WO2004048366A1

WO2004048366A1 - 2-oxoindoline derivatives

Info

Publication number: WO2004048366A1
Application number: PCT/JP2003/014736
Authority: WO
Inventors: Kiyohiro Samizu; Hiroyuki Hisamichi; Akira Matsuhisa; Yukitaka Ideyama; Sadao Kuromitsu
Original assignee: Yamanouchi Pharmaceutical Co., Ltd.
Priority date: 2002-11-22
Filing date: 2003-11-19
Publication date: 2004-06-10
Also published as: AU2003284572A1

Abstract

It is intended to provide vascular endothelial growth factor inhibitors useful as drugs, in particular, remedies for diseases in which angiogenesis participates, for example, solid cancer and diabetic retinopathy. Namely, 2-oxoindoline derivatives represented by the following general formula wherein a quinoline ring is directly bonded at the 2-position to the 3-position of an indoline ring and a double bond is isomerized have a favorable VEGF inhbitory effect, an angiogenesis inhibitory effect and an antitumor effect. Thus, these compounds are useful in regulating cancer, in particular, solid cancer and tumor such as angioma, and as preventives/remedies for diseases such as rheumatoid arthritis, psoriasis, scleriasis and carcinomatous ascitic/pleural effusion as well as preventives/remedies for retinal diseases such as diabetic retinopathy and ophthalmic diseases such as angiogenic glaucoma. wherein the substituents R1, R2, R3 and G are each as defined in claim 1.

Description

Specification

2-oxoindoline derivatives

The present invention has a vascular endothelial cell growth factor (VEGF) inhibitory activity and a cancer growth inhibitory activity, and is useful as a therapeutic agent for diseases involving angiogenesis, such as cancer and diabetic retinopathy. Oxoindoline derivatives. Background art

Some diseases are known to be associated with pathological angiogenesis that is closely linked to their symptoms and etiology. A typical disease is cancer, particularly solid cancer.In order for cancer tissue to grow beyond 1-2 mm in diameter, it is necessary for new blood vessels to extend from existing blood vessels to reach the cancer tissue. J. Natl. Cancer Inst, 82, 4 (1990)), and when blood vessels reach cancerous tissue, the growth of cancerous tissue is explosively accelerated. Also, diabetic retinopathy is associated with pathological neovascularization of the retina, which often causes blindness. Diseases such as rheumatoid arthritis, psoriasis, hemangiomas, scleroderma, and neovascular glaucoma also involve pathological angiogenesis, which is one of the main symptoms (N. Engl. J. Med. , 320, 121 1 (1989)). Therefore, substances that inhibit angiogenesis may be used for the treatment of solid cancers and the aforementioned diseases.

Vascular endothelial cells are cells that form the innermost layer of blood vessels. Angiogenesis is performed by proliferating vascular endothelial cells under stimulation from growth factors, physiologically active substances, or physical damage. Several growth factors that directly or indirectly stimulate the proliferation of vascular endothelial cells are known, but vascular endothelial cells are distinguished from other growth factors in that they act very specifically on vascular endothelial cells. Growth factors (VEGF) are known. That is, it has been reported that VEGF receptors are expressed only in very limited cells other than vascular endothelial cells and are selective for vascular endothelium (J. Clin. Invest., 89, 244-253 (1992)).

The following reports suggest a relationship between VEGF and cancer. Many cancer cells secrete VEGF (Biochem. Biophys. Res. Commun., 194, 1234 (1993)). When a cancer tissue section is stained with an anti-VEGF antibody, the cancer tissue and its surrounding new blood vessels are strongly stained (Exp. Med., 174, 1275 (1991), Cancer Res., 53, 4727 (1993)). One of the VEGF receptors In a genetically inactivated mouse, the growth of the transplanted cancer is suppressed (Nature, 367, 576 (1994)). Anti-VEGF neutralizing antibodies show antitumor activity against tumor-bearing mice (Nature, 362, 841 (1993), Biochem. Biophys. Res. Commun., 194, 1234 (1993)). These facts suggest that VEGF secreted by cancer cells plays a central role in tumor angiogenesis. VEGF is also known to be involved in the enhancement of vascular permeability, and is considered to be one of the factors that cause cancerous ascites and pleural effusion.

Two types of VEGF receptors are known in humans, Flt-1 and KDR / Flk-1 (FASEB III, 13, 9-22 (1999)). The results of these two gene disruptions indicate that Rt-1 is involved in normal endothelial cell differentiation and morphogenesis, and Flk-1 is involved in endothelial cell formation and proliferation.

(Nature, 376, 66-70 (1995), Nature, 376, 62-66 (1995)), Japanese Pharmacological Journal, 107, 119-131 (1996)). VEGF is thought to bind to the Flk-1 receptor and promote the proliferation of vascular endothelial cells via a signal transmission mechanism via tyrosine kinase (Proc. Natl. Acad. Sci. USA, 88, 9026-9030 ( 1991)). Furthermore, it has been shown that VEGF has an angiogenesis-inducing activity directly on endothelial cells in vitro (丄 Cell. Physiol, 149, 50-59 (1991)).

Therefore, VEGF inhibitors, which inhibit the binding of VEGF to VEGF receptors (especially Flk-1) or inhibit VEGF signal transmission, suppress angiogenesis and cancerous ascites, and are useful for the treatment of cancer, especially solid cancer. Is expected to be.

As VEGF inhibitors, anti-VEGF human monoclonal antibodies (JP-A-9-316099) and some polypeptides (JP-A-9-255700, JP-A-9-154588) have been reported. Recently, VEGF inhibitors such as SU6668 (Cancer Res., 60, 4152-4160 (2000)) and PTK787 / ZK222584 (Cancer Res., 60, 2178-2189 (2000)) represented by The following orally administrable low molecular weight compounds have been reported.

Further, as compounds useful as tyrosine kinase inhibitors and angiogenesis inhibitors, quinazoline-substituted oxyindole derivatives (W097 / 42187 and WO99 / 10349), Or pyrrolotriazine-substituted indoline-2-one derivatives (WO00 / 71129). However, no specific pharmacological data is disclosed.

On the other hand, as a 2-oxoindrin derivative, there is a report on a method for synthesizing 3-quinolin-2 (1H) -ylidenindrin-2-one (Annali di Chimica (Rome), 57 (6), 688) -97 (1967), Chemical & Pharmaceutical Bulletin, 18 (9), 1822-30 (1970), Chemical & Pharmaceutical Bulletin, 19 (8), 1669-80 (1971)). However, there is no disclosure of its pharmaceutical uses. Disclosure of the invention

There is still a strong need for the creation of vascular endothelial cell growth factor (VEGF) inhibitors, especially orally administrable drugs, that are useful as therapeutics for cancers involving angiogenesis, especially solid cancers and diseases such as diabetic retinopathy. I have.

The present inventors have conducted intensive studies on compounds that inhibit angiogenesis based on VEGF inhibitory action.As a result, the 2-position of the quinoline ring and the 3-position of the indolinone ring were directly bonded, and the double bond was isomerized. Quinoline-2 (1H) -ylidenindrin-2-one derivatives have good VEGF inhibitory activity and are useful as preventive or therapeutic agents for diseases associated with angiogenesis involving VEGF An application was filed earlier (PCT / JP02 / 05014; International Publication No. 02/94809 pamphlet). Furthermore, the present inventors have studied various compounds having the skeleton, found a specific compound having a good VEGF inhibitory action and a cancer growth inhibitory action, and completed the present invention.

That is, the present invention relates to 2-oxoindoline derivatives represented by the following formulas A to F or salts thereof. The present invention also relates to a pharmaceutical composition comprising a 2-oxoindoline derivative represented by the following formulas A to F or a salt thereof and a pharmaceutically acceptable carrier, in particular, a vascular endothelial cell growth factor inhibitor The present invention also relates to a pharmaceutical composition which is an angiogenesis inhibitor or an anticancer agent.

table 1

(In the table, ALK represents a C1-6 alkyl group, and R represents an H or a C1-6 alkyl group.) Among the compounds of the present invention, preferred compounds include the following 2-oxoindoline derivatives or salts thereof. No.

3- [6-[(4-methylbiperazin-1-yl) methyl] quinoline-2 (1H) -ylidene 2-oxoindolin-6-carbaldehyde 0- (pyridine-2-ylmethyl) oxime or a salt thereof,

3- [6-[(4-Ethylbiperazin-1-yl) methyl] quinoline-2 (1H) -ylidentate 2-oxoindolin-6-carbaldehyde 0- (pyridine-2-ylmethyl) oxime or a salt thereof,

3- [6-[(4-methylbiperazin-1-yl) methyl] quinoline-2 (1H) -ylident 2-oxoindolin-6-carbaldehyde 0- (1,3-thiazol-4-ylmethyl) oxym Or a salt thereof,

3- [6-[(4-Ethylbiperazine-1-ynole) methyl] quinoline-2 (1H) -ylidene 2-oxoindolin-6-carbaldehyde 0- (1,3-thiazol-4-ylmethyl) oxime or Its salt,

3- [2- (2-oxo-6-{[(pyridine-2-ylmethoxy) imino] methyl} indoline-3-ylide -1), 2-Dihydroquinoline-6-yl] propionic acid or a salt thereof,

3- (1,5-naphthyridine-2 (1H) -ylidene) -2-oxoindolin-6-carbaldehyde 0- (pyridin-2-ylmethyl) oxime or a salt thereof,

4- {1- [3- (2- {6-[(Methoxyimino) methylto 2-oxo-1,2-dihydro-3H-indole-3-ylidene} -1,2-dihydroquinoline-6-di L) propyl] piperidin-4-yl} ethyl butyrate or a salt thereof, 3- {1- [3- (2- {6-{(methoxyimino) methyl] -2-oxo-1,2-dihydro -3H-indole-3-ylidene} -1,2-dihydroquinoline-6-yl) propyl] piperidin-4-yl} propionic acid or

3- {1- [3- (2- {6-[(Methoxyximino) methyl] -2-oxo-1,2-dihydro-3H-indole-3-ylidene} -1,2-dihydroquinoline- 6-yl) propyl] piperidin-4-yl} methyl propionate or a salt thereof, and

3- [6- {3- [4- (3-Methoxy-3-oxopropyl) piperidin-1-yl] propyl} quinoline-2 (1H) -ylidene] -5-methyl-2-oxoindoline-6 -Methyl carboxylate or a salt thereof.

In the present invention, the C1-6 alkyl group is a linear or branched alkyl group having 1 to 6 carbon atoms, preferably an alkyl group having 1 to 4 carbon atoms, more preferably Methyl and ethyl groups.

The compound of the present invention has a plurality of tautomers or stereoisomers that are theoretically possible in a conjugated system linked from the nitrogen atom at position 1 of the quinoline ring to the nitrogen atom at position 1 of the indolinone ring. The invention includes a separated form or a mixture of these isomers.

The compound of the present invention may further have a geometric isomer or a tautomer depending on the type of the substituent, and the present invention includes a separated form or a mixture of these isomers. Further, the compound of the present invention may form a salt. Here, the salt is a pharmaceutically acceptable salt, and the acid addition salt is, specifically, an inorganic acid such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid and the like. Organic acids such as, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, tansulfonic acid, aspartic acid, and glutamic acid Examples of the salt with a base include inorganic bases containing metals such as sodium, potassium, magnesium, calcium, and aluminum, and methylamine, ethylamine, ethanolamine, lysine, orditin. And the like, salts with organic bases such as and the like, and ammonium salts. further, The present invention also includes various hydrates, solvates, and polymorphs of the compound of the present invention and salts thereof.

The compounds of the present invention also include physiologically acceptable prodrugs. The pharmacologically acceptable prodrug, substituents of the present invention by solvolysis or under physiological conditions, is a compound having a group which is converted into for example C0 ₂ H or the like. Prodrug-forming groups include those described in Prog. Med., 5, 2157-2161 (1985) and “Development of Drugs” (Hirokawa Shoten, 1990), Volume 7, Molecular Design 163-198. .

(Manufacturing method)

The compound of the present invention can be prepared by methods described in the literature, for example, Chem. Pharm. Bull., 18 (9), 1822-30 (1970), J. Am. Chem. Soc, 122 (7), 1360-70 (2000) And the like, or by applying a method known to those skilled in the art. Depending on the type of functional group, it may be effective in production technology to replace the functional group with an appropriate protecting group at the stage of raw materials or intermediates, that is, a group that can be easily converted to the functional group. is there. Thereafter, the desired compound can be obtained by removing the protecting group, if necessary. Examples of such a functional group include an amino group, a hydroxyl group, a carboxyl group, and the like. Examples of such a protective group include, for example, rprotective Groups in Organic Synthesis, by Greene and Wuts. The protecting groups described in the third edition can be mentioned, and these may be appropriately used according to the reaction conditions.

Hereinafter, a typical manufacturing method will be described.

(I)

(In the formula, R ⁴ represents a protecting group such as diethoxymethyl, P-toluenesulfonyl, and trimethylsilylethyl sulfonyl, and represents a leaving group applicable to the reaction such as halogen and sulfonate. The same applies hereinafter. )

First manufacturing method

The compound (I) of the present invention can be produced by reacting an indolinone (V) with a quinoline N-amine oxide compound (II) according to a conventional method. The reaction is carried out, for example, in Ann. Chim. (Rome), 57 (6), 188-97 (1967), Khim. Geterotsikl. Soedin., 10, 1371-3 (1970), Chem. Pharm. Bull., 18 ( 9), 1822-30 (1970), and the method described in Chem. Pharm. Bull., 19 (8), 1669-80 (1971). For example, an excess of either (II) and / or (V) or a corresponding amount of the compound (II) and / or (V) in an appropriate amount as an activator is used as an activator. ), A sulfonylating agent (such as P-toluenesulfonyl chloride), an alkylating agent (such as methane iodide) or a silylating agent (such as octatrimethylsilane) at room temperature or under heating, preferably at room temperature. It is advantageous to work at the reflux temperature. When acetic anhydride is used, it is advantageous to use it as a solvent, and it is preferable to use acetic anhydride at normal temperature to heating. Second manufacturing method

In the first step, a solvent inert to the reaction (for example, toluene, tetrahydrofuran (THF), etc.) according to the method described in J. Am. Chem. Soc "122 (7), 1360-70 (2000), etc. A palladium complex (e.g., palladium acetate, e.g., palladium acetate, Compound (III) can be produced by treating with palladium chloride, dibenzylideneacetone dipalladium, etc. If necessary, a ligand of a palladium complex

In some cases, the addition of (eg, BINAP, DPPF, Xantphos, etc.) favors the reaction. Next, in the second step, the compound was prepared according to a method described in W097 / 42187 and the like.

(III) is deprotected in the presence of an acid such as hydrochloric acid, or in the presence of a reducing agent (for example, tributyltin hydride) according to the method described in Tetrahedron, 56 (7), 979-988 (2000). By doing so, compound (I) can be produced.

Third manufacturing method

(In the formula, hal represents halogen, R ⁵ represents lower alkyl. The same applies hereinafter.)

The first step can be easily performed according to known reaction conditions (eg, Med. Chem., 42, 5120-5130 (1999) and the like). In a solvent inert to the reaction (eg, N, N-dimethylformamide (DMF), dimethylsulfoxide (DMSO), THF, etc.), excess amounts of the compounds (VIII) and / or (IX) in a reaction equivalent amount are added. The compound (X) can be produced by reacting the compound (X) in the presence of a base (eg, sodium hydride (NaH), sodium tert-butoxide, etc.) or an acid (eg, acetic acid) at room temperature or under heating. it can. Next, in the second step, the compound (X) is subjected to reduction according to a conventional reduction reaction, for example, according to the method described in Med. Chem., 42, 5120-5130 (1999), etc. (I) can be manufactured. If necessary, heating or pressurization may promote the reaction in an advantageous manner. During this time, it is also possible to convert the substituent of compound (X) by applying the usual conditions. For example, when R ¹ and R ² are aldehydes, ketones, etc., they can be converted to lyoxime compounds or the like by a condensation reaction or the like. Other manufacturing methods

The compound of the present invention can be produced by various known substituent modification reactions in addition to the above-mentioned production methods. For example, B. M. Trost: COMPREHENSIVE ORGANMC SYNTHESIS (Pergamon Press) (1991), R. C. Larock: COMPREHENSIVE ORGANIC

TRANSFORMATIONS (VCH Publishers) (1989), J. March: ADVANCED ORGANIC CHEMISTRY (John WILEY & SON) (1992) It can be easily manufactured by referring to the conditions described above. The main production methods are described below.

Compounds having a substituent containing an aminoalkyl group can be obtained from (1) a compound having a halogen-substituted alkyl group or an epoxide by a conventional amination reaction, and (2) a compound having an aldehyde or a ketone. By a conventional reductive amination reaction (for example, see Tetrahedron Lett., 31, 5595-5598 (1990), etc.), (3) a deprotection reaction from a compound having a protected aminoalkyl group (for example, , Tert-butoxycarponyl group

In the case of (Boc), it can be easily produced by treatment with hydrochloric acid or trifluoroacetic acid (TFA), and in the case of a phthalimide group, treatment with hydrazine or methylamine.

In the reductive amination reaction, when one of them is difficult to proceed with ketone or secondary amine or a combination of ketone and secondary amine, for example, 丄 Org. Chem., 55 (8), 2552 (1990 ) Is preferably produced by a method similar to that described in (1).

A compound having a carboxyl group can be produced from a compound having an ester group by a conventional hydrolysis reaction.

The compound having a substituent containing oxime can be produced from a compound having an aldehyde or ketone by a conventional dehydration condensation reaction using hydroxylamines or the like.

The N-oxide compound is produced by a well-known oxidation reaction, that is, a reaction with an oxidizing agent such as m-chloroperbenzoic acid or hydrogen peroxide in a solvent inert to the reaction (for example, chloroform or dichloromethane). be able to. Under the same oxidation conditions, it is also possible to convert sulfide to sulfoxide or sulfone. When a desired compound is obtained by performing a dehydroxylation reaction with a precursor compound having an N-hydroxyamide bond, a conventional reduction method is used. It can be easily carried out by passing through conditions (eg, reaction with metallic iron in acetic acid, hydrogenolysis reaction, etc.). The introduction of a heteroaromatic ring can be easily carried out by a method of introducing a substituent serving as a precursor thereof and then converting it into a heterocyclic ring by a conventional condensation reaction.

(Synthesis of starting compounds)

Some of the starting compounds of the compound of the present invention are novel compounds, and these compounds can be easily synthesized in the same manner as known starting compounds or by using methods known to those skilled in the art. Representative synthetic methods are shown below.

Synthesis method 1 (oxidation) Reference: Synthesis, 87-90 (1997), etc.

Synthetic method 2 Reference: 丄 Heterocyclic Chem., 15, 1425-1430 (1978), etc.

The quinolineacetic acid derivative (IX) can be prepared by converting compound (II) with ethyl acetate or malonic acid diester in the presence of a suitable acylating agent, sulfonylating agent, alkylating agent or silylating agent according to a conventional method. It can be manufactured by processing.

Synthesis method 3 (halogenation) References: 丄 Am. Chem. Soc, 77, 1054-1055 (1955), Tetrahedron, 54, 13655-13680 (1998), etc.

Method for synthesizing other starting compounds

Compounds having a substituent on the quinoline ring can be prepared, for example, by the method described in Heterocycles, 54, 105-108 (2001), 丄 Med. Chem., 26, 580-585 (1983), or Org. Synth. Col. Vol. 3, 272 (1955), Syn. Commun., 15, 125 (1995), etc. to produce 4-chloro quinoline derivatives Thereafter, it can also be produced by adapting a method of removing the chloro group through a reducing condition or the like by a conventional method.

The indolinone ring can be easily prepared by applying the conditions described in Synthesis, 51-53 (1993) or Eur. J. Med. Chem "15, 330-332 (1980).

The introduction of substituents on the indolinone ring involves the Suzuki Ichinomiyaura reaction with the halogen derivative, the Friedel Crafts reaction, and the conversion reaction to the heteroaromatic ring through the condensation reaction using the introduced -haloketones. It can be done by doing. For example, the method described in J. Med. Chem., 42, 5120-5130 (1999), Synthesis, 873-874 (1989), J. Org. Chem., 17, 1252-1255 (1952), etc. is applied. It is possible. Further, the primary amine on the introduced heteroaromatic ring can be removed, for example, by applying the method described in Med. Chem., 39, 834-841 (1996).

Further, if necessary, amination, iminoization, acylation, alkylation, amidation, sulfonamidation, esterification, ureation, halogenation, nitration, oxidation, reduction, protection, deprotection, etc. By subjecting the substituent to a known substituent modification reaction, a desired starting compound can be produced. These reactions can be performed, for example, by referring to the conditions described in the literature such as the above-mentioned “Chemical Chemistry Lecture”, 4th edition (Maruzen), edited by The Chemical Society of Japan.

Isolation and purification of the compound of the present invention thus produced is carried out by applying ordinary chemical operations such as extraction, concentration, distillation, crystallization, filtration, recrystallization, and various types of chromatography. Various isomers can be isolated by a conventional method using the difference in physicochemical properties between the isomers. For example, a racemic compound can be converted to an optically pure isomer by a general optical resolution method [for example, to a diastereomer salt with a general optically active acid (tartaric acid, etc.) and to perform optical resolution]. Can be. In addition, a mixture of diastereomers can be separated by, for example, fractional crystallization or chromatography. The optically active compound can also be produced by using an appropriate optically active raw material. A pharmaceutical composition comprising the compound of the present invention or a salt thereof and a pharmaceutically acceptable carrier is prepared by a method usually used using pharmaceutical carriers, excipients and the like usually used in the art. can do. For oral administration, such as tablets, pills, capsules, granules, powders, solutions, inhalants, etc., or injections such as intravenous and intramuscular injections, suppositories, eye drops, eye ointments, transdermal It may be in any form of parenteral administration using a liquid preparation, ointment, transdermal patch, transmucosal solution, transmucosal patch, or the like.

As a solid composition for oral administration, tablets, powders, granules and the like are used. In such a solid composition, the one or more active substances comprise at least one inert diluent, such as lactose, mannitol, glucose, hydroxypropylcellulose, microcrystalline cellulose, starch, polyvinylpyrrolidone, metasilicate. It is mixed with magnesium aluminate. The composition contains, in a conventional manner, additives other than inert diluents, for example, lubricants such as matanedium stearate, disintegrants such as calcium cellulose glycolate, stabilizers, and solubilizers. May be. If necessary, tablets or pills may be coated with sugar such as sucrose, gelatin, hydroxypropylcellulose, hydroxypropylmethylcellulose phthalate, or a film of a gastric or enteric substance. Liquid compositions for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, elixirs and the like, and commonly used inert diluents such as Contains purified water and ethanol. The composition may contain, in addition to the inert diluent, adjuvants such as wetting agents and suspending agents, sweetening agents, flavoring agents, fragrances, and preservatives.

Injections for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Aqueous solutions and suspensions include, for example, distilled water for injection and physiological saline. Examples of the water-insoluble solutions and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, alcohols such as ethanol, and polysorbate 80 (trade name). Such compositions may also contain adjuvants such as preserving, wetting, emulsifying, dispersing, stabilizing, and solubilizing agents. These are sterilized by, for example, filtration through a bacteria retaining filter, blending of a bactericide or irradiation. They can also be used to produce a sterile solid composition which is dissolved in sterile water or a sterile solvent for injection before use.

The dose of the compound of the present invention is generally about 0.001 to 50 mg Z kg per day for oral administration, preferably 0.0 "! To 1 O mg Z kg when administered intravenously. The appropriate daily dose is about 0.001 to 5 mg Z kg, which should be administered once or more times a day, depending on symptoms, age, sex, etc. Is determined as appropriate according to the individual case. Industrial applicability

INDUSTRIAL APPLICABILITY The compound of the present invention has a VEGF inhibitory effect, and is useful for improving the therapeutic effect of diseases and conditions associated with VEGF. In particular, as an inhibitor of angiogenesis caused by VEGF, it can be used to suppress the growth of tumors such as cancer, especially solid tumors and hemangiomas, to prevent and treat diseases such as rheumatoid arthritis, psoriasis and scleroderma, and to diabetes It is useful for the preventive treatment of retinal diseases such as retinopathy and ocular diseases such as neovascular glaucoma.

As shown in Test Examples described later, the compound of the present invention well inhibited the signal transmission of VEGF via tyrosine kinase, and had a favorable inhibitory activity on the proliferation of vascular endothelial cells induced by VEGF. Furthermore, in a cancer growth inhibition test using COLO205 (human colorectal cancer) -bearing nude mice, it was confirmed that the compound of the present invention strongly inhibited cancer growth even at a low dose of oral administration. It is useful as a newborn inhibitor and an anticancer agent.

Further, the compound of the present invention suppresses the increase in vascular permeability caused by VEGF, and is also useful as an agent for improving cancerous ascites / pleural effusion.

The effect of the compound of the present invention was confirmed by the following test examples.

Test example 1 KDR kinase inhibition test

1) Expression of KDR kinase domain

The KDR intracellular region (amino acids 790 to 1168) was amplified by PCR from cDNA prepared from Human umbilical vein endothelial cells (HUVEC). The gene in which the FLAG ™ (trademark of Sigma-Aldrich Co.) sequence (DYKDDDDK) was introduced at the C-terminus was cloned into the BamHI and Notl sites of pFASTBAC1 (formerly G-COBRL). Recombinant vaccum virus was prepared according to the manual of Bacto-To-Bac expression system (G formerly made by COBRL). Sf-9 cells were infected with the recombinant baculovirus for protein expression, and the cells were collected 72 hours later.

2) Purification of KDR kinase domain

Sf-9 cells expressing the KDR kinase domain were sonicated in a buffer solution (20 mM Tris, 150 mM NaCI, 1 mM PMSF (phenylmethanesulfonyl fluoride), 10 jig / ml aprotinin), and 10,000 rpm, 4 ° C, The supernatant was obtained after 30 minutes of centrifugation. After binding the KDR kinase domain in the supernatant to M2-agarose (manufactured by Sigma), it was eluted with 0.1 mg / ml of Flag peptide. Purified The KDR kinase domain was replaced with a storage buffer (20 mM Tris, 150 mM NaCI, 10% glycerol) by dialysis, and then stored at -80 ° C.

3) In vitro o KDR kinase assay by HTRF

Homogeneous, time-resolved fluorescence (HTRF) is applicable to the measurement of tyrosine kinase activity, the theory of which is reported in Clin. Chem. 41, 1391-1397 (1995). The test compound was adjusted to 50 times the final concentration with dimethyl sulfoxide (DMSO). 1 After adding the compound solution of 1 I to the well, the reaction buffer (50 mM HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid) pH7.5, 1 mM gCI ₂ , 4 mM MnCI ₂ 0.1 25 〃I of 100 ng of purified KDR kinase domain diluted with% BSA (bovine serum albmin) was added. The ATP solution stored at −20 ° C. at 100 mM was diluted to 2 μM with a reaction buffer, and the kinase reaction was started by adding 25 I to the well. After 20 minutes of reaction at room temperature, the reaction was stopped by adding 10M 0.5M EDTA. Subsequently, 6.5 ng of Cryptate-labeled anti-phosphorylated ticin antibody (PT66) (Cis Bio International) diluted with a detection antibody dilution buffer (50 mM HEPES pH 7.5, 0.1% BSA, 0.5 M KF) and 100 ng of XL665-labeled anti-FLAG ™ (M2) antibody (Cis Bio International) was added at 50/1, and the mixture was incubated at room temperature for 2 hours. The amount of phosphorylation was detected using Discovery (Packard). The ratio of the value measured by Discovery when DMSO was added was set to 100%, the ratio when ATP was not added was set to 0%, and the concentration at which the test compound inhibited by 50% was calculated as the ₅₀ value of the compound inhibitory activity.

Results: The results are shown in the table below. The compounds of the present invention successfully inhibited phosphorylation by KDR kinase. Therefore, it was confirmed that the compound of the present invention successfully inhibited signal transduction of VEGF via tyrosine kinase, and was useful as a VEGF inhibitor.

Table 2

Test Example 2 In vivo tumor growth inhibition test using human colon cancer-bearing nude mice

Test method: 3 to 4 × 10 ⁶ COLO205 cells, which are human colorectal cancer, were subcutaneously administered to the dorsal side of female Balb / c nude mice. Test compounds or when the tumor volume reached 50 to 150 mm ³ Orally once daily for 14 days. A 0.5% aqueous solution of methylcellulose was orally administered to the control group. The diameter of the tumor was measured using a caliper, the day after the final administration. The tumor capacity was calculated using the following formula.

Tumor volume = (minor axis ² X major axis) no 2

Results: In this test, the compounds of Examples 1, 2, 3, 4, 7, 8, and 9 of the present invention showed significant inhibition of cancer growth at 3 or 10 mg / kg / day orally after administration. It was confirmed to have antistatic activity.

Therefore, it was shown that the compound of the present invention has a favorable VEGF inhibitory activity and a cancer growth inhibitory activity, and is useful as an angiogenesis inhibitor and an anticancer agent. In particular, it has a good cancer growth inhibitory effect even when administered orally at a low dose, and thus is useful as an orally administrable anticancer agent. BEST MODE FOR CARRYING OUT THE INVENTION

Hereinafter, Production Examples of the compound of the present invention will be described in Examples, and Reference Production Examples of the compound of the present invention, Production Examples and Reference Examples of the starting compounds will be shown in Reference Examples.

Abbreviations of physicochemical properties described in Reference Examples and Tables below are: F +: FAB-MS (+ H) ⁺ ; F-: FAB-MS (MH)-; F: FAB-MS (M) +; N1: ¹ H-NMR (DMSO-d ₆₎ TMS internal standard) characteristic peak (5 ppm; N2: ¹ H-NMR (CDCI ₃ , TMS internal standard) characteristic peak S ppm Show.

Reference Example 1: Methoxylamine hydrochloride was added to a THF solution of 4-bromo-2-methyl-5-nitrobenzaldehyde, and the mixture was stirred at 50 ° C for 8 hours. The product was purified from the reaction solution to obtain colorless oily 4-bromo-2-methyl-5-nitrobenzaldehyde O-methyloxime. F-: 272,274. Reference Example 2: To a solution of 6- (2-bromoethoxy) quinoline in ethyl acetate was added 70% m-hydroxyperbenzoic acid, followed by stirring. The resulting precipitate was collected by filtration to obtain a pale yellow solid, 6- (2-bromoethoxy) quinoline N-oxide. F +: 268, 270.

Reference Example 3: To a mixed solution of 3-quinoline-6-ylpropan-1-ol in dichloromethane / DMSO, TEA and iodopyridine trioxide complex were added under ice-cooling, and the mixture was purified by stirring. 6-ylpropanal N2: 2.87-2.93 (2H, m), 3.15 (2H, t), 7.39 (1H, dd), 7.57 (1H, dd) _> 7.61 (1H, s) _! 8.05 ( 1 H, d) _l 8.08-8.12 (1 H, m), 8.87 (H, dd), 9.86 (1 H, t) o Reference Example 4: 4-Hokuguchi-2-Methoxy-5-nitrobenzoic acid was added to an ethanol solution of concentrated sulfuric acid, and the mixture was purified by stirring under reflux and purified as a colorless solid. This gave ethyl-5-nitrobenzoate.

F +: 260 _o

Reference Example 5: After dimethyl malonate was added dropwise to a suspension of 60% NaH in DMSO, the mixture was stirred at 100 ° C. After cooling to room temperature, N- (2,5-dichloro-4-nitrophenyl) acetamide was added, and the mixture was stirred at the same temperature and then at 100 ° C. After purification of the product, the product was recrystallized from ethyl acetate to obtain a colorless solid of [5- (acetylamino) -4-kuguchi-2--2-nitrophenyl] dimethyl dimethyl. F: 344.

Reference Example 6: Anhydrous lithium chloride and water were added to a DMSO solution of [5- (acetylamino) -4-kuguchi-2--2-nitrophenyl] malonate, and the mixture was stirred at 100 ° C. After cooling, the reaction solution was poured into a mixture of ethyl acetate and saturated saline, and extracted with ethyl acetate. The organic layer was washed, concentrated, and the resulting crude crystals were recrystallized from methanol to give methyl [5- (acetylamino) -4-chloro-2--2-nitrophenyl] acetate as a colorless solid. F: 286.

Reference Example 7: A 6M hydrochloric acid solution of getyl (4-formyl-2-ditrophenyl) mamate was stirred under reflux. After the reaction solution was cooled with ice, the resulting precipitate was collected by filtration and washed to obtain brown solid of (4-formyl-2-ditrophenyl) acetic acid. F +: 210.

REFERENCE EXAMPLE 8 Reduced iron was added to an acetic acid solution of methyl [5- (acetylamino) -4-kuguchi-2--2-nitrophenyl] acetate, and the mixture was stirred at 100 ° C. After cooling, the reaction solution was filtered through celite and washed with DMF. After concentrating the filtrate, water was added, and the resulting precipitate was collected by filtration and washed with water to obtain a colorless solid of N- (6-clomouth-2-oxoindoline-5-yl) acetamide. . F +: 225.

Reference Example 9: 3- [6- (2-Promoethoxy) quinoline-2 (1H) -ylidene] indolin-2-one (1.86 g) was suspended in acetonitrile (100 ml), and morpholine (2.11 g) was added. The mixture was stirred at 80 ° C for 4 hours. After cooling, a saturated aqueous solution of sodium hydrogencarbonate and saturated saline were added to the reaction solution, and the mixture was extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate, and the solvent was distilled off. The residue was purified by silica gel column chromatography (hereinafter abbreviated as SCC) (eluted with methanol-ethyl acetate-28% aqueous ammonia), and the obtained solid was recrystallized from ethanol to give a red solid 960 mg of [6- (2-morpholine-4-ylethoxy) quinoline-2 (1H) -ylidene] indrin-2-one was obtained. N1: 2.51 (4H, t), 2.78 (2H, t), 3.59 (4H, t), 4.16 (2H, t), 10.53 (1 H, s), 14.46 (1 H, s); F +: 390. Reference Example 10: A solution of 2- (2-oxoindoline-3-ylidene) -1,2-dihydroquinoline-6-carbaldehyde (0.5 g) and 2-morpholine-4-ylethylamine (0.91 mL) in dichloroethane (35 mL) was added. Then, acetic acid (0.99 mL) was added, and the mixture was stirred at room temperature for 2 hours. To the reaction solution was added sodium triacetoxyborohydride (1.1 g), and the mixture was stirred at room temperature for 13 hours. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted with dichloroethane. The organic layer was washed with water and saturated saline, dried over anhydrous magnesium sulfate, and the solvent was distilled off. The residue was purified by SCC (eluted with chloroform), the resulting solid was collected by filtration, washed with ethyl acetate, and the orange solid 3- (6-{[(2-morpholine-4- 168 mg of [ylethyl) amino] methyl} quinoline-2 (1H) -ylidene) indolin-2-one were obtained. N1: 3.55 (4H, t), 3.78 (2H, s), 10.56 (1 H, s); F +: 403.

Reference Example 11: 2- (2-oxoindoline-3-ylidene) -1,2-dihydroquinoline-6-carbaldehyde (0.6 g), dichloroethane of N- (2-methoxethyl) -N-methylamine (0.89 mL) (3 mL) Titanium tetraisopropoxide (0.68 mL) was added to the solution, and the mixture was stirred at room temperature for 1 hour. After cooling on ice, sodium triacetoxyborohydride (1.32 g) was added to the reaction solution, and the mixture was stirred at room temperature for 1.5 hours. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted with dichloroethane. The organic layer was washed with water and saturated saline, dried over anhydrous magnesium sulfate, and the solvent was distilled off. The residue was purified by SCC (eluted with chloroform), the resulting solid was collected by filtration, washed with ethyl acetate, and the red solid 3- (6-{[N- (2-methoxethyl)- There were obtained 114 mg of N-methylamino] methyl} quinolin-2 (1H) -ylidene) indolin-2-one. N1: 2.19 (2H, s), 2.25-2.60 (4H, m), 3.53 (2H, s), 10.57 (1 H, s), 14.38 (1 H, s); F +: 361.

Reference Example 12: To a solution of 6-methoxyindoline-2-one (200 mg) in acetic anhydride (10 mL) was added quinolin 1-oxide (214 mg), and the mixture was stirred at 50 ° C for 5 hours. After cooling, the mixture was poured into iced water and extracted with ethyl acetate. The organic layer was washed with a saturated aqueous solution of sodium hydrogen carbonate, water and saturated saline, dried over anhydrous magnesium sulfate, and the solvent was distilled off. The residue was purified by SCC (eluted with chloroform) and crystallized from getyl ether. The obtained crystals were collected by filtration, and washed with a filter form to obtain 30 mg of a red solid, 6-methoxy-3-quinolin-2 (1Η) -ylidenindolin-2-one. F-: 279.

Reference Example 13: 4- (2-Oxo-3-quinoline-2 (1H) -ylideneindrin-5-yl) ethyl ester (350 mg) was suspended in 6 M hydrochloric acid (15 mL) and refluxed. Stirred for hours. After allowing to cool, the resulting precipitate is collected by filtration, washed with water, and orange solid 4- (2-oxo-3-quinoline-2 (1H) -ylidenindrin-5-yl) 97 mg of butyric acid was obtained. F +: 347.

Reference Example 14: 3- [6-[(4-methylbiperazin-1-yl) methyl] quinoline-2 (1H) -ylidene] -2-oxoindoline-6-carbaldehyde 0-methyloxam (1.20 g ) In THF (15 mL) was added with formalin (5 mL) and concentrated hydrochloric acid (0.1 mL), and the mixture was stirred at room temperature for 24 hours. A 1 M aqueous solution of sodium hydroxide was added, and the mixture was extracted with a mixed solvent of formaldehyde-isopropanol (3: 1 V / V) and washed with saturated saline. After drying using anhydrous sodium sulfate, the mixture was concentrated, the residue was suspended in methanol (20 mL), a 28% aqueous ammonia solution (2 mL) was added, and the mixture was stirred at room temperature for 14 hours. Water was added, and the mixture was extracted with a mixed solvent of formaldehyde-isopropanol (3: 1 V / V), and washed with saturated saline. After drying over anhydrous sodium sulfate, the mixture was concentrated, and the residue was purified by SCC (eluted with methanol-chloroform) to give orange-colored 3- [6-[(4-methylbiperazin-1-yl) methyl ] Quinoline-2 (1H) -ylidene] -2-oxoindolin-6-carbaldehyde 881 mg was obtained. F +: 401.

Example 1: 3- [6-[(4-methylbiperazin-1-yl) methyl] quinoline-2 (1H) -yrident 2-oxoindoline-6-carbaldehyde (400 mg) in methanol (20 mL) 2-[(Aminoxy) methyl] pyridine (248 mg) and concentrated hydrochloric acid (1 drop) were added to the mixture, and the mixture was stirred at 50 ° C for 2 hours. After cooling, the concentrated residue was purified by SCC (eluted with methanol-chloroform), converted to a DMF (20 mL) solution, added with a 4 M hydrogen chloride / ethyl acetate solution (2 mL), and added at room temperature. Stirred for minutes. After concentration, the residue was recrystallized from water-ethanol to give a red solid, 3- [6-[(4-methylpiperazin-1-yl) methyl] quinoline-2 (1H) -ylidene] -2- 320 mg of oxoindoline-6-carbaldehyde 0- (pyridine-2-ylmethyl) oxime trihydrochloride was obtained.

In the same manner as in the above Reference Examples or Examples, Reference Example compounds described in Tables 3 and 4 below and Example compounds described in Table 5 were obtained. Tables 3 to 5 below show the structural formulas and physicochemical properties of each compound, respectively.

The abbreviations in the table are: Rex: Reference Example; Ex: Example; Str: Structure; Sy: Production method (indicating that the compound was produced by the same method as in the above Reference Example or Example). Dat: physicochemical properties; Me: methyl; and Et: ethyl. 61

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Claims

The scope of the claims

1. 2-oxoindoline derivatives represented by the formulas A to F in the following table or their salts _c

In the formula, ALK represents a C1-6 alkyl group, and R represents H or a C1-6 alkyl group. )

2.3- [6-[(4-Methylbiperazine-1-yl) methyl] quinoline-2 (1H) -ylidene] -2-oxoindoline-6-carbaldehyde O- (pyridine-2-ylmethyl) oxy Or salt thereof.

3. 3- [6-[(4-Ethylbiperazin-1-yl) methyl] quinoline-2 (1H) -ylidene] -2-oxoindoline-6-carbaldehyde 0- (pyridine-2-ylmethyl) oxime Or a salt thereof.

4. 3- [6-[(4-Methylbiperazine-1-yl) methyl] quinoline-2 (1H) -ylidentate 2-oxoindoline-6-carbaldehyde 0- (1,3-thiazol-4-ylmethyl) ) Oxime or a salt thereof.

5. 3- [6-[(4-Ethylbiperazin-1-yl) methyl] quinoline-2 (1H) -ylidentate 2-oxoindoline-6-carbaldehyde 0- (1,3-thiazol-4-ylmethyl) ) Oxime or a salt thereof.

6. 3- [2- (2-oxo-6-{[(pyridine-2-ylmethoxy) imino] methyl} indoline-3-ylidene) -1,2-dihydroquinoline-6-yl] propionic acid Or a salt thereof.

7. 3- (1,5-Naphthyridin-2 (1H) -ylidene) -2-oxoindoline-6-carbaldehyde 0- (pyridine-2-ylmethyl) oxime or a salt thereof.

8. 4- {1- [3- (2- {6-[(Methoxyximino) methyl] -2-oxo-1,2-dihydro-3H-indole-3-ylidene} -1,2-dihydroquinoline -6-yl) propyl] piperidine-4-yl} ethyl butyrate or a salt thereof.

9. 3- {1- [3- (2- {6-[(Methoxyimino) methyl] -2-oxo-1,2-dihydro-3H-indole-3-ylidene} -1,2-dihydroquinoline- 6-yl) propyl] piperidin-4-yl} propionic acid or a salt thereof.

10. 3- {1- [3- (2- {6-[(Methoxyimino) methyl] -2-oxo-1,2-dihydro-3H-indole-3-yrident 1,2-dihydroquinoline -6-yl) propyl] piperidine-4-yl} methyl propionate or a salt thereof.

1.1. 3- [6- {3- [4- (3-Methoxy-3-oxopropyl) pyridin-1-yl] propyl} quinoline-2 (1H) -yrident 5-methyl-2 -Kisoindoline -6-Methyl ruponate or a salt thereof.

12. A pharmaceutical composition comprising the 2-oxoindoline derivative or a salt thereof according to claim 1 and a pharmaceutically acceptable carrier.

13. The pharmaceutical composition according to claim 12, which is a vascular endothelial cell growth factor inhibitor.

14. The pharmaceutical composition according to claim 12, which is an angiogenesis inhibitor.

15. The pharmaceutical composition according to claim 12, which is an anticancer agent.