out under the reaction conditions described in 3). This anhydride is obtained by reacting compound 2 with an acid halide, for example, acid chloride, in a solvent that does not inhibit the reaction (for example, ethers such as tetrahydrofuran, halogenated hydrocarbons such as dichloromethane). The reaction temperature is 25 ° C to the reflux temperature. The reaction time is about 15 minutes to 10 hours. 5) Coexistence of Compound 2 with a silylating agent (eg, trimethylsilyl chloride, t-butyl-dimethylsilyl chloride and triethylamine), a solvent that does not inhibit the reaction (eg, halogenated hydrocarbons such as dichloromethane and chloroform, tetrahydrofuran, etc.) Ether). The reaction temperature is about 0 ° C to the reflux temperature. The reaction time is about 15 minutes to 16 hours.

[Amidation of Carboxylic Acid] TAN-166
General formula by subjecting 6B to an amidation reaction

[Chemical 10] [Wherein R ₁ ″ represents hydrogen and R ₂ ″ represents an amino group which may be substituted]. The amidation of carboxylic acid of TAN-1666B (Compound 2) can be carried out by a method known per se, for example, by converting Compound 2 into an acid halide, preferably an acid chloride (eg, trimethylacetyl chloride, ethyl chloroformate, benzyl chloroformate) or an acid. By synthesizing an acid anhydride of compound 2 with an anhydride (eg acetic anhydride, trifluoroacetic anhydride etc.) and then reacting with ammonia or amine (eg alkylamine, dialkylamine, aralkylamine, heterocyclic amine etc.) Done.
The above reaction is carried out in a solvent that does not inhibit the reaction (for example, halogenated hydrocarbons such as dichloromethane, ethers such as tetrahydrofuran, amides such as dimethylformamide). The reaction temperature is about 0 ° C to the reflux temperature. The reaction time is about 15 minutes to 16 hours. The production method and the physical properties of the thus obtained derivative are described in detail in Examples described later, and the structure thereof is shown in [Table 1].

[0023]

[Table 1]

Next, the biological activity test of TAN-1666 related compounds will be shown. (A) Substance P Radioligand Receptor (NK ₁ ) Binding Inhibitory Activity The method of R. Qirion and C. Pilapil [Neuropeptide
ide) Vol. 4, p. 32 (1984)]. Receptors were prepared from the brains of Wistar rats (male, 8 weeks old, Charles River). Rats were sacrificed by decapitation, the forebrain was removed, and each rat contained 30 ml of 120 mM sodium chloride and 5 mM potassium chloride.
Polytron homogenizer (Kinematika) in 50 mM Tris-HCl buffer (pH 7.4),
(Germany), followed by centrifugation at 40,000 × g for 20 minutes. 30 ml of 300 mM potassium chloride and 10 m of sediment
It is suspended in a 50 mM Tris-hydrochloric acid buffer solution (pH 7.4) containing M ethylenediaminetetraacetic acid, and gently stirred under ice cooling for 30 minutes. The suspension was centrifuged at 40,000 xg for 20 minutes and the precipitate was added to 30 ml of 50 mM Tris-HCl buffer (p
H7.4) and frozen as a receptor standard (-80
℃) preserved. A reaction buffer solution (50 mM Tris-HCl buffer solution (pH
7.4), 0.02% bovine serum albumin, 1 mM phenylmethylsulfonium fluoride, 2 μg / ml chymostatin, 40 μg / ml bacitracin, 3 mM manganese chloride) and 100 μl volume was used for the reaction. Add sample and ¹²⁵ I-BHSP (0.46 KBq),
The reaction was carried out in 0.2 ml of reaction buffer at 25 ° C for 30 minutes.
Substance P was added at a concentration of 2 × 10 ⁻⁶ M to determine the amount of non-specific binding. After reaction, cell harvester (290PHD, Cambridge Technology
The reaction was stopped by rapid filtration on a glass filter (GF / B, Whatman, USA) using Incorporation (Cambridge Technology, Inc., UK), and 250 μl of 50 mM Tris-HCl. Buffer solution (p
After washing 3 times with H7.4), the radioactivity remaining on the filter was measured by a gamma counter. The filter used was one that had been soaked in 0.01% polyethyleneimine for one day before use and then air-dried.

(B) Neurokinin A Radioligand Receptor (NK ₂ ) Binding Inhibitory Activity Paul L. M. van Guielsbergen (Paul)
LM VAN Giersbergen) et al. (Proc. Natl. Acad.
Sci. 88, 1661 (1991)) was modified and used. Receptors were prepared from the inner wall of beef rumen (purchased from Kyoto Central Livestock By-product Wholesale Cooperative). The inner wall of bovine abomasum stored at -80 ° C was shredded into 1 cm squares or less, and 3 liters of 120 mM sodium chloride, 5 mM potassium chloride, 0.02% bovine serum albumin (BSA) and 5% sucrose were added per 1 kg. Crush using a Polytron homogenizer in 50 mM Tris-HCl buffer (pH 7.4) containing 1000
It was centrifuged at × g for 10 minutes. The supernatant was further centrifuged at 45,000 xg for 20 minutes. 200 ml of 300 mM of the obtained sediment
Potassium chloride, 10 mM ethylenediaminetetraacetic acid, 0.
1 mM phenylmethylsulfonium fluoride and 0.
50 mM Tris-HCl buffer containing 02% BSA (pH
7.4), and gently stirred under ice cooling for 60 minutes. The suspension was centrifuged at 45,000 xg for 20 minutes, and the precipitate was added to 200 ml of 50 mM Tris-HCl buffer (pH 7.4).
The sample was washed with, and stored frozen (-80 ° C) as a receptor standard. This preparation was added to a reaction buffer (50 mM Tris-HCl buffer (pH 7.4), so that the protein concentration was 0.7 mg / ml,
Suspended in 0.02% bovine serum albumin, 4 mM manganese chloride) and used 100 μl volume for the reaction. Add sample and ¹²⁵ I-NKA (0.61 KBq), 0.2 ml
The reaction was carried out at 25 ° C. for 3 hours in the above reaction buffer. Neurokinin A was added at a concentration of 1 × 10 ⁻⁶ M to determine the amount of non-specific binding. After the reaction, the reaction was stopped by rapid filtration on a glass filter using a cell harvester, and 250 μl of 50 mM Tris-HCl buffer (pH
After washing 3 times with 7.4), the radioactivity remaining on the filter was measured with a gamma counter.

Receptor binding inhibitory activity was expressed as the concentration of the compound causing 50% inhibition (IC ₅₀ ). When the acute toxicity of TAN-1666A of the present invention was examined using mice, the acute toxicity was extremely weak, and no case of death was observed at a dose of 400 mg / kg by intraperitoneal or oral administration. Thus, the TAN-1666-related compound is a compound that inhibits the binding of tachykinin-type peptide receptors and has low toxicity to mammals, and is useful as a therapeutic or prophylactic agent for tachykinin-mediated diseases.

The compound of the present invention having a tachykinin receptor binding inhibitory action or a salt thereof is used for tachykinin-mediated diseases such as respiratory diseases (eg, dogs, pigs, cows, horses, monkeys, humans) Asthma, bronchitis, rhinitis, cough, sputum, etc.), eye diseases (eg conjunctivitis, spring catarrh etc.), skin diseases (eg contact dermatitis, atopic dermatitis, urticaria, other eczema-like dermatitis) , Inflammatory diseases (eg, rheumatoid arthritis, osteoarthritis, etc.), pain (eg, migraine, headache, toothache, cancer pain, backache, etc.), cardiovascular disease (eg, hypertension, angina, heart failure, thrombosis, etc.) ), And is useful as a compound having a tachykinin antagonism effective for the treatment or prevention of brain diseases (eg dementia, schizophrenia, etc.).

The tachykinin antagonist of the present invention comprises a compound represented by the general formula [I] or a salt thereof (hereinafter abbreviated as the compound of the present invention) in a pharmacologically acceptable carrier according to a method known per se. By mixing, it can be obtained as a parenteral or oral preparation. The tachykinin antagonist is in the form of parenteral preparations such as injections, drops, solutions, suspensions and suppositories (eg, rectal suppositories, vaginal suppositories), and oral preparations include, for example, It can be provided in a dosage form such as capsules, tablets, syrups, powders and granules, or in other dosage forms suitable as medicines. Parenteral preparations such as injections are prepared by dissolving the compound of the present invention in a conventional aqueous diluent to prepare a liquid preparation. Examples of the aqueous diluent include an aqueous glucose solution, physiological saline, Ringer's solution, and nutritional supplement solution. At this time, the compound of the present invention was added to a dispersant (eg, Tween 80 (manufactured by Atlas Powder Co., USA), HCO 6
0 (manufactured by Nikko Chemicals), polyethylene glycol, carboxymethylcellulose, sodium alginate, etc., isotonicity agent (eg glucose, sorbitol, mannitol, sodium chloride etc.), preservative (eg benzyl alcohol, chlorobutanol, parahydroxybenzoic acid) Methyl acid, methylparaben, propylparaben, etc.), anticoagulant (eg, dextran sulfate, heparin, etc.), solubilizer (eg, cyclodextrin, tween, etc.), stabilizer (eg, polyethylene glycol, polylactic acid, etc.) Etc. may be added. Further, the compound of the present invention can be made into an oily or aqueous solid, semisolid or liquid suppository. At this time, as the oily base used,
For example, glycerides of higher fatty acids [eg, cocoa butter, witepsols (manufactured by Dynamite Nobel), etc.], intermediate fatty acids [eg, migliols (manufactured by Dynamite Nobel), etc.] or vegetable oils (eg, sesame oil, soybean oil, cottonseed oil, etc.) ) And the like. Examples of the aqueous base include polyethylene glycols and propylene glycol, and examples of the aqueous gel base include natural gums, cellulose derivatives, vinyl polymers and acrylic acid polymers.

The oral agent is obtained by adding to the compound of the present invention, for example, an excipient (eg, lactose, sucrose, starch, etc.), a disintegrating agent (eg, starch, calcium carbonate, etc.), a binder (eg, starch,
Gum arabic, carboxymethyl cellulose, polyvinylpyrrolidone, hydroxypropyl cellulose, etc.)
Alternatively, a lubricant (eg, talc, magnesium stearate, polyethylene glycol 6000, etc.) is added, compression molding is performed, and then, if necessary, coating by a method known per se for the purpose of taste masking, enteric coating or sustainability. It is manufactured by Examples of the coating agent include hydroxypropyl methyl cellulose, ethyl cellulose, hydroxymethyl cellulose, hydroxypropyl cellulose, polyoxyethylene glycol, Tween 80, and Bluronic F 6
8. Dyes such as cellulose acetate phthalate, hydroxypropylmethylcellulose phthalate, hydroxymethylcellulose acetate succinate, Eudragit (manufactured by Rohm Co., Germany, methacrylic acid / acrylic acid copolymer) and red iron oxide are used.

These preparations are administered orally or parenterally. For example, when used in humans, the dose may vary depending on the type and degree of target disease, age of patient, etc.
The content of the compound of the present invention is about 0.1 per adult per day.
5 mg to 500 mg, preferably 1 mg to 200 mg, is used to treat the disease.

[0031]

The present invention will be described in more detail with reference to the following examples, which should not be construed as limiting the invention. The percentage (%) in the medium is expressed as weight / volume percentage unless otherwise specified. Other percentages (%) refer to volume / volume percent unless otherwise stated. The abbreviations used in the examples have the following meanings. s: singlet, br: broad, d: doublet,
dd: double doublet, t: triplet, q: quartet, m: multiplet, J: coupling constant, DMSO-d ₆ : heavy methyl sulfoxide Example 1 potato dextrose broth (manufactured by Difco, USA) 24 g, Aspergillus terreus FL-29694 strain cultivated at 28 ° C. for 7 days on a slant culture medium consisting of 20 g of agar and 1 liter of water, 2% glucose, 3% maltose, 0.75% raw soybean powder, 0.5% corn Steep Liquor (CSL), 0.25% peptone, 0.15%
500 ml containing yeast extract and 0.15% sodium chloride
To a seed medium (pH 6.0) and cultured in a 2-liter Sakaguchi flask at 28 ° C. for 48 hours on a reciprocal shaker to obtain a seed culture. 2 liters of the resulting seed culture solution
120 liters of 1.5% glycerol, 3.5% maltose, 2.0% raw soy flour, 1.0% CSL, 0.5% calcium carbonate, 0.
05% Act Call (Takeda Pharmaceutical Co., Ltd.), 0.02%
Transfer to a main fermentation medium containing silicon (pH 7.0) and
Culturing was carried out for 5 days under the conditions of 4 ° C., aeration rate, 120 liters / minute, stirring, 120 rpm, and internal pressure, 1 kg / cm ² .

Example 2 A culture solution (95 liters) was adjusted to pH 2.5, ethyl acetate (50 liters) was added, and the mixture was stirred for 30 minutes and then filtered using Radiolite 600 (manufactured by Showa Chemical Industry Co., Ltd.). The ethyl acetate layer (45 liters) was extracted with a 2% (weight) aqueous sodium hydrogen carbonate solution (20 liters × 2). T
The ethyl acetate layer containing AN-1666A (Compound 1) is
After washing with water (20 liters x 2), the mixture was concentrated to obtain the crude substance (12.3
g) was obtained. This is subjected to silica gel chromatography (5
00 ml, Kieselgel 60, 70-230 mesh, manufactured by A Merck, Germany, and developed with an eluent in which acetone was sequentially added to hexane to obtain a crude powder (6.20 g) from the 40% acetone elution fraction. ) Got. Further, after subjecting it to silica gel chromatography (300 ml) and developing with an eluent in which ethyl acetate was sequentially added to toluene, a crude substance of compound 1 (1.75 g) was obtained from a fraction eluted with 40% ethyl acetate.
Got Preparative HPLC (column: YMC Pack
S-363 I-15), and eluent (55% acetonitrile / 0.02M phosphate buffer pH 6.3, flow rate: 15 ml /
min) and collect peaks with retention times of 32 to 45 minutes, remove acetonitrile, and then remove ethyl acetate (100 ml).
It was extracted in × 2). The ethyl acetate layer is water (70 ml x 2)
After washing with water, dehydration with anhydrous sodium sulfate, and concentration were performed to obtain Compound 1 (1.35 g) as a white powder. TAN-1
A sodium hydrogen carbonate layer containing 666B (compound 2) (4
3 liters) was adjusted to pH 6.5, concentrated to 4.5 liters, and subjected to column chromatography with Diaion HP-20 (500 ml, manufactured by Mitsubishi Kasei Co., Ltd.).
After washing with water (2.0 liters) and 50% methanol water (2.0 liters), it was eluted with 70% methanol / 0.02N ammonia water (2.5 liters). Methanol and ammonia were concentrated and removed, and the pH was adjusted to 3.0, followed by extraction with ethyl acetate (300 ml x 2). The ethyl acetate layer was washed with water (200 ml x 2), dried over anhydrous sodium sulfate and concentrated to obtain a crude substance. (5.40 g) was obtained. Further, it was subjected to silica gel column chromatography (300 ml) and developed with an eluent obtained by sequentially adding methanol to dichloroethane, and the crude substance of Compound 2 (2.42 g) was obtained from the fractions eluted with 5% methanol and 20% methanol. ) Got. Preparative HPLC (column: YMC Pack S
-363 I-15), and eluent (35% acetonitrile / 0.02M phosphate buffer pH 6.3, flow rate: 15 ml /
min), and the peaks with a retention time of 16.0 minutes to 28.0 minutes were collected, acetonitrile was removed, and the pH was adjusted to 3.0, followed by extraction with ethyl acetate (100 ml × 2). The ethyl acetate layer was washed with water (70 ml × 2), dried over anhydrous sodium sulfate and concentrated, and the residue was crystallized from ethyl acetate to give compound 2 (1.16 g) as colorless crystals.

Example 3 Compound 2 (300 mg) was dissolved in a mixed solvent of ethyl acetate (100 ml) and methanol (20 ml), and diazomethane (ether solution) was added until the solvent became yellow. After confirming that the starting material, compound 2, has completely disappeared,
The reaction solution was concentrated, and the residue was crystallized from methanol to give compound 5 (271 mg) as colorless crystals. Its physical properties were as follows: Elemental analysis: as C ₃₁ H ₃₃ N ₃ O ₅ (%) Calculated value: C; 70.57, H; 6.30, N; 7.96 Measured value: C; 70.32, H; 6.29, N; 8.14 IRνmax (KBr, cm ^-1 ): 3280, 1740, 1650, 1620, 1550 ¹ H NMR δppm (300MHz, in DMSO-d ₆ ): 0.97 (3H, d, J
= 7.0Hz), 2.51 (1H, m), 2.79 (1H, dd, J = 3.7,13.6Hz), 2.91
(1H, dd, J = 9.8,13.6Hz), 3.10 (1H, dd, J = 5.2,13.6Hz),
3.64 (3H, s), 4.40 (1H, quintet, J = 7.2Hz), 4.53 (2H, m),
6.73 (1H, d, J = 15.8Hz), 7.08-7.54 (16H, m), 8.20 (1H, d, J =
7.3Hz), 8.24 (1H, d, J = 8.7Hz), 8.58 (1H, d, J = 8.2Hz) ¹³ C NMR δppm (75MHz, in DMSO-d ₆ ): 18.2
(Q), 36.9 (t), 37.6 (t), 4
8.2 (d), 51.9 (q), 53.3 (d),
53.4 (d), 121.8 (d), 126.1
(D), 126.5 (d), 127.4 (d),
127.4 (d), 127.8 (d), 127.8
(D), 128.1 (d), 128.1 (d), 1
28.8 (d), 128.8 (d), 129.1
(D), 129.1 (d), 129.1 (d), 1
29.1 (d), 129.3 (d), 134.8
(S), 137.0 (s), 137.6 (s),
138.9 (d), 164.5 (s), 170.8
(S), 171.8 (s), 171.9 (s)

Example 4 Compound 5 (137 mg) was dissolved in a 2M ammonia / methanol solution (100 ml) and left at room temperature overnight. The reaction solution was concentrated to dryness to obtain Compound 8 (82.6 mg) as a white powder. Elemental _{analysis: C 30 H 32 N 4 O} 4 · 1 / 4H 2 O as (%) Calculated: C; 69.68, H; 6.34 , N; 10.84 Found: C; 69.74, H; 6.23 , N; 10.81 IRνmax (KBr, cm ^-1 ): 3440, 3290, 1680, 1640, 1610,
1540 ¹ H NMR δppm (300MHz, in DMSO-d ₆ ): 1.00 (3H, d, J
= 7.0Hz), 2.52 (1H, m), 2.79 (1H, dd, J = 3.7,13.6Hz), 2.82
(1H, dd, J = 10.9,13.5Hz), 3.07 (1H, dd, J = 4.1,13.5Hz), 4.
36 (1H, quintet, J = 7.2Hz), 4.47 (2H, m), 6.75 (1H, d, J = 1
5.8Hz), 7.03-7.56 (18H, m), 8.20 (1H, d, J = 8.6Hz), 8.30
(1H, d, J = 7.2Hz ), 8.37 (1H, d, J = 8.7Hz) 13 C NMRδppm (75MHz, in _{DMSO-d 6): 18.1 (} q), 3
7.4 (t), 37.6 (t), 48.5 (d), 53.6 (d), 54.0 (d), 121.8
(d), 126.0 (d), 126.1 (d), 127.4 (d), 127.4 (d), 127.7
(d), 127.7 (d), 127.9 (d), 127.9 (d), 128.8 (d), 128.8
(d), 129.1 (d), 129.1 (d), 129.2 (d), 129.2 (d), 129.3
(d), 134.8 (s), 137.7 (s), 138.0 (s), 138.9 (d), 164.6
(s), 170.5 (s), 171.9 (s), 173.0 (s)

Example 5 Compound 2 (100 mg) was dissolved in DMF (5.0 ml),
Potassium carbonate (29.6 mg, 1.1 eq) and iodoethane (18.7 μl, 1.2 eq) were added, and the mixture was allowed to stand at room temperature for 20 minutes.
Stir for hours. After adding ethyl acetate (50 ml) to the reaction solution, a 2% (weight) aqueous solution of sodium hydrogencarbonate (20 m
l), washed with water (20 ml × 3), dried over anhydrous sodium sulfate, and concentrated to obtain compound 6 (81 mg) as a white powder. Elemental analysis: (%) (as C ₃₂ H ₃₅ N ₃ O ₅ ) Calculated value: C; 70.96, H; 6.51, N; 7.76 Measured value: C; 70.78, H; 6.61, N; 7.70 IRνmax (KBr, cm ^-1 ): 3280, 1730, 1640, 1620, 1550 ¹ H NMR δppm (300MHz, in DMSO-d ₆ ): 0.97 (3H, d, J
= 7.0Hz), 1.14 (3H, t, J = 7.1Hz), 2.52 (1H, m), 2.79 (1H, d
d, J = 3.8,13.5Hz), 2.91 (1H, dd, J = 9.6,13.6Hz), 3.07 (1H,
dd, J = 5.6,13.6Hz), 4.08 (2H, q, J = 7.1Hz), 4.39 (1H, quint
et, J = 7.2Hz), 4.51 (2H, m), 6.72 (1H, d, J = 15.8Hz), 7.09-
7.54 (16H, m), 8.21 (1H, d, J = 7.6Hz), 8.24 (1H, d, J = 9.1H
z), 8.55 (1H, d, J = 8.0Hz) ¹³ C NMR δppm (75MHz, in DMSO-d ₆ ): 13.9 (q), 1
8.2 (q), 37.0 (t), 37.6 (t), 48.2 (d), 53.3 (d), 53.5
(d), 60.5 (t), 121.8 (d), 126.1 (d), 126.4 (d), 127.4
(d), 127.4 (d), 127.7 (d), 127.7 (d), 128.1 (d), 128.1
(d), 128.8 (d), 128.8 (d), 129.1 (d), 129.1 (d), 129.1
(d), 129.1 (d), 129.3 (d), 134.8 (s), 137.0 (s), 137.8
(s), 138.9 (d), 164.4 (s), 170.8 (s), 171.3 (s), 171.9
(s) In the same manner as in the above method, compound 2 (100 mg), benzyl bromide (57.6 μl, 1.2 equivalents) to compound 7 (9
7.1 mg) was obtained. Elemental analysis: C ₃₇ H ₃₇ N ₃ O ₅ (%) Calculated value: C; 73.61, H; 6.18, N; 6.96 Measured value: C; 73.51, H; 6.23, N; 6.95 IRνmax (KBr, cm ^-1 ): 3280, 1730, 1650, 1620, 1550 ¹ H NMR δppm (300MHz, in DMSO-d ₆ ): 0.97 (3H, d, J
= 7.0Hz), 2.50 (1H, m), 2.77 (1H, dd, J = 3.6,13.6Hz), 2.94
(1H, dd, J = 9.6,13.6Hz), 3.12 (1H, dd, J = 5.5,13.6Hz), 4.4
1 (1H, quintet, J = 7.2Hz), 4.55 (2H, m), 5.12 (2H, d, J = 1.5
Hz), 6.73 (1H, d, J = 15.8Hz), 7.08-7.53 (21H, m), 8.23 (1
H, d, J = 7.2Hz), 8.26 (1H, d, J = 8.7Hz), 8.64 (1H, d, J = 8.1H
z) ¹³ C NMR δppm (75MHz, in DMSO-d ₆ ): 18.3 (q), 3
6.9 (t), 37.7 (t), 48.2 (d), 53.3 (d), 53.6 (d), 66.0
(t), 121.8 (d), 126.1 (d), 126.5 (d), 127.4 (d), 127.4
(d), 127.7 (d), 127.7 (d), 127.7 (d), 127.8 (d), 127.8
(d), 127.9 (d), 128.1 (d), 128.1 (d), 128.3 (d), 128.3
(d), 128.3 (d), 128.8 (d), 128.8 (d), 129.1 (d), 129.1
(d), 129.4 (d), 134.8 (s), 135.6 (s), 136.9 (s), 137.5
(s), 138.9 (d), 164.4 (s), 170.9 (s), 171.2 (s), 171.9
(s)

Example 6 Compound 2 (100 mg) was suspended in dichloromethane (10 ml) and benzylamine (25.6 μl, 1.2 eq), HOBT (1-hydroxybenzotriazole anhydrous) was added. Dojindo Laboratories) (31.6mg, 1.2eq), WSC (hydrochloric acid 1
-Ethyl-3- (3-dimethylaminopropyl) -carboimide
diimide hydrochloride) (made by Dojindo Laboratories) (44.
9 mg, 1.2 equivalents, triethylamine (32.4μ
(1, 1.2 equivalents) was added, and the mixture was stirred at 0 ° C. for 1 hr and further at room temperature for 16 hr. Ethyl acetate (5
(0 ml) was added, and the mixture was washed with 2% (weight) aqueous sodium hydrogen carbonate solution (20 ml) and water (20 ml × 3), and the ethyl acetate layer was concentrated to give compound 10 (100 mg) as a white powder. Elemental analysis: C ₃₇ H ₃₈ N ₄ O ₄・ 1 / 2H ₂ O (%) Calculated value: C; 72.65, H; 6.43, N; 9.16 Measured value: C; 72.65, H; 6.39, N; 9.14 IRνmax (KBr, cm ^-1 ): 3280, 1640, 1620, 1540 ¹ H NMR δppm (300MHz, in DMSO-d ₆ ): 0.98 (3H, d, J
= 7.0Hz), 2.50 (1H, m), 2.75 (1H, dd, J = 3.8,13.4Hz), 2.83
(1H, dd, J = 10.3,13.5Hz), 3.05 (1H, dd, J = 4.6,13.4Hz),
4.29 (2H, brt, J = 4.9Hz), 4.38 (1H, quintet, J = 7.1Hz),
4.55 (2H, m), 6.71 (1H, d, J = 15.8Hz), 7.03-7.54 (21H, m),
8.16 (1H, d, J = 8.5Hz), 8.22 (1H, d, J = 7.5Hz), 8.49 (1H, d, J
= 8.6Hz), 8.54 (1H, t, J = 5.9Hz) ¹³ C NMR δppm (75MHz, in DMSO-d ₆ ): 18.3 (q), 3
7.6 (t), 37.9 (t), 41.9 (t), 48.2 (d), 53.5 (d), 54.1
(d), 121.8 (d), 126.0 (d), 126.2 (d), 126.6 (d), 126.9
(d), 126.9 (d), 127.4 (d), 127.4 (d), 127.7 (d), 127.7
(d), 127.9 (d), 127.9 (d), 128.1 (d), 128.1 (d), 128.8
(d), 128.8 (d), 129.2 (d), 129.2 (d), 129.2 (d), 129.2
(d), 129.3 (d), 134.8 (s), 137.6 (s), 137.7 (s), 138.8
(s), 138.9 (d), 164.4 (s), 170.6 (s), 170.8 (s), 171.9
(s)

Example 7 Compound 2 (300 mg) was suspended in dichloromethane (30 ml), ethylamine hydrochloride (57.3 mg, 1.2 eq), HOBT (1-hydroxybenzotriazole anhydrous). (Manufactured by Dojindo Laboratories) (94.8 mg, 1.2 equivalents), WSC (1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride) (1-Ethyl-3- (3-dimethyl aminopropyl) -car
bodiimide hydrochloride) (manufactured by Dojindo Laboratories) (13
5 mg, 1.2 eq), triethylamine (162 μl,
(2.0 equivalents) was added, and the mixture was stirred at 0 ° C. for 1 hr, and further at room temperature for 16 hr. Ethyl acetate (50m
l) was added, the mixture was washed with 2% (weight) aqueous sodium hydrogen carbonate solution (20 ml) and water (20 ml), and the ethyl acetate layer was concentrated to give compound 9 (340 mg) as a white powder. Elemental analysis: As C ₃₂ H ₃₆ N ₄ O ₄ 1 / 2H ₂ O (%) Calculated value: C; 69.92, H; 6.79, N; 10.19 Measured value: C; 69.81, H; 6.69, N; 10.19 IRνmax (KBr, cm ^-1 ): 3270, 1640, 1620, 1540 ¹ H NMR δppm (300MHz, in DMSO-d ₆ ): 0.97 (3H, d, J
= 6.9Hz), 0.99 (3H, t, J = 7.1Hz), 2.47 (1H, dd, J = 10.2,13.
6Hz), 2.72 (1H, dd, J = 3.5,13.6Hz), 2.77 (1H, dd, J = 10.3,1
3.6Hz), 2.99 (1H, dd, J = 4.7,13.7Hz), 3.08 (2H, m), 4.38
(1H, quintet, J = 7.2Hz), 4.48 (2H, m), 6.71 (1H, d, J = 15.8H
z), 7.04-7.55 (16H, m), 7.98 (1H, t, J = 5.4Hz), 8.15 (1H,
d, J = 8.6Hz), 8.20 (1H, d, J = 7.6Hz), 8.42 (1H, d, J = 8.7Hz) ¹³ C NMR δppm (75MHz, in DMSO-d ₆ ): 14.4 (q), 1
8.3 (q), 33.3 (t), 37.5 (t), 38.0 (t), 48.2 (d), 53.6
(d), 54.0 (d), 121.9 (d), 126.0 (d), 126.1 (d), 127.4
(d), 127.4 (d), 127.7 (d), 127.7 (d), 127.9 (d), 127.9
(d), 128.8 (d), 128.8 (d), 129.2 (d), 129.2 (d), 129.2
(d), 129.2 (d), 129.3 (d), 134.8 (s), 137.6 (s), 137.8
(s), 138.8 (d), 164.4 (s), 170.4 (s), 170.5 (s), 172.0
(s)

Example 8 Compound 2 (100 mg) was suspended in dichloromethane (3.0 ml), and WSC (1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride) (1-Ethyl-3- ( 3-dime
thyl aminopropyl) -carbodiimide hydrochloride) (manufactured by Dojindo Laboratories) (45.7 mg, 1.2 equivalents), HOB
T (anhydrous 1-hydroxybenzotriazole (1-hydroxy
benzotriazole diluted)) (made by Dojindo Laboratories) (3
2.2 mg, 1.2 eq), N, N-dimethylpropanediamine (28 μl, 1.1 eq) were added and the mixture was allowed to stand at room temperature for 16
Stir for hours. Hexane (3.0 ml) and ethyl acetate (7.0 ml) were added to the reaction solution, followed by extraction with a 10% (weight) aqueous ammonium chloride solution (10 ml × 2). The ammonium chloride layer was adjusted to pH 8.3 with an aqueous sodium hydroxide solution and then extracted with ethyl acetate (10 ml × 2) and 5% isopropyl alcohol / ethyl acetate (10 ml), respectively. The organic layers were combined, washed with 2% (weight) sodium hydrogen carbonate and saturated brine, and dried over sodium sulfate.
The organic layer was concentrated to dryness to obtain Compound 11 (94 mg) as a white powder. Elemental analysis: C ₃₅ H ₄₃ N ₅ O ₄・ 1 / 4H ₂ O (%) Calculated value: C; 69.80, H; 7.28, N; 11.62 Actual value: C; 69.58, H; 7.30, N; 11.81 IRνmax (KBr, cm ^-1 ): 3280, 1640, 1620, 1540 ¹ H NMR δppm (in 300MHz, DMSO-d ₆ ): 0.99 (3H, d, J
= 7.0Hz), 1.50 (2H, quintet, J = 7.1Hz), 2.10 (6H, s), 2.18
(2H, t, J = 7.0Hz), 2.51 (1H, m), 2.75 (1H, dd, J = 3.5,13.3H
z), 2.79 (1H, dd, J = 10.2,13.7Hz), 3.01 (1H, dd, J = 5.7,13.
5Hz), 3.07 (2H, m), 4.38 (1H, quintet, J = 7.2Hz), 4.47 (1
H, m), 4.50 (1H, m), 6.72 (1H, d, J = 15.8Hz), 7.02-7.58 (16
H, m), 7.96 (1H, t, J = 5.5Hz), 8.16 (1H, d, J = 8.5Hz), 8.20
(1H, d, J = 7.5Hz), 8.39 (1H, d, J = 8.5Hz) ¹³ C NMR δppm (75MHz, in DMSO-d ₆ ): 18.3 (q), 2
6.7 (t), 36.8 (t), 37.5 (t), 37.9 (t), 44.9 (q), 44.9
(q), 48.3 (d), 53.6 (d), 54.1 (d), 56.4 (t), 121.9 (d),
126.0 (d), 126.2 (d), 127.4 (d), 127.4 (d), 127.7 (d), 1
27.7 (d), 127.9 (d), 127.9 (d), 128.8 (d), 128.8 (d), 12
9.2 (d), 129.2 (d), 129.2 (d), 129.2 (d), 129.3 (d), 13
4.8 (s), 137.6 (s), 137.8 (s), 138.8 (d), 164.5 (s), 17
0.6 (s), 170.6 (s), 172.0 (s) From the compound 2 (100 mg), N-methylpiperazine (24 μl, 1.1 equivalent) to the compound 1 in the same manner as above.
2 (61 mg) was obtained as a white powder. Elemental analysis: as _{C 35 H 41 N 5 O 4} · 1 / 4H 2 O (%) Calculated: C; 70.04, H; 6.97 , N; 11.66 Found: C; 69.84, H; 6.98 , N; 11.82 IRνmax (KBr, cm ^-1 ): 3430, 3300, 1650, 1630, 1540 ¹ H NMR δppm (300MHz, in DMSO-d ₆ ): 0.97 (3H, d, J
= 7.0Hz), 1.96 (1H, m), 2.08 (3H, s), 2.18 (3H, m), 2.52
(1H, m), 2.76 (1H, m), 2.78 (1H, m), 2.95 (1H, dd, J = 6.2,1
3.3Hz), 3.44 (4H, brs), 4.40 (1H, quintet, J = 7.1Hz), 4.5
3 (1H, td, J = 8.7,3.4Hz), 4.95 (1H, q, J = 6.8Hz), 6.72 (1H,
d, J = 15.8Hz), 7.12-7.54 (16H, m), 8.17 (1H, d, J = 6.8Hz),
8.21 (1H, d, J = 8.9Hz), 8.57 (1H, d, J = 8.5Hz) ¹³ C NMR δppm (75MHz, DMSO-d ₆ ): 18.3 (q), 3
7.7 (t), 37.8 (t), 41.3 (t), 44.8 (t), 45.4 (q), 48.2
(d), 49.4 (t), 53.5 (t), 54.0 (d), 54.3 (d), 121.9 (d),
126.0 (d), 126.3 (d), 127.4 (d), 127.4 (d), 127.7 (d), 1
27.7 (d), 128.0 (d), 128.0 (d), 128.8 (d), 128.8 (d), 12
9.2 (d), 129.2 (d), 129.3 (d), 129.4 (d), 129.4 (d), 13
4.9 (s), 137.3 (s), 137.7 (s), 138.9 (d), 164.4 (s), 16
8.9 (s), 170.3 (s), 171.8 (s)

Formulation Example Compound 9 (50 mg) obtained in Example 7 was used in an internal volume of 12
A sterile vial of ml was filled and then sealed.

[0040]

INDUSTRIAL APPLICABILITY According to the present invention, there is provided a novel compound having a tachykinin antagonistic effect and effective for treating or preventing a tachykinin-mediated disease.

[Brief description of drawings]

FIG. 1 ¹³ C NMR spectrum of TAN-1666A.

FIG. 2 ¹³ C NMR spectrum of TAN-1666B

Claims (5)

[Claims] 1. A general formula: [Wherein R ₁ represents hydrogen, R ₂ represents a bond together with a hydroxyl group, an optionally substituted hydrocarbon oxy group, an optionally substituted amino group or R ₁ ] Or a salt thereof. 2. The compound according to claim ₁ , wherein R ₁ is hydrogen and R ₂ is an alkylamino group. 3. The compound according to claim 2, wherein the alkylamino group is an ethylamino group. 4. A compound belonging to the genus Aspergillus and represented by the formula: Compound TAN-1666A represented by the formula: The compound TAN-1666A or B or a salt thereof, which comprises culturing a microorganism having the ability to produce the compound TAN-1666B represented by the formula (1) in a medium, allowing the compound to be produced and accumulated in the culture, and collecting the compound. Manufacturing method. 5. A tachykinin antagonist comprising the compound according to claim 1.