AU705798B2 - Novel heterocyclic derivatives - Google Patents

Description

1 SUBJECT MATTER DISCLOSED BUT NOT CLAIMED HEREIN IS DISCLOSED AND CLAIMED IN PATENT (PARENT) APPLICATION

NO.

35324/95 SERIAL NO. 693261 OF WHICH THIS APLICATION IS A DIVISIONAL

APPLICATION.

Technical Field Patent application no. 35324/95 relates to novel heterocyclic derivatives, a method of production thereof and pharmaceutical use thereof. More particularly, patent application no. 35324/95 relates to novel heterocyclic derivatives having an indoline ring or tetrahydroquinoline ring, a method of production thereof and pharmaceutical use Sthereof, specifically acyl-CoA: cholesterol acyltransferase (hereinafter referred to as ACAT) inhibitors and 15 lipoperoxidation inhibitors.

The present invention relates to novel intermediates used in the methods of production of the heterocyclic derivatives subject of patent application no. 35324/95.

The present invention provides a compound of the 20 general formula (X) SR1 11 R13 r wherein R 11

R

12 and R 13 may be the same or different and each is independently hydrogen atom, lower alkyl or lower alkoxy, R 14 is an amino-protecting group, and m is 1 or 2 and a related compound l-acetyl-5-bromo-4,6dimethylindoline.

A preferred compound of formula is l-acetyl-4,6dimethylindoline.

As indicated in patent application no. 35324/95, it is a well-known fact that arteriosclerosis is an extremely la important factor causing various circulatory diseases, and active studies have been undertaken in an attempt to achieve suppression of the evolution of arterial sclerosis or regression thereof. In particular, although the usefulness of a pharmaceutical agent which reduces cholesterol in blood or arterial walls has been acknowledged, an ideal pharmaceutical agent exhibiting positive clinical effects while causing less side-effects has not been realized.

In recent years, it has been clarified that cholesterol accumulated in arterial walls in the ester form ~thereof significantly evolves arteriosclerosis. A decrease in cholesterol level in blood leads to the reduction of accumulation of cholesterol ester in arterial walls, and is effective for the suppression of evolution of arteriosclerosis and regression thereof.

Cholesterol in food is esterified in mucous membrane small intestine, and taken into blood as chylomicron.

ACAT is known to play an important role in the generation 20 of cholesterol ester in mucous membrane of small intestine.

Thus, if esterification of cholesterol can be suppressed by inhibiting ACAT in mucous membrane of small intestine, .i absorption of cholesterol by mucous membrane and into blood .:oooi can be presumably prevented so as to ultimately result in lower cholesterol level in blood.

In arterial walls, ACAT esterifies cholesterol and causes accumulation of cholesterol ester. Inhibition of ACAT in arterial walls is expected to effectively suppress accumulation of cholesterol ester.

From the foregoing, it is concluded that an ACAT inhibitor will make an effective pharmaceutical agent for hyperlipemia and arteriosclerosis, as a result of suppression of absorption of cholesterol in small intestine and accumulation of cholesterol in i Conventionally, for example, there have been reported, as such

ACAT

inhibitors, amide and urea derivatives Med. Chem., 29 1131 (1986), Japanese Patent Unexamined Publication Nos. 117651/1990, 7259/1990, 32666/1993 and 327564/19921.

.However, creation and pharmacological studies of these compounds have been far from sufficient.

Meanwhile, peroxidation of low density lipoprotein (LDL) is also S highly responsible for accumulation of cholesterol ester in arterial walls. In addition, it is known that peroxidation of lipids in a iving body is deeply concerned with the onset of arteriosclerosis and S. cerebrovascular and cardiovascular ischemic diseases.

Accordingly, a compound having both ACAT inhibitory activity and lipoperoxidation inhibitory activity is highly useful as a pharmaceutical product, since it effectively reduces accumulation of cholesterol ester in arterial walls and inhibits lipoperoxidation in the living body, thereby preventing and treating various vascular diseases cqused thereby.

Parent patent application no. 35324/95 provides a compound having ACAT inhibitory activity and lipoperoxidation inhibitory activity, a method for production thereof and pharmaceutical use thereof, particularly as an ACAT inhibitory and lipoperoxidation inhibitor.

-2- As indicated in the parent patent application, it was found that certain heterocyclic derivatives having an indoline ring or tetrahydroquinoline ring have lipoperoxidation inhibitory activity in addition to strong ACAT inhibitory activity, and that these compounds have a strong anti-hyperlipemia effect and anti-arteriosclerosis effect.

Thus, parent patent application no. 35324/95 relates to a heterocyclic derivative of the formula (I)

R'

R2 (CH2).

S

I

9

R

3

N

R

4

I

R

wherein one of R 2

R

3 and R 4 is a group of the formula -NHCO-R 6 wherein R' is optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heterocycle, optionally substituted heterocyclic alkyl, -R^S03A, -RBPO3B where RA and R B are each alkylene and A and B are each alkali metal or hydrogen atom, -NR'R 8 where R' is optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl or optionally substituted arylalkyl and R 8 is hydrogen atom or lower alkyl, or -R'-OCOR'O where R 9 is alkylene and R 10 is optionally substituted alkyl, optionally substituted heterocycle or optionally substituted heterocyclic alkyl, and the remaining three may -3 be the same or different and each is independently a hydrogen atom, a lower alkyl or a lower alkoxy; Rs is an optionally substituted alkyl, an optionally substituted cycloalkyl, an optionally substituted cycloalkylalkyl, an optionally substituted aryl, an optionally substituted arylalkyl, an optionally substituted heterocycle, an optionally substituted heterocyclic alkyl, an alkenyl, an alkynyl, a dialkylaminoacyloxyalkyl, -RDS03D or -REP03E where R D and RE are each alkylene and D and E are each alkali metal or hydrogen atom, provided that when R 4 is -NHCO-R 6

R

5 and

R

6 optionally combinedly form a ring; and m is 1 or 2, S' [hereinafter this compound is also referred to as Compound and a pharmaceutically acceptable salt thereof.

The parent patent application also relates to a method for producing the above-mentioned heterocyclic derivative or a pharmaceutically acceptable salt thereof, which comprises a step of (D reacting an amine of the formula

(II)

R"

R 2 13 (CH2)m H2N

N

R

wherein

R'

2 and R' 3 may be the same or different and each is independently hydrogen atom, lower alkyl or lower alkoxy, and R 5 and m are as defined above [hereinafter also referred to as Compound and an isocyanate of the formula

(III)

R

7 NCO

(III)

wherein

R

7 is as defined above [hereinafter also referred to as Compound -4

(III)];

reacting Compound (II) and a halogen compound of the formula (IV)

R

6 -COX (Iv) wherein X is halogen atom.and

R

6 is as defined above [hereinafter also referred to as Compound reacting Compound (II) and a carboxylic acid of the formula (V)

R

6 COOH

(V)

wherein R 6 is optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heterocycle or optionally substituted heterocyclic alkyl [hereinafter also referred to as Compound or a reactive derivative thereof; reacting an isocyanate of the formula (VI)

I

R

OCN, v "N

RR

wherein R 5 R1 2 R13 and m are as defined above [hereinafter also referred to as Compound and an amine of the formula (VII)

HNR

7 Re

(VII)

wherein R' and R e are as defined above [hereinafter also referred to as Compound (VII)I; or reacting a compound of the formula (VIII)

R'

(CH2)m I

(VIII)

R R

N

H

wherein

R

2

R

3

R

4 and m are as defined above [hereinafter also referred to as Compound (VIII)], and a compound of the formula (IX)

R

5 X

(IX)

wherein R 5 and X are as defined above [hereinafter also referred to as Compound The parent patent application also relates to pharmaceutical compositions, ACAT inhibitors and lipoperoxidation inhibitors containing the above-mentioned heterocyclic derivative or a pharmaceutically acceptable salt thereof.

As indicated in the parent specification, each symbolshown above denotes the following.

Lower alkyl at R 2

R

3

R

4

R

8

R'

1

R

12 and R 13 may be linear or branched and preferably has 1 to 4 carbon atoms. Examples thereof include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl and the like.

Lower alkoxy at R 2

R

3

R

4

R'

2 and R 13 may be linear or branched and preferably has 1 to 4 carbon atoms. Examples thereof include methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, secbutoxy, tert-butoxy and the like.

Alkyl at R 5

R

6

R

6

R

7 and may be linear or branched and preferably has 1 to 12 carbon atoms. Examples thereof include methyl, Sethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, 1,1-dimethylpropyl, 1,1-dimethylbutyl, 1,1dimethylpentyl, 1,1-dimethylhexyl, 3,3-dimethylbutyl, 4,4-dimethylbutyl and the like.

Cycloalkyl at R 5

R

6 R" and R' preferably has 3 to 6 carbon atoms. Examples thereof include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like.

With regard to cycloalkylalkyl at R 5

R

6

R

6 and R 7 its cycloalkyl moiety preferably has 3 to 6 carbon atoms and alkyl moiety preferably has 1 to 3 carbon atoms. Examples of cycloalkylalkyl include cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, cyclopropylethyl, cyclopropylpropyl and the like.

Examples of aryl at R 5

R

6

R

6 and R 7 include phenyl, naphthyl and the like.

Arylalkyl at R 5

R

6

R"

6 and R 7 has an aryl moiety as exemplified -6- ~13 above and its alkyl moiety preferably has 1 to 4 carbon atoms. Examples of arylalkyl include benzyl, 1-phenylethyl, 2-phenylethyl, 1phenylpropyl, 2-phenylpropyl, 3-phenylpropyl and the like.

Heterocycle group at R 5

R

6

R

6 and is a monovalent group which occurs as a result of liberation of one hydrogen atom bonded to the ring of heterocyclic compound and may be aliphatic or aromatic.

Examples thereof include pyrrolidinyl, piperidyl, piperidino, morpholinyl, morpholino, piperazinyl, pyrrolyl, imidazolyl, pyridyl and the like.

Heterocyclic alkyl at R 5

R

6

R

6 and has a heterocyclic moiety as exemplified above and its alkyl moiety preferably has 1 to 8 carbon atoms. Examples thereof include (1-pyrrolidinyl)butyl, morpholinopropyl, 1,1-dimethyl-2-(1-pyrrolidinyl)ethyl, 1,1-dimethyl-2piperidinoethyl, 1,1-dimethyl-3-(imidazol-1-yl)propyl, (2,6dimethylpiperidino)methyl, (2,6-dimethylpiperidino)ethyl, (2,6dimethylpiperidino)propyl and the like.

The above-mentioned alkyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocycle and heterocyclic alkyl may be substituted.

Examples of the substituent include alkyl, amino, hydroxy, dialkylamino, aminoalkyl, alkoxy, carboxyl, alkoxycarbonyl, carboxyalkyl, acyloxy, phenyl, phenoxy, halogen atom and the like.

Alkyl in alkyl, dialkylamino, aminoalkyl and carboxyalkyi is exemplified by the above-mentioned lower alkyl. Alkoxy in alkoxy and alkoxycarbonyl is exemplified by the above-mentioned lower alkoxy.

Acyloxy may be linear or branched and preferably has 2 to 5 carbon atoms. Examples thereof include acetyloxy, propionyloxy, butyryloxy, valeryloxy, pivaloyloxy and the like. Halogen atom is exemplified by those to be mentioned later. Alkyl in dialkylamino may be substituted by phenyl.

Alkenyl at R 5 may be linear or branched and preferably has 2 to 8 carbon atoms. Examples thereof include ethenyl, propenyl, isopropenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, 3,3-dimethyl-2-propenyl and the like.

-7- Alkynyl at R' may be linear or branched and preferably has-2 to 8 carbon atoms. Examples thereof include ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, octynyl, 3 3 -dimethyl-2-propynyl and the like.

Alkyl moiety of dialkylaminoacyloxyalkyl at R 5 preferably has 1 to 8 carbon atoms, and its acyl moiety may be linear or branched and preferably has 2 to 5 carbon atoms. Examples thereof include acetyl, propionyl, butyryl, valeryl, pivaloyl and the like. The dialkylaminoacyloxyalkyl is specifically exemplified by N,Ndimethylaminoacetoxyethyl, N,N-dimethylaminoacetoxypropyl and the like.

Alkylene at RA, RB, RD RE and R' may be linear or branched S. and preferably has 1 to 8 carbon atoms. Examples thereof include methylene, ethylene, trimethylene, propylene, tetramethylene, pentamethylene, hexamethylene, 1,1-dimethylethylene, 2,2dimethylpropylene and the like.

Alkali metal at A, B, D and E is preferably sodium, potassium and the like.

Halogen atom at X is exemplified by chlorine atom, bromine atom, iodine atom and the like.

When R4 is -NHCO-R',

R

6 and R 5 may combinedly form a ring. The group (-R 6 formed by R' and R 5 in combination may be linear or branched and preferably has 2 to 12 carbon atoms. Examples thereof .i include alkylene such as 1,1 -dimethyltrimethylene, 1,1-dimethyltetramethylene, 2, 2 -dimethyltetramethylene, 1,1-dimethylpentamethylene, 2,2dimethylpentamethylene and the like, and alkylene having -OCO- bond, such as -C(CH3)2CH20CO(CH 2 3 -C(CH3)2CH20COC(CH 3 2

(CH

2 and the like.

The preferable Compound of the parent application includes, for example, 1-butyl-3-(1 -hexyl-4,6-dimethylindolin-5-yl)urea, 1-butyl-3-(1 -hexyl-4,6-dimethylindolin-7-yl)urea, N-(1 -hexyl-14, 6 -dimethylindolin-5-yl)-2,2-dimethylpropanamide, N-(1-hexyl-4, 6 -dimethylindolin-7-yl)-2,2-dimethylpropanamide, -8- N-(1-pentyl-4,6-dimethylindolin-7-yl)-2, 2 -dimethylpropanamide N-(1 -isobutyl-4,6-dimethylindolin-7-yl)-2, 2 -dimethylpropanamide, N-(1-hexyl-4,6-dimethylindolin-7-yl)-2, 2 -dimethylbutanamide N-(1lhexyl-4,6-dimethylindolin-7-yl)-2, 2 -dimethylpentanamide, N-(1-hexyl-4,6-dimethylindolin-7-yl)-cyclohexanamide, N-(1-hexyl-4,6-dimethylindolin-7-yl)-2, 2 -dimethyl-3-ethoxypropanamide, N-(1-ethoxypropyl-4,6-dimethylindolin-7-yl)-2, 2 -dimethylpropanamide, N-(1-hexyl-4,6-dimethylindolin-7-yl)-2, 2 -dimethyl-3-piperidinopropanamide, N-(1-piperidinopropyl-4,6-dimethylindolin-7-yl)-2, 2 -dimethylpropanamide, N-(l-hexyl-4,6-dimethylindolin-7-yl)-2,6-dimethylpiperidinopropanamide, and the like, and pharmaceutically acceptable salts thereof.

The Compound may be converted to a pharmaceutically acceptable salt thereof.

a The Compound may be converted to an acid addition salt, since it has a basic group, and the acid to form this acid addition salt includes, for example, an inorganic acid such as hydrochloric acid, S: sulfuric acid, phosphoric acid, nitric acid and the like; an organic acid such as oxalic acid, fumaric acid, maleic acid, citric acid, tartaric acid, methanesulfonic acid, toluenesulfonic acid and the like; and the like.

When Compound has an acidic group such as carboxyl, it can form an alkali metal salt such as sodium salt, potassium salt and the like; alkaline earth metal salt such as calcium salt, magnesium salt and the like; organic base salt such as triethylamine salt, dicyclohexylamine salt, pyridine salt and the like; and the like.

The Compound and pharmaceutically acceptable salts thereof can be produced, for example, by the following methods.

Production Method 1 Compound (II) and compound (III) are reacted.

This method produces a compound of the formula wherein R 6 is -NR7R 8 where Re is hydrogen atom.

This reaction generally proceeds in an inert solvent. Examples of the inert solvent include acetone, dioxane, acetonitrile, chloroform, benzene, methylene chloride, ethylene chloride, tetrahydrofuran, ethyl acetate, N,N-dimethylformamide, pyridine, water and the like, and mixtures thereof.

In addition, a base such as triethylamine, pyridine, 4dimethylaminopyridine, potassium carbonate and the like may be added.

The reaction temperature is generally from -10°C to 160 0

C,

preferably 20-100°C, and the reaction time is generally from 30 minutes to 10 hours.

The starting compound (II) can be prepared, for example, by the following method.

*I A nitro group is introduced into a compound of the general formula

(X)

R11 (CH2 J (x *1

I

4 wherein

R

1 2

R

13 and m are as defined above and R" is an aminoprotecting group [see J. Eric. Mordlander, et al., J. Org. Chem., 46, 778-782 (1981)], (introduction of nitro onto benzene ring) using nitric acid in a mixed solvent of acetic acid and sulfuric acid, and the amino-protecting group is eliminated. The compound thus obtained and compound (IX) are reacted, and nitro group is reduced using a catalyst such as palladium-carbon and the like to give starting compound

(II).

Examples of the amino-protecting group at include acyl such as formyl, acetyl, monochloroacetyl, dichloroacetyl, trichloroacetyl, trifluoroacetyl, propionyl, benzoyl and the like.

Said amino-protecting group is eliminated by a method known Per se.

For example, it is eliminated by the action of an acid hydrochloric acid, formic acid, trifluoroacetic acid and the like) or an alkali sodium hydroxide, potassium hydroxide, sodium carbonate, sodium hydrogencarbonate and the like), or other method.

Production Method 2 Compound (II) and compound (IV) are reacted.

This method produces a compound of the formula wherein

R

6 can be any one of those defined above.

.This reaction generally proceeds in an inert solvent. Examples of the inert solvent include acetone, dioxane, acetonitrile, chloroform, benzene, methylene chloride, ethylene chloride, tetrahydrofuran, ethyl acetate, N,N-dimethylformamide, pyridine, water and the like, and mixtures thereof.

In addition, a base such as triethylamine, pyridine, 4dimethylaminopyridine, potassium carbonate and the like may be added.

The reaction temperature is generally from -10°C to 100 0

C,

preferably 0-60 0 C, and the reaction time is generally from 30 minutes to 10 hours.

Production Method 3 Compound (II) and compound or a reactive derivative thereof are reacted.

VThis method produces a compound of the formula wherein R 6 is R6 This reaction generally proceeds in an inert solvent. Examples of the inert solvent include acetone, dioxane, acetonitrile, chloroform, benzene, methylene chloride, ethylene chloride, tetrahydrofuran, ethyl acetate, N,N-dimethylformamide, pyridine, water and the like, and S mixtures thereof.

In addition, a base such as triethylamine, pyridine, 4dimethylaminopyridine, potassium carbonate and the like may be added.

The reaction temperature is generally from -10°C to 100'C, preferably 0-60 0 C, and the reaction time is generally from 30 minutes to 10 hours.

Compound is subjected to said reaction as, for example, a free acid; a salt such as sodium, potassium, calcium, triethylamine, pyridine and the like; or a reactive derivative such as acid anhydride, mixed acid anhydride substituted phosphoric acid dialkylphosphoric acid), alkyl carbonate monoethyl carbonate) -1 1and the like], active amide amide with imidazole etc.), ester cyanomethyl ester, 4-nitrophenyl ester and the like), and the like.

When Compound .is used as a free acid or salt in this reaction, the reaction is preferably carried out in the presence of a condensing agent. Examples of the condensing agent include dehydrating agent such as N,N'-di-substituted-carbodiimides N,N'-dicyclohexylcarbodiimide), carbodiimide compounds 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide, N-cyclohexyl-N'-morpholinoethylcarbodiimide and Ncyclohexyl-N'-(4-diethylaminocyclohexyl)carbodiimide), azolide compounds N,N'-carbonyldiimidazole and N,N'-thionyldiimidazole) and the like. When these condensing agents are used, the reaction is considered to proceed via a reactive derivative of carboxylic acid.

Production Method 4 Compound (VI) and compound (VII) are reacted.

This method produces a compound of the formula wherein

R

6 is

-NR

7

R

8 This reaction generally proceeds in an inert solvent. Examples of the inert solvent include acetone, dioxane, acetonitrile, chloroform, Sbenzene, methylene chloride, ethylene chloride, tetrahydrofuran, ethyl acetate, N,N-dimethylformamide, pyridine, water and the like, and mixtures thereof.

In addition, a base such as triethylamine, pyridine, 4dimethylaminopyridine, potassium carbonate and the like may be added.

The reaction temperature is generally from -109C to 160°C, preferably 10-100°C, and the reaction time is generally from 30 minutes to 10 hours.

The starting compound (VI) can be produced, for example, by dissolving compound (II) in an inert solvent and bubbling in phosgene.

Production Method Compound (VIII) and compound (IX) are reacted.

This reaction generally proceeds in an inert solvent. Examples of the inert solvent include acetone, dioxane, acetonitrile, chloroform, -12benzene, methylene chloride, ethylene chloride, tetrahydrofuran- ethyl acetate, N,N-dimethylformamide, pyridine, water and the like, and mixtures thereof.

In addition, a base such as triethylamine, pyridine, 4dimethylaminopyridine, potassium carbonate, sodium hydride and the like may be added.

The reaction temperature is generally from -10 0 C to 100°C, preferably 0-60°C, and the reaction time is generally from 30 minutes to 10 hours.

The starting compound (VIII) can be prepared, for example, by the method wherein a nitro group is introduced into a compound of the formula (introduction of nitro onto benzene ring), and the nitro group is reduced using a catalyst such as palladium-carbon and the like to give a compound of the formula

(XI)

R 3 2 (CH2)m

(XI)

N

H2N I

R

wherein

R

12

R'

3 R14 and m are as defined above. Using this compound as a starting compound and according to Production Method 2, a compound of the formula

(XII)

R'

R2 (CH2)m

S(XII)

R 3

N

R!R I wherein

R

2

R

3 R4, R4 and m are as defined above, is obtained.

This compound is deprotected to give compound

(VIII).

The Compound of the parent application obtained as in the above can be purified by a method conventionally known, such as chromatography and recrystallization.

-13- This Compound can be converted to a pharmaceutically acceptable salt by a method known per se.

The Compound and pharmaceutically acceptable salts thereof of the parent application show superior ACAT inhibitory activity and lipoperoxidation inhibitory activity in mammals human, cow, horse, dog, cat, rabbit, rat, mouse, hamster and the like) and are useful as ACAT inhibitors and hyperlipemia inhibitors. To be specific, they are useful for the prevention and treatment of arteriosclerotic lesions such as arteriosclerosis, hyperlipemia and diabetes, as well as ischemic diseases of brain, heart and the like.

A pharmaceutical composition containing Compound

(I)

or a pharmaceutically acceptable salt thereof may contain an additive. Examples of the additive include excipients starch, lactose, sugar, calcium carbonate and calcium phosphate), binders starch, gum arabic, carboxymethylcellulose, hydroxypropylcellulose and crystalline cellulose), lubricants magnesium stearate and talc), disintegrators calcium carboxymethylcellulose and talc), and the like.

The above-mentioned ingredients are mixed, and the mixture can be formulated into an oral preparation such as capsule, tablet, fine granules, granules, dry syrup and the like, or a parenteral preparation such as injection, suppository and the like by a method conventionally known.

While the dose of Compound and pharmaceutically acceptable salts thereof varies depending on administration target, symptom and other factors, it is generally about 0.1-50 mg/kg body weight per dose for an adult patient with hypercholesterolemia by oral administration in about one to three times a day.

The present invention is described in more detail in the following by way of Examples, to which the present invention is not limited.

Example 1 1-butyl-3-(1-hexyl-4, 6 1-acetyl-4,6-dimethylindoline -14- 4,6-Dimethylindole (1.08 g) was dissolved in acetic acid (20 ml), and sodium cyanoborohydride (2.3 g) was added portionwise at 15°C. The mixture was stirred at said temperature for one hour and poured into ice water. Saturated aqueous sodium hydrogencarbonate was added to neutralize the mixture and the mixture was extracted with ethyl acetate.

The extract was washed with saturated brine and dried over sodium sulfate. The solvent was evaporated under reduced pressure. The residue was dissolved in benzene, and acetic anhydride (840 mg) was added, which was followed by stirring at room temperature for one hour.

The reaction mixture was washed with saturated aqueous sodium hydrogencarbonate and saturated brine, and dried over sodium sulfate.

9. The solvent was evaporated under reduced pressure. The residue as purified by silica gel column chromatography (developing 9: solvent chloroform-methanol=1:0 10:1) to give 1.3 g of the title compound 'H-NMR (CDCI3) 6 2.18 (6H, s, -CH3, -COCH3), 2.30 (3H, s, -CH3), 3.00 (2H, t, J=8.3Hz, C3-H2), 4.03 (2H, t, J=8.3Hz, C2-H2), 6.66 (1H, s, Cs-H), 7.89 (1H, s, CT-H) 1-acetyl-4,6-dimethyl-5-nitroindoline 1-Acetyl-4,6-dimethylindoline (2.6 g) was dissolved in acetic 9.i anhydride (35 ml), and nitric acid 0.92 ml) dissolved in acetic anhydride (15 ml) was added dropwise at OOC. The mixture was stirred at room temperature for one hour and poured into ice water. Saturated aqueous sodium hydrogencarbonate was added to neutralize the mixture, and the mixture was extracted with chloroform. The extract was washed with saturated brine and dried over sodium sulfate. The solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (developing solvent chloroform-methanol= 100:1) to give 2.4 g of the title compound 'H-NMR (CDCls) 6 2.17 (3H, s, -COCH3), 2.24 (3H, s, -CH3), 2.30 (3H, s, -CH3), 3.08 (2H, t, J=8.4Hz, C3-H2), 4.14 (2H, t, J=8.3Hz, C2-H2), 8.00 (1H, s, C7-H) 4,6-dimethyl-1-hexyl-5-nitroindoline 1-Acetyl- 4 ,6-dimethyl-5-nitroindoline (2.4 g) obtained in was dissolved in methanol.(25 ml) and 6N hydrochloric acid (20 ml) was added, which was followed by refluxing for 15 hours. After the completion of the reaction, the solvent was evaporated under reduced pressure. The residue was dissolved in chloroform, and the mixture was washed with saturated aqueous sodium hydrogencarbonate and saturated brine, and dried over sodium sulfate. The solvent was evaporated under reduced pressure. Indoline (1.8 g) thus obtained was dissolved in dimethylformamide (20 ml), and sodium hydride (abt. 60% in oil suspension, 457 mg) was added at 0C. The mixture was stirred at said temperature for 0.5 hour and hexyl bromide (1.8 g) was added to the reaction mixture, which was followed by stirring at room temperature for 3 hours. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The extract was washed with saturated brine and dried over sodium sulfate. The solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (developing solvent hexane-ethyl acetate= 1:0 10:1) to give 2.8 g of the title compound 'H-NMR (CDC13) 6 0.90 (3H, br-t, -CH3), 1.2 1.8 (8H, m, 2.17 (3H, s, -CH3), 2.30 (3H, s, -CH3), 3.00 (2H, t, J=8.4Hz, C3-H2), 3.09 (2H, t, J=7.2Hz, N-CH2), 3.51 (2H, t, J=8.3Hz, C2-H2), 5.99 (1H, s, C7-H) 1-butyl-3-(1-hexyl- 4 4,6-Dimethyl-l-hexyl-5-nitroindoline (1.0 g) obtained in was dissolved in benzene (40 ml) and 10% palladium-carbon (100 mg) was added to allow hydrogenation at 40'C. After the completion of the reaction, palladium-carbon was filtered off, and the filtrate was washed with saturated aqueous sodium hydrogencarbonate and saturated brine, and dried over sodium sulfate. The solvent was evaporated under reduced pressure. 5-Amino- 4 ,6-dimethyl-1-hexylindoline thus obtained was -1 6dissolved in chloroform (20 ml) and butyl isocyanate (400 mg) was added to the reaction mixture, which was followed by stirring at room temperature for 18 hours. Water was added to the reaction mixture, and the mixture was washed with saturated brine and dried over sodium sulfate. The solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (developing solvent chloroform-methanol=1:0 50:1) and recrystallized from ethanol to give 650 mg of the title compound 'H-NMR (CDC1l) 6 0.91 (6H, br-t, -CH3), 2.0 (12H, m, 2.09 (3H, s, -CH3), 2.19 (3H, s, -CH3), .2 .6 3.6 (8H, m, NH-CH2,

C

3

-H

2 N-CH2,

C

2

-H

2 4.29 (1H, br, NH), 5.45 (1H, br, NH) 6.18 (1H, s, C7-H) Example 2 N-(1-hexyl-4,6-dimethylindolin-5-yl)-2,2-dimethylpropanamide 5-Amino-4,6-dimethyl-l-hexylindoline (880 mg) was dissolved in chloroform (20 ml), and triethylamine (370 mg) and pivaloyl chloride (430 mg) were added, which was followed by stirring at room temperature for 1 hour. Water was added to the reaction mixture, and the mixture .i was washed with saturated brine and dried over sodium sulfate. The solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (developing solvent 50:1) and recrystallized from ethanol to give 650 mg of the title compound.

'H-NMR (CDCla) 6 0.84 (3H, br-t, 1.1 1.8 (8H, m, 1.33 (9H, s, 2.00 (3H, s, -CH3), 2.11 (3H, s, -CH3), 2.82 (2H, t, J=7.8Hz, C3-H2), 2.99 (2H, t, J=7.2Hz, N-CH2), 3.33 (2H, t, J=7.8Hz, C2-H2), 6.16 (1H, s, CT-H), 6.70 (1H, br, N-H) Eample 3 N-(1-hexyl-4, 6 -dimethylindolin-7-yl)-2,2 -dimethylpropan- -17amide 1-acetyl-5-bromo-4,6-dimethylindoline 1-Acetyl- 4 ,6-dimethylindoline (5.5 g) was dissolved in acetic acid (150 ml), and bromine (2.2 ml) was added dropwise at room temperature.

The mixture was stirred at room temperature for 1 hour, and poured into ice water. The precipitated solid was collected by filtration, and recrystallized from methanol to give 6.5 g of the title compound 'H-NMR (CDCl 3 6 2.19 (3H, s, -COCH3), 2.27 (3H, s, -CHa), 2.39 (3H, s, -CH3), 3.06 (2H, t, J=8.4Hz, C3-H 2 4.03 (2H, t, J=8.4Hz, C2-H2), 7.99 (1H, s, CT-H) 5-bromo-4,6-dimethyl-7-nitroindoline Concentrated sulfuric acid (25 ml) and nitric acid (d=1.56, 1.46 ml) were added to acetic acid (25 ml), and 1-acetyl-5-bromo-4,6dimethylindoline (6.5 g) obtained in was added while stirring the mixture at 0°C, which was followed by stirring at said temperature for 18 hours. The reaction mixture was poured into ice water, and the precipitated solid was collected by filtration and washed thoroughly with water. The obtained solid was suspended in ethanol (50 ml) and water (10 ml). Sodium hydroxide (20 g) was added and the mixture was refluxed for 3 hours. The solvent was evaporated under reduced pressure and chloroform was added. The mixture was washed with saturated brine and dried over sodium sulfate. The solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (developing solvent benzene) to give 4.8 g of the title compound 'H-NMR (CDCl) 6 2.29 (3H, s, C4-CHs), 2.65 (3H, s, C6-CHa), 3.10 (2H, t, J=8.4Hz, C3-H2), 3.82 (2H, t, J=8.4Hz, C2-H2), (1H, br, N-H) N-(1-hexyl-4,6-dimethylindolin-7-yl)-2, 2 -dimethylpropanamide 5-Bromo-4,6-dimethyl-7-nitroindoline (3.0 g) obtained in was dissolved in dimethylformamide (60 ml) and sodium hydride (abt. 60% in oil suspension, 530 mg) was added at OC, which was followed by -18stirring at said temperature for 0.5 hour. Hexyl bromide (1.8 g) was added to the reaction mixture and the mixture was stirred at room temperature for 12 hours. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The extract was washed with saturated brine and dried over sodium sulfate. The solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (developing solvent hexane-ethyl acetate= 10:1). The obtained solid was dissolved in benzene (40 ml) and palladium-carbon (100 mg) was added to allow hydrogenation at 40 0

C.

After the completion of the reaction, palladium-carbon was filtered off, and the filtrate was washed with saturated aqueous sodium hydrogencarbonate and saturated brine, and dried over sodium sulfate.

The solvent was evaporated under reduced pressure.

7-Amino-4,6-dimethyl-l-hexylindoline thus obtained was dissolved in chloroform (20 ml), and triethylamine (1.0 g) and pivaloyl chloride g) were added to the reaction mixture, which was followed by stirring at room temperature for 1 hour. Water was added to the reaction mixture, and the mixture was washed with saturated brine and dried over sodium sulfate. The solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (developing solvent chloroform-methanol=1:0 50:1). The obtained compound was dissolved in ethanol and 10N hydrochloric acid/ethanol ml) was added. The solvent was evaporated under reduced pressure.

The residue was recrystallized from ethanol to give 700 mg of hydrochloride of the title compound 'H-NMR (CDCls) 6 0.90 (3H, br-t, -CH3), 1.1 1.6 (8H, m, 1.41 (9H, s, 2.15 (3H, s, -CHs), 2.25 (3H, s, -CH3), 3.15 (4H, m, C3-H2, N-CH2), 3.70 (1H, m, C2-H), 4.00 (1H, m, C2-H), 7.12 (1H, s, Cs-H), 9.2 (1H, br, N-H) Example 4 1-butyl-3-(1-hexyl-4,6 -dimethylindolin-7-yl)urea 7-Amino-4,6-dimethyl-l-hexylindoline (800 mg) was dissolved in -1 9chloroform (20 ml) and butyl isocyanate (400 mg) was added, whieh was followed by stirring at room temperature for 18 hours. Water was added to the reaction mixture, and the mixture was washed with saturated brine and dried over sodium sulfate. The solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (developing solvent chloroform-methanol=1:0 50:1) and recrystallized from ethanol to give 450 mg of the title compound.

'H-NMR (CDCl3) 6 0.88 (6H, br-t, -CH3), 1.8 (12H, m, -(CH2) 2 2.13 (6H, s, -CH3X2), 2.83 (2H, t, J=8.3Hz, C3-H2), 3.20 (4H, N-CH2 X2), 3.43 (2H, t, J=8.3Hz, C2-H 2 4.80 (1H, br-t, NH-CH 2 5.52 (1H, br-s, CT-NH), 6.40 (1H, s, Cs-H) Examples 5-36 In the same manner as in any one of the above-mentioned Examples 1- 4, the compounds shown in Tables 1 and 2 were obtained.

S go* Table I Example R 2

R

-H -H

-CH

3 -C (CH 3 e6

-CU

3 -H

-CIT

3 -C(CH 3 7. Af'CTT -H-OH CH3 8 C3 -H -OCH 3 -C(CH 3

-CH

3 -H

-CH

3 -C(CH 3

-CU

3 -H

-CH

3 -C(CH 3

-CU

3 -H

-CU

3 -C(C11 3 14 LI J11 -H -(l 3 CO 12

-CU

3 -HU

-CU

3 -C(C11 3 )2 16 -CL

^TV-P

21 Table 2 R3

N

4

R

R 4

=-NHCO-RO

72, Example R 2

FR

R8 R 21 22 23 24 25 26 27 28 29 31 32 33 34 36

-CH

3

-CH

3

-CH

3

-CR

3 -CRs

-CH

3

-CH

3

-CH

3

-CR

3

-CH

3 -CRs -CH3

-CH

3

-CH

3

-CH

3 -CH 3

-H

-H

-H

-H

-H

-H

-H

-H

-H

-H

H

-H

-H

-H

-H

-CRs -CR 3 -CH3

-CR

3

-CR

3

-CH

3

-CH:

-CR-3 -CHl 3

-CR

3

-CH

3 -CUs

-OU

3 -CUs -CU 3

-(CU

2 3

CH

3

-C(CH

3 3 -C (CR 3 3

-C(CH

3 3

-C(CR

3 3 -C (CH 3

-C(CH

3 3

-C(CH

3 2

CH

2

OR

-C(CH

3 2

CH

2

OCOCH

3

-C(CH

3 )2 -C(CHs) 2CH 2

OCO

-C(CH

3 2

CH

2 0C 2

H

-CH

2

CH:

-(OH

2 2OCOCHs

-(CR

2 2 0CR 3

-(CR

2 2 0C 2 Hs

-C

5

R

11

-(CR

2

-CSH

13

-(CR

2 2CH(CH 3 2

-CU

2 CH1(H 3 2 -CHs

-C

3 H7 -C 3 -C(C11 3 )3 22 The 1 H-NMR data of the compounds of the above Examples 5-36 are shown in the following.

'H-NMR (CDcl 3 Example 5 a a Example 6 Example 7 Example 8 Example 9 Example 10: Example 11: Example 12: Example 13: :(hydrochloride) 0.91 (3H, br-t), 1.1 1.8 (8H1, in), 1.34 (9H1, S), 2.09 (3H, 2.93 3.48 (4IH, mn), 3.48 3.82 (1H, in), 3.82 41.35 (1H, in), 7.12 (OH, 7.32 (OH, d), 9.36 (OH, br-s) :0.93 (3H, br-t), 1.0 2.0 (19H1, in), 1.35 (611, s), 2.12 (3H, 2.19 (311, 2.98 (2H, 3.25 (2H, t), 3.65 (211, 6.80 (1H, 8.09 (OH, br-s) :0.87 (3H, br-t), 1.1 1.8 (811, in), 1.31 (9H, s), 2.841 (211, 3.16 (2H, 3.414 (2H, t), 3.74 (3H, 3.79 (311, 5.86 (OH, s) :0.92 (3H, br-t), 1.39 (9H1, 1.2 1.9 (411, in), 2.08 (3H, 2.15 O3H, 2.80 3.30 (4H, in), 3.60 (2H, 6.68 (1H, 7.78 (1H, br-s) 0.90 (3H, br-t), 0.92 O3H, br-t), 1.08 1.88 (10H, in),, 1.29 (611, s-)p 2.05 (3H, 2.12 (311, 2.81 (211, t), 3.13 (211, 3.110 (2H, 6.39 (11, 6.74 O1H, br-s) 0.63 1.05 (6H, in), 1.05 1.82 (1411, mn), 1.29 (611, s), 2.08 (3H, 2.10 (3H, 2.82 (2H, 3.13 (2H, t), 3.41 (211, 6.411 (1H, 6.77 O1H, br-s) 0.88 (3H, br-t), 1.01 1.87 (12H, mn), 1.35 (9H, s), 2.09 (3H, 2.14 (3H, 2.82 (211, 3.13 (2H, t), 3.52 (211, 6.60 (1H, br-s) 1.18 (3H, 1.35 (9H, 2.09 (311, 2.15 (3H, s), 2.92 (2H, 3.22 (2H1, 3.56 (2H1, 6.62 (1H, s), 7.57 OHi, br-s) 0.88 (3H, br-t), 1.08 (3H, 1.29 (6H, s), 1.1 1.9 (10H, in), 2.08 (3H, 2.22 (3H, s), 2.82 (211, 3.21 (2H, 3.33 (211, 6.43 O1H, s),I 6.84 (1H, br-s) 23 Example 14: 0.90 (OH, br-t), 0.92 (3H, br-t), 1.08 1.88 (14H,-m), Example 15 Example 16: Example 17: 4* 4* .4 *4 *4 4 4 4* *S4.

1 .29 3.13 0.63 2.11 3.60 0.89 2.11 2.6 0.87 1.32 2.7 7.83 0.87 2.17 3.10 6.8 0.87 1.20 2.91 6.56 0.85( 1.80- 2.25- (6H (2H 1.

(3H, (2H, (3H, (3H (3H, (3H, 3.4 (1 H,

(OH,

(6H, (2H, 8.2 (3H, (3H,9 2H, 1 H, 3H, ),2.05 ),3.40 05s (6H,m 2.17 6.72 br-t), 1 2.15 (4H, m) (3H, 2.12 (3H, 2.81 (2H, t), (2H, 6.39 (OH, 6.74 (1Hi, br-s) 1.05 1.82 (18H, mn), 1.34 (6H, s), (3H, 2.95 (2H, 3.13 (2H, t), (I1H, 7.8 (OH, br) 1.70 (8H, in), 1.81 (3H, s), (3H, 2.81 (2H, br-t), 6.39 (OH, 6.58 (1H, br), 6.72 (OH, br) .05 1.90 (8H, in), 1.25 (3H, s), (3H, 2.17 (3H, 2.67 (OH, mn), 3.59 (2H, 6.69 (OH, s), br-t), 1 2.11 (4H, m) Example 18: br-s) br-t), 1.05 2.48 (2H, 3.54 (2H, (OH, br-s), 8.

br-t), 1.05 1.35 (9H, 3.04 (2H, 7.25 (1H, br-t), 1.05 Example 19: 1.80 (8H, mn), 1.21 (6H, s), 2.55 (2H, 3.01 (2H, 6.70 (OH, s), .72 (OH, br-s) 1.80 mn), 1.17 (3H, t), 2.22 2.62 (4H, m), 3.48 (2H, br-t), br-s) 1.80 (8H, 1.27 44 4* *444..

Example 20: *2.25 in), 2.10 2.55 (2H, in), 2.90 (3H, 2.16 (3H, s), (2H, 3.04 (2H, t)v Example 21: Example 22: Example 23: 3.47 0.83 2.11 3.22 1.27 3.43 9.15 1 .33 3.38 (2H, (3H, (3H, (2H, (9H, (2H, (1 (9H, (3Hv 6.69 (1H, 8.49 (1H, br-t), 0.87 (3H, br-t), 1.1 2.16 (3H, 2.29 (1H, 3.55 (2H, 6.49 (1H, 2.04 (6H9 2.13 (3H, 3.75 (2H, 4.29 (2H, br-s) 2.04 (3H, 2.09 (3H-, 3.4 3.6 (4H, mn), 6.36 br-s), 8.95 (OH, br-s) 1.8 (11H, in), 2.85 (2H, t), 6.68 O1H, br) 3.06 (2H, t), 6.87 (OH, s), 2.8 2.9 (OH, 7.35 (4H, m) (1 H, br-s) 24 Example 24: Example 25: Example 26: Example 27: Example 28: Example 29: 1.16 2.82 0.88 2.08 3.55 1.1- 2.29 5.40 0.87 2.07 3.49 0.87 1.99 3.37 7.814 0.874 (3H, (2H, (311, (3H, (211, 1.9 (211, (11H, (3H, (311, (2H, (3H, O31, (2H, (111, 1.32 (9H, 2.04 (3H, 2.09 3.3 3.6 (8H, in), 6.53 (1H, 7.40 (OH, br-s) br-t), 1.35 (911, 1.2 1.9 (6H1, m), 2.15 2.92 (2H, 3.13 (211, t), 6.23 (OH, 7.60 (1H, br-s) (6H, mn), 1.39 (9H, 2.07 (3H, 2.13 (3H, s),o 2.84 (2H, 3.11 (2H, 3.43 (2H1, t), br), 6.52 (1H, 7.30 (11, br-s) br-t), 1.1 1.8 (10H, mn), 1.34 (9H, s)) 2.13 2.87 (2H, 3.13 (2H, t), 6.52 (1H, 7.20 (OH, br-s) br-t), 1.1 1.8 (811, mn), 1.26 (6H1, s), 2.08 (3H, 2.78 (2H, 3.10 (2H, t), 3.48 (2H, 4.00 (1H, br-s), 6.35 (1H, s), 3HP Example 30: Example 31: Example 32: Example 33: Example 34: 2.07 3.43 0.87 2.82 7.42 0.90 2.09 3.54 0.94 2.04 3.38 0.85 2.03 3.41 7.40 0.88 1 .27 (611 (2H, (311, (211, (2H, (6H, (3H, (2H, (6H, (3H, (2H, (31, (31, (2H, (211, (3H, (611, br-t), 1.1 2.12 (311, 4.20 (211, br-t) 1 .1 3.16 (2H, mn), 8.22 (OH, 1.3 1.8 2.16 (311, 6.65 (OH, 1.33 (911, 2.10 (311, 6.37 (11, br-t), 1.1 1 2.08 (311, 4.50 (211, in), 8.28 (11, Sn), 1.8

S),

Sn), 1.8 (8H1, mn), 1.28 (611, s), 2.84 (2H1, 3.14 (2H1, 0), 6.48 (1H, 7.35 O1H, s) (8H, in), 2.15 (611, s), 3.43 (2H, 6.43 O11, s), 8.75 O1H, mn), 9.16 (1H, br-s) in1.36 (9H1, s), 2.92 (211, 3.16 (2H, t), 7.65 O1H, br-s) 1.7 2.0 (11, mn), 2.82 (211, 2.94 (2H1, t), 6.70 (1H, br-s) (811, 1.44 (611, s), 2.80 (211, 3.14 (2H, t), 6.37 (1H, 7.04 (1H, br-a), 8.75 (11, in), 9.21 OH1, d) 2.07 (311, 2.10 (31, 2.81 (211, t), 25 3.18 6.37 Example 35: 0.88 .2.10 6.39 Example 36: 0.87 2.08 3.50 In the same 4, the compounds (2H, 3.41 (2H, 3.48 (2H, 3.55 (OH, 8.00 (OH, br-s) (OH, br-t), 1.1 1.8 (14tH, in), 1.24 (6H, s), (6H, 2.50.(2H, 2.90 (2H, 2.8 3.6 (8H, in), (OH, 7.00 (OH, br-s) (3H, br-t), 1.2 1.9 (2H, in), 1.35 (9H, s), (3H, 2.15 (3H, 2.93 (2H, 3.12 (2H, t), (2H, 6.30 (OH, 7.40 (1H, br-s) manner as in any one of the above-mentioned Examples 1shown in Tables 3 to 9 can be obtained.

a a a a. a.

a. a.

a a a.

a..

a a. a a.

a a. a a a.

a a 26 Table 3 R' =-NHCO-Ro Example

R'

a a. a a.

a

R

2 R3 RO 37 38 39 40 41 42 43 44 45 46 47 48 49 51 52 53 -0112 -0113

-CH

3 -CHs -0113 -CH3

-OH

3

-CII,

-CH3

-OH

3 -CHa -CIIs

-CH

2 -CIIs

-H

-H

-H

-HI

-H1

-H

-H

-H

-H

-H

-H

-H

-H

-H

-H

-H

-H

-CH3 -CHs.

-OH

2 -CHs -CHs

-OH

3 -CIIs

-OH

3

-CII,

-OH,

-OHs -0(0113) s -0(0113)2 -0(0113)3 -0o -0o

OH

2

NH

2 -0o -1OCH 2

CH

2 OO1 -O(CH3) 2 (0112) 4 N7J -C(CHs) 2 (0112)4- -C(CH3 2

CH

2 000H2N 1 0 -O(CH3)2-(11 2 2

C

-(CH

2 400(011:)3 -OH2-CH=C(OH3)2 -(0112) -(0112)3-Nf1CH -(112) sCN -(112)3

"C-C&

-(112)50113 -(112)50112 -(112) 5

COH

-(112) SCH3 -(0112)50113 -0112CH3 -(0112)2011 -(0112)30113 27 Table 4j R 4 =-NHCO-RI Example

S

S.

S *1 4" 54 56 57 58 59 60 61 62 63 64 66 67 -CH 3 -0113 -C11 3 -0113 -0113

-CH

3 -C11 3 -CHs

H

-H

-H

-H

-H

-H1 -CH3 -0113

-CH

3

-CH

3 -0113

-CH

3 -0113

-OH

3 -0C11 3 -OC11 3

-OCII(CH

3 2 -0011H3) -OCH(C1 3 2 -OC11(CH 3 2 -C(013) 2

-(CH

2 6

-N-CH

3 -0112C(CH 3 3 -C(C11 3 2 (0112) 2

SO

3 Na

-CII

2

C(CH

3 2 (0112) 2 SOSNa -C(0113) 3 -0(0113)3 -0(0113)3 -0(0112)3 -(0112)30113 -(0112)50113 -(0112)50113 -(0112)50113 -(112)50113 -(0112)50113 -(0112)50113 -(0112) 0H3 -(0112) 4C013 -(0112)50113

-(OH

2 )40113 -(0112)50113 -(0112)5011 1. 1 28 Table

R'

R 2 R 4

=-NHCO-R'

R

3

N

***Example R' 2R -Re-Rs- *68'

-CH

3 -H

-CH

2

-C(CH

2 2 (C11 2 2 69 -CH3 -H

-CH

3 -C .H 2

C'(CH

3 2

(CH

2 2

-CH

3 -H

-CH

3

-C(CH

3 2

CH

2 0CO(CH 2 )3- 71 -CH 3 H

-CH

3 -C(CHs) 2

CH

2 0COC(CH 3 2

(CH

2 3 29

R'

22> R 3 N 4 Table 6 R~ 4 -NICONH-R 7 T I I T Example i i 4 I_ __I .4* a a. a a a a. 0 0 *0 72 73 74 76 77 78 79 80 81 82 83 84 86 87 88 89 91 92

-H

-CH

3

-OCH

3

-CH

3

-CH

3

-CH

3

-CH

3 -CH3

-CH

3

-H

-H

-H

-H

-H

-H

-H

-H

-H

-H

-H

-H

-H

AH

-H

-H

-H

-H

-H

-H

-H

-H

-if

-H

-H

-H

-H

-H

-H

-H

-H

AH

-H

-CH

3 -CH3 -0013 -CHl2

-CH

3

-CH

3

-CH

3 -Cl 3 -CH3

-CH

3

-CH

3

-CH

3 -CH3 -CH3

-CH

3

-OCH

3 -OCH3 -OCHs

-OCH

3

-O-CH

3 -OCH3

-C(CHD

2 -0o

-C(CH

3 3

-C(CH

3 3 o

-C(CH

3 3 -C (CH 3 3

-Q

-(CH

2 5

CH

3

-OH

3

-(CH

2 3

CH

3

-(CH

2 5 CHs -Cls

-(OH

2 )s CH:

-(CH

2 5

CH

3 -Cls

-(CH

2

,CH,

-0C 2 5

CH

3 -Cls -(CH2) .CH 3 -(012) 5

CH

3

(CH

2 5

CH

3 -(CH2) 5

CH

3

-(CH

2 3

CH

3

(CH

2 3

CH

3 -(CH2) 7

CH

3

-CH

2

CH

3

-CH

2 Cl 3

-(CH

2 5

CH

3

H

-0C 2 sCH 3

-(OH

2 5 CHs

-(CH

2 7

CH

3

-(CH

2 7

CH

2

-(CH

2 7

CH

3

-CH

2

CH

3

-CH

2

CH

3

-CH

2

CH

3

-(CH

2 aCH3

-(CH

2 2

CH

3

-(CH

2 :01: L L 3 0- Table 7

R

2 =-NHCONH-R 7 *0 S S S

S.

S

Example 93 94 96 97 98 99 100 101 102 103

I

-H

-CH

3 -OC11 3

-CH

3 -CH3

-CH

3 -CH3

-CH

3

-CII:

-CH

3 -Cls RC 3

-CH

3

-CH

3

-CH:

-CH3

-CII:

-C11s -Cls -Cls

-CH

3 -C(C1 3 3

-C(CH

3 2

(CI

2 8 CH3

-C(CH

2 3

C

-C(CH

3 3

-C(CH

2 2 (C1 2 5

CH

3

-'(CH

2 3 Cl 3

-(CH

2 5

CH

3

-(CH

2 )sCH 3 -(0112)5 CH3 -(0112) 5 CH3

-(CH

2 3

CH

3 -(Cl 2 )30113

-(CH

2 2

CH

2

-(CH

2 7 CH3 -Cl 2

CII

-CH

2 CH3

-(CH

2 5

CH

3 -(0112)5013 S S

S.

S.SSS.

S

L

L

31 Table 8

R

2

=-NICO-R'

S

S. 5 5* S S S 5*

*S

5* 5 5S**

S

55**

S.

S

Example 104 105 106 107 108

RH

-H

-H

-H

-H

R

3

-OCH

3

-OCH

3

-OCH

3

-OCH

3 -OCH3

R-H

-H

-H

-H

-H

-CHs

-(C

2 )s 3

CH

-0C 2 iCH 3

-CH

3

-(OH

2 3

CH

3

-CH

2

CH

3

-CH

2

CH

3

-CH

2

)CH

3

-(CU

2 3

CH

3 32 Table 9 R 4 =-NHCO-R' Examplef R' R R 3 Re 0.

*0 0 *0 00 0 00 00 *5 0 *0 4 *00.

0 0000 *0 0 0 0 0* *000** 0 109 110 ill 112 113 114 115 116 117 118

-CH

3

-H

-CH

3

-OCH

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Experimental Example 1 ACAT inhibitory activity A high cholesterol feed [a feed added with cholesterol Clea Japan, Inc.] was fed to male Japanese white rabbits weighing 2-2.5 kg at 100 g per day and the rabbits were bred for 4 weeks. The rabbits were killed by bleeding under anesthesia and small intestine was removed. The mucous membrane of small intestine was peeled, collected and homogenated. The homogenate was centrifuged at 4°C and 10,000 rpm for 15 min. The obtained supernatant was further centrifuged at 1 °C and 41,000 rpm for 30 minutes to give microsomal fractions. Using this microsomal suspension as an enzyme sample, dimethyl sulfoxide (DMSO, El) or a test compound dissolved in DMSO (test compound solution, and a reaction substrate 1 -'C1-oleoyl CoA were added to a reaction buffer. After incubation at 37°C for 5 minutes, a chloroformmethanol mixture was added to stop the reaction. Water was added thereto and mixed, and chloroform layer was separated. The solvent was evaporated to dryness, and the residue was dissolved in hexane. The mixture was subjected to thin layer chromatography using a silica gel plate. The spots of cholesteryl oleate on the silica gel plate were scraped, and quantitatively assayed on a liquid scintillation counter.

The ACAT inhibitory activity of the test compound was expressed as a proportion of inhibition of cholesteryl oleate, namely, the proportion of inhibition of cholesteryl oleate production as compared to control.

The results are shown in Tables 10-11.

Experimental Example 2 serum total cholesterol reducing effect Male Wister rats weighing 180-200 g were bred under free access to a high cholesterol feed [added with cholesterol cholic acid and coconut oil Clea Japan, Inc.] for 3 days, during which period a test compound (10-100 mg/kg) suspended in 5% gum arabic solution was forcibly administered once a day orally for 3 days. Only gum arabic solution was administered to control animals. After final administration, the test animals were fasted for 5 hours and the blood was taken. The serum total cholesterol level was determined -34using a commercially available assay kit (cholesterol-E-Test Wako, Wako Pure Chemical Industries, Ltd.). The activity of the test compound was expressed as a proportion of reduction of serum total cholesterol level, namely, the proportion of reduction of serum total cholesterol as compared to control.

The results are shown in Tables 10-11.

Experimental Example 3 LDL peroxidation inhibitory effect Male Japanese white rabbits weighing 2-2.5 kg were bred on 100 g per day of a high cholesterol feed [added with cholesterol Clea ~Japan, Inc.] for 4 weeks. The blood was taken from carotid and plasma was obtained. Then, LDL was fractionated from the plasma by ultracentrifugation, dialyzed for one day and preserved at 4°C. LDL (400 g) and aqueous copper sulfate solution (final concentration M) were added to bufferized Ham F-10 medium (2 ml, GIBCO, USA).

DMSO

or a solution (20 Pl) of test compound dissolved in DMSO was added and the mixture was incubated at 37 0 C for 24 hours. After the completion of the incubation, LDL peroxide in medium was allowed to develop color by thiobarbituric acid method and assayed as malondialdehyde. The activity of the test compound was expressed as malondialdehyde production inhibitory ratio namely, the proportion of inhibition of production of malondialdehyde as compared to control.

The results are shown in Tables 10-11.

Experimental Example 4 plasma lipoperoxidation inhibitory effect The blood was taken from male Japanese white rabbits weighing 2-2.5 kg under anesthesia and heparinized plasma was separated by a conventional method. To the plasma (2.0 ml) was added DMSO or a solution (20 PlI, final concentration 10- 5 M) of test compound dissolved in DMSO, and aqueous copper sulfate solution (final concentration 5 mM) was added immediately thereafter. The mixture was incubated at 37'C for 3 hours. After the completion of the incubation, trichloroacetic acid was added to stop the reaction. Then, the mixture was centrifuged at 410, 4,500 rpm for 15 minutes. The lipoperoxide in the supernatant thus obtained was assayed as malondialdehyde upon color

M

development by thiobarbituric acid method. The activity of the -test compound was expressed as malondialdehyde production inhibitory ratio namely, the proportion of inhibition of production of malondialdehyde as compared to control.

The results are shown in Tables 10-11.

Table I I a a

S

a a a.

Test compound Example 1 Example 2 Example 3 Example 4 Example 6 Example 7 Example 8 Example 9 Example 10 Example 11 Example 12 Example 13 Example 14 Example 15 Example 17 Example 18 Example 20 Example 21 Example 22 Example 23 Result of Exp. Ex. 1 *1 76.6 59.9 97.5 94.9 99.8 96.2 99.7 81.3 98.7 99.2 71.9 96.0 96.6 93.1 96.7 69.6 22.7 94.4 78.5 88.8 Result of Exp. Ex. 2 *2 12.9 54.4 21.4 53.3 19.2 48.0 51.3 55.2 59.5 30.3 28.4 53.9 35.0 27.0 22.3 7.8 28.3 38.6 23.6 Result of Exp. Ex. 3 *3 99.2 99.5 93.3 59.5 24.8 86.6 91.9 93.7 18.3 91.6 90.3 82.4 88.3 12.3 89.6 34.0 67.8 92.1 40.6 54.8 Result of Exp. Ex. 4 *4 94.3 95.8 90.4 92.0 37.4 86.4 85.1 81.6 82.6 78.8 74.9 87.1 93.1 90.1 91.1 87.1 91.1 91.4 92.8 90.5 *1 ACAT inhibition (concentration 10-* M, 10 5

M)

*2 reduction of serum total cholesterol (dose 10 mg/kg/day, 30 mg/kg/day, 100 mg/kg/day) *3 LDL peroxidation inhibition (concentration 10- 5

M)

*4 plasma lipoperoxidation inhibition (concentration 10s

M)

-36- Table 11 Test compound Result of Exp Ex.

*1 Result of Exp. Ex. 2 Wz *2 I 4- 9* t.

a*.

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Example 24 Example 25 Example 26 Example 27 Example 28 Example 29 Example 30 Example 31 Example 32 Example 33 Example 34 Example 35 Example 36 97.1 96.8 26.5 95.5 79.4 86.6 83.0 93.8 93.9 82.2 91.0 84.6 95.9 92.3 3.4 28.3 50.5 37.4 56.9** 20.8 6.9 25.7 49.0 57.4 7.2 50.2 20.8 50.2 43.8 7.3* Result of Exp. Ex. 3 W% *3 42.2 92.2 43.3 92.2 92.2 91.9 94.1 90.3 85.3 87.0 82.9 71.9 83.2 0 89.4 Result of Exp. Ex. 4 W% *4 90.5 93.1 90.1 91.1 91.1 91.1 91.8 90.8 89.1 89.5 89.8 91.4 89.1 YM-750 Probucol 87.5 I. I J _I *1 ACAT inhibition (concentration 10-1 M, *:10-5 M) *2 :reduction of serum total cholesterol (dose *:10 mg/kg/day, mg/kg/day, 100 mg/kg/day) *3 LDL peroxidation inhibition (concentration 10-5 M) *4 plasma lipoperoxidation inhibition (concentration 10-5 M) YM-750 1 -cycloheptyl- luorenyl)methyl] 6-trimethylphenyl) urea Probucol 4,4' -isopropylidenedithiobis 6 -di-t--butylphenoi) 37 Formulation Example 1 Tablets having the following composition are prepared by a conventional method.

Compound of Example 3 25 mg Polyvinylpyrrolidone 20 mg Starch mg Magnesium stearate 2 mg Formulation Example 2 Capsules having the following composition are prepared by a conventional capsule packing method.

Compound of Example 6 100 mg Lactose 25 mg Magnesium stearate 1 mg The heterocyclic derivatives and pharmaceutically acceptable salts thereof of parent patent application no.

35324/95 show superior ACAT inhibitory activity and lipoperoxidation inhibitory activity, and are useful as ACAT inhibitors of hyperlipermia inhibitors. Specifically, they are useful for the prevention and treatment of arteriosclerotic lesions such as arteriosclerosis, hyperlipemia and diabetes, as well as ischemic diseases of brain, heart and the like.

-38- 39 THE CLAIMS DEFINING THE INVENTION ARE AS FOLLOWS: 1. l-Acetyl-5-bromo-4, 6 Odimethylindoline Dated this 24th day of March 1999 SANKYO COMPANY

LIMITED

By their Patent Attorney GRIFFITH

HACK

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