WO2002048096A1

WO2002048096A1 - Hydrazone derivatives and use thereof in medicines

Info

Publication number: WO2002048096A1
Application number: PCT/JP2001/010667
Authority: WO
Inventors: Masayuki Nakamura; Jun Inoue
Original assignee: Senju Pharmaceutical Co., Ltd.
Priority date: 2000-12-12
Filing date: 2001-12-06
Publication date: 2002-06-20
Also published as: JPWO2002048096A1; AU2002221066A1

Abstract

Medicines containing compounds of the general formula (I): (I) [wherein R1 is hydrogen or lower alkyl; and A is morpholino or a group of the general formula (II): (II) (wherein B is hydroxylated lower alkyl, carbamoyl, or glycyl whose amino group may be substituted)]. The compounds have potent inhibitory activity against cysteine proteases, particularly calpain, and proper water solubility for the use in medicines.

Description

Hydrazone derivatives and their pharmaceutical uses

The present invention relates to a novel hydrazone derivative having cysteine protease inhibitory activity, particularly calpain inhibitory activity. The present invention also relates to a medicament containing the novel hydrazone derivative. Background art

Biochemical studies have revealed that abnormal enhancement of cysteine protease is involved in various diseases. For example, one of the cysteine proteases, virupain, is one of the cytoplasmic proteases widely distributed in living organisms, and is activated by virucidium ions. At present, it is clear that this abnormal activation of calpain is involved in various diseases such as stroke, subarachnoid hemorrhage, Alzheimer's disease, ischemic disease, muscular dystrophy, cataract, platelet aggregation, and arthritis. [Trends in Pharmacological Sciences, Vol. 15, p. 412 (1994)] ₀ On the other hand, calpain inhibitors are effective in maintaining the lens transparency in an experimental cataract model by lens culture [Curr. Eye Res. , 10, 657-666 (1994)], and has been found to be useful as a therapeutic agent for cataract (WO93Z23032). Examples of cysteine protease inhibitors that have been reported include epoxy succinic acid peptide derivatives (Japanese Patent Publication No. 1-54348 (US Pat. No. 4,333,879), and JP-A-55-153778 (US Pat. No. 4,418,075). ], Peptide halomethane derivatives (Japanese Patent Publication No. 6-29229), peptide diazomethane derivative CBiochem. J., 253, 751-758 (1988), J. Med. Chem., 35, 216- 220 (1992)] and peptidyl aldehyde derivatives (Japanese Patent Laid-Open No. 10-147564 (USUS6,057,290, USP 6,214,800), etc.). Sex, stability, absorption into the body, It has problems such as stability against metabolism and has not yet been put to practical use. Disclosure of the invention

An object of the present invention is to provide a compound having a strong cysteine protease inhibitory activity, particularly a calpain inhibitory activity, and having an appropriate water solubility desired as a pharmaceutical. The present inventors have made intensive efforts to solve the above problems, and as a result, have created a compound having a strong calpain inhibitory activity, and have further studied to complete the present invention.

That is, the present invention

(1) General formula (I)

[Wherein, R ¹ represents hydrogen or a lower alkyl group, A is a morpholino group or a general formula (II)

-NH-B (I I)

(In the formula, B is a hydroxy lower alkyl group or a carbamoyl group, or a glycyl group optionally substituted with an amino group.) A compound represented by the formula:

(2) The compound of the general formula (I) is a compound of the general formula (I I I)

[Wherein, R ² is a lower alkyl group. In the above (1) which is a compound represented by The compound described,

(3) a medicine containing the compound according to (1) or (2) above,

(4) a calpain inhibitor containing the compound according to (1) or (2) above,

(5) The medicament according to the above (3), which is for preventing or treating a disease associated with calpain.

(6) Calpain-related diseases include ischemic disease, immune disease, Alzheimer's disease, osteoporosis, disease due to brain tissue disorder, cataract, glaucoma, chorioretinal disease, posterior ocular complications due to photocoagulation, and angiogenesis The medicament according to the above (5), which is an associated disease,

(7) a pharmaceutical composition comprising the compound according to (1) or (2) and a pharmaceutically acceptable carrier;

(8) The pharmaceutical composition according to the above (7), which is a calpain inhibitor,

(9) For the treatment of ischemic disease, immune disease, Alzheimer's disease, osteoporosis, disease caused by brain tissue disorder, cataract, glaucoma, retinochoroidal disease, posterior ocular complications due to photocoagulation, and for the treatment of angiogenesis The pharmaceutical composition according to (8),

(10) A method for treating a disease associated with calpain, which comprises administering to a mammal in need of treatment an effective amount of the compound according to (1) or (2),

(11) Calpain-related diseases include ischemic disease, immune disease, Alzheimer's disease, osteoporosis, disease due to brain tissue disorder, cataract, glaucoma, choroidal disease, posterior ocular complications due to photocoagulation, and angiogenesis The method according to the above (10), which is an associated disease,

(12) Use of the compound according to (1) or (2) for producing a calpain inhibitor, and

(13) For the treatment of ischemic diseases, immunological diseases, Alzheimer's disease, osteoporosis, diseases caused by brain tissue disorders, cataracts, glaucoma, retinochoroidal diseases, and post-ocular complications due to photocoagulation, and for the treatment of angiogenesis The method according to (12) above,

(14) The compound according to (1), which has a suitable water solubility desired as a pharmaceutical. In the above general formula (I), the lower alkyl group represented by R ¹ means a linear or branched alkyl group having 3 to 5 carbon atoms, such as propyl, isopropyl, butyl, isobutyl, tert-butyl, Pentyl, isopentyl, neopentyl, tert-pentyl and the like can be mentioned, preferably a C ₄ alkyl group, particularly preferably isoptyl. .

In the above general formula (II), the hydroxy lower alkyl group represented by B is a hydroxy-3 alkyl group, such as hydroxymethyl, hydroxyethyl, hydroxypropyl, etc., and preferably hydroxyethyl. is there.

In the above general formula (II), examples of the group which may be substituted by the amino group of the glycyl group represented by B include a lower alkyl group represented by R ² in the above general formula (III). The lower alkyl group means a linear or branched alkyl group having 1 to 6 carbon atoms, for example, methyl, ethyl, propyl, isopropyl, butyl, isoptyl, tert-butyl, pentyl, isopentyl, neopentyl, tert-pentyl. Hexyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, and 2-ethylbutyl. Preferably it is methyl.

In the above general formula (I), the fluorophenyl group can be appropriately converted into, for example, a phenyl group, a chlorophenyl group, a bromophenyl group, a tolyl group, a trifluoromethylphenyl group and the like.

The compounds of the present invention also include compound (I) and salts thereof, and various solvates and polymorphs thereof. The salt of the compound represented by the general formula (I) in the present invention is preferably a physiologically acceptable salt, such as a salt with an inorganic acid, a salt with an organic acid, or a salt with an acidic amino acid. Can be Preferred examples of the salt with an inorganic acid include, for example, salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, and the like. Preferred examples of salts with organic acids include, for example, formic acid, acetic acid, trifluoroacetic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, P— Salts with toluenesulfonic acid etc. No. Preferable examples of the salt with an acidic amino acid include, for example, salts with aspartic acid, glumic acid and the like.

The compound of the present invention has, for example, the following general reaction formula

(Wherein each symbol has the same meaning as described above.). That is, the compound represented by the general formula (IV) can be prepared by adding the compound represented by the general formula (IV) to an organic solvent or a mixture of organic solvent and water in the presence or absence of a base. ). And the desired compound represented by the general formula (I) can be obtained. The compound represented by the general formula (IV) is described in, for example, JP-A-10-147564 (USP 6,057,290, USP 6,214,800). It can be produced by the method described.

Examples of the organic solvent include a conventional solvent such as dioxane, ethyl acetate, methanol, ethanol, dichloromethane, chloroform, N, N-dimethylformamide, and tetrahydrofuran, or a mixed solvent thereof. Preferred are dioxane, a mixed solvent of dioxane and methanol, and ethyl acetate. Examples of the base include acetates such as sodium acetate and potassium acetate, trialkylamines such as triethylamine and diisopropylethylamine, organic bases such as pyridine, lutidine, picoline and 4-dimethylaminopyridine, and sodium hydrogen carbonate. And inorganic acid salts such as sodium carbonate, sodium hydroxide, sodium hydride, potassium hydrogen carbonate, potassium carbonate, and potassium hydroxide. Preferably it is sodium acetate. The reaction temperature is generally in a range from cooling to heating, and preferably in a range from about 0 ° C. to about 3 Ot :.

The compound of the present invention and a salt thereof are novel compounds that have not been published in the literature, and are shown in Test Examples described later. Since it has such excellent calpain inhibitory activity, it is useful as a medicament as a calpain inhibitor by combining them as active ingredients and, if necessary, a carrier described below.

Pharmaceuticals containing the compound of the present invention include, for example, ischemic diseases, immune diseases, Alzheimer's disease, osteoporosis, and brain in mammals such as mice, rats, rabbits, dogs, cats, mice, humans, and humans. Diseases due to tissue disorders (eg cerebral vasospasm, cerebral thrombosis, cerebral infarction, cerebral obstruction, intracerebral hemorrhage, subarachnoid hemorrhage, hypertensive encephalopathy, transient ischemic attack, multiple infarct dementia, cerebral artery Sclerosis, Huntington's disease, etc., cataract, glaucoma (eg, open-angle glaucoma, low-tension glaucoma, closed-angle glaucoma, etc.), retinochoroidal disease (eg, retinal vascular obstruction, peritoneal peritonitis, Ea1) es disease, retinal vascular abnormalities such as ischemic ocular syndrome and retinal arteriolar aneurysm, retinopathy due to hypertension or renal disease, diabetic retinopathy, retinal pigment epitheliosis, retinal dystrophy, macular dystrophy, choroid Membrane atrophy, chorioretinopathy, macular degeneration, macular edema, retinal pigment epithelial detachment, retinal detachment, degenerative retinal segregation, retinoblastoma, retinal pigment epithelial tumor, optic nerve capillar hemangioma, etc. As a prophylactic and therapeutic agent for posterior eye complications (for example, macular edema, retinal detachment, optic neuritis, visual field abnormalities, abnormal light perception, abnormal color vision, etc.) or as a preventive and therapeutic agent for angiogenesis, retinal detachment, etc. Useful.

The medicament containing the compound of the present invention is administered systemically or locally. In addition to systemic oral administration, parenteral administration such as intravenous injection, subcutaneous injection, and intramuscular injection is also possible. It is applied topically to the skin, mucous membranes, nose and eyes.

Pharmaceutical formulations containing the compound of the present invention include solid preparations such as powders, granules, tablets, capsules, suppositories and the like, and liquid preparations such as syrups, injections, eye drops, nasal drops and the like. Can be When manufactured as granules and tablets, for example, excipients (lactose, sucrose, glucose, starch, crystalline cellulose, etc.), lubricants (magnesium stearate, talc, stearic acid, calcium stearate, etc.), disintegrants (Starch, carmellose sodium, calcium carbonate, etc.), binder (starch paste solution, hydroxypropylcellulose solution, carmellose solution, gum arabic Solution, gelatin solution, sodium alginate solution, etc.) can be used to produce an arbitrary dosage form. For granules and tablets, suitable coating agents (gelatin, sucrose, gum arabic, carnapalow, etc.), enteric coating agents (eg, cellulose acetate fluorate, methacrylic acid copolymer, hydroxy Propylcellulose phthalate, carboxymethylethylcellulose, etc.).

In the case of capsules, suitable excipients, such as magnesium stearate, calcium stearate, talc, and light anhydrous anhydrous gay acid to improve fluidity and lubricity, and for pressurized fluidity In addition to the above-mentioned crystalline cellulose and lactose, and the like, the above-mentioned disintegrants and the like are added as appropriate, mixed uniformly, or filled with granules or coated with a coating agent suitable for granules, or A suitable capsule base (eg, gelatin) may be added to glycerin or sorbitol or the like to form a capsule with increased plasticity. If necessary, coloring agents and preservatives [sulfur dioxide, parabens (methyl paraoxybenzoate, ethyl, propyl ester), etc.] can be added to these capsules. The capsule can be an ordinary coated capsule, an enteric coated capsule, a gastric resistant capsule, or a controlled release capsule. When enteric coated capsules are used, ribosomes coated with an enteric coating agent can be filled into ordinary capsules, capsules themselves can be coated with an enteric coating agent, or molded based on enteric polymers. .

When used as a suppository, a suppository base (eg, cacao butter, macrogol, etc.) can be appropriately selected and used.

When used as a syrup, for example, a stabilizer (such as sodium edetate), a suspending agent (such as gum arabic or carmellose), a flavoring agent (such as simple syrup, dextrose), or a fragrance may be appropriately selected and used. it can.

In the case of injections, eye drops or nasal drops, pharmaceutically acceptable additives such as tonicity agents (sodium chloride, potassium chloride, glycerin, mannitol, sorbitol) , Boric acid, dextrose, propylene glycol, etc.), buffers (phosphate buffer, acetate buffer, borate buffer, carbonate buffer, citrate buffer, tris buffer, dal permic acid, ip E.g., silonaminocaproic acid), preservatives, laoxybenzoic acid esters, chlorobutanol, benzyl alcohol, benzalkonium chloride, sodium dehydroacetate, sodium edetate, boric acid, etc.), thickeners (hydroxymethylcellulose, Hydroxyethyl cellulose, hydroxypropyl cellulose, polyvinyl alcohol, polyethylene glycol, etc., stabilizers (sodium hydrogen sulfite, sodium thiosulfate, sodium edetate, sodium citrate, ascorbic acid, dibutylhydroxytoluene) Such) , It can be produced by dissolving or dispersing in a solution to which a pH adjuster (hydrochloric acid, sodium hydroxide, phosphoric acid, acetic acid, etc.) is appropriately added.

Since the compound of the present invention has appropriate water solubility desired as a pharmaceutical, it can be advantageously produced particularly in the above-mentioned preparations such as syrups, injections, eye drops and nasal drops.

The amount of additives in the above syrups, injections, eye drops, and nasal drops depends on the type and use of the additives to be added, but it is sufficient to add a concentration that can achieve the purpose of the additives. the isotonic agent is usually such that the osmotic pressure of about 229～ about 343mOsZ Kg · H ₂ 0, is added to about 0.5 5 to about 5. 0wZv%. The buffer is about 0.01 to about 2.0 /%, the thickener is about 0.01 to about 1.Ow / v%, and the stabilizer is about 0.001 to about 1.OwZv% Add about. The pH adjuster is appropriately added and usually adjusted to about pH 3 to about 9, preferably pH 4 to about 8. The dosage of the compound represented by the general formula (I) of the present invention varies depending on the target disease, symptom, administration subject, administration method, and the like. It is preferable to administer about 1 to about 10 Omg at a time. In the case of injection, for adults, it is preferable to administer about 0.1 to about 3 Omg once to three times a day. When used topically, ophthalmic solution adjusted to about 0.001 to about 1. Ow / v%, preferably about 0.01 to about 0.5%, is applied once to 20 to 50 times. 1, 1 day 2-6 It is good to give it twice.

Specific compounds of the present invention include, for example,

• N— (3-aza-1 3 -— (2- (dimethylamino) acetylamino) 1-1-1 (2-methylpropyl) prop-2-enyl) —2 — (((4-fluorophenyl) sulfonyl) amino 3-methylbutane amide (Compound 1),

• N— (3-aza—3-morpholine-1-ylprobut-2-enyl) 1-2 — (((4-fluorophenyl) sulfonyl) amino) -1-methylbutane amide (compound 2),

• N— (3-aza—3 — ((2-hydroxyethyl) amino) —1— (2-methylpropyl) prop—2-enyl) 1-2 — (((4-fluorophenyl) sulfonyl) Amino) 1-methylbutane amide (compound 3),

-N- (3—other 3— (2— (dimethylamino) acetylamino) prop— 2

—Enyl) — 2— (((4-fluorophenyl) sulfonyl) amino) 1-3-methylbutane amide (compound 4)

· N— (3-aza1-1— (2-methylpropyl) —3-morpholine-1-ylprop-2-enyl) 2 — (((4-fluorophenyl) sulfonyl) amino) 1-3 —Methylbutanamide (compound 5), and

• N— (3— (aminopropyl) amino) —3-aza-11- (2-methylpropyl) probut-2-enyl) —2-((((4-fluorophenyl) sulfonyl) amino) -1-3-methylbutane Namide (compound 6) and the like. Among these, N- (3-other 3- (2- (dimethylamino) acetylamino) -1- (2-methylpropyl) prop-2-ene-1-2-(((4-fluorophenyl) sulfonyl) amino) 3-Methylbutanamide (Compound 1) is excellent in water solubility.

BEST MODE FOR CARRYING OUT THE INVENTION

The present invention will be described in more detail with reference to the following examples, but the present invention is not limited thereto. Reference Example 1 N — ((4-Fluorophenyl) sulfonyl) —L—Varilu L—Mouth Isinal

Step 1: Dissolve valine (11.7 g, 10 Ommo 1) in 1 M aqueous sodium hydroxide solution (l O OmL), add purified water (15 OmL) and tetrahydrofuran (10 mL), and cool under ice-cooling. While stirring with, a 1M aqueous solution of sodium hydroxide (10 OmL) and a solution of 4-fluorobenzenesulfonyl chloride (17.5 g, 9 Ommo 1) in tetrahydrofuran (10 OmL) were simultaneously added dropwise. The solution was stirred at room temperature for about 18 hours to react. After completion of the reaction, the reaction solution was adjusted to pH 2-3 and extracted with ethyl acetate. The extract was washed with dilute hydrochloric acid and saturated saline, and then dried over anhydrous magnesium sulfate. Ethyl acetate is distilled off under reduced pressure, and the residue is mixed with a mixture of hexane and ethyl acetate (a solution in which hexane is mixed at a ratio of about 10 to about 20 volumes per 1 volume of ethyl acetate. Is the same as above.), And N-((4-fluorophenyl) sulfonyl) -L-parin (15.5 g,

56%) as white crystals.

Step 2: N-((4-fluorophenyl) sulfonyl) -L-parin (1 5. Og, 55 mmo 1) and N-hydroxysuccinimide (7.6 g, 66 mmol) in tetrahydrofuran (20 OmL) And stirred under ice-cooling while stirring 1-ethyl-3- (3-dimethylaminopropyl) carboimide hydrochloride (12.

A solution of 6 g, 66 mmo 1) in dichloromethane (20 OmL) was slowly added. The solution was stirred at room temperature for about 4 hours to react. After completion of the reaction, the solvent was distilled off under reduced pressure, and the residue was dissolved in ethyl acetate, washed with dilute hydrochloric acid, a saturated aqueous solution of sodium hydrogencarbonate, and saturated saline, and then dried over anhydrous magnesium sulfate. The ethyl acetate was distilled off under reduced pressure, and the residue was washed with a mixture of hexane and monoethyl acetate. N-(((4-fluorophenyl) sulfonyl) -L-parin N-hydroxysuccinimide ester (17.6 g, 87 %) As white crystals.

Step 3: N-((4-fluorophenyl) sulfonyl) -L-parin N-Hydroxysuccinimide (2.Og, 5.4 mmol) was converted to dichloromethan Dissolved in tan (50 mL) and added mouth isinol (0.82 g, 7. Ommol). The solution was stirred for about 2 hours to react. After completion of the reaction, the resultant was washed with dilute hydrochloric acid, a saturated aqueous solution of sodium hydrogencarbonate and saturated saline, and then dried over anhydrous magnesium sulfate. Dichloromethane was distilled off under reduced pressure, and the residue was washed with a mixture of hexane and monoethyl acetate, and N-((4-fluorophenyl) sulfonyl) 1 L-1 Paryl L-mouth isinol (1.9 g, 94%) was obtained in white. Obtained as crystals.

Step 4: N-((4-fluorophenyl) sulfonyl) -L-paryl-L-Mouth isinol (1.8 g, 4.8 mmol) was dissolved in dimethyl sulfoxide (20 mL) and dichloromethane (10 mL) and diisopropyl Ethiluamine (2.5 g, 19 mmo 1) was added. While stirring this solution at room temperature, a dimethyl sulfoxide solution (15 mL) of a sulfur trioxide pyridine complex (3.1 g, 19 mmo 1) was added, and the mixture was further stirred for about 40 minutes. After completion of the reaction, ethyl acetate was added, and the mixture was washed with dilute hydrochloric acid, a saturated aqueous solution of sodium hydrogencarbonate, and saturated saline, and then dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the residue was recrystallized from ethyl acetate. N-((4-fluorophenyl) sulfonyl) -L-valyl-L-mouth isinal (Reference compound 1) (1. lg, 60 %) As white crystals.

Melting point; 1 57

Ή— NMR (270MHz, DMSO-d ₆ ) d: 0.74 (d, 3H, J = 5.9Hz), 0.80 (d, 6H, / = 6.4Hz), 0.85 (d, 3H, / = 6.8Hz), 1.14 -1.46 (m, 3H), 1.81-1.93 On, 1H), 3.56-3.62 (dd, 1H, / = 6.6, 9.5 Hz), 3.80-3.88 (m, 1H), 7.33-7.42 (m, 2H), 7.79-7.86 On, 2H), 7.96 (d, 1H, / = 9.8 Hz), 8.27 (d, 1H, / = 7.3 Hz), 9.14 (s, 1H).

_{_{. Anal Calcd for C 17 H 25}} FN 2 0 4 S:. C, 54.82; H, 6.77; N, 7.52 Found: C '54.82; H, 6.76; N, 7.57.

[a] _D ²⁵ + 8.99 ° (c = 0.20, DMS0).

Reference Example 2 N— ((4-fluorophenyl) sulfonyl) 1 L

Step 1: N — ((4-fluorophenyl) sulfonyl) —L—parin Droxysuccinimide ester (4.0 g, llmmol) was dissolved in ethyl acetate (15 OmL), and aminoacetaldehyde dimethyl acetal (1.4 g, 13 mmol) was added. The solution was stirred at room temperature for about 24 hours. The reaction solution was washed with 1 M hydrochloric acid, a saturated aqueous solution of sodium hydrogen carbonate and saturated saline, dehydrated with anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was crystallized from a mixture of hexane and ethyl acetate, and N- (2,2-dimethoxyethyl) -2-((((4-fluorophenyl) sulfonyl) amino) -3-methylbutaneamide (3. lg, 80%) as colorless crystals.

Melting point: 1 06. 8- 107. 9

Ή-thigh (300MHz, D S0-d ₆ ) δ: 0.77 (d, 3H, / = 6.0 Hz), 0.79 (d, 3H, / = 6.6 Hz), 1.78 (m, 1H), 2.85 On, 1H) , 2.97 (m, 1H), 3.19 (s, 3H), 3.20 (s, 3H), 3.48 (dd, 1H, / = 9.3, 7.2 Hz), 4.10 (t, 1H, / = 5.4 Hz), 7.34- 7.42 (m, 2H), 7.77-7.84 (m, 2H), 7.86 (d, 1H, / = 9.3 Hz), 7.93 (t, 1H,; = 5.9 Hz).. Anal Calcd for C 15 H 23 N 2 0 ₅ SF: C, 49.71; H, 6.40; N, 7.73. Found: C, 49.8; H, 6.48; N, 7.97.

[CK] _d ²⁵ + 25.2 ° (c = 0.20, DMS0).

Step 2: N- (2,2-dimethoxyethyl) 1-2-(((4-fluorophenyl) sulfonyl) amino) _3-methylbutaneamide obtained in Step 1 (3.1 g, 8.6 mmo 1 ) Was dissolved in tetrahydrofuran (2 OmL) and water (1 OmL), and trifluoroacetic acid (1 OmL) was added. The solution was stirred at about 60 for about 5 hours. The reaction solution was diluted with ethyl acetate, washed with 1M hydrochloric acid, saturated aqueous sodium hydrogen carbonate solution and saturated saline, dehydrated with anhydrous magnesium sulfate, and concentrated under reduced pressure. The obtained white solid was crystallized from hexane to give N-((4-fluorophenyl) sulfonyl) -L-valylglycineal (Reference Compound 2) (2.lg, 78%) as colorless crystals. Was.

Melting point; 1 12.8-1 13.8 ° C

Ή-NM (300MHz, DMS0-d ₆ ) δ: 0.81 (d, 3H, / = 6.6 Hz), 0.82 (d, 3H, / = 6.6 Hz), 1.83 (m, 1H), 3.55 (m, 1H), 3.67 (d, 2H,; = 5.4 Hz), 7.35-7.41 (m, 2H), 7.78-7.84 (m, 2H), 7.97 On, 1H), 8.36 (t, 1H, J = 5. Hz), 9.19 (s, 1H).

_{_{. Anal Calcd for C 13 H 17}} N 2 0 4 SF:. C, 49.36; H, 5.42; N, 8.86 Found: C, 49.48; H, 5.64; N, 8.66.

[o!] _D ²⁵ + 22.2 ° (c = 0.20, DMSO).

Example 1 N— (3-aza—3— (2- (dimethylamino) acetylamino) -1- (2-methylpropyl) prop-2-enyl) —2 — (((4-fluorophenyl) sulfonyl) amino ) —3-Methylbutanamide (Compound 1)

Reference Compound 1 (0.5 g, 1.3 mmo 1) was dissolved in a mixture of dioxane (2 OmL) and water (1 OmL), and N, N-dimethyldaricin hydrazide hydrochloride (0.28 g, 1 .5 mmo1) and sodium acetate (0.55 g, 6.7 mmo1) were added. The solution was stirred at room temperature for about 30 minutes. The reaction solution was diluted with ethyl acetate, washed with a saturated aqueous solution of sodium hydrogencarbonate and saturated saline, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The residual white solid was recrystallized from ethyl acetate to obtain the desired compound 1 (0.27 g, 43%) as colorless crystals.

Melting point: 162.8-163.3 ° C

_{Ή-NMR (300MHz, CDC1 3} ) δ: 0.84 (d, 3Η, / = 5.1 Hz), 0.85 (d, 3H, / = 6.9 Hz), 0.88 (d, 3H, / = 4.8 Hz), 0.96 (d , 3H, / = 6.9 Hz), 1.33-1.35 (m, 3H), 2.15 (m, 1H), 2.32 (s, 6H), 3.07 (s, 2H), 3.58 (m, 1H), 4.51 (m, 1H), 5.63 (m, 1H), 6.59 (d, 1H, / = 7.5 Hz), 7.11-7.18 (m, 2H), 7.60 (d, 1H, / = 3.6 Hz), 7.85-7.90 (m, 2H ), 9.95 (s, 1H).

Anal.Calcd for C ₂₁ H ₃₄ N ₅ 0 ₄ SF-0.2H ₂ 0: C, 53.08; H, 7.30; N, 14.74. Found: C, 52.88; H, 7.23; N, 14.76.

[o;] _D ²⁵ + 10.4 ° (c = 0.20, DMSO).

Example 2 N- (3-aza-1-3-morpholine-4-ylprop-2-enyl) -2-(((4-fluorophenyl) sulfonyl) amino) -3-methylbutane amide (Compound 2)

Reference compound 2 (0.30 g, 0.94 mmol) was dissolved in ethyl acetate (2 OmL), and N-amino morpholine (0.12 g, 1. lmmol) was added. The solution was stirred at room temperature for about 30 minutes. The reaction mixture was diluted with ethyl acetate, washed with a saturated aqueous solution of sodium hydrogencarbonate and brine, dehydrated with anhydrous magnesium sulfate, and concentrated under reduced pressure. The obtained solid was crystallized from ethyl acetate to obtain Compound 2 (0 20 g, 53%) as colorless crystals.

Melting point: 165.7-166.0 ° C

Ή-NR (300MHz, DMS0-d ₆ ) δ: 0.78 (d, 3Η, / = 6.9 Hz), 0.81 (d, 3H,; = 6.9 Hz), 1.80 (m, 1H), 2.81-2.84 (m, 4H), 3.45 (m, 1H), 3.53-3.57 (m, 2H), 3.68-3.71 (m, 4H), 6.54 (t, 1H, / = 4.7 Hz), 7.35-7.42 (m, 2H), 7.79 -7.83 (m, 2H), 7.89 (s, 1H), 8.10 (t, 1H, / = 5.4 Hz).

_{_{. Anal Calcd for C 17 H 25}} N 4 0 4 SF:. C, 50.99; H, 6.29; N, 13.99 Found: C, 51.04; H, 6.40; N, 13.33.

[Q!] _D ²⁵ +33.2. (c = 0.20, DMSO).

Example 3 N— (3-aza-3-((2-hydroxyethyl) amino) —1- (2-methylpropyl) probut-2-enyl) —2 — (((4-fluorophenyl) sulfonyl) Amino) -3-Methylbutane amide (Compound 3)

Reference compound 1 (0.80 g, 2. lmmol) was dissolved in ethyl acetate (20 mL), and hydrazinoethanol (0.20 g, 2.6 mmol) was added. This solution was stirred at room temperature for about 30 minutes. The reaction mixture was diluted with ethyl acetate, washed with a saturated aqueous solution of sodium hydrogen carbonate and saturated saline, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The obtained solid was crystallized from ethyl acetate to obtain Compound 3 (0.55 g, 60%) as colorless crystals.

Melting point: 112. 5-114. 9 ° C

Ή-NMR (300MHz, DMSO- d ₆ ) δ: 0.71 (d, 3H, / = 4.8 Hz), 0.76-0.82 (m, 9H), 0.99-1.20 (m, 3H), 1.80 (m, 1H), 2.90-2.95 On, 2H), 3.40-3.48 (m, 3H), 4.08 On, 1H), 4.49 (m, 1H), 6.36 (m, 1H), 6.63 (m, 1H), 7.33-7.39 (m, 2H), 7.80-7.84 (m, 4H).

_{_{. Anal Calcd for C 19 H 31}} N 4 0 4 SF:. C, 53.00; H, 7.26; N, 13.01 Found: C, 52.76; H, 7.08; N, 13.09.

[a] _D ²⁵ -3.4 (c = 0.21, DMSO).

Example 4 N- (3-aza-3- (2- (dimethylamino) acetylamino) probut-2-enyl) -12-(((4-fluorophenyl) sulfonyl) amino) -3-methylbutynamid (compound Four)

Reference compound 2 (0.4 g, 1.3 mmo 1) was dissolved in dioxane (2 OmL) and water (1 OmL), and N, N-dimethylglycine hydrazide hydrochloride (0.26 g, 1.4 mmo 1) was added. Sodium acetate (1.0 g, 13 mmo 1) was added. The solution was stirred at room temperature for about 30 minutes. The reaction mixture was diluted with ethyl acetate, washed with a saturated aqueous solution of sodium hydrogencarbonate and brine, dehydrated with anhydrous magnesium sulfate, and concentrated under reduced pressure. The obtained solid was crystallized from ethyl acetate to obtain Compound 4 (0.27 g, 51%) as colorless crystals.

Melting point: 1 31. 0-132.7

_{Ή-NMR (300MHz, CDC1 3} ) 6: 0.87 (d, 3H, J = 6.9 Hz), 0.89 (d, 3H, / = 6.9 Hz), 2.03 (m, 1H), 2.35 (s, 6H), 3.12 (s, 2H), 3.65 (m, 1H), 3.93-3.97 (m, 2H), 5.87 (bs, 1H), 7.09-7.19 (m, 2H), 7.24 (m, 1H), 7.40 (t, 1H , / = 3.9 Hz), 7.85-7.91 (m, 2H), 10.08 (bs, 1H).

[a] _D ²⁵ + 29.4 ° (c = 0.20, DMSO).

Example 5 N— (3-aza-1- (2-methylpropyl) -13-morpholine-4-ylprop-2-enyl) -12-(((4-fluorophenyl) sulfonyl) amino) — 3-Methylbutane amide (Compound 5)

Reference compound 1 (1.0 g, 27 mm 01) was dissolved in dioxane (2 OmL), methanol (5. OmL) and water (5. OmL), and N-amino morpholine (0.33 g) was dissolved. , 3.2 mmo1) and sodium acetate (2.2 g, 27 mmo1). The solution was stirred at room temperature for about 30 minutes. The reaction solution was diluted with ethyl acetate, washed with a saturated aqueous solution of sodium bicarbonate and brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The obtained solid is crystallized from ethyl acetate, Compound 5 (0.50 g, 41%) was obtained as colorless crystals.

Melting point: 1 38.4-138.9 ° C

Ή-NMR (300MHz, DMS0-d ₆ ) δ: 0.70 (d, 3Η, / = 5.4 Hz), 0.75-0.82 (m, 9H), 1.05-1.22 (ι, 3H), 1.82 (m, 1H), 2.7-2.82 (i, 4H), 3.48 (dd, 1H, / = 8.9, 6.8 Hz), 3.65 (t, 4H, / = 4.8 Hz), 4.14 (m, 1H), 6.6 (d, 1H, / = 4.8 Hz), 7.32-7.38 (, 2H), 7.78-7.86 (in, 4H).

_{_{. Anal Calcd for C 21 H 33}} N 4 0 4 SF:. C, 55.24; H, 7.29; N, 12.27 Found: C, 55.18; H, 7.04; N, 12.08.

Example 6 N- (3- (aminopropyl) amino) -3- (11- (2-methylpropyl) prop-2-enyl) -2-(((4-fluorophenyl) sulfonyl) amino) 1 3-Methylbutane amide (Compound 6)

Reference compound 1 (1.0 g, 2.7 mmo 1) was dissolved in dioxane (2 OmL), methanol (5. OmL) and water (5. OmL), and semicarbazide hydrochloride (0.33 g, 3. 2mmo1) and sodium acetate (2.2g, 27mmo1) were added. The solution was stirred at room temperature for about 30 minutes. The reaction solution was diluted with ethyl acetate, washed with a saturated aqueous solution of sodium hydrogen carbonate and a saturated saline solution, dehydrated with anhydrous magnesium sulfate, and concentrated under reduced pressure. The obtained solid was crystallized from ethyl acetate to obtain Compound 6 (0.85 g, 74%) as colorless crystals.

Melting point: 100. 4— 102.4 ° C

Ή-NMR (300MHz, DMS0-d ₆ ) δ: 0.70 (d, 3Η, / = 8.4 Hz), 0.76-0.81 (m, 9H), 1.00-1.23 (m, 3H), 1.74-1.86 (m, 1H ), 3.54 (m, 1H), 4.19 (m, 1H), 6.22 (s, 2H), 6.99 (d, 1H, /=4.2 Hz), 7.32-7.38 (m, 2H), 7.79-7.83 (m, 2H), 7.90-7.93 (i, 2H), 9.85 (s, 1H).

_{_{. Anal Calcd for C 18 H 28}} N 5 0 4 SF · 0.35H 2 0:. C, 49.61; H, 6.64; N, 16.07 Found: C, 49.99; H, 6.69; N, 15.69.

MALDITOF-MS [M + H] ⁺ Calcd 430.192, Found 430.186; [M + Na] ⁺ Calcd 452.174, Found 452.191; [M + K] ⁺ Calcd 468.148, Found 468.139.

Test Example 1 Measurement of ^ -calpain inhibitory activity

—The inhibitory activity of calpain was measured according to the method described in the literature CAnal. Biochei. Vol. 208, 387-392 (1993)]. That is, 200 L of a reaction solution containing 0.5 mg ZmL casein, 50 mM Tris-HC1 (pH 7.4), 2 OmM dithiothreitol, and 0.03 enzyme unit ^ -calpain, and dimethyl containing various concentrations of the test drug. Sulfoxide solution 2.5 iL and 2 OmM salt ^: 50 ah of calcium aqueous solution was added to a 96-well plate. After reacting at about 30 ° C for about 60 minutes, transfer 100 L of the reaction solution to another 96-well plate, add 5 L of purified water and 100 L of 50% Coomassie Brilliant Blue solution, and add about 15 minutes at room temperature. After standing for 5 minutes, the absorbance was measured at 595 nm. The control value was measured after the same treatment without the test drug, and the blank value was obtained by adding lmM EDTA instead of 2 OmM calcium chloride aqueous solution. The amount required for inhibition (IC ₅₀ ) was determined.

Inhibition rate = {1— (measured value—blank value) / (control value—blank value)} × 100 The results are shown in Table 1. Table 1. -Calpain inhibitory activity of the compounds of the present invention

-Calpain inhibitory activity

IC ₅₀ (liM)

Compound 1 0 37

Compound 3 0 19

Compound 5 0 79

Compound 6 068 As a result, the compound of the present invention was found to have a strong calpain inhibitory effect.

Test Example 2 Solubility

5-: L Omg of each test compound is added to 2 mL of buffer solution (pH 4, 5, 6 and 7) to form a suspension. After shaking at about 25 ° C for about 5 hours, a membrane filter (0.45 m ). The concentration of each test compound in the filtrate was quantified using HP LC under the conditions shown below, and the solubility was calculated.

HPLC conditions

Column: SHI SE I DO CAPCELL PAK C 18 UG 120 (4.6 X 100 mm)

Mobile phase: acetonitrile / 1 OmM phosphate buffer aqueous solution (pH7) = 30/70 Column temperature: 40 ° C

Detection wavelength: 250 nm

Flow rate: 1. OmL / min

Sample solution: The filtrate ImL was adjusted to 1 OmL with the mobile phase, and 10 L of this solution was analyzed by HPLC.

Standard solution: Approximately 1 Omg of each test compound was precisely weighed, adjusted to 1 OmL with the mobile phase, and 10 L of this solution was analyzed by HPLC.

Test result 2

The results are shown in Table 2. Table 2. Solubility of the compound of the present invention

Test compound solubility (mg / mL)

pH4 pH5 pH6 pH7 Compound 1 4.33 1.98 0.702 0.213 Compound 2 1.26 0.84 0.6692 0.6667 Compound 4> 5> 5> 5> 5 Reference compound 0.10 0.10 0. 10

Reference compound 1 has a stronger calpain inhibitory activity than that of the compound of the present invention, but its solubility was 0.1 OmgZmL irrespective of pH, but the compound of the present invention is practically usable as a pharmaceutical preparation. It was found that the water solubility was improved at a certain pH of 4 to 7.

In particular, it was revealed that Compound 1 has high calpain inhibitory activity and high water solubility near pH 4, which is a practical area for pharmaceutical preparations.

Formulation Example 1 Tablet

Compound 35 Omg

Lactose 8 Omg

17mg starch

Magnesium stearate 3mg

Microcrystalline cellulose 10mg

The above ingredients are used as a tablet material, mixed uniformly, and formed into tablets by a conventional method. Sugar coating may be applied as necessary.

Formulation Example 2 Injection

Compound 1 0.2 g

0.9 g of sodium chloride

Sodium hydroxide / hydrochloric acid qs (pH 5.0) Distilled water for injection Total volume l O OmL The above components are mixed and dissolved by a conventional method to prepare an injection.

Formulation Example 3 Eye drops

Compound 1 l O Omg

Boric acid 700mg

Borax suitable amount

Sodium chloride 500mg

Hydroxymethyl cellulose 500mg

Sodium edetate 0.05 mg

Benzalkonium chloride 0.0005mg

Sodium hydroxide / hydrochloric acid qs (pH 5.0) Sterilized purified water total volume l O OmL

The above components are aseptically mixed and dissolved by a conventional method to produce eye drops.

Since the compound represented by the formula (I) of the present invention has an excellent calpain inhibitory activity and has an appropriate water solubility desired as a pharmaceutical, diseases involving calpain include ischemic disease, immune disease, and Alzheimer's disease. It can be advantageously used for osteoporosis, diseases caused by brain tissue disorders, cataracts, glaucoma, retinochoroidal diseases, posterior segment complications due to photocoagulation, and diseases involving angiogenesis. While a number of specific embodiments of the present invention have been described in detail, those skilled in the art will recognize that specific embodiments illustrated therein can be made without departing from the novel teachings and advantages of the present invention. Since it is possible to make various modifications and changes, all such modifications and changes are intended to be included within the spirit and scope of the invention as defined by the following claims.

This application is based on a patent application No. 2000-377582 and a patent application No. 2001-149683 filed in Japan, the contents of which are incorporated in full herein. It is.

Claims

The scope of the claims

General formula (I)

-NH-B (I I)

(Wherein B is a hydroxy lower alkyl group, a carbamoyl group, or a glycyl group which may be substituted with an amino group.) ] The compound represented by these.

2. The compound of the general formula (I) has the general formula (I I I)

[Wherein, R ² is a lower alkyl group. The compound according to claim 1, which is a compound represented by the formula:

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

4. A calpain inhibitor comprising the compound according to claim 1 or 2.

5. The medicament according to claim 3, which is used for prevention or treatment of a disease associated with calpain.

6. Diseases involving calpain include ischemic disease, immune disease, Alzheimer's disease, osteoporosis, disease due to brain tissue disorder, cataract, glaucoma, chorioretinal disease, posterior ocular complications due to photocoagulation, and angiogenesis 6. The medicament according to claim 5, which is an associated disease.

7. A pharmaceutical composition comprising the compound according to claim 1 or 2 and a pharmaceutically acceptable carrier.

8. The pharmaceutical composition according to claim 7, which is a calpain inhibitor.

9. For the treatment of ischemic disease, immunological disease, Al-Hachi-Ima disease, osteoporosis, disease due to brain tissue disorder, cataract, glaucoma, retinochoroidal disease or posterior ocular complications due to photocoagulation or for the treatment of angiogenesis 9. The pharmaceutical composition according to claim 8, which is:

10. A method for treating a calpain-related disease, comprising administering to a mammal in need of treatment an effective amount of the compound according to claim 1 or 2.

1 1. Calpain-related diseases include ischemic disease, immune disease, Alzheimer's disease, osteoporosis, brain tissue disorder, cataract, glaucoma, chorioretinal disease, posterior ocular complications due to photocoagulation, and angiogenesis 10. The method according to claim 10, wherein the disease is an associated disease.

12. Use of the compound according to claim 1 or 2 for producing a calpain inhibitor.

1 3. Claims for the treatment of ischemic disease, immunological disease, Alzheimer's disease, osteoporosis, disease due to brain tissue disorder, cataract, glaucoma, retinochoroidal disease or posterior ocular complications due to photocoagulation or for the treatment of angiogenesis. The method according to range 1 to 2.