WO2001083429A1

WO2001083429A1 - Aniline derivatives

Info

Publication number: WO2001083429A1
Application number: PCT/JP2001/003721
Authority: WO
Inventors: Toshio Honda; Toshihiko Makino
Original assignee: Chugai Seiyaku Kabushiki Kaisha
Priority date: 2000-04-28
Filing date: 2001-04-27
Publication date: 2001-11-08
Also published as: AU2001252648A1

Abstract

Compounds represented by the formular (A), tautomers thereof, or medicinally acceptable salts of both exhibit an NOS-inhibitory activity and are useful as drugs in the treatment of various diseases.

Description

Specification

Aniline derivatives Technical field

TECHNICAL FIELD The present invention relates to an aniline derivative having an inhibitory effect on nitric oxide synthase (nitricoxidedesynthaise, NOS). Background art

It is said that there are at least three types of NOS isoforms. NNOS (type 1), which is constitutively present in nerve cells and is calcium-dependent, eNOS (type 3), which is constitutively present in vascular endothelial cells and is calcium-dependent, and In many other cells, iNOS (type 2) is apparently calcium-independent iNOS (type 2), which is induced and synthesized by the stimulation of cytokines and in vivo trace toxins (1 ipopolysaccharide, LPS) (FASEB J. 16, 3051- 3064, 1992; Mo 1. Chem. Neurop athol. 26, 107-157, 1995).

Many compounds have been reported that selectively inhibit one of the three isoforms, such as iNOS and nNOS. These compounds are expected to be useful as selective inhibitors of nNOS for the treatment of cerebrovascular disorders and the like, and selective inhibitors of iNOS for the treatment of septic shock and the like (WO 96 / 18608 publication).

Among the compounds having NOS inhibitory activity, isourea derivatives, isothiourea derivatives, or guanidine derivatives are described in WO95 / 09619, WO96Z09286, WO96Z18608, WO98 / 42667, WO99 / No. 23069, and the like.

However, no compound has been reported that is sufficiently effective clinically, and NOS inhibitors that can be expected to have therapeutic utility are still desired. Disclosure of the invention An object of the present invention is to provide an aniline derivative having NOS inhibitory activity and useful as a medicament. Another object of the present invention is to provide an aniline derivative having a selective inhibitory activity on nNOS and useful as a medicament.

The present inventors have conducted intensive studies to solve such problems, and as a result, have found that an aniline derivative having a specific structure, that is, an isourea derivative, an isothiourea derivative, or a guanidine derivative has an excellent NOS inhibitory action. The present inventors have found that they have, or have a strong nNOS inhibitory activity, or have a selective inhibitory activity on nNOS, and completed the present invention based on such findings.

That is, the present invention provides the following compound group (1)

Compound group (1):

Or a possible tautomer thereof, or a pharmaceutically acceptable salt thereof. The present invention also provides a medicine, a NOS inhibitor, a selective nNOS inhibitor, or a therapeutic agent for cerebrovascular disease containing the above compound. Furthermore, the present invention provides various disease states containing the above compound, which can be expected to have therapeutic effects by selectively inhibiting nNOS, for example, head trauma, spinal injury, pain, Parkinson's disease, Alzheimer's disease, convulsions And a therapeutic agent for morphine tolerance or dependence.

In the present invention, the “NOS inhibitor” refers to a compound having an inhibitory effect on at least one isoform of three N ァ S isoforms. The “n NS selective inhibitor” has a weak effect on eNOS and iNOS among three isoforms of NOS and has a selective inhibitory effect on nNOS. Refers to a compound. The compounds included in the present invention show good NOS inhibitory activity or good nNOS-selective inhibitory activity, as described in the test examples described later. The types of cerebrovascular disease include cerebral hemorrhage, subarachnoid hemorrhage, transient ischemic attack (TIA), cerebral edema, and cerebral infarction. Subtypes of cerebral infarction include atherothrombotic infarction, lacunar infarction, or cardiogenic embolism.

In addition, when the compound of the present invention is used as an analgesic, the target pain types include, for example, headache. Also, subtypes of headache include migraine, tension-type headache, or cluster headache or chronic paroxysmal headache.

Further, the compounds in the compound group (1) may have tautomers depending on the surrounding environment of the compounds, and these tautomers are all included in the present invention. BEST MODE FOR CARRYING OUT THE INVENTION

The compound of the present invention selected from the compound group (1) can be obtained, for example, by the method described in Examples described later.

The pharmaceutically acceptable salt of the compound of the present invention is not particularly limited as long as it is a pharmaceutically acceptable salt. Examples thereof include inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, and hydroiodic acid. And salts with organic acids such as formic acid, acetic acid, fumaric acid and tartaric acid.

The compound of the present invention or a salt thereof may be a suitable excipient, scavenger, lubricant, preservative, disintegrant, buffer, binder, stabilizer, wetting agent, emulsifier, coloring agent, flavor or fragrance. Orally or parenterally in the form of tablets, granules, fine granules, powders, capsules, syrups, elixirs, suspensions, emulsions, injections, etc. Can be. In the hyperacute phase (immediately after a seizure), acute phase (seizures to 2-3 days), and subacute phase (2-3 days to 2 weeks after a seizure) of cerebrovascular disorder, administration is mainly by intramuscular or intravenous injection There is expected. In addition, if the patient is ingested during the chronic phase (after the third week after the attack), oral administration may be considered.

The dose of the compound of the present invention or a salt thereof depends on the patient's body type, age, physical condition, and degree of disease. Although it can be selected as appropriate depending on the time elapsed after the onset, etc., it is expected to be 0.01 to 100 mg O body per day. In general, even if the same dose is administered, the blood concentration may vary greatly from patient to patient, so it is ideal to determine the optimal dose of the drug for each patient while monitoring the blood concentration of the drug. It is.

When formulated as an oral solution, for example, as a carrier for the formulation, lactose, sucrose, sorbitol, mannitol, potato starch or starch such as corn starch, or a starch derivative, a cellulose derivative or gelatin can be used normally. Auxiliaries are suitable, and at the same time lubricants such as, for example, magnesium stearate, carpo-wax or polyethylene glycol can be added, and these mixtures are made into granules, tablets, capsules, etc. in a conventional manner. Can be done.

When formulated as an aqueous preparation, for example, an effective amount of the main component is dissolved in distilled water for injection, and if necessary, an antioxidant, a stabilizer, a solubilizing agent, a buffer, a preservative, and the like are added. After complete dissolution, filtration, filling, sealing, and sterilization by a high-pressure steam sterilization method or a dry heat sterilization method can be performed to obtain an injection.

When formulated as a lyophilized agent, an aqueous solution in which the main component is dissolved in distilled water for injection may be lyophilized by an ordinary method.If necessary, mannitol may be used as an excipient that can be easily lyophilized. And sugars such as inositol, lactose, maltose, and sucrose, or sugar alcohols, or glycine and the like, and then freeze-dried in the usual manner to prepare the preparation. Example

Hereinafter, production of the compound of the present invention will be described in detail with reference to Examples.

Further, in order to show the usefulness of the present invention, test results on the inhibitory effects of the compounds included in the compound group (1) on various NOS are shown in Test Examples. Example 1a

Synthesis of t-butyl N- (5-nitro-1- (2-phenylethoxy) phenylmethyl) carbamate A mixture of 2-phenylethyl alcohol (0.478 ml), sodium hydride (content 60%, 4 Omg) and dimethylformamide (10 ml) was stirred under ice cooling for 10 minutes. Then, di-tert-butyl N- (2-fluoro-5-nitrophenylmethyl) iminodicarbonate (37 Omg) was added, and the mixture was stirred for 8 hours under ice cooling. The reaction mixture was concentrated under reduced pressure, water was added to the residue, and the mixture was extracted with ethyl acetate. The organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (developing solution; n-hexane: ethyl acetate = 8: 2) to obtain 334 mg of the title compound (yield 90%).

One NMR (CDC 1 ₃₎

<5: 8.15-8.11 (2H, m), 7.38—7.26 (5H, m), 6.89-6.86 (lH, m), 4.80 (1H, brs) ), 4.33—4.28 (4H, m), 3.16 (2H, t, J = 6.6Hz), 1.46 (9H, s) Example 2a, 3a, 4a 5a

In the same manner as in Example 1a, the compounds shown in Table 1 were obtained using the corresponding reactants.

Example 6a

Synthesis of N- (2-Ethyl-5-ditrophenylmethyl) -N- (2-phenylethyl) amine

To a solution of 3-chloromethyl-4-ethylnitrobenzene (1.11 g) in dimethylformamide (10 ml) was added diisopropylethylamine (2.83 ml) and 2-phenylethylamine (2.1 ml), and the mixture was brought to room temperature. And stirred overnight. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent; n-hexane: ethyl acetate = 2: 1) to give 1.54 g of the title compound. (98% yield).

^ -NMR (CDC 1 ₃₎

(5: 8.21 (d, J = 2.2Hz, 1H), 8.04 (dd, J = 2.4, 83Hz, 1H), 7.50-7.2.0 (m, 6H), 3.85 (s, 2H), 303-2.80 (m, 4H), 2.73 (q, 'J = 7.6Hz, 2H), 144 (brs, 1H), 1. 22 (t, J = 7.3Hz, 3H) Example 7a, 8a

In the same manner as in Example 6a, the compounds shown in Table 2 were obtained using the corresponding nitro compound and 3-phenylpyruamine as a reactant. Table 2

Example 6b

Synthesis of N- (2-ethyl-5-nitrophenylmethyl) -N- (2-phenylethyl) carboxamic acid

A mixture of the compound obtained in Example 6a (1.54 g), getyl ether (10 ml) and tetrahydrofuran (10 ml) was mixed with a 2N aqueous solution of potassium hydroxide (20 ml), and di-butyl dicarbonate (1.54 g). ) Was added. After stirring at room temperature for 1 hour, the reaction mixture was extracted with ethyl acetate, and the organic layer was dried over anhydrous sodium sulfate. After drying, the mixture was concentrated under reduced pressure. The obtained residue is purified by silica gel

(Developing solution; hexane: ethyl acetate = 5: 1) to give 2.06 g of the title compound (yield 99%).

(5: 8.05 (dd, J = 2.4, 83 Hz, 1H),

7.92 (brs, 1H), 7.40 7.10 (m, 6H) 4.50-4.

26 (m, 2H), 3.65-3.43 (m, 2H), 2.92-2.73 (m, 2H), 2.64 (q, J = 7.6 Hz, 2H), 1 . 49 (brs, 9H), 1.20 (t, 3 = 7.6 Hz, 3H) Example 7b, 8b

The compounds shown in Table 3 were obtained in the same manner as in Example 6b. Table 3

Example 9a

Synthesis of 2-methoxycarbonyl 2- (3-ditrophenyl) indane

.3-Methyl nitrophenylacetate (4. '7 g), o-xylene dichloride (5 To a mixture of 48 g) and dimethyl sulfoxide (800 ml), potassium butoxide (5.4 g) was added little by little at room temperature. After stirring the reaction mixture at room temperature for 1 hour, water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed successively with dilute hydrochloric acid and saturated saline, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (developing solution; n-hexane: ethyl acetate = 4: 1), and then recrystallized and purified using a mixed solvent of ethyl acetate-n-hexane. 1.94 g of the compound was obtained (yield 27%).

δ: 3.37 (2H, d, J = 15.5Hz), 3.63 (3H, s), 4.03 (2H, d, J = 15.5Hz), 7.18-7.29 (4H, m), 7.5 1 (1H, t, J = 7.9Hz), 7.74 (1H, d, J = 7.9Hz), 8.13 (1H, d, J = 7. 9Hz), 8..28 (1H, t, J = 2.0Hz) Example 9b

Synthesis of t-butyl N- (2- (3-ditrophenyl) indan-2-yl) caprate

To a solution of the compound (1.94 g) obtained in Example 9a in methanol (30 ml) was added sodium hydroxide (1.04 g), and the mixture was stirred at room temperature for 3 hours. The reaction mixture was acidified by adding 2N hydrochloric acid, and extracted with chloroform. The organic layer was washed sequentially with water and saturated saline, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. To the obtained residue were added t-butyl alcohol (20 ml), diphenylphosphoric azide (1.80 g) and triethylamine (66 Omg), and the mixture was heated and stirred at 90 for 18 hours. To the reaction mixture was added a 10% aqueous solution of cunic acid, and the mixture was extracted with ethyl acetate. The organic layer was washed successively with water and saturated saline, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (developing solution; n-hexane: ethyl acetate = 4: 1), and then purified by recrystallization with a mixed solvent of ethyl acetate-n-hexane to give the title compound. 1.13 g was obtained (yield 49%).

- NMR (CDC 1 ₃₎

δ: 1.34 (9Η, s) ', 3.51 (4H, s), 5.28 (1H, brs), 7.21—7.26 (4H, m), 7.47 (1H, t, J = 8.0Hz), 7.73 (1H, d, J = 8.0Hz), 8.09 (1H, dd, J = 1.3 Hz and 8.0 Hz), 8.32 (1H, t, J = 1.3 Hz) Example 10a

Synthesis of N-phenyl 2- (t-butoxycarbonyl) amino-3- (4-ditrophenyl) propionamide

In a solution of aniline (0.4 g) in methylene chloride (20 ml) under ice cooling, 4-dimethylaminopyridine (0.79 g), 2- (t-butoxycarbonyl) amino-3- (4-nitrophenyl) A solution of propionic acid (2.0 g) in methylene chloride (20 ml) and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (1.24 g) were added sequentially, and the reaction mixture was brought to room temperature. And stirred for 18 hours. The reaction mixture was washed with saturated saline, and the organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: n-hexane: ethyl acetate = 6: 4) to obtain 1.8 g of the title compound (yield 73%).

_{'H-NMR (CDC 1 3} )

6: 8.16 (2H, d, J = 8.6Hz), 8.02 (1H, brs), 7.44-7.10 (7H, m), 5.10 (1H, brs), 4.50 (lH, m), 3.36 (1H, dd, J = 6.9, 14.2 Hz), 3.16 (1H, dd, J = 7.3, 14.2 Hz), 1.42 (9H, s) Example 16a

Synthesis of 1- (t-butoxycarponyl) amino-6-nitroindane

To a mixture of 1-aminoindan (5 g) and sulfuric acid (20 ml) was added potassium nitrate (3.8 g) at 0 ° C. After the reaction mixture was stirred at room temperature for 2 hours, it was added to a mixture of 50% aqueous sodium hydroxide solution and ice, and extracted with chloroform. The organic layer was concentrated under reduced pressure, and the obtained residue was subjected to silica gel column chromatography (development solution; chloroform: methanol = 95: 5) to give 1-amino-1-412. A mixture of loindan and 1-amino-6-nitroindan was obtained. To the resulting mixture were added 1,4-dioxane (34 ml), aqueous sodium hydroxide solution (2N, 34 ml) and di-tert-butyl dicarbonate (8.83 g). After the reaction mixture was stirred at room temperature for 3 hours, a saturated aqueous solution of sodium chloride and ethyl acetate were added. The organic layer was washed with a saturated aqueous solution of sodium chloride, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent; n-hexane: ethyl acetate) to give 7.65 g of the title compound (yield 82%).

(5: 8.15-8.11 (lH, m), 8.10 (1H, d d, J = 8.6, 1.

8Hz), 7.37 (1H, d, J = 8.6Hz), 5.31—5.14 (1H, m), 4.96-4.84 (1H, m), 3.04 ( 1H, ddd, J = 17.

1, 9.2, 3.1 Hz), 2.91 (1H, ddd, J = 17.1, 8.6,

8.6), 2.73-2.59 (1H, m), 1.96—1.82 (lH, m), 1.50 (9H, s) Example 1b

Synthesis of t-butyl N- (5-amino-2- (2-phenylethoxy) phenylmethyl) carbamate

A mixture of the compound (330 mg) obtained in Example 1a, 10% palladium on carbon (70 mg) and ethanol (10 Oml) was stirred at room temperature under a hydrogen atmosphere for 16 hours. After filtering the reaction mixture, the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (developing solution; n-hexane: ethyl acetate = 1: 1) to obtain 27 Omg of the title compound (89% yield).

δ: 7.35-7.23 (5Η, m), 6.67—6.51 (3H, m), 4.80 (1Η, brs), 4.15-4.10 (4H, m) , 3.39 (2H, brs), 3.07 (2H, t, J = 6.6 Hz), 1.43 (9H, s) Example 2b, 3b, 4b, 5b, 6c, 7c, 9c, 10b, 16b

The compounds shown in Tables 4 and 5 were obtained using the corresponding nitro compounds in the same manner as in Example 1b ((1) under the reaction conditions in the table was 10% Pd-C, H2, EtOH , (2) indicates 5% Pd— (:, H ₂ , MeOH).

ετ

Halla

IZLZO / lOdT / lDd 6 £ £ 8/10 OAV (so

's—mouth y—9

¾y ^ o) Ma /:, ψ <^ -i-^^^ a-z 's—Mouth 4 = 1-19-1 (3)

B g I

Halla lZL £ 0 / lOd £ / ∑Jd 6 £ £ 8/10 OAV Pyridine (1.18 ml) and hydroxylamine hydrochloride (678 mg) were added to the mixture. After the reaction mixture was stirred at room temperature for 10 hours, a saturated aqueous solution of potassium hydrogen sulfate was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated saline, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (developing solution; n-hexane: ethyl acetate = 8: 1) to obtain 447 mg of the title compound (yield 42%).

- NMR (CDC 1 ₃₎

(5: 8.80 (1H, s), 8.45 (1H, d, J = 2.4 Hz), 8.19 (1Η, dd, J = 7.9, 2.4 Hz), 7.38 (1Η, d, J = 7.9Hz), 3.03 (2H, s), 1.60 (6H, s) Example 15b

Synthesis of 1,6-diamino-2,2-dimethylindane

A mixture of the compound (325 mg) obtained in Example 15a, platinum oxide (65 mg) and ethanol (10 ml) was stirred at room temperature and 5 atm under a hydrogen atmosphere for 10 hours. The platinum oxide was removed by filtering the reaction mixture, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (developing solution; port form: methanol = 4: 1) to obtain 22 lmg of the title compound (yield: 85%).

^X H-NMR (CD ₃ OD)

δ: 6.99 (1H, d, J = 7.9 Hz), 6.82 (1H, d, J = 2.4 Hz), 6.70 (1H, dd, J = 7.9, 2.4 Hz) ), 4.11 (1H, s), 2.82 (1H, d, J = 15.3Hz), 2.63 (1H, d, J = 15.3Hz), 1.20 (3H, s) s), 1.15 (3H, s) Example 15 c

Synthesis of 6-amino-11- (t-butoxycarbonyl) amino-2,2-dimethylindane

Of the compound obtained in Example 15b (10 Omg) and methanol (2 ml) To the mixture were added an aqueous potassium carbonate solution (1M, 1.14 ml) and di-t-butyl dicarbonate (149 mg). After the reaction mixture was stirred at room temperature for 5 hours, it was concentrated under reduced pressure, and 10% methanol-containing chloroform and saturated aqueous sodium chloride solution were added to the obtained residue. The organic layer was washed with saturated saline, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: n-hexane: ethyl acetate = 2: 1) to obtain 53 mg of the title compound (yield: 33%).

δ: 6.92 (1H, d, J = 7.9Hz), 6.56-6.5 1 (2H, m), 4.79 (1H, d, J = 9.8Hz), 4. 63 (1H, brd, J = 9.8 Hz), 3.53 (2H, brs), 2.64 (1H, d, J = 15.3 Hz), 2.55 (1H, d, J = 15.3 Hz), 1.49 (9H, s), 1.22 (3H, s), 0.91 (3H, s) Example c

Synthesis of t-butyl N- (2- (2-phenylethoxy) -5-thioperidophenylmethyl) carbamate

Compound obtained in Example 1 b (90 mg) and 4-dimethylaminopyridine

Tiophosgene (0.028 ml) was added dropwise to a solution of (9 Omg) in methylene chloride (8 ml), and the mixture was stirred at room temperature for 5 minutes. 28% aqueous ammonia solution (4m

1) was added and the mixture was stirred at room temperature for 16 hours. The reaction mixture was neutralized with 2N hydrochloric acid and extracted with ethyl acetate. The organic layer was washed successively with water and saturated saline, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (developing solution; n-hexane: ethyl acetate = 1: 1) to quantitatively obtain the title compound.

δ: 7.97 (1Η, brs), 7.36—6.83 (8H, m), 6.10 (2H, brs), 4.84 (1H, brs), 4.24-4 .1 9 (4H, m), 3.12 (2H, t, J = 6.6Hz), 1.42 (9H, s)

Synthesis of t-butyl N- (5-((S-ethylisothioperido))-2- (2-phenylethoxy) phenylmethyl) carbamate

To a solution of the compound (10 Omg) obtained in Example 1c in acetonitrile (1 Oml) was added iodide thiol (0.04 ml), and the mixture was heated under reflux for 2 hours. The reaction mixture was concentrated under reduced pressure, a saturated aqueous sodium hydrogen carbonate solution was added to the residue, and the mixture was extracted with ethyl acetate. The organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (developing solution; chloroform: methanol = 9: 1) to obtain 76 mg of the title compound (yield 71%).

δ: 7.36-7.24 (5H, m), 6.84-6.78 (3H, m), 4.80 (1H, brs), 4.20—4.15 (4H, m), 3. 12—3.00 (4H, m), 1.43 (9H, s), 1.34 (3H, t, J = 7.3 Hz) Example 2d, 3d, 4d, 5d

In the same manner as in Example 1d, the compounds shown in Table 7 were obtained using the corresponding thiourea forms.

Table 7

Example 9 e

N- (2- (3- (S-Ethylisothioureido) phenyl) _indane-1-2-yl) Synthesis of t-butyl t-butylhydroiodide

To a solution of the compound (21 mg) obtained in Example 9d in acetonitrile (4 ml) was added iodide tyl (17 mg), and the mixture was heated under reflux for 4 hours. The reaction mixture was concentrated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (eluent; dichloromethane: methanol = 20: 1) to give 2 Omg of the title compound (yield 68%).

^{_{X H-NMR (CDC 1 3}} )

δ: 7.26-7.17 (5H, m), 7.08-7.03 (2H, m), 6.82-6.77 (lH, m), 5.15 (1H, brs), 4.13 (2H, brs), 3.48 (4H, s), 3.03 (2H, brs), 1.32 (9H, s), 1.26 (3H, t, J = 7.3Hz) Example 6d

Synthesis of t-butyl N- (5- (N, 1-t-butoxycarbo-N, N ,, ethylethylanidino) -1-ethylphenylemethyl) -1-N- (2-phenylethyl) carbamate

1-Ethyl-3- (3-dimethylamino) was added to a mixture of the compound (10 Omg) obtained in Example 6c, N-ethyl-N'-t-butoxycarboxylthiodrea (69 mg) and dimethylformamide (3 ml). (Propyl) carbodiimide · hydrochloride (65 mg) was added, and the mixture was stirred at room temperature for 16 hours. The reaction mixture was diluted with ethyl acetate, washed with saturated saline, and the organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent; n-hexane: ethyl acetate = 7: 3) to obtain 12 Omg of the title compound (81% yield).

- NMR (CDC 1 ₃₎

δ: 7.28-6.90 (8Η, m), 4.35 (2H, brs), 3.39-3.32 (4H, m), 2.79 (2H, br), 2.55 ( 2H, q, J = 7.6), 1.53 (9H, s), 1.47 (9H, s), 1.17 (3H, t, J = 7.6Hz), 1.03 (3H, s) br) Example 12a, 7d, 10c, 15d, 16c, 17a

In the same manner as in Example 6d, the compounds shown in Table 8 were obtained using the corresponding aniline derivatives.

Example 13a

Synthesis of T-butyl N- (2-ethyl-1-5- (O-ethylisoureide) phenylmethyl) -1-N- (2-enylethyl) carbamate

Example 8c N- (5-((Ethylisoperido) -1-2-methoxyphenylmethyl) -N- (3-phenylpropyl) Synthesis of t-butyl rubbamate

Using the compound obtained in Example 8b, t-butyl N- (5-amino-2-methoxyphenylmethyl) -N- (3-phenylpropyl) carbamate was prepared in the same manner as in Example lb. Obtained. The title compound was obtained in the same manner as in Example 13a using the obtained compound.

^ -NMR (CDC 1 ₃₎

δ: 7.35-7.10 (5H, m), 6.85-6.70 (3H, m), 4.43 (2H, s), 4.25 (2H, q, J = 7 3Hz,), 4.15—3.80 (2H, m), 3.76 (3H, s), 3.40—3.13 (2H, m), 2.57 (2H, t, J = 7.8 Hz), 1.96—1.72 (2H, m), 1.44 (9H, s), 1.32 (3H, t, J = 7.3 Hz) Example 1 e

N—— (5— _ (S—ethylisothioperide) 2 (2-phenylethoxy) -phenylmethyl) amine Synthesis of dihydrochloride

A mixture of the compound (72 mg) obtained in Example 1d and trifluoroacetic acid (10 ml) was stirred at room temperature for 1 hour, and then concentrated under reduced pressure. After dissolving the residue in ethanol, a 1,4-dioxane solution of hydrogen chloride (4N) was added, and the mixture was stirred at room temperature for 2 minutes, and the reaction solution was concentrated under reduced pressure. The residue was dissolved in water, washed with ethyl acetate, and the aqueous layer was lyophilized to give the title compound quantitatively.

XH-NMR (D ₂₀ )

(5: 7.46-7.11 (8H, m), 4.47 (2H, t, J = 6.3Hz), 4.01 (2H, s), 3.25-3.17 (4H , M), 1.41 (3H, t, J = 7.3 Hz) Example 2 e, 3 e, 4 e, 5 e, 9 f, 12 b, 6 e, 7 e, 10 d, 13 b, 8d, 14b

In the same manner as in Example 1e, the results are shown in Tables 10 to 12 using the corresponding protected Boc. The compound was obtained ((1) in the reaction conditions in the table indicates trifluoroacetic acid and 1,4-dioxane solution of hydrogen chloride (4N), and (2) indicates trifluoroacetic acid and 1N hydrochloric acid.). Table 10

(ZHI · 1 = ['H £) il

Η ί 'b' HZ) Si 'I' (in

'H) 06 -Ζ-09' δ '(ΐπ ΉΖ)

SO 'to OS' f '(ZHS, i = i "b

'HZ)' (πι ΉΑ) 0Ζ 'i

-8 'L' (s' HI) 93 'L' (m

'HZ) 00' 6-OS '6' (πι ΉΖ) 'ョ Ογ ^^ ^ ~ li d

09 '6-08' 6 '(s' HI) il · Π Κ „ ^Ά '.

: Q (9P-0SKa) N-Hl (2) q ε I

(ZHS 'i = I Ή £) 0Z Ί' ((a

'HS) Z' 2-22 'S' (ZHS

• 9 '6 · ει = ί.,' Pp 'HI)

'(ZHS' 9 'S' 8 = 1 'PP'HI)

Z £ 'f' (ω 'Η6).' L 'χ χ:

: (Οζα) 丽-HI (I) O T

(ω 'Η9) 12 Ί-IS "ΐ

'(ΉΖ) 60' Ζ-Η '1' (

'Η) 11 -Z-S8' I '(ZHS

■ i = I ΉΖ ΉΖ) ί 'ε' (ΖΗ £ 'L

= 1 'b' HZ S, S '(s' ΕΖ)

η '(in' m) iz 'ί-ζζ' ζ,: o? —

: 9 (OZQ) MN-HI (Τ)

(m Ή9) 9Ζ-Ί '(ζ

Η6 · £ = ί '&' ΗΖ) 6L '1' (2

HS 'i = f Ί' ΗΖ) 6ί

'L = t' b 'HZ) 0 * S' (ZHS

= f Ί * HZ) S9 2 '(s' ΗΖ)

η · '(ω Ή8) εε'-Ί

: 9 (OZO N-Ht 'Λχ

(Τ) 3 9

(ZHS * Ζ = ί

Ί Ή8) ΙΖ '(m

'Ht)' I-88 'I' (ZHS "i,

= Γ Ί 'HZ)' I '(ZHS Ά = ί

ΉΖ) 02 · ε '(«ι' Ht) 9I

-0Z '' (ω Ή8) II "l-O 'L

: S> (οζα) Hiring-Hi (ΐ)

lZ.CO/TOdT/X3d Table 12

Example 11a

Synthesis of N— (5— (N, ethylethylanidino) —2 -— (3-phenyl-1-propoxy) phenylmethyl) amine'dihydrochloride

Using the compound obtained in Example 2b, in the same manner as in Example 6c, N— (5- (Ν′-t-butoxycarporinol N ′, ethylethylanidino) -2- (3- Phenyl -1-propoxy) phenylmethyl) rubamic acid! : One butyl was obtained. The title compound was obtained in the same manner as in Example 1e using the obtained compound.

H-NMR (D ₂ 〇)

δ: 7.41-7.09 (8H, m), 4.18 (2H, t, J = 6.3Hz), 4.07 (2H, s), 3.29 (2H, q, J = 7 3 Hz), 2.85 (2 H, t, J = 7.3 Hz), 2. 24-2.15 (2H, m), 2.12 (3 H, t, J = 7.3 Hz) Example 15 e

1-Amino-2,2-dimethyl-1-6- (N, 1-ethylidanidino) Arrest of Indan

A mixture of the compound (8 Omg) obtained in Example 15d, trifluoroacetic acid (0.28 ml) and methylene chloride (3 ml) was stirred at room temperature for 3 hours, and then concentrated under reduced pressure. To the resulting residue were added chloroform and 25% aqueous ammonia solution. The aqueous layer was extracted with 10% methanol-containing chloroform, combined with the previous organic layer, washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained residue was purified by column chromatography on amino-modified silica gel (eluent; chloroform: methanol = 9: 1) to give 4 lmg of the title compound (93% yield).

- NMR (CDC 1 ₃₎

d: 7.12-7.04 (2H, m), 6.94-6.99 (1H, m), 3.79 (1H, s), 3.28 (2Η, q, J = 7 3Hz), 2.63 (2H, s), 1.93 (2H, s), 1.22 (3H, t, J = 7.3Hz), 1.17 (3H, s), 0.82 (3H, s) Example 16 d, 17 b

In the same manner as in Example 15e, the compounds shown in Table 13 were obtained using the corresponding protected Boc.

Table 13

The structures of the example compounds disclosed in the present specification are summarized in Table 147.

Table 14

Ph.

ocHN.

BocHN, BocHN. B

N0 _{_2,} N0 ₂ NO, Example la Example 2a Example 3a

Example 15a

Example lb Example 2b Example 3b

P

WO 01/83429

Table 16

Table 17

Example 15e Example 16d Example] 7b

Test example

Test example 1

The inhibitory effect of the compound of the present invention on three types of NS isoforms known so far was compared with existing NS inhibitors.

As a reference compound,

L -NA ( ^NG -nitr oLa rginine),

LM IN (S- me t hy l - L- thiocitru 1 1 i ne), L- NAME (N G - nitr oLa rginine me t hy les ter),

It was used.

Each crude enzyme preparation was prepared according to the following procedure (Neuror eport 6, 1541-1545, 1995).

A crude enzyme sample of nNOS was prepared by the following procedure. Untreated male Sprague Daw 1 ey (SD) rats (body weight 300-400 g) were decapitated, the whole brain was quickly removed, and the cerebral cortex was collected on ice. Next, a 5-fold volume of a 50 mM Tris-HC1, ImM DTT (pH 7.4) solution was added, homogenized for 3 minutes, and centrifuged at 1,000 X g for 10 minutes. The obtained supernatant was centrifuged at 100,000 × g for 60 minutes, and the soluble cytoplasmic fraction of the finally obtained supernatant was used as a crude enzyme preparation of nNOS.

A crude enzyme sample of eNOS was prepared by the following procedure.ゥ Pulmonary artery vascular endothelial cell line (CPAE) was cultured in MEM medium containing 20% FBS. A few days later, this was detached from the flask with a 0.25% trypsin, ImM EDTA solution, added with an appropriate amount of FBS, and centrifuged at 1,000 rpm for 10 minutes. An appropriate amount of a phosphate buffer solution containing no calcium and magnesium (; H7.4) was added to the cells in the sediment, and the mixture was centrifuged at 1,000 rpm for 10 minutes. After the same procedure was repeated to wash the cells, 50 mM Tris-HC1 (pH 7.4) containing 1% TritonX-100 and ImM DTT was added, and the mixture was left on ice for 1 hour. Subsequently, after homogenizing for 3 minutes, the mixture was left on ice for 30 minutes while repeating stirring. Finally, the supernatant obtained by centrifugation at 100, OOOXg for 60 minutes was used as a crude enzyme sample of eNOS.

A crude enzyme preparation of iNOS was prepared by the following procedure. LPS (1 Omg / kg) was intraperitoneally administered to the rats, and after 6 hours, the rats were perfused with 1 OUZml of heparin-containing saline, and the lungs were removed. Then, a 5-fold dose of a 5 OmM Tris-HC1, ImM DTT (pH 7.4) solution was added, homogenized for 3 minutes, and centrifuged at 1,00O Xg for 10 minutes. The obtained supernatant was then centrifuged at 100, OOOXg for 60 minutes, and the soluble cytoplasmic fraction of the finally obtained supernatant was used as a crude enzyme sample of iNO.S.

NO S activity, according to the method basically literature, is one of the substrate L- ^[3 H] is one of the reaction products from arginine L- ^[3 H] was measured by quantifying the amount of conversion into citru 1 1 ine (B rain Edema IX, 60, p 285- 288, 1994;. Neur or epo rt 6, 154 1—1545, 1995).

The reaction solution was 100 nM L— [ ³ H] arginine, crude enzyme preparation (10-30 zg / ml protein), 1.25 mM Ca C12, 1 mM EDTA, 10 g / m 1 calmodu lin, ImM NADPH, 100 M tet ahyd r ob i op terine, 10 μ, Μ FAD, 10 U FMN, 5 OmM Tr is—HC 1 (pH 7.4), to which the compound of the present invention or A control compound was added.

The reaction was started by addition of L- ^[3 H] arginine, after 10 minutes the ink Yubeshiyon at 37 ° C, 5 OmM Tr is -HC l (pH5. 5), ImM E DTA was added 2 ml, placed in an ice To stop the reaction. The reaction solution cation exchange resin column (Dowex AG50WX- 8, Na + fo rm, 3. 2ml) through a substrate L- ^[3 H] arginine remaining unreacted and is the reaction product ^L-[3 H ] Citru 1 1 ine was separated. And this eluate, further a certain amount of distilled water The eluate obtained through the column to Minipaiaru were collected ^{L- [3 H] cit ru l} 1 i ne. Thereafter, scintillator was added and the radioactivity was measured by liquid core guides laser Chillon counter to quantify ^{L- [3 H] citru 1 1} ine.

The activities of nNOS and eNOS were determined by subtracting the activity detected in the absence of CaC12 and calmodulin from the activity detected in the presence of CaCl2 and calmodulin. The activity of iNOS was detected in the absence of CaCl2 and calmodulin. 'The protein concentration in the crude enzyme preparation was determined using a Microassay kit manufactured by Piolad. All experiments were performed in duplicate. Table 18 shows the IC50 value (concentration required for 50% activity inhibition) of the test compound for each NOS isoform and the ratio of each IC50 value as an index indicating selectivity. Further, in Table 19, when using a test compound 10_ ⁶ M, and displays the inhibitory activity against NN_〇 S or e NO S in the above reaction conditions. Table 18

Note) “n. D.” Means not measured, and Γ-j means uncalculated, Table 19

Inhibitory activity ''

Example compound

nNOS eNOS

(Type 1) (Type 3)

1 0d 89% 29%

1 1a 82% 15%

1 2b-82% 16% Industrial applicability

The compound of the present invention has an excellent NOS inhibitory action. In addition, the compound of the present invention has a strong nNOS inhibitory activity or a selective inhibitory effect on nN〇S, and various disease states in which a therapeutic effect can be expected by selectively inhibiting nNOS, such as cerebral blood It is useful as a therapeutic agent for vascular disorders, head trauma, spinal injury, pain, Parkinson's disease, Alzheimer's disease, convulsions, morphine resistance or dependence. Alternatively, the compound of the present invention can be expected to have a sufficiently excellent effect as compared with existing NOS inhibitors, or can be expected to have a clinically useful effect.

Claims

1. The following compound group (1) Compound group (1):

B

Or a possible tautomer thereof, or a pharmaceutically acceptable salt thereof.

2. A medicament containing the compound according to claim 1.

3. A NOS inhibitor comprising the compound of claim 1.

4. The agent according to claim 3, wherein the NOS inhibitor is an nNS selective inhibitor.

5. A therapeutic agent for cerebrovascular disease, comprising the compound according to claim 1.

6. The agent according to claim 5, wherein the cerebrovascular disorder is cerebral hemorrhage.

7. The agent according to claim 5, wherein the type of cerebrovascular disorder is subarachnoid hemorrhage.

8. The agent according to claim 5, wherein the type of cerebrovascular disorder is transient ischemic attack (TIA).

9. The agent according to claim 5, wherein the type of cerebrovascular disorder is cerebral edema.

10. The agent according to claim 5, wherein the cerebrovascular disorder is cerebral infarction.

11. The agent according to claim 10, wherein the subtype of cerebral infarction is atherothrombotic infarction, lacunar infarction, or cardiogenic embolism.

12. A therapeutic agent for head trauma comprising the compound according to claim 1.

13. A therapeutic agent for spinal injury containing the compound according to claim 1.

14. An analgesic containing the compound according to claim 1.

15. The agent according to claim 14, wherein the type of pain is headache.

16. The agent according to claim 15, wherein the subtype of headache is migraine, tension headache, or cluster headache or chronic paroxysmal headache.

17. A therapeutic agent for Parkinson's disease containing the compound according to claim 1.

18. A therapeutic agent for Alzheimer's disease containing the compound according to claim 1.

19. An anticonvulsant comprising the compound according to claim 1.

20. A therapeutic agent for morphine tolerance or dependence comprising the compound of claim 1.