WO2001094311A1

WO2001094311A1 - Cytoprotectors

Info

Publication number: WO2001094311A1
Application number: PCT/JP2001/004796
Authority: WO
Inventors: Atsuyuki Ashimori; Satoshi Horie; Shinichi Takanashi
Original assignee: Mitsubishi Pharma Corporation
Priority date: 2000-06-08
Filing date: 2001-06-07
Publication date: 2001-12-13
Also published as: AU2001262711A1

Abstract

Novel cytoprotectors are provided which contain as the active ingredient compounds of the general formula (I) or pharmaceutically acceptable salts thereof wherein Y is CH2, S, O, CO, or NR4; n is an integer of 1 to 10; R?1 and R2¿ are each H, alkyl, OH, alkoxy, halogeno, nitro, amino, or trifluoromethyl; R3 is H or alkyl; and R4 is H, alkyl, or aralkyl.

Description

Description Cell protective agent Technical field

The present invention relates to a cytoprotective agent, and more particularly to a cytoprotective agent, particularly an agent useful for protecting brain cells, nerve cells, and kidney cells. Background art

Recently, apoptosis has been drawing attention as a concept of cell death. Apoptosis (apoptosis, also known as apoptosis) means the suicide or suicide of a cell. This apoptosis, unlike necrosis (necrosis), which is a pathological cell death, is considered to be the active death of the cell itself incorporated as genetic information. In other words, it is thought that a signal that induces apoptosis is activated by some external or internal factor, and the cell itself actively breaks down, leading to death. In particular, attention has been paid to the relationship between neuronal (brain) cell death and apoptosis (Japanese Unexamined Patent Publication No. Hei 8-27272). Disclosure of the invention

An object of the present invention is to provide a drug having an excellent cell-protecting (apoptosis-suppressing) action.

In addition, the present inventors have further studied in consideration of the above circumstances, and as a result, have found that a group of compounds having a specific structure has an excellent cytoprotective action, and completed the present invention. The present invention relates to a cytoprotective agent comprising a compound represented by the general formula (I) or a salt thereof as an active ingredient.

(Wherein, Y is CH2, S, 0, CO or NR4, n is an integer from !! to 10, Ri and R2 are each H, alkyl, OH, alkoxy, nodogen, nitro, (Toro, amino or trifluoromethyl, R 3 represents H or alkyl, and R ⁴ represents H, alkyl or aralkyl)

The details will be described below. BEST MODE FOR CARRYING OUT THE INVENTION

The active ingredient of the cytoprotective agent of the present invention is a compound represented by the general formula (I) or a pharmaceutically effective salt thereof.

(I)

(Wherein, Y ′ is CH ₂ , S, 〇, CO or NR ⁴ , n is an integer from: to 10, and R ¹ R ² is H, alkyl, OH, alkoxy, nitrogen, nitrogen, collected by filtration, and § Mi Roh or Application Benefits Furuoromechiru, R ^3, and is H or alkyl, R ⁴ is H, alkyl or Ararukiru) is a general formula (I) good Ri preferred correct compound in Y is C H2 or S, n is an integer of 4 to 8, and R ¹ and R ² are H compounds. Alkyl may be linear or branched having 1 to 6 carbon atoms, such as methyl, ethyl, n-propyl, iso-propynole, n-butynole, iso-butynole, and tert-alkyl. Petinole, n-pentyl, n-hexyl. Alkoxy may be straight-chain or branched-chain having 1 to 6 carbon atoms. For example, methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, iso-butoxy, tert_butoxy, n-pentoxy, n-hexyloxy. Aralkyl includes an aryl portion and an alkyl portion, and the aryl portion includes an aryl having 6 to 10 carbon atoms. For example, phenyl, naphthyl and the like. The alkyl part has the same meaning as the above-mentioned alkyl. Halogen includes chlorine, bromine, iodine, fluorine and the like. An example of a method for synthesizing the compound of the present invention is shown below. Synthesis method 1

(II) (1-1)

'(Wherein, Y ^a is S, O, CO or NR ^4, R ³ is an alkyl, X is a leaving group, n, Ri, R 2, R is as defined above.) Compound (III one 1) in the alkyl of R ^{3 a} is as defined above. X represents a leaving group. Specific examples include a halogen and a sulfonyloxy group. Halogen is as defined above. In addition, examples of the snolephoninoleoxy group include methancenorleonyloxy, tonoleens / lehoninolexy, and benzenes nolehoninoleoxy. This reaction is a step of alkylating an amino group. That is, the compound (II) is reacted with the compound (III-11) in the presence of a base to synthesize the compound (I-11). Bases used in this reaction include alkali metal hydroxides (such as sodium hydroxide and hydroxide hydroxide) and alkaline hydroxide metals (such as magnesium hydroxide and calcium hydroxide). ), Alkali metal carbonate (sodium carbonate, potassium carbonate, etc.), alkaline earth metal (Magnesium carbonate, calcium carbonate, etc.), inorganic bases such as alkali metal bicarbonate (sodium bicarbonate, potassium bicarbonate, etc.), triethylamine, diisopropylethylamine, 2,2,6, Organic bases such as 6-tetramethylbiperidine, pyridine, 4-dimethylaminoviridine, 1,5-diazabicyclo [5.4.0] ndene. The reaction is usually performed in an inert solvent, and examples of the inert solvent used include acedone, chloroform, benzene, toluene, tetrahydrofuran, N, N-dimethylformamide. And dimethyl sulfoxide. Although the reaction temperature is not particularly limited, it is usually carried out at room temperature or under heating. Synthesis method 1:

(In the formula, Υ &, _n , R i, and R 2 have the same meanings as described above.) By hydrolyzing the compound (I-11), a free carboxylic acid (I-12) is synthesized. be able to. For example, lithium hydroxide, Ability to perform basic hydrolysis using sodium hydroxide or calcium hydroxide A known method such as acidic hydrolysis using hydrochloric acid or sulfuric acid is used. Synthesis method 1:

_/ X- (CH ₂ ) _n -COOH

(||) _: ^ (1-2)

(111-2)

(In the formula, X and n have the same meanings as described above.) The compound (II) and the compound (III-2) are reacted under the same conditions as in the synthesis method 11 to directly give a free compound. Carboxylic acid (I-2) can also be synthesized. , Synthesis method 1 4:

(In the formula, n, Ri, and R2 have the same meanings as described above.) The ketone group of the 2,3-dioxoindole (1-3) is reduced to synthesize the 2-oxoindole (1-4). This reduction can be performed by a known method, for example, catalytic hydrogen reduction using a catalyst such as palladium carbon, platinum oxide, and Raneykel, and sodium borohydride in the presence of Louis acid. Reduction, silane reduction in the presence of Lewis acid or trifluoroacetic acid, and Wo1ff-Kishhner reduction using hydrazine and the like can be used. Synthesis method 5

(I-5) (I-6)

(Wherein, Y, n, R ¹ , R ² , and R ^3a have the same meanings as described above.) A free carboxylic acid (1-5) is reacted with an alcohol (IV) to form an ester (1 — Combine 6). Esterification itself is known, and is converted into an active derivative such as an acid halide and then reacted, or is converted into an ester form by the reaction in the presence of a suitable condensing agent.

Known condensing agents can be used for this reaction. Concrete Specifically, dicyclohexylcarbodiimide, N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide, carbonyldiimidazole, 2-ethoxy_1-ethoxycarbone 1,2 —Dihydroquinoline, Jesyl phosphoryl cyanide, etc. The esterification reaction is usually performed in an inert solvent, and the solvent used is not particularly limited as long as it is nonprotonic. Specifically, there are acetonitrile, dichloromethan, chlorofonolem, N, N-dimethylformamide and the like.

Synthesis Method 1 6

Oxidation

(I-3)

(Wherein, X, n, Ri, and R 2 are as defined above.)

In the first step of this synthesis method, compound (V) and compound (VI) are reacted under the same conditions as in synthesis method 11 to synthesize compound (VII). In the second step, the compound (VII) It oxidizes the coal moiety to synthesize free carboxylic acid (1-3). Known methods can be used for the oxidation reaction. Specifically, TEM PO (2,2,6,6—tetramethylbiperidine 1—oxyl, free radical) / hypochlorite, permanganate sphere, chromium oxide, ruthenium oxide Z It can be performed using periodate or the like. These compounds can also be prepared according to a known method, as described in (11 3 932, 950, 53, 969, 61, JP-A-56-972668). The same pharmacologically effective salt in the present invention is an alkaline earth metal salt of the above compound, for example, Alkali metal salts such as tritium salts and calcium salts, for example, calcium salts, salts with amino compounds, for example, 1, dis (hydroxymethyl) aminomethane, N-methyldalcamin and so on. These salts can be prepared according to a known method. For example, the compound of the present invention having a free carboxylic acid at the 1-position may be a hydroxide for forming a salt (eg, hydrolyzed sodium, calcium hydroxide, calcium hydroxide, etc.) Can be formed by coexisting in a solution. Further, the compound of the present invention may be a hydrate. The compound of the present invention can be formulated according to a known method. For example, solid preparations and liquid preparations can be prepared. Examples of the solid preparation include powders, tablets, capsules and the like. Liquids include ethanol, cyclodextrin clathrates, ribosomes, and fat emulsions. And the like. Since the compound of the present invention has an excellent apoptosis inhibitory action, it suppresses apoptosis of cells, particularly brain cells, nerve cells or kidney cells, and is useful as a cytoprotective agent. Thus, these compounds may be associated with neurological disorders, diseases involving neurodegeneration, Alzheimer's disease, Parkinson's disease, Huntington's chorea, amyotrophic lateral sclerosis, spinal canal stenosis, or nephritis, renal failure, It is useful for prevention and treatment of renal diseases such as glomerulonephritis and nephrotic syndrome.

The route of administration may be oral or parenteral. The dosage can be adjusted appropriately according to the sex, age, medical condition, weight, etc. of the patient. Usually, 0.1 to 100 mg per day per adult, preferably 0.1 to 100 g per day. In order to explain the present invention in more detail, the following Examples, Compound Production Examples, Formulation Examples, and Pharmaceutical Tests will be given, but the present invention is not limited thereto. is not. The percentages shown in Examples 1 to 12 are yields (unless otherwise specified). ^_0/0 Gurutaruarudehi de soluble solution shown embodiment is shown in 1-7. Ethanolate Ichiru water I% of volume liquid and Example 2 1 is [nu / [nu%. The apoptosis induction suppression rate shown in Example 21 is also shown. / ₀ is the percentage of the measured result, where 0% indicates that all cells induced apoptosis, and 100% indicates that all cells did not induce apoptosis. , Example 1

6-Go to (2-3-Jihi _ Doro 1-2) _ii PT / JPO 1/04796

Synthesis of xanic acid

(1) To a mixture of isatin (2.16 g, 14.7 mmol), calcium carbonate (6.15 g, 44.5 mmol) and DMF (21 mL) At room temperature, ethyl 6-bromohexanoate (3.14 mL, 17.6 mmo, 1) was added dropwise. After the obtained mixture was stirred at room temperature for 47 hours, water was added, and the mixture was extracted with ethyl acetate. The extract was washed with saturated saline, dried, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (ethyl hexane monoacetate), and 6- (2,3-dihi, dro-1,2,3-dioxoindole-1-yl) ethyl hexanoate was purified. 4.29 g (100%) of an orange oily substance was obtained.

(2) A solution of the compound obtained in (1) (4.29 g, 15.2 mmo 1) in ethanol (59 mL) was dissolved in a 0.5: M sodium hydroxide aqueous solution (45 mL). ml) was added dropwise under ice cooling. The resulting mixture was stirred at room temperature for 6 hours, cooled with ice, adjusted to about pH 2 with hydrochloric acid, and extracted with a chloroform port. The extract was washed with saturated saline, dried, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (black-mouthed methanol), and 6- (2,3-dihydro-1,2,3-dioxoindole_1-yl) hexanoic acid was converted to orange. 1.68 g (4.3%) of a solid was obtained.

(3) Hydrazine monohydrate (13 mL) was added to the compound (1.68 g, 6.43 mmo 1) obtained in (2), and the mixture was heated under reflux for 50 minutes. The reaction solution was ice-cooled, acidified with concentrated hydrochloric acid, and then extracted with a black form. The extract is washed with saturated saline, dried, and concentrated under reduced pressure. Was. The residue was separated and purified by silica gel column chromatography (form-form methanol) and recrystallized from form-form hexane to give 974 mg (61 mg) of the title compound as white crystals. %) Obtained. . mp 1 3 6 5~ 1 3 7. 5 ° C ( hexane click b Rohonoremu ^{one) 1 H - NMR (4 0} 0 MH z, CDC 1 3) δ: 1. 4 3 (m 2 H), 1.70 (m, 4 H), 2.36 (t, J = 7.3 Hz 2 H), 3.5 3 ″, 2 H), 3.7 2 (t, J = 7.3 Hz 2 H), 6.8 3 (d, J = 7.9 Hz, 1 H), 7.03 (t J = 7.3 Hz, 1 H ), 7.20 to 7.32 (m, 2H)

M S (FAB +) m / z: 2 4 8 (MH +),

About C 14 Hi 7 N O 3

Theoretical value: C, 68.000; H, 6.93; N, 5.66

Measured value: C, 67.64; Η, 6.98; ,, 5., 62 Example 2

Synthesis of 7 -_ (2, _3 dihydrochloride 2-oxoinone) 1-butanol

7-Bromoheptanoic acid (17.7 g, 74.7 mmo 1) except that 11.3 g of the title compound was obtained as pale yellow crystals by the same reaction as in Example 1. (Total yield 66%). mp 56 to 57 ° C (ethyl ethyl isopropyl acetate) ¹ H-NM R (400 MHz, CDC ") δ: 1.25 to: 1.50 (m, 4H), 1 50 0-: 1.80 (m, 4Η), 2.32 (t, J = 7.3 Hz, 2H), 3.51 (s, 2H), 3.68 (t J = 7.3Hz, 2H), 6.80 (d, J = 7.9 Hz, 1 H), 7.01 (t, J = 7.8 Hz, 1 H), 7.20 to 7.35 (m, 2H)

MS (EI) m / z: 26 1 (M ⁺ )

About C i 5 H i 9 N O 3

Theoretical value: C, 68.904; H, 7.33; N, 5.36

Measured value: C, 68.95; H, 7.29; N, 5.35 Example 3

5 — (2, 3 — Synthesis of dihydro 2-oxo-d-one) 1 _ —-inole) -pentanoic acid

The title compound was converted to pale yellow crystals by a reaction similar to that of Example 1 except that ethyl 5-bromopentanoate (8.53 g, 40.8 mmo 1) was used. (Total yield: 61%). mp 98-99 ° C (ethyl acetate-hexane)

. 1 H - NM R (4 0 0 MH z, CDC 1 3) δ: 1 6 0~: L: 8

5 (m, 4H), 2.42 (t, J = 7Hz, 2H), 3.53 (s, 2H), 3.74 (t, J = 7Hz, 2 H), 6.8 3 (d, J =

8Hz, 1H), 7.03 (t, J = 8Hz, 1H), 7.20-

7.4.5 (m, 2 H)

MS (EI) m / z: 2 3 3 (M ⁺ )

For C ₁₃ H i 5 N 0 ₃

Theoretical value: C, 66.94; H, 6.48; N, 6.0%

Obtained values: C, 66.71; H, 6.39; N, 5.88 Example 4

4 Synthesis of 1, 2, 3-dihydro-1-oxoindole 1-butanoic acid

The title compound was converted to white crystals by the same reaction as in Example 1 except that 4-bromobutanoic acid ethinolate (2.43 mL, 17.Ommol) was used. (Total yield 49%). m p 12 9 to 13 0 ° C (Cross-mouth form-hexane)

. i H - NMR (4 0 0 MH z, CDC 1 3) δ: 2. 0 2 (m, 2 H), 2 4 7 (t, J = 7. 1 Η ζ, 2 Η), 3. 5 5 (s, 2 Η), 3.8 1 (t, J = 7.1 Η, 2 Η), 6.9 1 (d, J = 7.8 Η, 1 Η), 7.05 (t, J = 7.4 Η 1, 1 Η), 7.2 2 to 7.3 4 (m, 2 Η)

M S (F A B +) m / z: 2 20 (ΜΗ +)

About C i 2 Η i 3 Ν Ο 3

Theoretical value: C, 65.74; H, 5.98; N, 6.39

Actual value: C, 65.46; H, 5.95; N, 6.36 Example 5

3 — (2, 3 — dihydro — 2 — oxoindole) Synthesis of pionic acid

(1) To a mixture of isatin (2.09 g, 14.2 mmol), calcium carbonate (5.94 g, 43.Ommol) and DMF (20 mL) Ethyl acrylate (2.40 mL, 22.1 mmo 1) was added, and the mixture was stirred at 80 to 85 ° C. for 2.5 hours. The reaction solution was ice-cooled, water was added, and extracted with ethyl acetate. Wash the extract with saturated saline, After drying, the mixture was concentrated under reduced pressure. The residue was separated and purified by silica gel chromatography (hexane-ethyl acetate), and 3- (2,3-dihydro2,3-dioxoindole-1-yl) ethyl propionate was purified. Was obtained as an orange oily substance (45 lmg, 13%).

(2) The title compound was obtained by the same reaction as in Examples 1 (2) and (3) using the compound (449 mg, 1.82 mmo 1) obtained in (1). 215 mg (total yield 33%) were obtained as pale yellow crystals. m p 15 3 to 15 55 ° C (Kurohonolem-Hexane)

. 1 H - NMR (4 0 0 MH z, CDC 1 3) δ: 2. 7 7 (t, J = 7. 3 H z, 2 H), 3 5 4 (s, 2 H), 4. 0 3 (t, J = 7.3Hz, 2H), 6.92 (d, J = 7.8Hz, 1H), 7.05 (t, J = 7.6Hz, 1 H), 7.18 to 7.35 (m, 2 H)

M S (F A B "m / z: 206 (MH +)

About C i i H i i N O 3

Theoretical value: C, 6 4.38; H, 5.40; N, 6.83

Measured value: C, 6 4.39; Η, 5.46; ,, 6.84 Example 6

8 — (2,3 — Dihydro 2 — oxoindole 1 -yl) Synthesis of octanoic acid

(1) To a mixture of isatin (2.07 g, 14.1 mmol), calcium carbonate (5.88 g, 42.5 mmol) and DMF (20 mL) 8-Bromo_1-octanol (3.6 lmL, 21.1 mmo1) was added, and the mixture was stirred at room temperature for 13 hours. Add water to the reaction solution And extracted with ethyl acetate. The extract was washed with saturated saline, dried, and concentrated under reduced pressure. The residue is purified by silica gel column chromatography.

Separation and purification using hexane monoethyl acetate, 8— (2,3—dihydro-1,2,3-dioxoindole-11-yl) octane-111-ol was converted to a dark orange solid. 2.71 g (70%) was obtained.

: (2) The compound obtained in (1) (2.61 g, 9.46 mmo 1), Shiojiri trioctylmethylammonium (26 mg, 0.064 mmo 1) 1) Dichloromethane (57 mL). An aqueous solution of potassium bromide (226 mg, 1.90 mmol) (4.7 mL) and T EMPO (25 mg, 0.12 mmo 1) was added. To the resulting mixture was added an aqueous solution of sodium hypochlorite (1.37 M, 15.9 mL) and sodium hydrogen carbonate (2.44 g, 29.30 mL) under ice-cooling. A solution consisting of 0 mmo 1) and water (15.9 mL) was added dropwise. The resulting mixture was stirred for 50 minutes under ice cooling, acidified with 1 M hydrochloric acid, and extracted with dichloromethane. The extract was washed with brine, dried and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (closed-form-methanol), and 8 — (2,

1.48 g (54%) of 3—dihydro-1,3-dioxoindole) octanoic acid was obtained as an orange solid.

(3) Using the compound (1.48 g, 5.11 mmo1) obtained in (2), the title compound was converted to pale yellow by the same reaction as in Example 1 (3). 470 mg (33%) were obtained as crystals. mp 76 to 80 ° C (ether) ^{. 1 H - NM R (4} 0 0 MH z, CDC 1 3) δ: 1, 3 6 (brs, 6 H), 1 6 5 (m, 4 H), 2. 3 4 (t, J = 7 3 Hz, 2H), 3.53 (s, 2H), 3.70 (t, J = 7, 3Hz, 2H), 6.83 (d, J = 7. 9 Hz, 1 H), 7, 03 (t, J = 7.6 Hz, 1 H), 7.2 2 to 7.32 (m, 2H) MS (FAB +) m / z : 2 7 6 (MH +)

About C i 6 H ₂ l NO 3

Theoretical value: C, 69.79; H, 7.67; N, .5.09

Actual value: C, 69.47; H, 7.68; Ν, 5.11 Example 7

9— (2 ^ 3—Dihydro-2-oxoindole 1yl) Synthesis of nonanoic acid

9—Promote 1—Nonanol (3.20 mL, 17.3 mmo

By the same reaction as in Example 6 except that 1) was used, 1.10 g (total yield 26%) of the title compound was obtained as slightly yellow 'crystals. m p 62 to 63 ° C (ether)

^{.. 1 H - NM R (} 4 0 0 MH z, CDC 1 3) δ: 1 3 3 (m 8 H), 1 6 4 (m, 4 H), 2. 3 4 (t, J = 7. 6 Hz 2 H), 3.53 (s, 2 H), 3.70 (t, J = 7.3 Hz 2 H), 6.83 (d, J = 7.8 Hz , 1H), 7.03 (t = 7.4 Hz, 1H), 7.20 to 7, 32 (m, 2H)

M S (FAB +) m / z: 290 (MH +)

About C i ₇ H ₂ 3 NO 3

Theoretical value: C, 70.56; H, 8.01; N, 4.84 Measured value: C, 70..26; H8; N, 4, 79 Example 8

7 — (2,3-Dihidro 5 —Methoxy 2 —Oxoindole 1-11yl) Synthesis of heptanoic acid,

B art lett et al.'S method (J. Am. Chem. Soc.,

The same reaction as in Example 2 was carried out using 5-methoxyisatin (1.70 g, 9.60 mmo1) synthesized from 80, 126 to 1336, 1958). As a result, 8993 mg (total yield: 32%) of the title compound was obtained as pale yellow crystals. m p 1 16 to 1 17 ° C (chloro honolem hexane)

.... 1 H - NM R (4 0 0 MH z, CDC 1 3) δ 1 3 0~: 1 4, 0 (m, 4 H), 1 5 5~: 1 7 0 (m, 4 H ), 2.33 (t, J = 7.3Η, 2Η), 3.48 (s, 2Η), 3.65 (t, J = 7.3Η, 2Η) , 3.77 (s, 3 Η), 6.70 (d, J = 8.3 7, 1 Η), 6.7.7 (dd, J = 8.3 Η, J '= 2 .4Η, 1 1), 6.86 (d, J = 2.4 4, 2Η) MS (FAB "m / z: 29 1 (ΜΗ +)

About C i 6 Η ₂ l Ο Ο 4

Theoretical value: C, 65.96; Η, 7.27; Ν, 4.81

Actual measured value: C, 65.73; H, 7.18; N, 4.77 Example 9

7- (2,3—Dihydro-5—Hydroxy-1-2—Oxoin-d-yl-1-yl) Synthesis of heptanoic acid — At 78 ° C, boron tribromide (1 mL) was added to a dichloromethane-containing methane solution (30 mL) of the target compound of Example 8 (500 mg, 1,72 mmo 1). In addition, after the temperature was raised to room temperature, the mixture was stirred at the same temperature for 3 hours. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The extract was washed with water and saturated saline, and dried. The solvent was distilled off under reduced pressure, and the obtained residue was recrystallized from acetone-hexane to give 400 mg of the title compound as pale yellow crystals (yield 84%). ) Obtained. mp 13 0 to 13 1 ° C (acetone / hexane)

... 1 H - NM R (4 0 0 MH z, DMSO- d 6) δ: 1 2 6 (. Br s, 4 H), 1 4 0~: L 5 5 (m, 4 H), 2 16 (t, 1 = 7.3 Hz, 2 H), 3.44 (s, 2 H), 3.55 (t, J = 7.3 Hz, 2 H), 6. 6 (d, J = 8.3 Hz, 1 H), 6.71 (s, 1 H), 6.76 (d, J = 7.6 Hz, 2 H), 9.0 2 (s, 1 H), 1 1.96 (s, 1 H)

MS (FAB +) m / z: 27 7 (MH ⁺ )

About C 1 5 Hi 9 N O 4

Theoretical value: C, 64.97; H, 6.91; N, 5.05

Actual measured value: C, 64.80; Η, 6.88; Ν, 4.90 Example 10

Synthesis of 7- (2,3-Dihydro-1-2-oxobenzothiazole-3-yl) heptanoic acid

2 - Hydro carboxymethyl benzothiazole (.. 1 0 0 g, 6 6 2 mmo 1) DMF solution (2 0 m L) of, 0 ° C in 6 0 ^_0/0 hydrogen Kana Application Benefits um (6 6 2 mg, 16.6 mmo 1) A DMF solution (10 mL) of moheptanoic acid (1,66 g, 7.94 mmo 1) was added dropwise. After the reaction solution was stirred at 70 to 80 ° C. for 1.5 hours, the reaction solution was returned to room temperature, and 1 M hydrochloric acid (100 mL) was added. This mixed solution was extracted with ethyl acetate, and the obtained organic layer was washed with water and saturated saline, and then dried. The solvent was concentrated under reduced pressure, purified by silica gel force chromatography (hexane-ethyl acetate), and recrystallized from ethyl acetate-diisopropyl ether-hexane. 477.7 mg (26%) of the compound were obtained as pale yellow crystals. mp 73 to 74 ° C (ethyl acetate isopropyl ether)

¹ H one NMR (4 0 0 MH z, CDC 1 3) δ:.... 1 3 0 ~ 1 5 0 (m, 4 H), 1 6 0 ~ 1 7 0 (m, 2 H), 1 70 to 1.80 (m, 2H), 2.35 (t, J = 7.4Hz, 2H), 3.94 (t, J = 7.3Hz, 2 H), 7.04 (d, J = 7.8 Hz,, 1H), 7.16 (t, J = 7.8 Hz, 1H), 7, 32 (t, J = 7.8 Hz, 1 H), 7.43 (d, J = 7.8 Hz, 1 H)

M S (E I) m / z 2 7 9 (M +)

About C 1 4 H ₁₇ NO ₃ S

Theoretical value: C, 60.19; H, 6.13; N, 5.01

Measured value: C, 60.37; H, 6.05; N, 4.6 2 Example 11

7 — (2, —3 — dihydro 2 —oxobenzoxazole-3 ノレ) Synthesis of heptanoic acid

The title compound was converted to colorless crystals by the same reaction as in Example 10 except that 2-dioxybenzoxoxazonole (1.0 g, 7.41 mmo 1) was used. Thus, 562 mg (yield 29%) was obtained.

mp 70 to 72 ° C (ethyl acetate-hexane)

^{.... 1 H - NMR} (4 0 0 MH z, CDC 1 3) δ: 1 3 5~ 1 5 5 (m, 4 H), 1 5 0~: L 7 0 (m, 2 H), 1.70-: L.85 (m, 2H), 2.33 (t, J = 7.3 Hz, 2H), 3.80 (t, J = 7.3 Hz) , 2H), 6.95 (d, J = 7.8 Hz, lH), 7.05 to 7.30 (m, 3H)

M S (E I) m / z 26 3 (M +)

About C _x 4 Hi ₇ NO 4

Theoretical value: C, 63.87; H, 6.51; N, 5.32

Actual measured value: C, 63.74; H, .6.44; N, 5.19 Example 12

7— (2,3—Dihydro-1-2-oxobenzoimidazole-1—yl) Synthesis of heptanoic acid

2-Hydroxybenzimidazole (1.00 g, 7.46 mm 01) was reacted in the same manner as in Example 2 to give the title compound as colorless crystals (356 mg, total). Yield 18%). m p 109 to 110 ° C (ethyl acetate-diisopropanol-hexane)

.... 1 H - NMR (4 0 0 MH z, CDC 1 3) δ: 1 2 6 (. Br s, 4 H), 1 3 5~: L 5 0 (m, 2 H), 1 5 0 to: 1.6 5 (m, 2H), 2.16 (t, J = 7.3 Hz, 2H), 374 (t, J = 7.3 Hz, 2H), 6.95 to 7.00 (m3H), 7.09 (d, J = 6.8 Hz, 1H), 10.0.78 (s1H), 11.996 (s, 1H )

M S (E I) m / z: 26 2 (M +)

About C i 4 H i 7 NO 4-1/5 H ₂ 0

Theoretical value: C, 63.24; H, 6.97; N, 10.54 Actual value: C, 63.41; H, 6.99; N, 10.26 The following compounds were synthesized in the same manner as in the above examples. Example 13

7- (5-Buguchi-mo 2,3 —dihidroh 2 —oxoin-donore-1-yl) heptanoic acid '1 H-NMR (400 MHz, CDC ") S: 1.3 1 -1 4 4 (m, 4 Η), 1.57-: 1.73 (m, 4 Η), 2.35 (t, J = 7.3 Η, 2 2), 3.5 2 (s, 2 Η), 3.68 (t J = 7.3 5, 2 Η), 5.90 (brs, 1 Η), 6.70 (d, J = 8.3 ζ) , 1 Η), 7.3 3 〜 7.44 (m, 2 Η) C ₁₅ Η 18 Ν Β r Ο 3

Theoretical value: C, 52.96; H, 5.33; N, 4.12 Actual value: C, 52.97; H, 5.34; N, 4.46 14

7 — — (2, — 3 ■ dihydrofluoro-2-oxoindole) ^{.... 1 H - NMR} (4 0 0 MH z, CDC 1 3) δ 1 3 2~: 1 4 4 (m, 4 H), 1 5 9~ 1 7 2 (m, 4 H), 2 3.5 (t, 1 = 7.3Hz, 2H), 3.52 (s, 2H), 3.69 (tJ = 7.3Hz, .2H), 6. 7 3 (m, 1 H), 6.93 to 7.04 (m, 2 H)

About C is His N F Os

Theoretical value: C, 64.50; H, 6.50; N, 5.01 Actual value: C, 64.35; Η, 6.61; ,, 5.26

Example 15

7- (2,3-Dihydro-5-methyl-2-oxoindole-1-yl) heptanoic acid

1 Η -. NM R (4 0 0 MH z, CDC 1 3) δ 1 3 0~ 1.

4 3 (m, 4H), 1.57-: L.73 (m, 4H), 2.33 (s, 3Η), 2.34 (t, J = 7.3 =) ζ, 2 Η), 3.5 0 (s

2 Η), .3.6. 8 (t, J = 7.4 Η 2, 2 Η), 6.7.2 (d

J = 8.3 Η 1, 1 Η), 7.0 2 to 7.1 1 (m, 2 Η), 8

9 8 (brs, 1Η)

About C ₁₆ Η ₂ ι Ν 0 ₃ :

Theoretical value: C, 69.79; Η, 7.69; Ν, 5.09 Actual value: C, 69., 62; Η, 7.55; Ν, 5.29 Example 1 6

7 ~ (2,3 dihydrodro-one-done)

^{1 Η - NMR (4 0 0} MH z, CDC 1 3) δ 3 2 48 (m, 4H), 1.48-: L.80 (m, 4H), 2.36 (t, J = 7.3Hz, 2H), 3.63 ( s, 2H), 3.76 (t, J = 7.3Hz, 2H), 6.90 (d, J = 8.7Hz, 1H), 8.15 (s , 1 H), 8. 2 6 (d, J = 8. 3 H z, 1 H) for C is His N ₂ 0 _5:

Theoretical value: C, 5'8.82; H, 5.92; N, 9.15 Actual value: C, 58.89; H, 5.84; N, 9.04 each Table 1 shows the relationship between the compounds synthesized in the examples and Υn, RR2, and R3 defined in the general formula (I).

[Table 1] Example Υ n R ¹ R ² R ³

1 CH ₂ 5 HHH

2 CH ₂ 6 HHH

3 CH ₂ 4 HHH

4 CH ₂ 3 HHH

5 CH ₂ 2 HHH

6 CH ₂ 7 HHH

7 GH ₂ 8 HHH

8 CH ₂ 6 5-MeO HH

9 CH ₂ 65-OH HH

10 S 6 H H H

11 0 6 H H H

12 NH 6 HHH 13 GH ₂ 65-Br HH

14 CH ₂ 6 5-FHH

15 CH ₂ 6 5-Me HH

16 CH ₂ 6 5-N0 ₂ HH Example 1 7

Formulation Example 1 (Cyclodextrin inclusion)

Example 1 In a solution prepared by dissolving 1,7 mg of each of the compounds of the present invention prepared in Examples 1 to 16 in 0.2 mL of ethanol] 35.7 mg of cyclodextrin was heated to 6 mL of water The solution prepared by dissolution was added, mixed at 45 ° C., and then returned to room temperature to precipitate a precipitate. This was allowed to stand overnight at 0 ° C., filtered, washed with a 50% aqueous solution of edanol, and sterilized and dried to obtain a cyclodextrin clathrate. Example 18

Formulation Example 2 (Libosome conversion)

After dissolving 60 mg of yolk phosphatidylcholine and 11 mg of oleylamine in 5 mL of clog form, 30 μg of each compound of the present invention prepared in Examples 1 to 16 was added. A solution dissolved in 100 L of ethanol was added, the mixture was placed in an eggplant-shaped flask, and the solvent was distilled off with a rotary evaporator. To this was added 0.1 M isotonic phosphate buffer solution (pH 5) lmL, and the mixture was shaken, sonicated (so-gate) and centrifuged, and the supernatant was collected with 0.2 μπι medium. The solution was filtered through a membrane filter to obtain a ribosome preparation. Example 19 P TJP01 / 04796

26

Formulation Example 3 (Ethanol solution)

Ethanol solution was obtained by dissolving 500 μg of each compound of the present invention prepared in Examples 1 to 16 in ImL of ethanol. When used, dilute with physiological saline or glucose solution before use. '' Example 20

Formulation Example 4 (Fat emulsion)

To 30 g of purified soybean oil, 5.4 g of highly purified egg yolk phospholipid, 1.5 mg of each of the compounds of the present invention prepared in Examples 1 to 16 and 0,072 g of oleic acid were added. The mixture was heated and dissolved at 40 to 75 ° C. Distilled water (20 OmL) was added thereto, and then 7.5 g of glycerin of the Japanese Pharmacopoeia was mashed, and the total amount was brought to 300 111 with distilled water at 20 to 40 °, The mixture was roughly emulsified with a homogenizer. Furthermore, high-pressure emulsification was carried out with a manton-gaurine homogenizer to obtain a homogenized fine fat emulsion. The average particle size of this emulsion was 0.115 to 0.4 μπι, and it did not contain particles of 1 μm or more. Example 2 1

Efficacy test 1: Inhibitory effect of the compound of the present invention on induction of apoptosis by amyloid beta peptide ''

1) Culture of rat cerebral cortex-euron

The cortex was removed from the cerebrum of a 17-day-old rat embryo from the cerebrum under ice-cooling, shredded, and the cells were dispersed using a nerve cell dispersion (SUMILON). Thereafter, the cells were seeded at a density of about 0.7 × 10 5 cells / cm ² in a culture flask previously polyethylene-coated. 4796

27

After culturing for 4 days, it was subjected to the next test. The culture solution was B27 sublimation (150 volumes), 2-mercaptoethanol (27.5 μM), L-glutamic acid (25βΜ) and glutamate. Neurobasa 1 culture medium (manufactured by Gibco) supplemented with 0.5 mM (0.5 mM) was used.

2) Induction of apoptosis by amyloid beta peptide and addition of the compound of the present invention

Amyloid beta peptide 25-35 (Aj825-35) was dissolved in distilled water to a concentration of lmM, incubated at 37 ° C for about 1 week, and Aged-A / 325-35 were prepared. Induction of apoptosis in nerve cells was performed by exchanging with the above culture medium (excluding L-glutamate) containing 10 Aged-A | 825-35. Further, after dissolving the compound of the present invention in DMSO, Aged-AjS

25-35 was added to the culture at the same time. Its final concentration is 0.

1 or 1M.

3) Detection of apoptosis

Twenty-four hours after the induction of apoptosis, the cells were washed with PBS (-) and fixed with a 1% dartal aldehyde solution (PBS solution) for 30 minutes at room temperature. Next, after washing twice with PBS (-), the cells were reacted with a 1 mM solution of 333333 solution (PBS solution) for about 2 minutes. After that, the morphology of nuclear chromatin was observed in any of four to six fields under a fluorescence microscope, and the number of normal cells and the number of apoptosis-positive cells (cells showing chromatin fragmentation or aggregation) were counted. The apoptosis induction suppression rate (%) was calculated. Table 2 shows the results. Table 2, JP0104796

28

The compound number in the shows the number of the example of the synthesis of the compound,

[Table 2]

Example 22

Drug efficacy test 2

After suspending the compound of the present invention prepared in Examples 1 to 16 in hydroxypropylmethylcellulose to a concentration of 30 mg / 2 mL, a normal rat (female, weight of about 200 g) was used. ) Was orally administered at a dose of 30 mg / kg body weight, but no fatal cases could be observed. Example 2 3

Tablets were prepared using the following ingredients.

10 g of the compound of the present invention prepared in Examples 1 to 16, 10 g of fine particles for direct injection No. 209 (manufactured by Fuji Chemical Co., Ltd.) 110 g, crystalline cellulose 60 g, CMC calcium 18 g, magnesium stearate 2 g.

The above components (other than magnesium stearate) are mixed using a mixer, and magnesium stearate is added to the homogeneously mixed powder and mixed for a short time (30 seconds). Was tableted to give a tablet of 200 mg per tablet. Example 2 4

A capsule was prepared using the following raw materials.

50 g of the compound of the present invention prepared in Examples 1-16, lactose 930 g, magnesium stearate 20 g.

The above components were uniformly mixed, and the mixed powder was filled into hard gelatin capsules in 20 mg portions. Industrial applicability

The present invention relates to a cytoprotective agent, and more particularly to a cytoprotective agent, particularly an agent useful for protecting brain cells, nerve cells, and kidney cells.

Claims

The scope of the claims

1. A cytoprotective agent comprising a compound represented by the general formula (I) or a pharmaceutically effective salt thereof as an active ingredient.

(Wherein, Y is CH ₂ , S, 0, C〇 or NR ⁴ , n is an integer of from! To 10, and R i and R 2 are each H, alkyl, OH, alkoxy, nodogen, nitro, Toro, amino or trifluoromethyl, R 3

H or alkyl; R ⁴ represents H, alkyl or aralkyl)

2. —The compound of formula (I), wherein Y is CH ₂ or S, n is an integer of 4-8, and R 1 and R ² are each H.

3. The compound of the general formula (I) is 6- (2,3-dihydro-2-oxoin, dol-1-yl) hexanoic acid, 7- (2,3-dihydro-2-oxo-y) N-dole 1 yl) heptanoic acid, 5 — (2, 3 — dihydro 2 — oxoindole 1 — yl) pentanoic acid, 8 — (2, 3 — dihydro 2 — oxoindole 1 1) Le) octanoic acid, 9- (2,3-dihydro-2—oxoindole-1-yl) nonanoic acid and 7— (2,3-dihidr-2-2-oxobenzothiazol-3-yl) heptanoic acid The cytoprotective agent according to claim 1, which is selected from the group.

4. The cytoprotective agent according to any one of claims 1 to 3, wherein the pharmaceutically effective salt of the compound is an alkaline earth metal salt.

5. The cytoprotective agent according to any one of claims 1 to 4, which is useful for protecting brain cells, nerve cells, and kidney cells.

6. The cell protection agent according to any one of claims 1 to 5, which is a solid preparation or a liquid preparation.

7. Dosage is 0.:! Up to 10 O mg

: The cytoprotective agent according to any one of! To 6.

8.5- (2,3—Dihidro 2 —Oxoin-Donore 1—Innole) pentanoic acid, 6 — (2,3 —Dihidro 2 —Oxoindole 1-yl) hexanoic acid, 7 — ( 2,3—dihydro-1-2-oxoindole-11-yl) heptanoic acid, 8— (2,3—dihydro-1-2-oxo-indole-1-yl) octanoic acid, 9-1 (2,3) —Dihydro 2-oxo-dole 1 —yl) Nonanoic acid and 7 —

(2,3-Dihydro-1-oxobenzothiazole_3-inole) A compound selected from the group consisting of heptanoic acid, or a pharmaceutically effective salt thereof. ,

9. A method for protecting cells comprising administering a compound represented by the general formula (I) in claim 1 or a pharmaceutically effective salt thereof to a patient.

10. Use of a compound represented by the general formula (I) in claim 1 or a pharmaceutically effective salt thereof for preparing a cytoprotective agent.