WO1996016038A1 - 2-[2-(indol-3-yl)ethylamino]-1-phenylethanol derivative - Google Patents

Abstract

A 2-[2-(indol-3-yl)ethylamino]-1-phenylethanol derivative represented by general formula (I) and a pharmaceutically acceptable acid-addition salt thereof, wherein R1 stands for halogen or trifluoromethyl; R2 stands for hydrogen, lower alkyl or fluorinated lower alkyl; and R3 stands for hydrogen, halogen, trifluoromethyl, nitro or cyano. This compound has a highly selective activity of stimulating β3 adrenaline receptors and is usable for treating or preventing diabetes, obesity, irritable bowel syndrome, acute or chronic diarrhea and so forth, and also for ameliorating syndromes accompanying peptic ulcer, acute or chronic gastritis, and so forth, such as abdominal pain, nausea, vomiting and epigastric discomfort.

Description

 Specification

2— [2— (indole-3-yl) ethylamino] 1-111

 Phenylethanol derivatives Technical field

The present invention, 5 ₃ A Dorenari emissions receptor highly selective, ₃ A Dorenarin novel 2 having a receptor stimulating effect [2- (Indoru one 3-I le) Echirua amino] one 1 one-phenylalanine ethanol Related to derivatives.

Conventional technology

There are two subtypes of sympathetic receptors, 3! And ₂ ; the former is predominantly located in the heart, and the latter has long been thought to be present in bronchial and vascular smooth muscle [L ands, AM et al .: Nature, 214, 597-598 (1967)]. Currently,;! 5 Adorenari emissions receptor agonists as cardiac function enhancers or vasopressors, beta ₂ Adorenarin receptor agonists have been clinically used respectively as a bronchodilator.

Recently, 3 adrenergic receptors have been isolated as a third subtype different from the two subtypes described above [E morine. L. J. et al .: Science, 5, 1118-1121 (1989). ]. These _3- adrenergic receptors are present in the digestive tract, adipose tissue and skeletal muscle, and are thought to be involved in energy consumption based on lipolysis, promotion of glycogen degradation, and relaxation of intestinal smooth muscle. 3 ₃ as a drug which selectively actuated adrenergic phosphorus receptors, e.g. SR- 5 8 6 1 1 A [(R S.) -. N- (7- E Bok Kin carbonyl main Tokishi 1 2, 3, 4 Tetrahydric naphthol 2- (yl) -12- (3-chlorophenyl) -12-hydroxyethaneamine hydrochloride: Japanese Patent Application Laid-Open No. Sho. 255415 specification] and BRL 35135 C (R * R *)-(±)-[4-1 [2- [3- (3-chlorophenyl) -12-hydroquinethylamino] propyl] phenoxy ] Methyl acetate hydrobromide; Japanese Patent Publication No. 63-26744 and European Patent No. 23385] are known. SR—586

11A has an excellent inhibitory effect on spontaneous movement of the isolated rat colon.CBrit. J. Pharmacol., 100, 831-839 (1990)]

35135 has been reported to have an antiobesity effect and a blood glucose lowering effect by oral administration [Drugs of the Future, 16, 797-800 (1991)].

 'On the other hand, there are several known 2- [2- (indole-1-yl) ethylamino] -11-phenylethanol derivatives. Jackman, GB et al., Cited 21- (5- (5-cycloindole-13-yl) ethylamino) -111-phenylethanol [J. Pharm. Pharmacol., 17, 742-746 (1965)]. And Biniecki, S. et al., 2- [2- (indole-3-inole) ethylamino] -1-phenylethanol [Acta Polon. Pharm., 38, 407-410 (1981) (Chem. Abstr., 96. 142543k (1982))], but does not mention any of its pharmacological effects.

Also, in British Patent No. 861428, an indole derivative represented by the following formula (VI) and a pharmaceutically acceptable non-toxic acid addition salt thereof have a bronchodilator effect and a central nervous system. It is disclosed to show valuable effects on the system (antiemetic and hypnotic enhancing effects). This specification mentions 3- [2- [2-hydroxy-2- (3.4-dihydroxyphenyl) ethylamino] propyl] indole as the most preferred compound. H

 (In the formula, Alk represents a straight-chain or branched-chain alkylene group having 2 to 4 carbon atoms-Alk 'represents a straight-chain or branched-chain alkylene group having 2 to 4 carbon atoms or a hydroquinone alkylene group. X represents a hydrogen atom or a hydroxyl group, and Y represents a hydroxyl group or a methoxy group.)

 In addition, Sarkisyan, A.B. et al. Reported that the N-indolylalkylaminol-1-aryl-substituted alkynol derivative represented by the following formula (K) has a moderate and ^ adrenergic receptor blocking action. Chem. Abs tr., 109. 128763η (1988)].

α — π- CH ₂ CHR ¹ NHCH ₂ CHROH

 Η

(In the formula, R represents a phenyl group, an m-ditrophenyl group or a p-ditrophenyl group, and R ¹ represents a hydrogen atom or a methyl group.)

Disclosure of the invention

An object of the present invention is to provide a 2- [2- (indole-13-yl) ethylamino] 1-1-phenyl having an excellent ^ _3- adrenergic receptor stimulating effect having a chemical structure different from that of a conventional known compound. Provided are an ethanol derivative and an acid addition salt thereof, and a method for producing the ethanol derivative and an acid addition salt thereof. Is to provide the law.

BEST MODE FOR CARRYING OUT THE INVENTION

 According to the present invention, a 2- [2- (indole-13-yl) ethylamino] -11-phenylethanol derivative represented by the following formula (I) and a pharmaceutically acceptable acid addition salt thereof are provided. Provided.

(I)

 H

(In the formula, means a halogen atom or a trifluoromethyl group,

R ₂ represents a hydrogen atom, a lower alkyl group or a fluoro lower alkyl group, and R ₃ represents a hydrogen atom, a halogen atom, a trifluoromethyl group, a nitro group or a cyano group. )

 Examples of the pharmaceutically acceptable acid addition salt of the compound represented by the formula (I) include inorganic acids such as hydrochloride, hydrobromide, hydroiodide, sulfate, phosphate and the like. Salts and organic acid salts such as oxalate, maleate, fumarate, lactate, malate, citrate, tartrate, benzoate and methanesulfonate. Since the compound represented by the formula (I) and a pharmaceutically acceptable acid addition salt thereof may exist in the form of a hydrate or a solvate, these hydrates and solvates may also be used. Included in the compounds of the present invention.

The compound represented by the formula (I) has at least one asymmetric carbon violet atom, that is, a carbon atom to which a hydroxy group is bonded in the formula (I). Further, when R ₂ is a lower alkyl group or a fluoro lower alkyl group, the carbon atom to which this group is bonded is also an asymmetric carbon atom. Therefore, equation (I) In the formula, when R ₂ is a hydrogen atom, there may be two kinds of stereoisomers, and when R ₂ is a lower alkyl group or a fluoro lower alkyl group, there may be four kinds of stereoisomers. These stereoisomers, their mixtures and racemates are included in the compounds of the present invention.

 The terms used in this specification are described below.

The term "lower alkyl group" means one having 1 to 3 carbon atoms, and includes methyl, ethyl, propyl and isopropyl, with methyl being particularly preferred. "Fluoro-lower alkyl group" refers to an alkyl group having 1 to 3 carbon atoms in which a hydrogen atom has been replaced with a fluorine atom, and examples thereof include fluoromethyl, trifluoromethyl, and 2-fluoroethyl. Particularly preferred. “Halogen atom” means fluorine, chlorine, odor violet, and iodine violet, but fluorine, chlorine, and bromine are preferable, and fluorine and chlorine are particularly preferable. When R ₃ is an atom or a group other than a hydrogen atom, the bonding position of R ₃ may be any of the 4, 5, and 6.7 positions of the indole ring, but the 6 position is particularly preferred.

Among the compounds of the present invention, preferred are those in the above formula (I), wherein is a chlorine atom, R ₂ is a hydrogen atom, a methyl group or a fluoromethyl group, and R ₃ is a hydrogen atom, a halogen atom or a trifluoromethyl group. Compounds that are methyl groups and pharmaceutically acceptable acid addition salts thereof.

Further suitable compounds are the formula is R! Is a chlorine atom in (I), R ₂ is a hydrogen atom or a methyl group, Ku R ₃ is Wakashi fluorine atoms of the hydrogen atom or 6-position is a chlorine atom Compounds and pharmaceutically acceptable acid addition salts thereof. Particularly preferred compounds are those wherein R 1 is a chlorine atom, R ₂ is a methyl group and R ₃ is a water atom or a fluorine atom or a chlorine atom at the 6-position in the formula (I), Is an acid addition salt that is acceptable.

In the above formula (I), the compound represented by R ₂ or a group other than a hydrogen atom has two asymmetric groups. Since it has carbon atoms, there are four stereoisomers, but the (R, R) configuration is preferred.

 Particularly preferred compounds include, for example, the following compounds and pharmaceutically acceptable acid addition salts thereof.

 2-[(1R) -2- (indole-3-yl) -1-methylethylamino] 1 (1R) -11- (3-chlorophenyl) ethanol, and

 2- [2- (6-fluoroindole-1-yl) -1-methylethylamino] 1-1- (3-chlorophenyl) ethanol

 Specific examples of the compounds included in the present invention include the following compounds, their stereoisomers, and pharmaceutically acceptable acid addition salts thereof, in addition to the compounds of Examples described later. '

 2- [2- (6-fluoroindole-1-yl) ethylamino] 1-1 (3-trifluoromethylphenyl) ethanol,

 2- [2- (6-fluoroindole-3-inole) -1-methylethylamino] 1-11- (3-trifluoromethylphenyl) ethanol,

 2- [2- (6- (1-methyl) indole-3-yl) ethylamino] -1- (3-trifluoromethylphenyl) ethanol,

 2- [2- (6- (methyl) indole-1-yl) -1-methylethylamino] 1-1-1 (3-trifluoromethylphenyl) ethanol

 2- [2- (indole-3-yl) ethylamino] 1-1-1 (3-trifluoromethylphenyl) ethanol,

 2- [2- (indole-1-yl) -1-methylethylamino] 1-1-1 (3-trifluoromethylphenyl) ethanol,

2- (2- (4-chloroindole-1-yl) -1-methylethylamino) 1-1- (3-chlorophenyl) ethanol, 2- [2- (6- (trifluoromethylindole-1-3-yl) ethylamino] 1-11- (3-chlorophenyl) ethanol,

 2- [2- (6- (trifluoromethylindole-1-3-yl) -1-1-methylletylamino] 1-1-1 (3-chlorophenyl) ethanol,

 2- [2- (6-cyanoindole-1-yl) ethylamino] 1-1 (3-chlorophenyl) ethano

 2- [2- (6-cyanoindol-3-inole) -1-methylethylamino] 1-1-1 (3-chlorophenyl) ethanol,

 2- [2- (6- (2-nitroindole-3-yl) ethylamino] -1-1 (3-chlorophenyl) ethanol,

 2- [2-((6-12 troindole-1-3-inole) -1-1-methylethylamino] -11- (3-chlorophenyl) ethanol,

 2- [2- (6-fluoroindole-3-inole) -1-1-fluoromethylethylamino] 1-1-1 (3-chlorophenyl) ethanol,

 2- (2- (6-chloroindole-1-3-inole) -1-1-fluoromethylethylamino) -11- (3-chlorophenyl) ethanol, and

 2- [2- (indole-3-yl) -11-fluoromethylethylamino] 1-1-1 (3-chlorophenyl) ethanol

 The compound of the present invention can be produced, for example, by the following method.

Production method (a):

 The compound of the present invention represented by the formula (I) is represented by the following formula (Π)

(式中、 は前掲に同じものを意味する。 ）

(In the formula, means the same as above.)

And a compound represented by the following formula (m)

H ₂ N— CH— CH ₂

 (I ")

H

(In the formula, R ₂ and R ₃ mean the same as above.)

By reacting with a compound represented by the formula:

The reaction between the compound represented by the formula (π) and the compound represented by the formula (m) is carried out in a suitable solvent or without a solvent. The three solvents to be used should be appropriately selected according to the type of the raw material compounds, etc., for example, alcohols such as methanol, ethanol, isopropyl alcohol, ketones such as acetone, methylethylketone, methylene chloride, Halogenated hydrocarbons such as black form, ethers such as dimethyl ether, tetrahydrofuran, dioxane, aromatic hydrocarbons such as benzene and toluene, ethyl acetate, N, N-dimethylform Amides, dimethyl sulfoxide and the like can be mentioned, and these solvents are used alone or in combination of two or more. The compound of the formula (m) can be used in the form of an acid addition salt. Examples of these acid addition salts include inorganic acid salts such as hydrochloride and hydrobromide, oxalate, and maleic acid. Organic acid salts such as acid salts and fumarate salts. When such an acid addition salt is used, the reaction is carried out in the presence of a base, and specific examples of the base include sodium bicarbonate, alkali bicarbonate such as potassium bicarbonate, sodium carbonate, and the like. Carbonic acid such as sulfuric acid or triethylamine, tributylamine, diisopropylethylamine, N-methylmorpholine Such organic bases are mentioned. The reaction temperature varies depending on the type of the starting compound used and the like, but is usually about 20 ° C to about 150 ° C, preferably about 25 ° C to about 100 ° C.

 In this production method, when the starting compound has an asymmetric carbon atom, the configuration of the asymmetric carbon atom is retained in the product of the compound of the formula (I).

For example, a compound of formula ([pi) are racemic compounds of formula (I) is a racemate obtained from compounds of formula R ₂ is a hydrogen atom (m), R ₂ is other than hydrogen atoms The compound of formula (m) which is a group gives the compound of formula (I) which is a diastereomer mixture.

 From the compound of formula (π) having a specific configuration and the compound of formula (m), a compound of formula (I) having the same configuration is obtained.

The enantiomer of the compound of the formula (Π) can be obtained, for example, by the method of Bloom, JD et al. [J. Med. Chem., 3δ, 3081-3084 (1992)] or the method of Eliel, EL and Delmonte, DW [J. Org. Chem., 21. 596-597 (1956)]. The enantiomer of a compound in which R ₂ is a group other than a hydrogen atom in the above formula (ΙΠ) can be obtained, for example, by the method of Repke, DB and Ferguson, WJ [J. Heterocycl. Chem., 13, 775-778 (1976)] It can be manufactured according to.

Production method (b):

In the above formula (I), the compound wherein R ₃ is a hydrogen atom, a halogen atom or a trifluoromethyl group is represented by the following formula (IV)

(式中、 は前掲に同じものを意味する。 ）

(In the formula, means the same as above.)

And a compound represented by the following formula (V)

)

(In the formula, R ₂ means the same as described above, and R ₃ means a hydrogen atom, a halogen atom or a trifluoromethyl group.)

Can be produced by reacting with a compound represented by the following formula under reducing conditions.

 The term “under normal conditions” in the present production method means conditions under the presence of a reducing agent capable of reducing the imine moiety without affecting the liponyl group or under the conditions of contact under normal conditions.

 Examples of the reducing agent that can pass through the imine moiety without affecting the carbonyl group include, for example, sodium cyanoborohydride. This reaction is carried out in an appropriate solvent, and suitable solvents include alcohols such as methanol and ethanol. The reaction temperature is usually about 20 ° C to about 80 ° C.

When this production method is carried out under the conditions of contact flow, palladium, platinum oxide or the like is used as a catalyst. Preferred solvents include alcohols such as methanol and ethanol. The reaction temperature is usually about 10 ° C. to about 25 ° C. _c The compound of formula (IV) and the compound of formula (V) Instead of reacting under normal conditions, a catalytic amount of an acid is added to both compounds, and the reaction is carried out with a device such as Dean Stark while removing the generated water, whereby the following formula (X) is obtained.

(Wherein, R!, R ₂ and R _3, means the same thing supra.)

After producing the compound represented by the formula, the product can also be reduced to reduce the product.

 The step of producing the compound of the formula (X) is carried out in a suitable solvent, and p-toluenesulfonic acid, pyridin p-toluenesulfonic acid and the like are used as the acid. As the solvent, aromatic hydrocarbons such as benzene and toluene are preferable, and the reaction temperature is usually about 80 ° C to about 150 ° C. The step of passing the compound of the formula (X) is performed under conditions suitable for the transfer of the imine moiety, and the reduction during the reaction between the compound of the formula (W) and the compound of the formula (V) is performed. The conditions can be adopted as they are. This reduction step is also suitably performed using sodium borohydride as a base agent. Preferred solvents include alcohols such as methanol and ethanol, and the reaction temperature is usually about It is from 10 ° C to about 25 ° C.

Production method):

In the above formula (I), the compound wherein R ₃ is a hydrogen atom, a halogen atom or a trifluoromethyl group is also represented by the following formula (VI)

(式中、 R ,、 R ₂及び R ₃ ,は前掲に同じものを意味する。 ）

(In the formula, R, R ₂ and R ₃ , mean the same as described above.)

Can also be produced by reacting the compound represented by the formula (1) with an appropriate base agent.

 As the base agent to be used, diborane, lithium aluminum hydride and its alkoxy complex or a transition metal salt, aluminum chloride, boron trifluoride, phosphorus oxychloride or hydrogen added with a carboxylic acid (eg, acid, trifluoroacetic acid) And sodium borohydride. The main reaction is carried out in an ether solvent such as diethyl ether, tetrahydrofuran, dimethoxetane, dioxane, diglyme, and the reaction temperature varies depending on the type of the reducing agent, and is generally about 0 to about 0. One hundred sixty.

 In this production method, the configuration of the compound of the formula (VI), which is the starting compound, with respect to the asymmetric carbon atom is retained in the product.

 The starting compound represented by the formula (VI) is a novel substance, for example, the following formula (XI)

(In the formula, means the same as above.)

And the following formula (dish)

H (In the formula, R ₂ and R ₃ , mean the same as described above.)

 By reacting with a compound represented by the formula:

 The reaction between the compound of the formula (XI) and the compound of the formula (SO) is carried out by the reaction of benzotriazole-N-oxytris (dimethylamino) phosphoniumhexafluorophosphate, dicyclohexylcarbodiimide, 1-ethyl-3- (3-di The reaction can be carried out in the presence of a condensing agent such as methylaminopropyl) carbodiimide hydrochloride, N, N'-carbodilimidazole, 1-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline. The reaction is carried out in an appropriate solvent, and the solvent used may be the same as the specific examples described in the production method (a), and the compound of the formula () may be the same as that described in the production method (a). Can also be used in the form of an acid addition salt, in which case the reaction of triethylamine, tributylamine, diisopropylethylamine, N-methylmorpholine Carried out in the presence of Una organic base. The reaction temperature is usually about 20 ° C to about 50 ° C.

 The configuration of the compound of the formula (XI) and the compound of the formula (ΧΠ) with respect to the asymmetric carbon atom is the same as described for the reaction of the compound of the formula (Π) with the compound of the formula (m) in the production method (a). In the product [compound of formula (VI)]. The enantiomers of the compound of formula (XI) can be found, for example, in Collet, A. and

 It can be produced according to the method of Jacques, J. [Bull. Soc. Chim. France, 3330-3334 (1973)] or the method described in Reference Examples 1 and 2 below. (S) — (+) — or (R) — (1-) 1-1,2-diphenylethylamine in place of (+)-or (-1) -1-phenethylamine in Reference Examples 1 and 2. An optically active amine commonly used as an optical resolving agent can be used.

The enantiomer of the compound in which R ₂ is a group other than a hydrogen atom in the above formula (ΧΠ) can be produced, for example, according to the method described in JP-A-63-225δ9. can do.

 The product obtained by each of the above production methods can be isolated and purified by a conventional method such as chromatography, recrystallization, and reprecipitation.

 The product obtained by each of the above processes is in the form of an acid addition salt or a free base depending on the reaction conditions. These products can be converted into the desired acid addition salts or free bases by conventional methods.

 When the compound of the present invention or the starting compound obtained by each of the above-mentioned production methods is a racemate or a diastereomer mixture, each stereoisomer can be obtained by a conventional method, for example, a method described in European Patent Application Publication No. 455006. Can be separated.

Action effect

The results of pharmacological tests of representative compounds of the present invention are shown below. Test Example 1 was performed in comparison with an existing adrenergic receptor agonist. The existing 3 Adorenari emissions receptor agonist, was used salbutamol as non-selective agonists as (1) one isoproterenol no le or [delta] ₂ selective agonist. Test Examples 1 to 3 were performed in accordance with the method of Bianchetti, A. and Manara, L. [Brit. J. Pharma col., 100. 831-839 (1990)].

Test Example 1 Rat ^ ₃ adrenergic receptor stimulating action: Suppressive action on spontaneous contraction of the isolated rat colon:

The proximal colon was excised from an SD male rat weighing 350-400 g, and a 2.5-3 cm long specimen was prepared. The specimen was Krebs-Henseleit (118 mM NaCl, 4.75 mM NaCl, 4.75 mM KC1.2.5 mM containing pentramin (10 M), desmethylimibramine (0.5 M) and hydrocortisone (30 / M). _{CaCl 2. 1.2mM KH 2 P0 4} . 1.2mM MgS 0 4. 25mM NaHC0 3. 10 mM glucose) was filled with Ma Appended to Gunusu tube (1 Om 1) in 95% 0 ₂ - and kept at a mixed gas of 5% C0 ₂ through the gas while 37 hands. After a resting tension lg was applied and the spontaneous contraction stabilized, the test compound was added cumulatively. The concentration at which spontaneous contraction was reduced by 50% (IC ₅₀ value) was calculated from the dose-effect curve by the least squares method, assuming that the spontaneous contraction without the test compound was 100%. The results are shown in Table 1. Table 1

* Refers to the compound of Example 1 (the same applies hereinafter).

Test Example 2 Rat S ₂ adrenergic receptor stimulating effect: Suppressive effect on spontaneous contraction of the isolated rat uterus:

The uterus was excised from a non-pregnant Wistar female rat and a specimen was prepared by a conventional method. Specimens, full We Roh carboxymethyl mouth click containing benzal Min (12〃M) (Locke:. 15 7mM NaCl 5.6mM KC1, 2.2mM CaCl 2, 18mM NaH C0 3, 5.6mM glucose) Magnus tube filled with liquid (20 m 1) was vertical suspended in 95% 0 ₂ - 5% C0 ₂ c resting tension lg of the mixed gas was kept in aeration 37 ° C for loaded, after spontaneous contraction was stabilized, the test compound Cumulatively Was added. With IC ₅₀ values of test compounds were calculated in the same manner as in Test Example 1 <Example 1 to 4, I Ji ₅ of the compound of Example 7 and Example 9 ₍₎ value is 10 · ⁷ Micromax a stone more Yes, the effect was extremely weak compared to the ^ ₃ adrenergic receptor stimulating effect. In particular, ^ _receptor stimulating action of the compound of Example 2 and Example 9 was filed approximately 100 fold selective compared to beta ₂ receptor stimulating action. Test Example 3 Guinea pig ^, adrenergic receptor stimulatory effect: Right guinea pig isolated right atrial rhythm increase effect:

An isolated right atrial specimen with a pacemaker was prepared from a dd モ ル male guinea pig by a conventional method. Specimens, modified click Repusu containing full Nokishibenzamin (12 iM) (Krebs:. 122.2mM NaCl, 4.2mM KC1, 2.5mM Ca Cl 2, ImM MgCl 2, 1.2mM NaH 2 P0 4, 15.5mM NaHC0 3 11.5mM Glucose ) was suspended in Magnus tube (20 m 1 in :) filled with 95% 0 ₂ - was incubated at 37 ° C for while passing 5% C0 ₂ gas mixture. A resting tension lg was applied and after 30 minutes the test compound was added cumulatively. 100% up增加the right atrium rhythm number by the test compound was calculated by the least squares method concentrations showing the 50% (EC ₅₀ value) from the dose first working curve.

Both are not observed effect even high 'concentrations of 10 · ⁵ Micromax compounds of Examples 1 to 4 and Examples ReiRyo to 10, ₃ A Dorenari emissions receptor stimulation work for these compounds, beta Ryo Dorenarin receptor It is 200 times or more intense than the body stimulating effect, and has high selectivity for adrenergic receptors.

Test Example 4 Mouse small intestinal charcoal powder transport ability inhibitory action (in vivo small intestinal motility inhibitory action):

Test compounds were orally administered to 18-hour fasted mice (7 to 11 mice per group). One hour later, 0.2 ml of a 5% charcoal powder suspension suspended in a 10% aqueous solution of gum arabic was orally administered per mouse, and 20 minutes later, the gastrointestinal tract was removed. The transfer rate of the tip of the charcoal powder to the entire small intestine was calculated. The compound of Example 2 showed a significant inhibitory effect on charcoal powder transport at a dose of lmg / kg.

As is clear from the results of Test Example 1-4, the compounds of the invention have been superior in selectivity / 3 ₃ Adorenarin receptor stimulating effect.

Further also examined for work for human / 3 ₃ Ryo Dorenarin receptor representative compounds of the present invention. First, a method for preparing a cell line that highly expresses human; S ₃ -adrenergic receptor is described. Then, Test Examples 5 and 6 performed using the cell line and the results thereof are shown below.

Method for preparing a cell line highly expressing human ₃ adrenergic receptor

(1) Preparation of expression vector:

Expression vector pKCRH2 for animal cells [Mishina et al., Nature: 604-608 (1984)] was digested with the restriction enzyme SalI and blunt-ended with a DNA Blunting Kit (Takara Shuzo). Next, another expression vector pS V2-neo for animal cells [S outhern and Berg, J. Mol. Appl. Genet. 1: 327-341 (1982)] was digested with restriction enzymes Accl and Aatl I, and DNA Blunting was performed. The tip was made blunt by Kit. These were ligated using a DNA Ligation Kit (Takara Shuzo), introduced into Escherichia coli HB101 by a conventional method, and the transformants were selected on an LB agar medium containing 100 g / m1 of ampicillin and 25 ug / m1 of kanamycin. The plasmid DNA was extracted from this transformant, digested with the restriction enzyme Pstl, and then electrophoresed on a 2% gel of NuSieve 3: 1 Agarose (Takara Shuzo) to obtain about 3.8 kbp. 2.2 kbp. 1.4 kbp and 0.9 kbp. The clone from which the kbp fragment was obtained was selected. This plasmid DNA was designated as pKCNO. This plasmid pKCNO was digested with the restriction enzyme Hindi II, and ligated with the synthetic adapter 1 using the DNA Ligation Kit. The sequence of Synthetic Adapter 1 is as follows. 5 * -AGCTCCTGCAGGCGCGCCGATATCTCGAGCGGCCGCGGTACCA-3 '

 3 * -GGACGTCCGCGCGGCTATAGAGCTCGCCGGCGCCATGGTTCGA-5 'After introducing this into E. coli HB101 by a conventional method, transformants were selected on an LB agar medium containing 100 g / ml of ampicillin. Plasmid DNA was extracted from this transformant, digested with restriction enzymes Dral and Hindi II, and electrophoresed on a 2% gel of Nu Sieve 3: 1 Agarose to cut pKCN0 with the same restriction enzyme. A clone was selected that yielded a fragment of about 43 Obp instead of the fragment of about 380 bp that was sometimes obtained. This plasmid DNA was used as the expression vector pKCNl.

(2) Preparation of expression plasmid:

 RNA was extracted from human glioblastoma SK-N-MC (ATCC HTB 10) by a conventional method, and cDNA was synthesized using Superscript Systems (Life Technologies). The cDNA was amplified by GeneAmp PCR Kit (Perkin-Elmer) using oligonucleotides 1 and 2 as primers. When performing the PCR reaction, 10% dimethyl sulfoxide was added to the reaction solution. The sequences of oligonucleotides 1 and 2 are as follows.

5 "-CCACCTGCAGGTGATTTGGGAGACCCC-3 '—— oligonucleotide 1 5'-TTCTCGAGCCGGGGAATCCCATGGGAC-3' —— oligonucleotide 2 After purifying the reaction product by SpinBind System (Takara Shuzo), Approximately 1.4 kbp fragment was recovered and purified by electrophoresis using 2% gel of Nu Sieve 3: 1 Agarose.This fragment and the expression vector pKCN1 digested with restriction enzymes Sse8387I and EcoRV were DNA-ligated. After ligation using Kit and introduction into E. coli HB101 by a conventional method, transformants were selected on LB agar medium containing ampicillin 100 / ml, and plasmid DNA was extracted from the transformants. Restriction As a result of examining the base sequence of a fragment of about 1.3 kbp obtained by digestion with enzymes Sse83871 and XhoI, this sequence was reported by Lelias et al. [FEB S Lett. 324: 127-130 (1994)]. The sequence was identical to that of the human drainage receptor cDNA. The plasmid DNA expressing the human drainage receptor was designated as pKREX10.

 (3) Preparation of highly expressing cell line:

Human ^ ₃ adrenergic receptor expression plasmid pKREX 10 was introduced into Chinese hamster ovary cells CHO—K1 (ATCC CCL 61) by calcium phosphate method, and the transformant was transformed to 600 ig / ml G—418. Selection was performed using MEM-Dulbecco medium (ICN Biomedicals) containing (Lif e Technologies). The medium was supplemented with 10% fetal calf serum and 11.5 g / ml proline. After removing the medium from the 69 G-418 resistant clones, the cells were detached by leaving them to stand at 37 ° C for 10 minutes in phosphate buffered saline containing 0.5 mM ethylenediaminetetraacetic acid (EDTA). Collect more cells to centrifugation and suspended at about 5 XLO ^e cells / m 1 in 10 mM Tris-HCl slow銜液(p H 7.5) containing ImM EDTA. This suspension was prepared by adding 201 11.5 nM (-) 3- [ ^125I ] iodocyanopindolol (Amersham¾: 1% serum albumin, 0.1% NaN ₃ and 2 OmM HEPES buffer (pH 7.4). The mixture was mixed in RPMI-1640 medium (UCN Biomedicals) 200 1 containing, and allowed to stand at 4 for 2 hours. Using a bio-dot device (Bio-Rad Laboratories), the filter was carefully washed with a glass filter GF / C (Whatman) pre-soaked in 0.3% polyethyleneimine, and the radioactivity on the filter was measured using a radiometer. It measured using. The highest clone radioactivity selection Bikore human; was 5 ₃ adrenoceptor highly expressing cell line CHO / pK REX 1 0- 36. Test Example 5 human / S ₃ Adorenarin Receptor Binding Test:

The above as in the human / 3 ₃ Adorenarin receptor withers expressing cell line CH O / pKREXl 0- 36 produced 10% © shea calf serum, of 11.5 / m 1 proline, and 200 ng / m 1 of G- After culturing in MEM-Dulbecco medium containing 418 for 3 days, the medium was removed, and the cells were detached by leaving still at 37 ° C for 10 minutes in phosphate buffered saline containing 0.5 mM EDTA. The cells were collected by centrifugation, and suspended in 1 OmMTris-HC 1 buffer (pH 7.5) containing 1 mM EDTA to a concentration of about 5 × 10 ⁶ cells / m]. The test compound was mixed with this suspension 201 in RPMI-1640 medium 100 ^ 1 containing 1% serum albumin, 0.1% NaN ₃ and 2 OmM HEPE S buffer (pH 7.4). Then, the mixture was allowed to stand for 30 minutes at 4, and then 0.2 nM (-) 3- [ ^125I ] iodocyanopindolol 100 ^ 1 was added thereto, and the mixture was allowed to stand for further 4 hours. Using a biodot device, the glass filter was pre-washed with a glass filter GF / C soaked in 0.3% polyethyleneimine in advance, and the radioactivity on the filter was measured using an X-ray detector. The amount of binding with or without imM (—)-alprenolol was 100% inhibition and 0% inhibition, respectively. The concentration that inhibits the binding amount by 50% (IC ₅₀ value) is the minimum from the concentration-inhibition ratio curve. It was calculated by the square method. Table 2 shows the results.

Table 2

 * Taste the compound of Example 1 (the same applies hereinafter).

Test Example 6 Human S ₃ adrenergic receptor stimulating action (cyclic AMP accumulation action):

The human 5 ₃ Adorenarin receptor withers expressing cell line CH O / pKREXl 0- 36 10% © shea fetal serum was prepared in the manner of, proline of 11.5〃 g / m 1, and 200 g / m 1 of G — MEM containing 418 — After culturing in Dulbecco medium for 3 days, CHO / pKREXl O-36 is collected by the same method as in Test Example 5 and contains 1 mM ascorbic acid and 1 mM 3-isobutyl-1-methylxanthine The cells were suspended in Hanks' balanced salt solution (ICN Biooraedicals) at about 2 × 10 ⁶ cells / ml. This suspension 1001 and the test compound were mixed in the same equilibrium salt solution 5001, and allowed to react at 37 ° C for 30 minutes, followed by boiling for 5 minutes to stop the reaction. After centrifuging the reaction solution, the amount of cyclic AMP in the supernatant was measured using the cAMP EIA System (Amershara). 10 - ⁵ M (-) Saikuri' of over I source when added to pro tele Knoll or without the addition time The concentration (EC ₅₀ ) that caused the accumulation of ₅₀ % cyclic AMP was calculated from the concentration-response curve by the least-squares method with the amount of AMP being 100% and 0%, respectively. Table 3 shows the results.

Table 3

 * Refers to the compound of Example 1 (the same applies hereinafter).

As is clear from the results of Test Examples 5 and 6, the compound of the present invention also has a stimulating effect on the human ^ ₃ adrenergic receptor.

The compounds of the present invention are useful as) S ₃ adrenergic receptor agonists as preventive and therapeutic agents for obesity, diabetes, irritable bowel syndrome, acute or chronic diarrhea, and the like. The compounds of the present invention can also be used for improving symptoms such as abdominal pain, nausea, vomiting, and upper abdominal discomfort associated with peptic ulcer, acute or chronic gastritis, biliary dyskinesia, cholecystitis, etc. it can.

When the compound of the present invention represented by the above formula (I) and a pharmaceutically acceptable acid addition salt thereof are used as a ^ ₃ adrenergic receptor agonist, any of oral administration, parenteral administration and rectal administration can be used. Oral administration is preferred, but the preferred dosage is the method of administration, the patient's symptom 'age, the form of treatment (prevention or treatment) Usually, it is 0.01 to 2 O mg / kg / day, preferably 0.1 to 1 O mg / kg /.

 The compound of the present invention is usually administered in the form of a preparation prepared by mixing with a preparation carrier. As the pharmaceutical carrier, a substance that is commonly used in the pharmaceutical field and does not react with the compound of the present invention is used. Specifically, for example, lactose, glucose, mannite, dextrin, starch, sucrose, magnesium aluminate metasilicate, synthetic aluminum gaterate, crystalline cellulose, sodium carboxymethyl cellulose, hydroxypropyl starch, carboxymethyl cellulose Calcium, ion exchange resin, methylcellulose, gelatin, gum arabic, hydroxypropylcellulose, low-substituted hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, polyvinylalcohol, light gay anhydride, magnesium stearate , Talc, carboxyvinyl polymer, titanium oxide, sorbitan fatty acid ester, sodium lauryl sulfate, glycerin, fatty acid glycerin ester, Examples include purified lanolin, glycerinated gelatin, polysorbate, macrogol, vegetable oil, wax, nonionic surfactant, propylene glycol, water and the like.

Dosage forms include tablets, capsules, granules, powders, syrups, suspensions, suppositories, gels, injections and the like. These preparations are prepared according to a conventional method. In the case of liquid preparations, they may be in the form of being dissolved or suspended in water or other appropriate medium at the time of use. Tablets and granules may be coated by a known method. In the case of injections, it is prepared by dissolving a pharmaceutically acceptable acid addition salt of the compound represented by the above formula (I) in water, and dissolving it in an isotonic agent if necessary. PH adjusting agents, buffers and preservatives may be added. These preparations can contain the compound of the present invention in an amount of 0.01% or more, preferably 0.05 to 70%. These formulations may also contain other therapeutically active ingredients.

 Hereinafter, the present invention will be specifically described with reference to Reference Examples, Examples, and Formulation Examples, but the present invention is not limited to these Examples and the like. The compounds were identified by elemental analysis values, mass spectra, IR spectra, NMR spectra, and the like.

Reference example 1

 Production of (R)-(1-)-1-chloromandelic acid:

 28 g of 3-chloromandelic acid was dissolved in 100 ml of acetone, and a solution of 18.6 g of (+)-phenethylamine in acetone (20 ml) was added thereto. The precipitated solid was collected and recrystallized twice from ethanol to obtain 9.9 g of (R)-(1-)-13-chloromandelic acid (+)-monophenylamine salt. To 9.6 g of this salt was added 25 ml of 10% hydrochloric acid, and the mixture was extracted with ethyl ether. The getyl ether layer was washed with saturated saline and dried over anhydrous magnesium sulfate, and then the solvent was distilled off under reduced pressure to obtain 5.9 δg of (R)-(-1) -13-chloromandelic acid.

[] D ²⁷ = — 116.4 ° (c = l, acetone)

Reference example 2

 Preparation of (S)-(+) — 3-chloromandelic acid:

 Perform the same operation as in Reference Example 1 using 10.2 g of crude (S)-(+)-3-chloromandelic acid recovered from the mother liquor of Reference Example 1 and 6.78 g of (1-)-α-phenethylamine. As a result, 3.9 g of a pure product of (S)-(+)-3-chloromandelic acid was obtained.

[Na]] ²⁷ = + 115.5 ° (c = l, acetone) Example 1

 Preparation of 2- [2- (6- (Fluoroindole-1-yl) ethylamino] -1-1 (3-chloro-2-phenyl) ethanol:

 To 15 ml of methanol was added 0.54 g of (3-chlorophenyl) oxysilane, 1.5 g of 3- (2-aminoethyl) -16-fluoroindole hydrochloride, and 1.9 ml of triethylamine. Stirred for hours. After the solvent was distilled off under reduced pressure, chloroform was added to the residue, and the mixture was washed with saturated aqueous sodium bicarbonate, water and saturated saline in this order, and dried over anhydrous sodium sulfate. After evaporating the solvent under reduced pressure, the residue was subjected to silica gel column chromatography and eluted with chloroformumethanol (15: 1), and the fraction containing the desired product was concentrated under reduced pressure to obtain 1 g of an oil. To this was added getyl ether, which was solidified and then recrystallized from ethanol to obtain 0.25 g of the desired product.

137-139 ° C

lH NMR spectrum _{(CDC 1 3, ό ppm)} : 1.9 δ (2Η, br, HO-C HCH 2 NH), 2.67 (1 H, dd, J = 12Hz, 9Hz), 2.84-3.1 3 ( 5H, m), 4.63C1H, dd.J = 9Hz, 4Hz, CHOH), 6.82 to 6.95 (lH, m), 6.97 to 7.11 (2H, m) .7.36 (1H, s, indole 21H) , 7.42 ~ 7.56 (lH, m) '7.97 (lH, s, indole NH)

Examples 2 to 7:

第 4表 The reaction and treatment were carried out in the same manner as in Example 1 using the corresponding indoles instead of 3- (2-aminoethyl) -16-fluoroindole in Example 1, and the product was recrystallized from a suitable solvent. Thus, the compounds shown in Table 4 were obtained. The recrystallization solvent was ethanol for the compound of Example 4 and ethanol for all the remaining compounds.

Table 4

Example 8

 Preparation of 2- [1-ethyl-2- (indole-3-yl) ethylamino] 1-1- (3-chlorophenyl)

A solution of 0.63 g of (3-chlorophenyl) oxylan and 1.5 g of 3- (2-aminobutyl) indole in 10 ml of methanol was stirred with 20 pieces for 42 hours. After evaporating the solvent under reduced pressure, the residue was subjected to silica gel column chromatography, eluted with chloroform-methanol (12: 1), and the fraction containing the desired product was concentrated under reduced pressure to give the desired product, 0.1 g to 6 g. Was obtained as an oil. Liquid secondary ion mass spectrometry (SI MS) m / z: 343 (MH ⁺ )

Example 9

2 — [(1R) —2— (indole-3-yl) -1-methylethylamino] Preparation of 1 (1R) -1 1- (3-chlorophenyl) ethanol

 First step:

 3.73 g of (R) -3- (2-aminobutylpyr) indole and 3.48 g of (R)-(1-)-13-chloromandelic acid prepared according to the method described in JP-A-63-22559. To a solution of N, dimethylformamide (40 ml) was added 8.84 g of benzotriazole-N-oxytris (dimethylamino) phosphonium hexafluorophosphine, and 4.9 ml of triethylamine was added dropwise. The reaction mixture was stirred for 2 hours, ethyl acetate was added to the reaction solution, washed with water, 1N hydrochloric acid, water, 2N aqueous sodium hydroxide solution, water, and saturated saline in this order, and dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure to obtain 7.9 g of N-[(1R) -2- (indole-3-inole) -1-methylethyl] -1- (R) -3-chloromandelic acid amide as a yellow solid. Was.

126-129 ° C (recrystallized from ethyl acetate-n-hexane).

[N] _D ²⁸ = — 26.3 ° (c = 0.5. Methanol)

Second step:

 At 20 ° C., a solution of 1.2 g of the amide derivative obtained in the first step in tetrahydrofuran (3 Oml :) was added dropwise to a 20 ml solution of the 1M borane-tetrahydrofuran complex at 20 ° C. After the reaction solution was heated to reflux for 4 hours, 1 Oml of methanol was added dropwise under ice cooling. After passing through for 1 hour to decompose excess borane, the solvent was distilled off under reduced pressure. Ethyl acetate was added to the residue, washed with water and saturated saline in this order, and dried over anhydrous magnesium sulfate. After evaporating the solvent under reduced pressure, the residue was subjected to silica gel column chromatography and eluted with chloroform-methanol (12: 1) to obtain 0.67 g of the desired product as a solid.

113-115 ° C (recrystallized from hydrous isopropyl alcohol).

[A] _D ²⁸ = — 46.4 ° (c = 0.5, methanol) The optical purity of the product was determined to be 99% ee or more by high performance liquid chromatography (HPLC) under the following conditions (retention time: 14.7 minutes).

 HP LC measurement conditions:

 1. Column: ULTRON E S-OVMC4.6 ιππΐίό x 150 mm: Shinwa Ka)

2. Move layer: 20 mM KH ₂ P0 ₄ aqueous solution (pH 3.6) Ba methanol: § Seto Nitrile = 3: 1) = 92: 8,

 3. Flow rate: 1. Oml / min,

 4. Detection wavelength: 220 nm.

- NMR spectrum _{(CD C 1 3, ippm)} : 1.15 (3 H, d, J = 6 H z, CH 3) .2.5 (2H, br, OH, NH), 2.64 (1 H, dd, J = 12Hz, 6Hz), 2.85C2H, d, J = 6Hz), 2.88 (1H, dd, J = 12Hz, 4Hz), 2.98 to 3.11 (lH, m), 4.51C1H.dd, J = 9Hz, 4 Hz, CHOH), 7.0 to 7.62 (9H.m, Ar-H), 8.04 (1H.s, indole NH)

Examples 10 to 12:

 The following compound was obtained by reacting and treating in the same manner as in Example 8 using a starting compound having a specific configuration of 3- (2-aminobutyric pill) indole and 3-chloromandelic acid. . The optical purity of the compound of the example was measured according to the method described in Example 9.

(Example 10)

 2-[(1R)-2- (indole-3-inole) -1-methylethylamino] 1 (1S) -11- (3-chlorophenyl) ethanol:

Optical purity: 99% ee or more (retention time: 27.7 minutes) (Example 11)

 2-[(lS) -2 -— (indole-3-yl) -1-methylethylamino] 1 (1R) -11- (3-chlorophenyl) ethanol

Optical purity: 99% e. E. Or more (retention time: 16.4 minutes)

 (Example 12)

 2-[(1S) -2- (indole-3-yl) -11-methylethylamino] 1- (1S) -11- (3-chlorophenyl) ethanol;

Optical purity: 99% e.e. or more (retention time: 18.3 minutes)

Formulation example (tablet manufacturing method)

 According to a conventional method, the following components are mixed, granulated, and compression-molded to prepare 1000 tablets of 10 Omg per tablet. '

 2- [2- (indole-3-yl) -11-methylethylamino] -11- (3-chlorophenyl) ethanol · oxalate: 5 g,

 Corn starch: 25 g,

Lactose: 54 g,

Microcrystalline cellulose: 11 g :,

Pill cellulose in the mouth of hydroxy: 3 g,

Light gay anhydride: 1 g,

Magnesium stearate: 1 g.

Industrial applicability

Compounds obtained by the present invention, / S ₃ has a highly selective ₃ Adorenarin receptor stimulating effect on Adorenarin receptor, diabetes, obesity, irritable bowel syndrome, as a preventive and therapeutic agent, such as acute or chronic diarrhea It can also be used to improve symptoms such as abdominal pain, nausea, vomiting, and upper abdominal discomfort associated with peptic gallbladder, acute or penetrating gastritis, and the like.

Claims

The scope of the claims 1. 2- [2- (indole-13-yl) ethylamino] -11-phenylethanol derivative represented by the following formula (I) or a pharmaceutically acceptable acid addition thereof <(I)

(In the formula, means a halogen atom or a trifluoromethyl group, R ₂ represents a hydrogen atom, a lower alkyl group or a fluoro lower alkyl group, and R ₃ represents a random atom, a halogen atom, a trifluoromethyl group, a nitro group or a cyano group. ) 2. The method according to claim 1, wherein R i is a chlorine atom, R ₂ is a hydrogen atom, a methyl group or a fluoromethyl group, and R ₃ is a hydrogen atom or a fluorine atom or a chlorine atom at the 6-position. Compound. 3. The compound according to claim _{2, wherein} R ₂ is a methyl group, and R ₃ is a hydrogen atom or a fluorine atom or a chlorine atom at the 6-position.  4. 2-[(1 shaku) -2- (indole-13-inole) -11-methylethylamino] -1 (1R) -11- (3-chlorophenyl) ethanol or its pharmaceutically acceptable Acid addition salts.  5. 2- [2- (6-Fluoroindole-3-inole) -1-1-methylethylamino] -1-1 (3-chlorophenyl) ethanol or a pharmaceutically acceptable acid addition thereof. 6. The following formula (Π) (II)

(Wherein, represents a halogen atom or a trifluoromethyl group.) And a compound represented by the following formula (ΠΙ) H ₂ N—

 H (Wherein, R ₂ represents a hydrogen atom, a lower alkyl group or a fluoro lower alkyl group, and R ₃ represents a hydrogen atom, a halogen atom, a trifluoromethyl group, a nitro group, or a cyano group.) Reacting with a compound represented by the following formula (I):

(In the formula, R !, R ₂ and R ₃ mean the same as described above.) A compound represented by or a manufacturing method _c of its pharmaceutically acceptable acid addition salt thereof 7. (a) The following formula (IV)

(In the formula, represents a rho atom, a logen atom or a trifluoromethyl group.) A compound represented by the following formula (V) 0 ( _v)

(In the formula, R ₂ means a hydrogen atom, a lower alkyl group or a fluoro lower alkyl group, and R ₃ means a hydrogen atom, a halogen atom or a trifluoromethyl group.) Reacting with the compound represented by the following conditions, or (b) The following formula (VI) One CO one NH one CH one CH ₂ ~ jj ~ _{n (V} |)

To R ₂  H (Wherein R !, R ₂ and R ₃ mean the same as above.) Wherein the compound represented by the following formula is reacted in the presence of a reducing agent. (νπ)

Or a pharmaceutically acceptable acid addition salt thereof