KR100274005B1 - 4-terminal substituted-alkylamino)-quinoline-2-carboxylic acid derivatives acting as NMDA receptor antagonists - Google Patents

Abstract

PURPOSE: Provided are the novel NMDA receptor antagonists, 4-(end-substituted-alkylamine)-quinoline-2-carboxylic acid derivatives and the preparation method thereof. CONSTITUTION: The novel derivatives are represented by the chemical formula 1, where n is 0-10, Nu is either appropriately substituted or unsubstituted arylthio, alkylphosphonate, arylsulfonyl, thiourea, non-aromatic thio, heterocyclic thio, arylphosphonate, heterocycle, alkyl, aryl, cyano, hydroxyl, halogen, arylphosphonate alkylsulfonyl, arylsulfonyl, carbamate, amide, urea, acylurea, sulfonylurea, acylthiourea, sulfonylthiourea, thiocarbamate, amidine, guanidine, imidate, thioimidate, phosphorylamide, sulfonamide or amine groups. The synthetic process contains the following steps: alkylating 5,7-dichlorokynurenic acid methylester with 1,3-dibromopropane; eliminating N-tosyl group with sulfuric acid and substitution of the bromine with NaN3 to make azidomine; reducing with Pd/C to amine compound; converting to the corresponding thioisocyanate and finally hydrolysing the ester group to make the compound of formula 1. The synthetic process of the compound of formula 1, where Nu is substituted arylthio or alkyphosphonate group, comprises the steps of: alkylating 5,7-dichlorokynurenic acid methylester with 1,3-dibromopropane; treating the resultant bromine compound either with arylthio compound to make sulfides or with alkylphosphonates to obtain phosphonate compound; treating these compounds with sulfuric acid to eliminate N-tosyl group and finally hydrolysing them with base to obtain the compound of formula 1. They are effective in the treatment and prevention of the chronic neurodegenerative disease such as epilepsy, Alzheimer's disease, Huntington's and Parkinson's disease and also effective as an analgesic, tranquilizer, treatment agents for depression and psychotic diseases.

Description

4- (terminal substituted-alkylamino) -quinoline-2-carboxylic acid derivatives acting as NMDA receptor antagonists}

The present invention relates to 4- (terminal substituted-alkylamino) -quinoline-2-carboxylic acid derivatives of formula (1) which act as antagonists of excitatory amino acids, methods for their preparation and use as therapeutic agents for neurological diseases.

In particular, the compounds of the present invention antagonize the excitatory activity of NMDA receptors and damage the central nervous system resulting from anemia or hypoxia such as stroke, hypoglycemia, ischemia, cardiac arrest, trauma. It is particularly useful for reducing.

In addition, the compounds of the present invention are useful for preventing chronic neurodegenerative diseases including epilepsy, Alzheimer's disease, Huntington's disease and Parkinson's disease. The compounds of the present invention are also used as anticonvulsants, analgesics, antidepressants, anti-anxiety agents and antischizophrenia.

L-glutamate is the most important excitatory neurotransmitter in mammalian central nervous system neurotransmission. Nearly all central nervous system neurons are excited by L-glutamate, which acts on several receptor-ion channel complexes on the surface of neurons to open ion channels and induce cations such as sodium or calcium. To excite nerve cells by inducing polarization between nerve cell membranes.

These CNS neurons are non-NMDA not excited by NMDA receptors and NMDA excited by glutamate analogue N-methyl-D-aspartate (abbreviated as "NMDA"). It contains a receptor.

NMDA receptors are cell surface protein complexes that are widely distributed in the brain or spinal cord and are known to transmit neuronal excitability at the synapse of neurons and to be closely involved in neuronal growth.

One of the characteristics of this NMDA receptor is that its activity is completely blocked by Mg ⁺⁺ when the cells are at rest. However, this blockade is voltage-dependent and the block is released when the cell is activated by non-NMDA receptors or when other excitatory substances are partially nonpolarized. As such, the mechanism of action of NMDA receptors depends on conditions and plays an important role in learning and memory processes.

Another property of NMDA receptors is that the cells die when the receptor is overly excited. This may be due to excessive accumulation of Ca ⁺⁺ in cells. Recently, many studies have been conducted on cell death by excitatory toxicity of NMDA receptors.

After culturing neurons in vitro, glutamate treatment causes cells to swell and cytotoxicity. This excitatory toxicity is dependent on the presence of Ca ^{++. When} a large amount of Ca ⁺⁺ enters neurons through the ion channel of glutamate receptors, normal cell activity is destroyed and the release of glutamate is accelerated by feedback. Excitatory toxicity occurs. In this process, proteases and lipases are activated to destroy the membranes of neurons and produce free radicals that kill the cells [JW Mcdold, MV Johnson, Brain Res. Reviews 15, 41 (1990)]. As such, excitatory toxicity caused by neurotransmitters kills neurons, and research has been found to be an important cause of neurodegenerative diseases and stroke associated with brain cell death.

Recently, research has focused on the mechanism by which L-glutamate is excessively secreted by brain injury, spinal cord injury, stroke, ischemia, or hypoglycemia resulting in permanent damage to the central nervous system. It has been found that the damage caused by is prevented by NMDA receptor antagonists.

NMDA receptor antagonists prevent many clinical diseases, including ischemic symptoms and epilepsy, and prevent chronic neurodegenerative diseases such as Alzheimer's disease, Huntington's disease and Parkinson's disease [G. Johnson, Annu. Rep. Med. Chem. 24, 41 (1989); G. Johnson and CF Bigge, ibid. 26 , 11 (1991); Werling et al., J. Pharmacol. Exp. Ther. 255 , 40 (1990). Accordingly, several NMDA receptor antagonists have recently been used to treat acute stroke or brain damage.

There are also reports that NMDA receptor antagonists are useful for improving memory and learning, such as 1-aminocyclopropanecarboxylic acid methyl esters, D-cycloserine and R-(+)-3-, which are specific glycine site ligands. Amino-1-hydroxypyrrolidin-2-one- (HA-966) [Bliss, TV P, Collingride, GL, Nature 345 , 347 (1990); GB 2231048A (1990).

Recent reports suggest that NMDA receptor antagonists have analgesic, antidepressant, antipsychotic and anti-anxiety effects [Dickenson, AH and Aydar, E., Neuroscience Lett. 121 , 263 (1990); R. Trullas and P. Skolnick, Eur. J. Pharmacol. 185 , 1 (1990); JH Kehne, et al., Eur. J. Pharmacol. 193 , 283 (1991); PH Hutson, et al., Br. J. Pharmacol. 103 , 2037 (1991).

Non-competitive NMDA receptor antagonists such as ketamine and dextrometopan are useful for the treatment of growth disorders, autism and the like in children. NMDA receptor antagonists are also useful as protective agents in laser neurosurgery.

The fact that NMDA receptors play an important role in the synaptic degeneration of neurons, which is thought to be one of the causes of these diseases, is more evident by recent studies on their physiological function and subunit structure. Kumar KN, et al., Nature 354 , 70-73 (1991); Nakanishi, S., et al., Nature 354 , 31-37 (1991); Monyer, H., et al., Science 256 , 1217-1221 (1992). The NMDA receptor has at least five distinct substrate binding sites, including (a) a binding site for binding to the neurotransmitter L-glutamate, (b) an allosteric modulator site for binding to glycine, and (c) in the channel Sites that bind fencyclidine and related compounds, (d) binding sites that bind Mg ⁺⁺ , and (e) binding sites that bind to the divalent cation Zn ⁺⁺ and exhibit inhibitory activity [Lynch, DR, et al., Mol. pharmacol. 45 , 540-545 (1994); Kuryatov, A., et al., Neuron 12 , 1291-1300 (1994); Nakanishi, S., Science 256 , 1217-1221 (1992).

Looking at the physiological action of subunits of NMDA receptors, the NMDA receptors are activated by glutamate and glycine or their agonists. In addition, the associated Ca ⁺⁺ permeable channel is physiologically blocked by Mg ⁺⁺ in a voltage-dependent manner, and Zn ⁺⁺ with its own regulatory site reduces the activity of synapses of NMDA receptors [Lynch, DR, et al. , Mol. pharmacol. 45 , 540-545 (1994); Kuryatov, A., et al., Neuron 12 , 1291-1300 (1994); Nakanishi, S., Science 256 , 1217-1221 (1992).

Pharmacological interest has focused on the NMDA receptors over the past two decades, and agonists and antagonists that depend on each of the binding sites described above have been explored as candidates for useful drugs. Among them, the most promising method for regulating NMDA receptor activity was the development of an allosteric modulator of glycine binding site.

In 1987, Johnson and Ascher discovered glycine binding sites in NMDA receptors. Subsequent studies have revealed that glycine binding sites and glutamate binding sites in NMDA receptors are present in the same protein and glycine binding sites in NMDA receptors. It was found that there is a negative allosteric coupling between and glutamate binding sites.

Since the presence of negative allosteric coupling, binding of agonists at glutamate recognition sites reduces glycine affinity for glycine recognition sites, and antagonist binding at glutamate recognition sites is enhanced by glycine site antagonists and vice versa [ Beneveniste, M., et al. J. Physiol. 428 , 333 (1990); Leser, RA; Tong, G. and Jahr, CE, J. Neurosci. 13 , 1088 (1993); Clements, JD; Westbrook, GL, Neuron. 7 , 605 (1991).

In addition, recent in vivo microdialixis (dialysis) studies have found that large amounts of glutamate are released in the ischemic brain region but little release of glycine in the rat ubiquitous ischemic model [Globus, MY T, et. al., J. Neurochern. 57 , 470-478 (1991).

As a result of the experiment, it can be seen that the glycine antagonist is a very powerful neuroprotective drug that can reduce excessive excitability of neurons when neurons, neurons, are excessively excited by glutamate to cause cytotoxicity.

Glycine antagonists act non-competitively with glutamate to prevent the NMDA channels from opening, thus overcoming the high concentrations of endogenous glutamate released in the ischemic brain region, unlike competitive NMDA antagonists that compete competitively with glutamate. There is no need to do it. Thus, NMDA receptors are regulated rather than completely inhibited by glycine site antagonists, and these regulatory actions may be much more physiological than receptor blockade where receptor function is completely inhibited (compared to channel blockers), thus glycine antagonists Has fewer side effects than other antagonists. In fact, glycine antagonists can be injected directly into the brains of rodents without side effects [Tricklebank, MD, et al., Eur. J. Pharmacol. 167 , 127 (1989); Koek, W., et al., J. Pharmacol. Exp. Ther. 245 , 969 (1989); Willets and Balster, Neuropharmacology 27 , 1249 (1988).

As such, glycine site antagonists are noncompetitive antagonists that regulate rather than block NMDA receptors, and thus have fewer side effects from NMDA receptor blockade than other forms of NMDA receptor antagonists. It has emerged as a target substance.

Glycine site antagonists have emerged as central nervous system agonist candidates because they have a wide therapeutic window between the desirable effects of neuroprotection and the side effects of hyperactivity commonly found in competitive glutamate antagonists or channel blockers.

On the other hand, the biggest problem in developing glycine-ligand compounds that react with NMDA-associated glycine sites was that most of these compounds did not pass through the blood-brain barrier (abbreviated as "BBB"). . As a result of our efforts to develop glycine-site ligands that can cross the blood-brain-membrane, we have shown that kynureric acid derivatives (2-carboxyquinoline), 2-carboxyindole derivatives, quinoxaline derivatives and 2- Quinolone derivatives and the like have been developed. These are selective noncompetitive antagonists that act on NMDA receptors and are also active upon oral administration [McOuaid, LA, et al., J. Med. Chem. 35 , 3423 (1992); Leeson, PD, et al., J. Med. Chem. 36 , 3386 (1993); Kulagowski, JJ, et al., J. Med. Chem. 37 , 1402 (1994); Cai, SX, et al., J. Med. Chem. 39 , 4682 (1996); and 39, 3248 (1996); EP 489,458; EP 459,561; EP 685,466 A1; WO 94/20470; WO 93/10783, EP 685,466 A1 and EP 481,676 A1.

Recently, it has been discovered that the chineurenic acid derivative, compound No. 1 in Formula 2, exhibits selectivity for NMDA antagonistic activity. That is, the endogenous product of the tryptophan metabolic pathway is a blockade of the glycine regulatory sites of the NMDA receptor and thus has selective NMDA antagonistic activity [Kessler, M., et al., J. Neurochem. 52 , 1319 (1989); Kemp, JA, et al., Proc. Natl. Acad. Sci. USA 85 , 6547 (1988).

In most existing Structure Activity Relationship (SAR) studies of NMDA antagonists, the hydroxyl groups at the C-4 position of the chineurenic acid interact with the H-binding of the receptor and their spatial orientation It has been found to be very important for binding activity. In other words, when the electron-rich substituent is appropriately introduced at the C-4 position of the kynurenic acid mother nucleus, binding affinity to the NMDA receptor is increased.

In particular, Harrison et al. Recently found that compounds 2 and 3 of the following Chemical Formula 2, in which the hydroxyl group at the C-4 position of the kynurenic acid are substituted with heteroatoms linked with heteroatoms, are more effective and selectable than the kynurenic acid itself. Was found in Harrison BL, et al., J. Med. Chem. 33 , 3130 (1990). However, these compounds do not penetrate BBB due to the high polarity of the carboxyl group, and thus have a disadvantage of lacking in vivo activity.

Therefore, the inventors of the present invention have been trying to prepare an excellent in vivo activity of the kynurenic acid derivatives, and the hydroxyl group at the C-4 position is substituted with the alkylamino group which is substituted with the alkylamino group, and the structural structure of the conventional kynurenic acid The present invention was completed by finding out the excellent in vivo activity.

It is an object of the present invention to provide a chineurenic acid derivative substituted with a C-4 position which acts as an antagonist of an excitatory amino acid and a method for preparing the same.

In order to achieve the above object, the present invention provides a 4- (terminal substituted-alkylamino) -quinoline-2-carboxylic acid derivative and a method for easily preparing these compounds.

Hereinafter, the present invention will be described in detail.

The present invention relates to 4- (terminal substituted-alkylamino) -quinoline-2-carboxylic acid derivatives of the general formula (1), including tautomeric isomers or pharmaceutically acceptable salts thereof.

Formula 1

In Chemical Formula 1,

n is selected from 0 to 10;

Nu is arylthio, alkylphosphonate, arylsulfonyl, thiourea, non-aromatic thio, heterocyclic thio, arylphosphonate, heterocycle, alkyl, aryl as appropriately substituted or unsubstituted , Cyano, hydroxyl, halogen, alkylphosphonate, arylphosphonate, alkylsulfonyl, arylsulfonyl, carbamate, amide, urea, acylurea, sulfonylurea, acylthiourea, sulfonylthiourea, Thiocarbamate, amidine, guanidine, imidate, thioimidate, phosphorylamide, sulfonamide or amine and the like.

Arylthio, which is appropriately substituted in Nu, may be represented by the following Chemical Formula 3;

In Formula 3, R 'is halogen, alkyl, aryl, alkylamino, arylamino, alkoxy, aryloxy, fused heterocycle, guanidine, imidate, phosphorylamide, sulfonamide, urea, halogenated alkyl, sia Furnace or carbocycle and the like.

Non-aromatic thios in Nu also include alkylthio, aralkylthio, carbocyclic thio or soluble bicyclic thio.

In the form of formula (1), preferred compounds are those wherein n is selected from 0 to 3, and when Nu is an arylthio group of formula (3), when R 'is m-CH ₃ or p-NH ₂ , and Nu is alkylphosphonate , Arylsulfonyl or thiourea.

Alkyl groups are C ₁ -C ₂₀ alkyl groups, preferably C ₁ -C ₄ alkyl groups and include methyl, ethyl, propyl, isopropyl, butyl, secondary-butyl and tert-butyl groups.

The aryl group is preferably an aryl group having _{6 to 12} carbon atoms and includes phenyl, naphthyl, phenanthryl, anthracyl, indenyl, azulenyl, biphenyl, biphenylenyl and fluorenyl groups.

Heterocyclic groups include C ₃ -C ₇ heterocycloalkyl, C ₃ -C ₇ heterocycloalkyl (C ₁ -C ₆ ) alkyl, heteroaryl and heteroaryl (C ₁ -C ₆ ) alkyl; Suitable heterocycloalkyl groups include piperidyl, piperazinyl and morpholinyl groups; Suitable heteroaryl groups include thiophenyl, furyl, pyrrolyl, indolyl, thiazolyl, benzothiazolyl, oxazolyl, benzooxazolyl, imidazolyl, tetrazolyl, triazolyl, pyridyl, pyrimidinyl and phthalimidyl groups Include.

For use in medicine, the salts of the compounds of formula 1 should be non-toxic and pharmaceutically acceptable salts. Various salts may be used to prepare the compounds of the invention or their toxic and pharmaceutically acceptable salts.

Pharmaceutically acceptable salts of compounds of formula 1 include alkali metal salts such as lithium, sodium, or potassium salts, alkaline earth metal salts such as calcium or magnesium salts, and suitable organic ligands. Salts formed, for example, quaternary ammonium salts. Acid addition salts can be prepared by mixing a solution of a compound of the invention with a solution of a pharmaceutically acceptable non-toxic acid such as hydrochloric acid, fumaric acid, maleic acid, succinic acid, acetic acid, citric acid, tartaric acid, carbonic acid or phosphoric acid. have.

The present invention includes prodrugs of compounds of formula 1 within the scope of the present invention. In general, such prodrugs are functional derivatives of the compound of formula (I), and should be able to be readily changed to the compound required to enter the body and exhibit efficacy. Conventional procedures for the selection and preparation of suitable prodrug derivatives are described in the existing literature [Design of Prodrug, ed. H. Bundgaard, 1985].

If the compound corresponding to the present invention has at least one asymmetric center, an enantiomer may be present, and if the compound corresponding to the present invention has two or more asymmetric centers, the diastereoisomer May be present. Such isomers and mixtures thereof are included within the scope of the present invention.

Particularly preferred compounds in the present invention include the following compounds. However, the following compounds are illustrative of the present invention and the present invention is not limited to the following compounds.

1) 5,7-dichloro-4- [3- (3-phenylthioureido) propylamino] -quinoline-2-carboxylic acid,

2) 5,7-dichloro-4- [3- (4-methoxyphenylthioureido) propylamino] -quinoline-2-carboxylic acid,

3) 5,7-dichloro-4- [3- (m-tolylsulfanyl) propylamino] -quinoline-2-carboxylic acid,

4) 5,7-dichloro-4- [3- (4-aminophenylsulfanyl) propylamino] -quinoline-2-carboxylic acid,

5) 5,7-dichloro-4- [3- (diethoxyphosphoryl) propylamino] -quinoline-2-carboxylic acid,

6) 5,7-dichloro-4- [3- (toluene-3-sulfinyl) propylamino] -quinoline-2-carboxylic acid,

7) 5,7-dichloro-4- [3- (toluene-3-sulfonyl) propylamino] -quinoline-2-carboxylic acid

The pharmaceutical compositions of the invention are preferably in unit dosage form such as tablets, capsules, powders, granules, sterile solutions or suspensions, or suppositories for oral, intravenous, parenteral or rectal administration. To prepare solid compositions such as tablets, the main active ingredient is a pharmaceutical carrier, e.g., the conventional tableting ingredients cornstarch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, di A solid preformulation composition comprising a homogeneous mixture of a compound of the invention or a pharmaceutically acceptable non-toxic salt thereof, in admixture with calcium phosphate or gums and other pharmaceutical diluents such as water Form. The homogeneous preliminary solid composition allows the active ingredient to be dispersed evenly throughout the composition so that the composition can be easily divided into effective unit dosage forms containing the same amount such as tablets, pills and capsules. This preliminary solid composition is further subdivided into unit dosage forms of the abovementioned type comprising from 0.1 to 500 mg of the active ingredient of the invention. Tablets or pills of the novel compositions may be coated or synthesized to provide an advantageous dosage form when sustained release is required. For example, a tablet or pill may be composed of an internal dosage ingredient and an external dosage ingredient, in which the external dosage ingredient surrounds the internal dosage ingredient. The two components can be separated by an enteric membrane, which prevents the digestion of the stomach and allows the internal components to pass through the duodenum or delay the release of the internal components intact. A variety of materials are used as such enteric membranes or coating materials, which include a plurality of polymeric acids and mixtures of polymeric acids and materials such as shellac, cetyl alcohol, cellulose acetate. The liquid form of the novel compositions of the present invention prepared for oral or injectable administration is an emulsion consisting of an aqueous solution, a properly flavored syrup, an aqueous or oily suspension and an edible oil such as cottonseed oil, sesame oil, coconut oil or peanut oil. , Elixirs and similar pharmaceutical vehicles. Suitable dispersing or suspending agents for making aqueous suspensions include synthetic or natural gums such as tragacanth, acacia, alginate, dextran, sodium carboxymethylcellulose, methylcellulose, polyvinylpyrrolidone or gelatin. ). In the treatment of neurodegeneration, a suitable dosage is 0.01-250 mg / kg per day, preferably 0.05-100 mg / kg per day, more preferably 0.05-5 mg / kg per day. The compound may be conveniently administered by intravenous injection.

Methods for preparing the compound of Formula 1, which is a compound of the present invention, are shown in Schemes 1 and 2.

That is, the manufacturing process of the compound of Chemical Formula 1 according to the present invention may be described by dividing into Scheme 1 and Scheme 2 according to the kind of Nu which is a substituent of an alkyl amino group.

Scheme 1 shows that Nu is of Formula 1 thiourea, carbamate, amide, urea, acylurea, sulfonylurea, acylthiourea, sulfonylthiourea, thiocarbamate, amidine, guanidine, imidate, thiimidate, As a manufacturing method which can be applied in the case of phosphorylamide, sulfonamide or amine, in particular the case where Nu is thiourea;

1) alkylating the compound of formula 31 with 1,3-dibromopropane to obtain a bromide compound of formula 32 (first step);

2) sulfuric acid treatment of the compound of formula 32 obtained in the first step to remove N-tosyl groups (-NTs), and then substituted bromine with NaN ₃ to obtain an azido compound of formula 33 (second step);

3) catalytic hydrogenation of the compound of formula 33 obtained in the second step and reaction with an appropriate thioisocyanate to obtain the thiourea compound of formula 34 (third step); And

4) basic hydrolysis of the compound of formula 34 prepared in the third step to obtain a compound of formula 1 which is the target compound of the present invention (fourth step)

Is made of.

X in Scheme 2 represents Nu of Formula 1, which corresponds to the case where Nu is an appropriately substituted arylthio or alkylphosphonate or the like;

1) treating the compound of formula 32 with an arylthiol (HSAr) compound to prepare a sulfide compound of formula 36 or treating with triethylphosphite [P (OEt) ₃ ] to produce a phosphonate of formula 37 Stage 1); And

2) removing the N-tosyl group (-NTs) by sulfuric acid treatment of the compound of formula 36 or 37 obtained in the first step and basic hydrolysis to obtain the compound of formula 1 which is the target compound of the present invention (second step)

Is made of.

In this case, when Nu is arylthio in Scheme 2, di-oxidized aryl of Structural Formula 40 by reacting and oxidizing m-chloroperbenzoic acid (m-CPBA) in addition to the second step. Sulfonyl and mono-oxidized arylsulfinyl compounds can be prepared, which is shown in Scheme 3 below.

The manufacturing method of the present invention will be described in more detail.

The compound of formula 31, which is a starting material in Scheme 1, may be easily prepared by a conventional method by reacting the kynurenic acid methyl ester with p-tolylsulfonyl isocyanate.

In the first step, the starting material is reacted with dibromopropane in the presence of K ₂ CO ₃ and refluxed to obtain a bromide compound alkylated with the C-4 position of the kynurenic acid, wherein the reaction solvent used is acetonitrile. Etc.

In the second step, the compound prepared in the first step is treated with concentrated sulfuric acid to remove N-tosyl groups (-NTs), and then bromine is substituted with NaN ₃ to obtain an azido compound. For example, DMF.

The third step is a process of producing a thiourea by hydrogenation of the azide compound in the presence of Pd-C catalyst, and reaction with an appropriate thioisocyanate in the presence of triethylamine (Et ₃ N),

In the fourth step, a conventional basic hydrolysis of the compound prepared in the third step is carried out to obtain a compound of Formula 1, which is a final target compound. The methyl ester prepared in the third step is hydrolyzed under basic conditions to prepare a carboxylic acid. It's a step.

In Scheme 2, the first step is to convert the bromide to sulfide by treating the compound of formula 32 prepared in the first step of Scheme 1 with an arylthio compound and Na ₂ CO ₃ in anhydrous tetrahydrofuran (THF) solution, Converting bromide to phosphonate by simply refluxing the compound of formula 32 in the presence of triethylphosphite.

In the second step, the compound prepared in the first step is treated with concentrated sulfuric acid to remove N-tosyl groups (-NTs), and conventional basic hydrolysis is performed to obtain a compound of formula 1 as a final target compound.

Furthermore, in addition to the above process, when Nu of the final compound of interest is sulfide, mono-oxidized sulfinyl compound and di- by reacting the compound with m-CPBA in methylene chloride solvent and performing oxidation The process of preparing an oxidized sulfonyl compound as an inseparable mixture may be further performed.

Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are merely to illustrate the invention and the present invention is not limited by the examples.

Example 1 5,7-dichloro-4- [3- (3-phenylthioureido) propylamino] -quinoline-2-carboxylic acid

(Step 1) 4-[(3-Bromopropyl)-(toluene-4-sulfonyl) -amino] -5,7-dichloroquinoline-2-carboxylic acid methyl ester

To acetonitrile (30 mL) was added suspension of 1,3-dibromopropane (Suponamide (0.42 g, 1.0 mmol) and K ₂ CO ₃ (0.55 g, 4.0 mmol), a compound of formula 2 in Scheme 1). 4.0 mL, 30 mmol) was added and the mixture was refluxed with heating for 3 hours. The remaining insoluble solid was filtered off and the filtrate was evaporated on a rotary evaporator to give a brown residue. This residue was purified by flash column chromatography (EtOAc: hexane = 1: 6) to give the title compound as pale yellow crystals (0.33 g, yield 60%).

mp 189-190 ° C; ¹ H-NMR (CDCl ₃ ) δ 2.04-2.29 (m, 2H, CH ₂ ), 2.48 (s, 3H, ArCH ₃ ), 3.36-3.98 (m, 2H, CH ₂ Br), 3.37 (m, 2H, SO ₂ NCH ₂ ), 4.05 (s, 3H, OCH ₃ ), 7.33 (d, 2H, J = 8.0 Hz, ArH), 7.50 (d, 2H, J = 8.8 Hz, ArH), 7.52 (s, 1H, ArH), 7.78 (d, 1H, J = 2.0 Hz, ArH), 8.23 (d, 1H, J = 2.0 Hz, ArH); MS (EI) m / e 546 [M ⁺ ], 513 [M ⁺ -CH ₃ OH], 482 [M ⁺ -SO ₂ ], 391 [M ⁺ -SO ₂ C ₆ H ₄ CH ₃ ], 331, 251 , 155.

(Step 2) 4- (3-azidopropyl) -5,7-dichloroquinoline-2-carboxylic acid methyl ester

Quinolonic sulfonamide (10 mL, 90% in water), the title compound of (Step 1), was added in small portions to a solution of sulfuric acid (1.50 g, 2.75 mmol) in an ice bath, and then the mixture was added to 2 Stir for hours. At this time, the reaction temperature should be maintained below 4 ℃. Slowly pour the resulting mixture, stirring for 1 hour on crushed ice. An aqueous solution containing an insoluble solid was basified with an aqueous 8N NaOH solution at pH 12. The remaining solid was filtered, washed with water (50 mL) and dried in vacuo to yield the intermediate bromide compound as a pale yellow solid (0.96 g, 92% yield).

¹ H-NMR (CDCl ₃ ) δ 2.31 (m, 2H, CH ₂ ), 3.54 (t, 2H, J = 6.8 Hz, CH ₂ Br), 3.56 (t, 2H, J = 6.8 Hz, NCH ₂ ), 4.03 (s, 3H, OCH ₃ ), 7.23 (s, 1H, ArH), 7.39 (br t, 1H, J = 4.4 Hz, NH), 7.42 (d, 1H, J = 2.2 Hz, ArH), 8.05 ( d, 1H, J = 2.2 Hz, ArH).

The suspension prepared above with bromide (0.90 g, 2.38 mmol) and NaN ₃ (0.62 g, 9.52 mmol) prepared in DMF (20 mL) was heated at 50 ° C. for 3 hours. After cooling to room temperature, the resulting mixture was poured into water (50 mL) and the aqueous layer was extracted with methylene chloride (50 mL × 2). The organic layer was evaporated under reduced pressure and the remaining residue was purified by column chromatography (silica gel, 90% EtOAc in n-hexane) to give the title compound pure azido compound as a pale yellow solid (0.80 g, 99% yield).

¹ H-NMR (CDCl ₃ ) δ 2.04 (m, 2H, CH ₂ ), 3.42 (dt, 2H, J = 6.9, 4.0 Hz, NCH ₂ ), 3.50 (t, 2H, J = 6.9 Hz, CH ₂ N ₃ ), 4.01 (s, 3H, OCH ₃ ), 7.18 (s, 1H, ArH), 7.38 (d, 1H, J = 2.2 Hz, ArH), 7.41 (br t, 1H, J = 4.4 Hz, NH) , 8.03 (d, 1H, J = 2.2 Hz, ArH).

(Step 3) 5,7-dichloro-4- [3- (3-phenylthioureido) propylamino] -quinoline-2-carboxylic acid methyl ester

To a mixture of ethyl acetate-methanol (4: 1, 20 mL) was added azido methyl ester compound (0.80 g, 2.35 mmol) prepared in the above (step 2) and a 10% palladium catalyst using activated carbon as a carrier. It was vigorously stirred in a hydrogen atmosphere (using an inflatable flask). After stirring overnight, the reaction mixture was filtered through a pad of celite and the filtrate was evaporated under reduced pressure to afford the crude amino compound as a colorless syrup (0.68 g, yield 92%). This crude syrup was diluted with methylene chloride anhydride (20 mL), then phenylthioisocyanate (0.38 g, 2.85 mmol) and triethylamine (0.40 mL, 2.85 mmol) were added carefully. The reaction mixture was stirred at rt for 6 h and diluted with 100 mL of methylene chloride, then washed with 1N HCl (100 mL), H ₂ O (100 mL × 2) and brine solution (100 mL). The solvent was evaporated under reduced pressure to give a sticky residue. The residue was purified by column chromatography (80% EtOAc in hexanes) to give the title compound as a colorless solid with unidentified compound (0.32 g) (0.54 g, 50% yield).

¹ H-NMR (CDCl ₃ ) δ 1.95 (m, 2H, CH ₂ ), 3.54 (m, 2H, NCH ₂ ), 3.89 (m, 2H, CH ₂ N), 4.03 (s, 3H, OCH ₃ ), 6.22 (t, 1H, J = 4.5 Hz, NH), 6.68 (t, 1H, J = 4.5 Hz, NH), 7.12 to 7.72 (m, 2H, ArH), 7.28 (s, 1H, ArH), 7.30 to 7.68 (m, 2H, ArH), 7.82 (br s, 1H, NH), 8.02 (d, 1H, J = 6.4 Hz, ArH), 8.14 (d, 1H, J = 6.2 Hz, ArH);

Unidentified Compounds:

¹ H-NMR (CDCl ₃ ) δ 1.92 (m, 2H, CH ₂ ), 3.52 (m, 2H, NCH ₂ ), 3.86 (m, 2H, CH ₂ N), 4.01 (s, 3H, OCH ₃ ), 6.18 (t, 1H, J = 4.1 Hz, NH), 6.92 (t, 1H, J = 4.1 Hz, NH), 7.10-7.18 (m, 2H, ArH), 7.23 (s, 1H, ArH), 7.28- 7.40 (m, 2H, ArH), 7.45 (d, 1H, J = 2.1 Hz, ArH), 7.82 (br s, 1H, NH), 7.98 (d, 1H, J = 6.6 Hz, ArH), 8.11 (d , 1H, J = 2.1 Hz, ArH).

(Step 4) 5,7-Dichloro-4- [3- (3-phenylthioureido) propylamino] -quinoline-2-carboxylic acid

The urea methyl ester mixture (0.25 g, 0.54 mmol) obtained in (step 3) above was treated with NaOH (86 mg, 2.16 mmol) in 10 mL of THF-H ₂ O (1: 1) mixed solvent for 2 hours at room temperature. Saponified. This mixture was diluted with 20 mL of water and extracted with methylene chloride (20 mL × 2). The aqueous layer was acidified with 1N hydrochloric acid at pH 3. The resulting white precipitate was filtered, washed with water and dried in vacuo to afford the title compound, the target compound of the invention, as a white solid with an unidentified solid (0.18 g, yield 75%).

¹ H-NMR (DMSO-d ₆ ) δ 2.12 (m, 2H, CH ₂ ), 3.61 to 3.82 (m, 4H, NCH ₂ & CH ₂ N), 7.14 to 7.56 (m, 5H, ArH), 7.85 ( d, 1H, J = 7.6 Hz, ArH), 8.32 (s, 1H, ArH), 8.48 (br s, 1H, NH), 8.68 (d, 1H, J = 7.6 Hz, ArH), 9.42 (br s, 1H, NH), 10.04 (br s, 1H, NH);

Unidentified Compounds:

¹ H-NMR (DMSO-d ₆ ) δ 2.14 (m, 2H, CH ₂ ), 3.64 to 3.82 (m, 4H, NCH ₂ & CH ₂ N), 7.14 to 7.56 (m, 4H, ArH), 7.76 ( m, 1H, ArH), 8.04 (m, 1H, ArH), 8.38 (m, 1H, ArH), 8.45 (br s, 1H, NH), 8.70 (m, 1H, ArH), 9.47 (br s, 1H , NH), 10.06 (br s, 1 H, NH).

Example 2 5,7-dichloro-4- [3- (4-methoxyphenylthioureido) propylamino] -quinoline-2-carboxylic acid

(Step 1) 4-[(3-Bromopropyl)-(toluene-4-sulfonyl) -amino] -5,7-dichloroquinoline-2-carboxylic acid methyl ester

The title compound was prepared in the same manner as in (Step 1) of <Example 1>.

(Step 2) 4- (3-azidopropyl) -5,7-dichloroquinoline-2-carboxylic acid methyl ester

The title compound was prepared in the same manner as in (Step 2) of <Example 1>.

(Step 3) 5,7-dichloro-4- [3- (4-methoxyphenylthioureido) propylamino] -quinoline-2-carboxylic acid methyl ester

The title compound was prepared with an unidentified compound (0.28 g) by carrying out the reaction in the same manner as in (Step 3) of <Example 1>, except that 4-methoxyphenylthioisocyanate was used (0.72). g, yield 62%).

¹ H-NMR (CDCl ₃ ) δ 1.89 (m, 2H, CH ₂ ), 3.62 (m, 2H, NCH ₂ ), 3.80 (s, 3H, OCH ₃ ), 3.86 (m, 2H, CH ₂ N), 4.02 (s, 3H, OCH ₃ ), 5.99 (br t, 1H, J = 4.2 Hz, NH), 6.74 (br t, 1H, J = 4.2 Hz, NH), 6.88 (d, 2H, J = 8.0 Hz , ArH), 7.04 (d, 2H, J = 8.0 Hz, ArH), 7.14 (br s, 1H, NH), 7.18 (d, 1H, J = 2.0 Hz, ArH), 7.20 (s, 1H, ArH) , 8.10 (d, 1H, J = 2.0 Hz, ArH);

Unidentified Compounds:

¹ H-NMR (CDCl ₃ ) δ 1.89 (m, 2H, CH ₂ ), 3.50 (m, 2H, NCH ₂ ), 3.78 (s, 3H, OCH ₃ ), 3.84 (m, 2H, CH ₂ N), 4.00 (s, 3H, OCH ₃ ), 6.12 (br t, 1H, NH), 6.86 (d, 1H, J = 8.1 Hz, ArH), 6.91 (br t, 1H, NH), 7.08 (d, 2H, J = 8.1 Hz, ArH), 7.14 (s, 1H, ArH), 7.42 (d, 1H, J = 2.1 Hz, ArH), 7.96 (br s, 1H, NH), 8.06 (d, 1H, J = 2.1 Hz, ArH).

(Step 4) 5,7-Dichloro-4- [3- (4-methoxyphenylthioureido) propylamino] -quinoline-2-carboxylic acid

Using the title compound (0.35 g, 0.71 mmol) in (Step 3) of <Example 2> and carrying out the reaction in the same manner as (Step 4) of <Example 1>, together with the title compound, which was not identified Was obtained (0.28 g, yield 82%).

¹ H-NMR (DMSO-d ₆ ) δ 2.14 (m, 2H, CH ₂ ), 3.62 to 3.78 (m, 4H, NCH ₂ & CH ₂ N), 3.78 (s, 3H, OCH ₃ ), 6.94 (d , 2H, J = 8.2 Hz, ArH), 7.28 (br s, 1H, NH), 7.42 (d, 2H, J = 8.2 Hz, ArH), 7.80 (d, 1H, J = 6.7 Hz, ArH), 8.41 (s, 1H, ArH), 8.79 (d, 1H, J = 6.7 Hz, ArH), 9.74 (br s, 1H, NH), 9.96 (br s, 1H, NH);

Unidentified Compounds:

¹ H-NMR (DMSO-d ₆ ) δ 2.12 (m, 2H, CH ₂ ), 3.60 to 3.80 (m, 4H, NCH ₂ & CH ₂ N), 3.82 (s, 3H, OCH ₃ ), 6.96 (d , 2H, J = 8.7 Hz, ArH), 7.28 (br s, 1H, NH), 7.25 (m, 1H, ArH), 7.30 (d, 2H, J = 8.7 Hz, ArH), 7.95 to 8.12 (m, 2H, ArH), 8.36 (m, 1H, ArH), 8.58 (m, 1H, ArH), 9.19 (br s, 1H, NH), 9.68 (br s, 1H, NH).

Example 3 5,7-dichloro-4- [3- (m-tolylsulfanyl) propylamino] -quinoline-2-carboxyl acid

(Step 1) 4-[(3-Bromopropyl)-(toluene-4-sulfonyl) -amino] -5,7-dichloroquinoline-2-carboxylic acid methyl ester

The title compound was prepared in the same manner as in (Step 1) of <Example 1>.

(Step 2) 5,7-Dichloro-4- [3- (m-tolylsulfanyl) -propyl- (4-toluenesulfonyl) -amino] -quinoline-2-carboxylic acid methyl ester

To a solution of the bromide (1.50 g, 2.75 mmol) of (step 1) in THF (30 mL) was added m-thiocresol (0.41 g, 3.30 mmol) and K ₂ CO ₃ (0.76 g, 5.50 mmol). . The mixture was refluxed for 4 hours and then cooled to room temperature. The solution was diluted with methylene chloride (100 mL) and washed with brine (50 mL × 2) and water (50 mL × 2) and dried over magnesium sulfate (MgSO ₄ ). The organic layer was concentrated in vacuo and the residue was purified by column chromatography (30% EtOAc in hexane) to give the title compound as a white solid (1.39 g, yield 82%).

¹ H-NMR (CDCl ₃ ) δ 1.62-2.06 (m, 2H, CH ₂ ), 2.28 (s, 3H, ArCH ₃ ), 2.49 (s, 3H, ArCH ₃ ), 2.85 (m, 2H, CH ₂ S ), 3.62 (m, 1H, NC H H), 3.98 (m, 1H, NC H H), 4.07 (s, 3H, OCH ₃ ), 4.07 (s, 3H, OCH ₃ ), 6.92-7.12 (m, 4H, ArH), 7.29 (d, 2H, J = 8.5 Hz, ArH), 7.45 (s, 1H, ArH), 7.46 (d, 2H, J = 8.5 Hz, ArH), 7.76 (d, 1H, J = 2.2 Hz, ArH), 8.28 (d, 1H, J = 2.2 Hz, ArH).

(Step 3) 5,7-dichloro-4- [3- (m-tolylsulfanyl) propylamino] -quinoline-2-carboxylic acid

Sulfonamide (1.0 g, 1.70 mmol), the title compound of step (2), was added portionwise to a solution of sulfuric acid (3 mL, 90% in water) in an ice bath, and the mixture was stirred for 2 hours. It was. At this time, the reaction temperature should be maintained below 4 ℃. Slowly pour the resulting mixture, stirring for 1 hour on crushed ice. An aqueous solution containing an insoluble solid was basified with an aqueous pH 4N NaOH solution. The mixture was extracted with methylene chloride (50 mL × 3) and the organic layer was washed with water (100 mL × 2) and brine (50 mL) and concentrated in vacuo to afford the crude amino quinoline acid methyl ester as a pale yellow solid. Obtained (0.85 g).

¹ H-NMR (CDCl ₃ ) δ 1.71-1.98 (m, 2H, CH ₂ ), 2.28 (s, 3H, ArCH ₃ ), 2.84 (t, 2H, J = 7.1 Hz, CH ₂ S), 3.38 (dt , 2H, J = 4.2, 7.1 Hz, NCH ₂ ), 4.02 (s, 3H, OCH ₃ ), 6.72-7.18 (m, 4H, ArH), 7.24 (s, 1H, ArH), 7.64 (d, 1H, J = 2.3 Hz, ArH), 8.22 (d, 1H, J = 2.3 Hz, ArH), 8.64 (br t, 1H, J = 4.2 Hz, NH).

Without further purification, the crude methyl ester prepared above was dissolved in THF (20 mL) and treated with aqueous 0.4N NaOH solution (17 mL, 6.80 mmol) for 2 hours at room temperature. The mixture was concentrated under reduced pressure until the total volume reached 20 mL, washed with 20 mL of methylene chloride, and then acidified with concentrated hydrochloric acid at pH 3. The resulting precipitate was collected, washed with water (5 mL × 3) and dried in vacuo to afford the title compound as a white solid carboxylic acid (0.57 g, 83% yield).

¹ H-NMR (DMSO-d ₆ ) δ 2.12 (m, 2H, CH ₂ ), 2.32 (s, 3H, ArCH ₃ ), 3.01 (m, 2H, CH ₂ S), 3.65 (m, 2H, NCH ₂ ), 6.92-7.40 (m, 4H, ArH), 7.26 (s, 1H, ArH), 7.78 (d, 1H, J = 2.4 Hz, ArH), 8.04 (br s, 1H, NH), 8.24 (d, 1H, J = 2.4 Hz, ArH).

Example 4 5,7-dichloro-4- [3- (4-aminophenylsulfanyl) propylamino] -quinoline-2-carboxylic acid

(Step 1) 4-[(3-Bromopropyl)-(toluene-4-sulfonyl) -amino] -5,7-dichloroquinoline-2-carboxylic acid methyl ester

The title compound was prepared in the same manner as in (Step 1) of <Example 1>.

(Step 2) 5,7-Dichloro-4- [3- (4-aminophenylsulfanyl) -propyl- (4-toluenesulfonyl) -amino] -quinoline-2-carboxylic acid methyl ester

The reaction was carried out in the same manner as in (Step 2) of <Example 3> using 4-aminothiophenol (0.34 g, 2.75 mmol) and K ₂ CO ₃ (0.63 g, 4.58 mmol). After treatment, purification by flash column chromatography (30% EtOAc in hexanes) gave pure sulfide as a white solid (1.11 g, yield 82%).

¹ H-NMR (CDCl ₃ ) δ 1.58-1.96 (m, 2H, CH ₂ ), 2.46 (s, 3H, ArCH ₃ ), 2.66 (t, 2H, J = 7.2 Hz, CH ₂ S), 3.54 (m , 1H, NC H H), 3.68 (br s, 2H, NH ₂ ), 3.97 (m, 1H, NC H H), 4.06 (s, 3H, OCH ₃ ), 6.45 (d, 2H, J = 8.6 Hz , ArH), 7.01 (d, 2H, J = 8.6 Hz, ArH), 7.28 (d, 2H, J = 8.4 Hz, ArH), 7.44 (s, 1H, ArH), 7.46 (d, 2H, J = 8.4 Hz, ArH), 7.74 (d, 1H, J = 2.1 Hz, ArH), 8.26 (d, 1H, J = 2.2 Hz, ArH).

(Step 3) 5,7-dichloro-4- [3- (4-aminophenylsulfanyl) propylamino] -quinoline-2-carboxylic acid

Using the methyl ester (1.0 g, 1.66 mmol) prepared in this Example (step 2), the reaction was carried out in the same manner as in (Step 3) of <Example 3>, to obtain the title compound as a white solid (0.53). g, yield 77%).

¹ H-NMR (DMSO-d ₆ ) δ 2.12 (m, 2H, CH ₂ ), 3.22 (t, 2H, J = 7.0 Hz, CH ₂ S), 3.88 (m, 2H, NCH ₂ ), 7.38 (d , 2H, J = 8.3 Hz, ArH), 7.40 (s, 1H, ArH), 7.56 (d, 2H, J = 8.3 Hz, ArH), 8.06 (d, 1H, J = 2.4 Hz, ArH), 8.42 ( d, 1H, J = 2.4 Hz, ArH), 9.38 (br s, 1H, NH).

Example 5 5,7-Dichloro-4- [3- (diethoxyphosphoryl) propylamino] -quinoline-2-carboxylic acid

(Step 1) 4-[(3-Bromopropyl)-(toluene-4-sulfonyl) -amino] -5,7-dichloroquinoline-2-carboxylic acid methyl ester

The title compound was prepared in the same manner as in (Step 1) of <Example 1>.

(Step 2) 5,7-Dichloro-4- [3- (diethoxyphosphoryl) -propyl- (4-toluenesulfonyl) -amino] -quinoline-2-carboxylic acid methyl ester

The solution of the title compound (1.25 g, 2.29 mmol) of (step 1) in triethyl phosphite (10 mL) was refluxed for 8 hours. This mixture was concentrated under reduced pressure to give crude compound. This crude compound was purified by flash column chromatography (60% EtOAc in hexane) to give the title compound pure phosphonate compound as a colorless solid (1.30 g, 94% yield).

¹ H-NMR (CDCl ₃ ) δ 1.05-1.34 (m, 6H, OCH ₂ CH ₃ ), 1.64-2.04 (m, 4H, CH ₂ & CH ₂ P), 2.44 (s, 3H, ArCH ₃ ), 3.54 (m, 1H, NC H H), 3.91 (m, 1H, NC H H), 3.98-4.15 (m, 4H, 20CH ₂ ), 4.02 (s, 3H, OCH ₃ ), 7.30 (d, 2H, J = 8.0 Hz, ArH), 7.46 (d, 2H, J = 8.0 Hz, ArH), 7.48 (s, 1H, ArH), 7.75 (d, 2H, J = 2.2 Hz, ArH), 8.28 (d, 1H, J = 2.2 Hz, ArH).

(Step 3) 5,7-Dichloro-4- [3- (diethoxyphosphoryl) propylamino] -quinoline-2-carboxyl acid

The title compound as a white solid was prepared by carrying out the same procedure as in (Step 3) of <Example 3> with the methyl ester (1.0 g, 1.66 mmol) prepared in this Example (Step 2) (0.57). g, yield 81%).

Unprotected aminopropyl phosphonate (intermediate):

¹ H-NMR (DMSO-d ₆ ) δ 1.20-1.42 (m, 6H, OCH ₂ CH ₃ ), 1.84-2.08 (m, 4H, CH ₂ & CH ₂ P), 3.54 (m, 2H, NCH ₂ ) , 4.08 (s, 3H, OCH ₃ ), 4.16 (m, 4H, 2OCH ₂ ), 7.18 (s, 1H, ArH), 7.65 (br s, 1H, NH), 7.76 (d, 1H, J = 2.3 Hz , ArH), 8.01 (d, 1H, J = 2.3 Hz, ArH);

Title compound:

¹ H-NMR (DMSO-d ₆ ) δ 1.24 (m, 6H, OCH ₂ CH ₃ ), 1.80-2.02 (m, 4H, CH ₂ & CH ₂ P), 3.78 (m, 2H, NCH ₂ ), 4.05 (m, 4H, 20CH ₂ ), 7.43 (s, 1H, ArH), 7.92 (d, 1H, J = 2.4 Hz, ArH), 8.37 (d, 1H, J = 2.4 Hz, ArH), 9.18 (br t , 1H, J = 7.6 Hz, NH).

Example 6 5,7-dichloro-4- [3- (toluene-3-sulfinyl) propylamino] -quinoline-2-carboxyl acid; 5,7-dichloro-4- [3- (toluene-3-sulfonyl) propylamino] -quinoline-2-carboxylic acid

To a solution of Example 3 (0.30 g, 0.77 mmol) in acetic acid (10 mL) was added H ₂ O ₂ (2.20 eq, 35 wt% aqueous solution) and stirred overnight. The solvent acetic acid was evaporated under reduced pressure, and the resulting solid was washed with water (10 mL × 2) and dried in vacuo to give the title compound mono-oxidized compound and di- as a pale yellow solid. Di-oxidized compounds were obtained as an unseparated mixture.

5,7-dichloro-4- [3- (toluene-3-sulfonyl) propylamino] -quinoline-2-carboxylic acid:

¹ H-NMR (DMSO-d ₆ ) δ 2.05 (m, 2H, CH ₂ ), 2.39 (s, 3H, ArCH ₃ ), 3.01 (m, 4H, NCH ₂ & CH ₂ SO ₂ ), 7.18 (s, 1H, ArH), 7.75 (m, 3H, ArH);

5,7-dichloro-4- [3- (toluene-3-sulfuryl) propylamino] -quinoline-2-carboxylic acid:

¹ H-NMR (DMSO-d ₆ ) δ 2.05 (br m, 2H, CH ₂ ), 2.45 (s, 3H, ArCH ₃ ), 3.45 (br m, 4H, NCH ₂ & CH ₂ SO ₂ ), 7.18 ( m, 1H, ArH), 7.75 (m, 3H, ArH).

I. In vitro screening for binding activity

1) Preparation of Synaptic Membrane

Male Sprague-Dawley rats (300-400 g) were obtained from a laboratory animal laboratory at Korea Research Institute of Chemical Technology. Laboratory animals were treated with water and standard in an air-conditioned room (22 + 1 ° C .; relative humidity, 60 + 5%) in a slightly darkened condition (brightness at 8 am) during the preparation period of 4-10 days prior to use. The laboratory food was fed and bred. Synaptic membranes for receptor binding studies are modified methods of Foster and Fagg [Eur. J. Pharmacol. 133, 291 (1987) and the modified method by Murphy et al. [Br. J. Pharmacol. 95, 932 (1988). In summary, male Sprague-Dawley rats were killed, brain cortex and hippocampus (chopocampus) were chopped with a scalpel and homogenized five times with 10 volumes of 0.32 M sucrose using a Teflon-fiber equalizer. Centrifuged for 10 minutes at 1000x g for 10 minutes with a Beckman J2 / 21 centrifuge (roto: JA20), and the supernatant was collected and centrifuged for 20 minutes at 20,000x g. The supernatant was removed and homogenized with a pellet homogenizer (5 times setting, 30 seconds).

After incubation at 4 ° C. for 30 minutes, the membrane suspension was centrifuged at 39,800 × g for 25 minutes with a Beckman L8-M ultracentrifuge. The pellet was left overnight at -70 ° C. The next day the pellet is dissolved at room temperature for 10 minutes and then resuspended in 20 volumes of 50 mM tris-acetone (pH 7.1, 4 ° C.) containing 0.04% Triton X-100, incubated at 37 ° C. for 20 minutes and 20 minutes. Centrifuge as above at 39,800 × g. The pellet was washed three times by centrifugation as above with 20 volumes of 50 mM tri-acetate at pH 7.1 without detergent. The final pellet was suspended in 50 mM Tris-Acetate, and protein concentration was measured using Bio-Rad agent (Bradford, 1976). The resuspension buffer was adjusted to a membrane protein concentration of 1 mg / ml and stored at -70 ° C.

2) [ ³ H] glycine binding measurement

[ ³ H] glycine binding measurements were performed as described in Baron et al., 1991. For [ ³ H] glycine saturation binding assay, reaction of the synaptic membrane (100 μg membrane protein) with a final volume of 0.5 ml containing 50 mM tris-acetate buffer solution at pH 7.1, 5-500 nM of [ ³ H] glycine The reaction was carried out at a temperature of 4 ° C. for 30 minutes in a borosilicated glass tube containing the mixture. For drug potential measurements, the synaptic membrane (100 μg of membrane protein) was reacted as above in a reaction mixture containing 50 mM tris-acetate buffer, pH 7.1, 50 nM of [ ³ H] glycine, and various concentrations of experimental compounds. I was.

The reaction was then terminated by the addition of 2.5 mL of 50 mM Tris-Acetate buffer solution at ice-cold pH 7.1 and a Whatman GF / B glass fiber filter pre-soaked in 0.3% polyethyleneimine in assay buffer. Bound radioactivity was attached by detachment by rapid filtration with a used Brandel cell harvester (Brandel M-12R). The used filter was washed twice in 10 seconds with 2.5 mL of cold buffer solution and the radioactivity of the filter was filtered using a liquid scintillation counter using Beck mL LS with 3 mL of Luma gel at 50-55% counting efficiency. 6000TA] Non-specific binding was measured in the presence of 1 mM glycine.

All experimental compounds were dissolved in dimethylsulfoxide (DMSO) and diluted sequentially at various concentrations for binding measurements. The final concentration of DMSO in the assay mixture was 2% and it was evident that this concentration did not affect the binding of radioligand.

Table 1 shows the results of the percentage values (% inhibition) indicating the degree of competition with [ ³ H] glycine when the concentration of the compounds prepared in the present invention is 100 μM.

Compound number % Inhibition at 100 μM Example 1 77.9 Example 2 75.1 Example 3 50.0 Example 4 76.2 Example 5 58.1 Example 6 71.3

II. In vivo screening for anticonvulsive activity

1) NMDA (i.c.v.)-Induced spasm experiment

Based on the method of Chugai, 40-160 ng of NMDA per mouse was calculated using i.c.v. Obtained for induced spasms after administration. A 28 gauge injection needle attached to a 50 μl syringe was inserted through the bregma 1 mm right cartilage on the parietal suture and the test solution was inserted using a manipulator. The insertion volume was 5 μl per rat. Convulsions in response to NMDA occurred within 5 minutes, 1) rough running or jumping, 2) myoclonic seizures; (clonic seizures; loss of direction and repetitive movements of the limbs at the same time). Animals are i.c.v. Observation was performed for 10 minutes after injection and the seizure seen when myoclonus occurred. In order to investigate the NMDA-antagonist properties of the experimental drug, the following procedure was performed. The experimental drug (injection dose, 1 ml / 100 g) was i.p. 15 min after injection, 160 μl of NMDA was injected at 5 μl per horse. i.c.v. The experimental drug and NMDA solution were injected at the same time for investigation by administration. Ten rats were used for the same dose. The dose was increased until antagonistic NMDA reached a limit (eg solubility, lethality) that induces convulsions or can no longer be tested. NMDA was dissolved in 0.9% sodium chloride solution, and AP5,5,7-dichloroquinernic acid and 7-chloroquinernic acid were dissolved in a minimum amount of 0.1 N sodium hydroxide with 0.9% physiological saline.

2) Permanent Ligation of the Middle Celebral Atery (MCA)

Midbrain artery ligation is performed by Nagfuji et al. [Neurosci. Lett. 147, 159 (1992), Mol. Chem. Neuropathol. 26, 107 (1995), Neuro Report. 6, 1541 (1995). In summary, male Sprague Dawley rats were anesthetized with 2% halotan and then maintained at 1.5% halotan in nitrous oxide: oxygen (70:30) using a halotan evaporator. After the animal was lying on its side, the left temporal muscles were cut and partially separated from the iliac arc with bipolar coagulant. Small bilateral incisions were performed using a micro-drill cooled with cold saline in front of the hole of the mandible nerve under the surgical microscope. The medial end of the olfactory nerve and the middle cerebral artery were seen through the thin brain dura. The midbrain artery close to the lenticulostriate artery (LSA) was exposed and ligated using a miniclip. The midbrain artery and the lenticular striatum behind the miniclip were corroded with bipolar coagulant. During surgery, the temperature of the right temporal muscles and rectum was maintained at about 37 ° C. using a heating pad. The contracted temporal nerves were returned to place and sutured. The animals were placed in cages and kept warm using a heating lamp overnight. Twenty four hours after midbrain artery ligation, the animal's neck was cut quickly, soaked in cold saline, and sliced at 2 mm intervals from the frotal pole. The slices were freshly prepared in salt and incubated in 2% TTC solution preheated to 37 ° C. Slices chopped for 30 minutes at 37 ° C. were fixed with 4% formalin solution. An infarct region of each slice was measured with an image analyzer using a stereomicroscope. Infarct size in the left hemisphere was measured by subtracting the left hemisphere infarct size from the right hemisphere infarct size to rule out the effect of edema formation.

The 4- (terminal-substituted-alkylamino) -quinoline-2-carboxylic acid derivative of the present invention is a potent and unique antagonist that acts on the trikinin non-sensitive glycine binding site in the NMDA receptor complex, and has a strong antagonist action on the NMDA receptor. It penetrates well into the central nervous system and has high solubility.

The compounds of the present invention are useful for the treatment or prevention of neurodegenerative diseases, and are particularly useful for reducing damage to the central nervous system caused by anemia or hypoxia such as heart attack, hypoglycemia, ischemia, cardiac arrest, trauma.

In addition, the compounds of the present invention are not only used for the prevention and treatment of chronic neurodegenerative diseases including epilepsy, Alzheimer's disease, Huntington's disease and Parkinson's disease, but also have an effect as antispasmodic, analgesic, antidepressant, anti-anxiety and antischizophrenic agents. have.

Claims (16)

4- (terminal substituted-alkylamino) -quinoline-2-carboxylic acid derivative having the structure of Formula 1, tautomeric isomers thereof and pharmaceutically acceptable salts thereof. Formula 1 In Chemical Formula 1, n is selected from 0 to 10; Nu is arylthio, alkylphosphonate, arylsulfonyl, thiourea, non-aromatic thio, heterocyclic thio, arylphosphonate, heterocycle, alkyl, aryl as appropriately substituted or unsubstituted , Cyano, hydroxyl, halogen, alkylphosphonate, arylphosphonate, alkylsulfonyl, arylsulfonyl, carbamate, amide, urea, acylurea, sulfonylurea, acylthiourea, sulfonylthiourea, Thiocarbamate, amidine, guanidine, imidate, thioimidate, phosphorylamide, sulfonamide or amine and the like. The 4- (terminal substituted-alkylamino) -quinoline-2-carboxylic acid derivative, tautomer isomer thereof and pharmaceutically acceptable thereof, according to claim 1, wherein the appropriately substituted arylthio is a substituent of the formula (3) salt. Formula 3 In Formula 3, R 'is halogen, alkyl, aryl, alkylamino, arylamino, alkoxy, aryloxy, fused heterocycle, guanidine, imidate, phosphorylamide, sulfonamide, urea, halogenated alkyl, sia Furnace or carbocycle and the like. The 4- (terminal substituted-alkylamino) -quinoline-2-carboxylic acid derivative, tautomer isomer thereof and pharmaceutically acceptable compound according to claim 2, wherein R 'is m-CH ₃ , or p-NH ₂ . Possible his salts. The 4- (terminally substituted-alkylamino) -quinoline-2-carboxylic acid derivative according to claim 1, wherein the non-aromatic thio is alkylthio, aralkylthio, carbocyclicthio or soluble bicyclic thio. Tautomeric isomers thereof and pharmaceutically acceptable salts thereof. The alkyl group is an alkyl group having _{1 to 20} carbon atoms, The aryl group is an aryl group having a carbon number of C ₆ ~C _12, Heterocyclic groups are characterized in that C ₃ -C ₇ heterocycloalkyl, C ₃ -C ₇ heterocycloalkyl (C ₁ -C ₆ ) alkyl, heteroaryl and heteroaryl (C ₁ -C ₆ ) alkyl -(Terminal substituted-alkylamino) -quinoline-2-carboxylic acid derivatives, tautomeric isomers thereof and pharmaceutically acceptable salts thereof. The C1-C20 alkyl group of claim 5, wherein the C ₁ -C ₂₀ alkyl group comprises methyl, ethyl, propyl, isopropyl, butyl, secondary-butyl and tert-butyl groups, Aryl groups having _{6 to 14} carbon atoms include phenyl, naphthyl, phenanthryl, anthracyl, indenyl, azulenyl, biphenyl, biphenylenyl and fluorenyl groups; Suitable heterocycloalkyl groups in the heterocyclic group include piperidyl, piperazinyl and morpholinyl groups; Suitable heteroaryl groups include thiophenyl, furyl, pyrrolyl, indolyl, thiazolyl, benzothiazolyl, oxazolyl, benzooxazolyl, imidazolyl, tetrazolyl, triazolyl, pyridyl, pyrimidinyl and phthalimidyl groups 4- (terminal substituted-alkylamino) -quinoline-2-carboxylic acid derivative, tautomer isomer thereof and pharmaceutically acceptable salt thereof. The arylthio of claim 1, wherein n is selected from 0 to 3, and Nu is R 'is m-CH ₃ or p-NH ₂ ; Alkyl phosphonates; Arylsulfonyl; Or 4- (terminal substituted-alkylamino) -quinoline-2-carboxylic acid derivative, tautomer isomer thereof and pharmaceutically acceptable salt thereof, characterized in that it is thiourea. The compound of claim 1 wherein the compound is 5,7-dichloro-4- [3- (3-phenylthioureido) propylamino] -quinoline-2-carboxylic acid, 5,7-dichloro-4- [3- (4-methoxyphenylthioureido) propylamino] -quinoline-2-carboxylic acid, 5,7-dichloro-4- [3- (m-tolylsulfanyl) propylamino] -quinoline-2-carboxylic acid, 5,7-dichloro-4- [3- (4-aminophenylsulfanyl) propylamino] -quinoline-2-carboxylic acid, 5,7-dichloro-4- [3- (diethoxyphosphoryl) propylamino] -quinoline-2-carboxylic acid, 5,7-dichloro-4- [3- (toluene-3-sulfinyl) propylamino] -quinoline-2-carboxylic acid and 5,7-dichloro-4- [3- (toluene-3-sulfonyl) propylamino] -quinoline-2-carboxylic acid 4- (terminal substituted-alkylamino) -quinoline-2-carboxylic acid derivative, tautomer isomer thereof and pharmaceutically acceptable salt thereof. An antagonist of excitatory amino acids to the NMDA receptor containing the compound of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient. A therapeutic agent for damage to the central nervous system caused as a result of anemia or hypoxia such as stroke, hypoglycemia, ischemia, cardiac arrest, trauma containing the compound of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient. A prophylactic and therapeutic agent for neurodegenerative diseases containing the compound of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient. A therapeutic agent for epilepsy, cerebral hemorrhage, Alzheimer's disease, Huntington's disease and Parkinson's disease containing the compound of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient. An anticonvulsant, analgesic, antidepressant, anti-anxiety and antischizophrenic agent comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient. 1) alkylating the compound of formula 31 with 1,3-dibromopropane to obtain a bromide compound of formula 32 (first step); 2) sulfuric acid treatment of the compound of formula 32 obtained in the first step to remove N-tosyl groups (-NTs), and then substituted bromine with NaN ₃ to obtain an azido compound of formula 33 (second step); 3) catalytic hydrogenation of the compound of formula 33 obtained in the second step and reaction with an appropriate thioisocyanate to obtain the thiourea compound of formula 34 (third step); And 4) basic hydrolysis of the compound of formula 34 prepared in the third step to obtain a compound of formula 1 which is the target compound of the present invention (fourth step) Nu of the formula (1), characterized in that consisting of thiourea, carbamate, amide, urea, acylurea, sulfonylurea, acylthiourea, sulfonylthiourea, thiocarbamate, amidine, guanidine, imidate, thio A process for preparing 4- (terminal substituted-alkylamino) -quinoline-2-carboxylic acid derivative of the present invention applied to imidate, phosphorylamide, sulfonamide or amine. Scheme 1 1) treating the compound of formula 32 with an arylthiol (HSAr) compound to prepare a sulfide compound of formula 36 or treating with triethylphosphite [P (OEt) ₃ ] to produce a phosphonate of formula 37 Stage 1); And 2) removing the N-tosyl group (-NTs) by sulfuric acid treatment of the compound of formula 36 or 37 obtained in the first step and basic hydrolysis to obtain the compound of formula 1 which is the target compound of the present invention (second step) A method for producing 4- (terminal substituted-alkylamino) -quinoline-2-carboxylic acid derivative of the present invention, which is applied when Nu in Formula 1 is an arylthio, alkylphosphonate or the like, which is suitably substituted. Scheme 2 In Scheme 2, X is the same as Nu in the general formula (1). 16. The sulfonyl and sulfide of formula 40 according to claim 15, wherein in the case where Nu of formula 1 is sulfide, in addition to the second step, the target compound is reacted with m-chloroperbenzoic acid (m-CPBA) and oxidized A process for preparing the 4- (terminal substituted-alkylamino) -quinoline-2-carboxylic acid derivative of the present invention, which comprises preparing a sulfinyl compound. Scheme 3