KR19990071057A - 4- (Terminal Substituted-alkylamino) -quinoline-2-carboxylic acid derivatives which act as NMDA receptor antagonists - Google Patents

Abstract

The present invention relates to a 4- (terminally substituted-alkylamino) -quinoline-2-carboxylic acid derivative of formula 1, a process for its preparation, and a potent and specificity for the strychinine non- As an < / RTI > antagonist. The compounds of the present invention are useful for the treatment and prevention of neurodegenerative diseases. In particular, the compounds of the present invention are useful for reducing central nervous system damage caused as a result of anemia such as stroke, hypoglycemia, ischemia, cardiac arrest, trauma, or hypoxia. In addition, the compounds of the present invention are useful for the prevention and treatment of chronic neurodegenerative diseases including epilepsy, Alzheimer's disease, Huntington's disease and Parkinson's disease. In addition, the compounds of the present invention are used as anticonvulsants, analgesics, antidepressants, antianxiety agents, and antipsychotic drugs.

(Wherein Nu is as defined in the specification).

Description

4- (Terminal Substituted-alkylamino) -quinoline-2-carboxylic acid derivatives which act as NMDA receptor antagonists

The present invention relates to a 4- (terminally substituted-alkylamino) -quinoline-2-carboxylic acid derivative of formula (I) which acts as an antagonist of excitatory amino acids, a process for its preparation and its use as a therapeutic agent for neurological disorders.

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

In addition, the compounds of the present invention are useful for the prevention of 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, antianxiety agents and antipsychotic agents.

The most important excitatory neurotransmitter in mammalian central nervous system neurotransmission is L-glutamate. Almost all of the central nervous system nerve cells are excited by L-glutamate, which acts on a variety of receptor-ion channel complexes present on the surface of neurons, opening ion channels and introducing cations such as sodium or calcium To stimulate nerve cells by inducing polarization between the nerve cell membranes.

These central nervous system neurons express NMDA receptors excited by N-methyl-D-aspartate (N-methyl-D-aspartate, hereinafter abbreviated as NMDA) and non-NMDA receptors that are not excited by NMDA .

The NMDA receptor is a cell surface protein complex distributed widely in the brain or spinal cord. It is known to transmit neuronal excitability in neuronal synapses and is closely related to neuronal cell growth.

One of the characteristics of this NMDA receptor is that it is completely blocked by Mg ⁺⁺ when the cell is at rest. However, this blockage is voltage-dependent and the block is released if the cell is activated by a non-NMDA receptor or another excitable material is introduced and becomes partially depolarized. Thus, the mechanism of action of NMDA receptors plays an important role in the learning and memory process because it depends on conditions.

Another characteristic of the NMDA receptor is that if the receptor is excessively excited, the cell will die. This seems to be due to excessive accumulation of Ca ⁺⁺ in the cells. Recently, many studies have been conducted on the apoptosis caused by excitotoxicity of the NMDA receptor.

When neurons, neurons, are cultured in vitro and then treated with glutamate, the cells are swollen and cytotoxic. This excitatory toxicity is dependent on the presence of Ca ^{++. When} a large amount of Ca ⁺⁺ enters the neuron through the ion channel of the glutamate receptor, normal cell activity is destroyed and the release of glutamate is accelerated again by feedback Excitatory toxicity occurs. In this process, proteases and lipases are activated, resulting in the destruction of nerve cell membranes and the production of free radicals [JW McDold, MV Johnson, Brain Res. Reviews 15, 41 (1990). Thus, excitotoxicity caused by neurotransmitters kills neurons, and this research continues to reveal that this is a major cause of neurodegenerative diseases and stroke associated with brain cell death.

In recent years, the focus has been focused on the mechanisms by which excessive damage to the central nervous system occurs due to excessive secretion of L-glutamate by brain damage, spinal cord injury, stroke, ischemia or hypoglycemia. Studies have shown that central nervous system Lt; RTI ID = 0.0 > NMDA < / RTI > receptor antagonist.

NMDA receptor antagonists prevent many clinical disorders including ischemic 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)]. Recently, several NMDA receptor antagonists have been used to treat acute stroke and brain damage.

It has also been reported that NMDA receptor antagonists are useful for improving memory and learning, examples being 1-aminocyclopropanecarboxylic acid methyl ester, a specific glycine ligand, D- cycloserine and R - (+) - 3- Amino-1-hydroxypyrrolidin-2-one- (HA-966) [Bliss, TV P, Collingride, GL, Nature 345 , 347 (1990); GB 2231048A (1990)].

A recent report suggests that NMDA receptor antagonists have analgesic, antidepressant, antipsychotic, and anxiolytic 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)).

Noncompetitive NMDA receptor antagonists such as ketamine, dextromethorphan, and the like are useful for the treatment of childhood growth disorders, autism, and the like. 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 considered to be one of the causes of these diseases is more evident by recent studies on their physiological functions and the structure of subunits [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)]. There are at least five distinct substrate binding sites in the NMDA receptors, including (a) a binding site that binds to the neurotransmitter L-glutamate, (b) an allosteric modulator site that binds to glycine, (c) (D) a binding site that binds to Mg ⁺⁺ and (e) a binding site that binds to the divalent cation Zn ⁺⁺ and is known to inhibit [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 actions of the subunits of NMDA receptors, NMDA receptors are activated by glutamate and glycine or their agonists. Also, the channel of associated Ca ⁺⁺ permeability is blocked by Mg ⁺⁺ in a physiologically voltage-dependent manner, and Zn ⁺⁺ , with its own regulatory site, reduces the activity of the NMDA receptor synapses [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)].

Over the past two decades pharmacological interest has been focused on NMDA receptors, and efficants and antagonists that rely on each of the binding sites described above have been identified as candidates for useful drugs. Among them, the most promising method for controlling NMDA receptor activity was the development of an allosteric modulator of glycine binding sites.

In 1987, Johnson and Ascher discovered a glycine binding site in the NMDA receptor, and subsequent studies have shown that the glycine binding site and the glutamate binding site of the NMDA receptor are present in the same protein and the glycine binding site of the NMDA receptor Lt; RTI ID = 0.0 > allosteric < / RTI >

Since there is a negative allosteric coupling, binding of the agonist at the glutamate recognition site reduces the affinity of glycine for the glycine recognition site, and antagonist binding at the glutamate recognition site is enhanced by the glycanside antagonist, 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 microdialysis (dialysis) studies have found that a large amount of glutamate is released in the ischemic brain region in the mouse ubiquitous ischemic model, but release of glycine is scarce [Globus, MY T, et al., J. Neurochern. 57 , 470-478 (1991).

As a result of the above experiment, it can be seen that the glycine antagonist is a very potent neuronal cell protective agent that can reduce excessive excitement of neurons when the neuron, which is a neuron, is excessively excited by glutamate and causes cytotoxicity.

Glycine antagonists can prevent NMDA channels from opening by acting in a non-competitive manner with glutamate, thereby overcoming the high concentration of endogenous glutamate released in the ischemic brain regions, as opposed to competitive NMDA antagonists that compete with glutamate You do not have to. As a result, NMDA receptors are regulated by glycine antagonists rather than completely suppressed, and this regulatory action may be more physiological than receptor blockade of receptor function (as compared to channel blockers), so glycine antagonists Have fewer side effects than other antagonists. In fact, glycine antagonists can be injected directly into the brain of a rodent 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)].

Thus, glycine antagonist is a noncompetitive antagonist that regulates NMDA receptors rather than blocks them. As a result, NMDA receptor blockade results in less side effects than other forms of NMDA receptor antagonists, leading to the development of new central nervous system agents, It has emerged as a target substance.

Glycine antagonists are emerging candidates for central nervous system drugs because they have a broad therapeutic window between the desired 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 is that most of these compounds do not pass through the blood-brain barrier (BBB). As a result of intensive efforts to develop a glycine-site ligand capable of passing through the blood-brain membrane, kynurenic acid derivatives (2-carboxyquinoline), 2-carboxyindole derivatives, quinoxaline derivatives and 2- Quinolone derivatives and the like have been developed. They are also selective antagonists acting on NMDA receptors as antagonists that act 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; WO94 / 20470; WO93 / 10783, EP 685,466 A1 and EP 481,676 A1].

It has recently been found that the quinrenic acid derivative, which is the compound No. 1 in the following formula (2), exhibits selectivity for NMDA antagonistic activity. That is, an endogenous product of the tryptophan metabolic pathway, has selective NMDA antagonistic activity by blocking the glycine regulatory site of the NMDA receptor [Kessler, M., et al., J. Neurochem. 52, 1319 (1989); Kemp, JA, et al., Proc. Natl. Acad. Sci. USA 85 , 6547 (1988)].

In the structure activity relationship (SAR) study on most of the existing NMDA antagonists, the hydroxyl group at the C-4 position of the quinrenic acid interacts with the H-bond of the receptor and its spatial orientation Lt; RTI ID = 0.0 > binding activity. ≪ / RTI > That is, when an electron-rich substituent is appropriately introduced at the C-4 position of the kinesinic acid parent nucleotide, the binding affinity to the NMDA receptor increases.

Recently, Harrison et al. Have found that compounds 2 and 3 of the following formula 2, in which the hydroxyl group at the C-4 position of the quinolenic acid is replaced by acetic acid with a heteroatom, are more effective and selective than kynurenic acid itself (Harrison BL, et al., J. Med. Chem. 33 , 3130 (1990)). However, these compounds have a disadvantage in that they lack in vivo activity because they can not penetrate BBB well due to high polarity of carboxyl group.

Accordingly, the present inventors have made efforts to produce a kynurenic acid derivative having excellent in vivo activity, and a kynrenic acid derivative in which a hydroxyl group at the C-4 position is substituted with an alkylamino group substituted at the terminal is structurally different from a conventional kynurenic acid And thus the present invention has been completed.

It is an object of the present invention to provide a quinrenic acid derivative substituted at the C-4 position, which acts as an antagonist of an excitatory amino acid, and a process for producing the same.

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

Hereinafter, the present invention will be described in detail.

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

Formula 1

In Formula 1,

n is selected from 0 to 10;

Nu is an optionally substituted or unsubstituted arylthio, alkylphosphonate, arylsulfonyl, thiourea, non-aromatic thio, heterocyclic thio, arylphosphonate, heterocycle, alkyl, aryl An alkylthio group, an alkylsulfonyl group, an alkylsulfonyl group, an arylsulfonyl group, an arylsulfonyl group, an arylsulfonyl group, an arylsulfonyl group, an arylsulfonyl group, an arylsulfonyl group, Thiocarbamate, amidine, guanidine, imidate, thioimidate, phosphorylamide, sulfonamide or amine.

The appropriately substituted arylthio of Nu may be represented by the following formula (3);

Wherein R 'is selected from the group consisting of halogen, alkyl, aryl, alkylamino, arylamino, alkoxy, aryloxy, fused heterocycle, guanidine, imidate, phosphorylamide, sulfonamide, urea, Furnace or carbocycle.

The non-aromatic thio of said Nu also includes 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 the above formula (3), when R 'is m-CH ₃ or p-NH ₂ and Nu is an alkylphosphonate , Arylsulfonyl or thiourea.

The alkyl group is a C ₁ -C ₂₀ alkyl group, preferably a C ₁ -C ₄ alkyl group, and includes methyl, ethyl, propyl, isopropyl, butyl, sec-butyl and tert-butyl groups.

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

The heterocyclic group includes 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, benzoxazolyl, imidazolyl, tetrazolyl, triazolyl, pyridyl, pyrimidinyl and phthalimidyl groups .

For use in medicine, the salt of the compound of formula (I) is not toxic and should be a pharmaceutically acceptable salt. A variety of salts may be used to prepare the compounds of the present invention or their non-toxic, pharmaceutically acceptable salts.

The pharmaceutically acceptable salts of the compounds of formula (I) include alkali metal salts such as lithium, sodium or potassium salts and include alkaline earth metal salts such as calcium or magnesium salts and suitable organic ligands For example, quaternary ammonium salts. Acid addition salts can be prepared by mixing solutions of the compounds of the invention with solutions of pharmaceutically acceptable non-toxic acids such as hydrochloric acid, fumaric acid, maleic acid, succinic acid, acetic acid, citric acid, tartaric acid, have.

The present invention includes within its scope prodrugs of the compounds of formula (I). In general, such prodrugs are functional derivatives of the compound of formula (I) and should be readily convertible into the compound required to enter the organism and exhibit its drug 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 according to the present invention has at least one asymmetric center, if an enantiomer can be present and the compound according to the present invention has two or more asymmetric centers, a diastereoisomer, Lt; / RTI > 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 exemplify 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) 5,7-dichloro-4- [3- (4-methoxyphenylthioureido) propylamino] -quinoline-

3) 5,7-Dichloro-4- [3- (m-tolylsulfanyl) propylamino] -quinoline-

4) 5,7-Dichloro-4- [3- (4-aminophenylsulfanyl) propylamino] -quinoline-

5) 5,7-Dichloro-4- [3- (diethoxyphosphoryl) propylamino] -quinoline-

6) 5,7-Dichloro-4- [3- (toluene-3-sulfinyl) propylamino] -quinoline-

7) 5,7-Dichloro-4- [3- (toluene-3-sulfonyl) propylamino] -quinoline-

The pharmaceutical compositions of the present invention are preferably unit dosage forms such as tablets, capsules, powders, granules, sterile solutions or suspensions, or suppositories for oral, intravenous, parenteral or rectal administration. In order to prepare solid compositions such as tablets, the major active ingredient is mixed with a pharmaceutical carrier such as conventional tableting ingredients such as corn starch, 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 a suitable excipient, such as, for example, calcium phosphate or gums and other pharmaceutical diluents, . Once the preliminary solid composition is homogenized, the active ingredient is evenly dispersed throughout the composition, allowing the composition to be easily split into effective unit dosage forms containing the same amount as tablets, pills, and capsules. This preliminary solid composition is further subdivided into unit dosage forms of the type mentioned above containing from 0.1 to 500 mg of the active ingredient of the present 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 can be comprised of an inner dosage component and an outer dosage component, wherein the outer dosage component surrounds the inner dosage component. These two components can be separated by an enteric membrane, which prevents degradation in the stomach and allows the internal components to pass through the duodenum without delay and delay the release of internal components. A variety of materials are used as such enteric coatings or coating materials, including a number of polymeric and polymeric acids and mixtures of materials such as shellac, cetyl alcohol, and cellulose acetate. The liquid form of the novel compositions of the present invention made for oral administration or administration by injection may be in the form of aqueous solutions, suitably syrups, aqueous or oily suspensions, and emulsions of edible oils such as cottonseed oil, sesame oil, coconut 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 an appropriate dosage is 0.01 to 250 mg / kg per day, preferably 0.05 to 100 mg / kg per day, more preferably 0.05 to 5 mg / kg per day. The present compounds may conveniently be administered by intravenous injection.

The process for preparing the compound of the formula (1), which is a compound of the present invention, is shown in the following Reaction Schemes 1 and 2.

That is, the process for preparing the compound of formula (1) of the present invention can be explained by the reaction scheme 1 and the reaction scheme 2 according to the kind of Nu which is a substituent of the alkylamino group.

Scheme 1 shows that Nu of Formula 1 is a thiourea, carbamate, amide, urea, acylurea, sulfonylurea, acylthiourea, sulfonylthiourea, thiocarbamate, amidine, guanidine, imidate, thioimidate, A phosphonamide, a sulfonamide, an amine, or the like, and particularly exemplifies the case where Nu is thiourea;

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

2) a step of treating the compound of the formula 32 obtained in the first step with sulfuric acid to remove the N-tosyl group (-NTs), followed by replacement of the bromine with NaN ₃ to obtain an azido compound of the formula 33 (second step);

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

4) a step of basic hydrolysis of the compound of the structural formula 34 prepared in the third step to obtain the compound of the formula 1 (the fourth step)

.

In Scheme 2, X represents Nu in Formula 1, where Nu is an appropriately substituted arylthio or alkylphosphonate;

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

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

.

When Nu is arylthio in the above reaction scheme 2, m-chloroperbenzoic acid (m-CPBA) is reacted and oxidized in addition to the second step to convert di-oxidized aryl Arylsulfonyl and mono-oxidized arylsulfinyl compounds can be prepared, which is illustrated in Scheme 3 below.

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

The compound of formula 31, which is the starting material in Scheme 1, can be readily prepared by conventional methods by reacting the kynurenic acid methyl ester with p-toluenesulfonyl isocyanate.

The first step is a step of obtaining a bromide compound in which the starting material is alkylated at the C-4 position of the kynurenic acid by reacting and refluxing in the presence of dibromopropane in the presence of K ₂ CO _{3. As} the reaction solvent used, acetonitrile .

The second step is a step of treating the compound prepared in the first step with concentrated sulfuric acid to remove the N-tosyl group (-NTs), followed by substituting bromine with NaN ₃ to obtain an azido compound, DMF is available.

The third step is a step of hydrogenating the azide compound in the presence of a Pd-C catalyst and reacting with an appropriate thioisocyanate in the presence of triethylamine (Et ₃ N) to prepare thiourea,

In the fourth step, the compound prepared in the third step is subjected to ordinary basic hydrolysis to obtain the compound of formula (1) as the final target compound, and the methyl ester prepared in the third step is hydrolyzed under basic conditions to prepare a carboxylic acid .

In Scheme 2, the first step is the treatment of the compound of formula 32 prepared in the first step of Scheme 1 with an aryl thio compound and Na ₂ CO ₃ in an anhydrous tetrahydrofuran (THF) solution to convert the bromide to a sulfide, Is the step of dialing the bromide to the phosphonate by simply refluxing the compound of formula 32 in the presence of triethyl phosphite.

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

In addition to the above process, when the Nu of the final target compound is sulfide, the reaction is carried out by reacting the compound with m-CPBA in a methylene chloride solvent to oxidize the mono-oxydized sulfinyl compound and di -Oxidized sulfonyl compound into an inseparable mixture can be further carried out.

Hereinafter, the present invention will be described in detail with reference to examples. The following examples illustrate the invention and are not to be construed as limiting the invention thereto.

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

(0.42 g, 1.0 mmol) and K ₂ CO ₃ (0.55 g, 4.0 mmol) were added to acetonitrile (30 mL) and 1,3-dibromopropane 4.0 mL, 30 mmol) was added and the mixture was refluxed with heating for 3 h. The remaining insoluble solids were filtered off and the filtrate was evaporated on a rotary evaporator to give a brown residue. The 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 캜; _{^{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 = 8.0Hz, ArH), 7.50 (d, 2H, J = 8.8Hz, 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

Quinolinicsulfonamide (10 mL, 90% in water), which is the title compound of step 1 above, was added portionwise to a solution of sulfuric acid (1.50 g, 2.75 mmol) in an ice bath, Lt; / RTI > The reaction temperature should be kept below 4 ° C. The resulting mixture is poured slowly over crushed ice for 1 hour while stirring. An aqueous solution containing insoluble solids was basified with 8N aqueous NaOH solution at pH 12. The remaining solid was filtered, washed with water (50 mL) and dried in vacuo to give 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 = 6.8Hz, CH 2 Br), 3.56 (t, 2H, J = 6.8Hz, 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 = 2.2Hz, ArH), 8.05 ( d, 1 H, J = 2.2 Hz, ArH).

The bromide (0.90 g, 2.38 mmol) and _{NaN 3 (0.62 g, 9.52 mmol} ) prepared above in DMF (20 mL) into the suspension was heated for 3 hours at 50 ℃ temperature. 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 x 2). The organic layer was evaporated under reduced pressure and the residue was purified by column chromatography (silica gel, 90% EtOAc in n-hexane) to give the pure azido compound as a pale yellow solid (0.80 g, yield 99%).

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

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

To the mixed solution of ethyl acetate-methanol (4: 1, 20 mL) was added a suspension of the azodomethyl ester compound (0.80 g, 2.35 mmol) prepared in the above step 2 and 10% palladium catalyst Under a hydrogen atmosphere (using a balloon-type flask). After stirring overnight, the reaction mixture was filtered through a pad of celite and the filtrate was evaporated under reduced pressure to give the crude amino compound as a colorless syrup (0.68 g, 92% yield). This crude syrup was diluted with methylene chloride anhydride (20 mL) and then phenylthioisocyanate (0.38 g, 2.85 mmol) and triethylamine (0.40 mL, 2.85 mmol) were carefully added. The reaction mixture was stirred at room temperature for 6 hours, diluted with 100 mL of methylene chloride and washed with 1N HCl (100 mL), H ₂ O (100 mL x 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 (0.32 g) as a colorless solid (0.54 g, yield 50%).

_{^{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), 2H, ArH), 7.28 (s, 1H, ArH), 7.30-7.22 (t, 1H, J = 4.5 Hz, NH) 7.68 (br, 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), 2H, ArH), 7.23 (s, 1H, ArH), 7.28 (t, 1H, J = 1H), 7.98 (d, 1H, J = 6.6Hz, ArH), 8.11 (d, 1H, , ≪ / RTI > 1H, J = 2.1 Hz, ArH).

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

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 a mixed solvent of 10 mL of THF-H ₂ O (1: 1) Lt; / RTI > The mixture was diluted with 20 mL of water and extracted with methylene chloride (20 mL x 2). The aqueous layer was acidified with 1N hydrochloric acid at pH 3. The resulting white precipitate was filtered, washed with water and then dried under vacuum to give the title compound of the present invention as a white solid (0.18 g, yield 75%) with an unidentified solid.

^{1 H-NMR (DMSO-d} 6) δ 2.12 (m, 2H, CH 2), 3.61~3.82 (m, 4H, NCH 2 CH 2 N), 7.14~7.56 (m, 5H, ArH), 7.85 (d 1H, J = 7.6 Hz, ArH), 8.32 (s, 1H, ArH), 8.48 (br s, , ≪ / RTI > NH), 10.04 (br s, 1H, NH);

Unidentified compounds:

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

Example 2 Synthesis of 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 reaction was carried out in the same manner as in step 1 of Example 1 to give the title compound.

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

The reaction was carried out in the same manner as in step 2 of Example 1 to give the title compound.

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

The reaction was carried out in the same manner as in the step 3 of Example 1 except that 4-methoxyphenylthioisocyanate was used to prepare the title compound (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 = 4.2Hz, NH), 6.74 (br t, 1H, J = 4.2Hz, NH), 6.88 (d, 2H, J = 8.0Hz 1H, ArH), 7.04 (d, 2H, J = 8.0 Hz, ArH), 7.14 (br s, , 8.10 (d, 1 H, 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 = 8.1Hz, ArH), 6.91 (br t, 1H, NH), 7.08 (d, 2H, 1H, J = 2.1 Hz, ArH), 7.96 (br s, 1H, NH), 8.06 (d, Hz, ArH).

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

The title compound was obtained (0.28 g, 0.71 mmol) in the same manner as in step (4) of Example 1 using the title compound of step 2 of Example 2 (0.35 g, 0.71 mmol) g, yield 82%).

^{1 H-NMR (DMSO-d} 6) δ 2.14 (m, 2H, CH 2), 3.62~3.78 (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 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), 3.60~3.80 (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, IH, NH), 7.25 (m, IH, ArH), 7.30 (d, 2H, J = 8.7 Hz, ArH) , 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-carboxylic acid

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

The reaction was carried out in the same manner as in step 1 of Example 1 to give the title compound.

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

M-thiocresol (0.41 g, 3.30 mmol) and K ₂ CO ₃ (0.76 g, 5.50 mmol) were added to a solution of the above bromide (1.50 g, 2.75 mmol) in THF (30 mL) . The mixture was refluxed for 4 hours and then cooled to room temperature. The solution was diluted with methylene chloride (100 mL), washed with brine (50 mL x 2) and water (50 mL x 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 hexanes) to give the title compound as a white solid (1.39 g, 82% yield).

_{^{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, 1H, ArH), 7.46 (d, 2H, J = 8.5 Hz, ArH), 7.76 (d, 2.2Hz, ArH), 8.28 (d, 1H, J = 2.2Hz, ArH).

(Step 3) 5,7-Dichloro-4- [3- (m-tolylsulfanyl) propylamino] -quinoline-

Sulfonamide (1.0 g, 1.70 mmol), the title compound from step 2 above, 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 h Respectively. The reaction temperature should be kept below 4 ° C. The resulting mixture is poured slowly over crushed ice for 1 hour while stirring. An aqueous solution containing insoluble solids was basified with aqueous 4 N NaOH at pH 10. The mixture was extracted with methylene chloride (50 mL x 3) and the organic layer was washed with water (100 mL x 2) and brine (50 mL) and then concentrated in vacuo to afford crude aminoquinolinic acid methyl ester as a pale yellow solid (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 = 7.1Hz, CH 2 S), 3.38 (dt , 2H, J = 4.2, 7.1Hz , 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 0.4 N aqueous NaOH (17 mL, 6.80 mmol) at room temperature for 2 h. The mixture was concentrated under reduced pressure to a total volume of 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 x 3), and dried in vacuo to give the title compound (0.57 g, 83% yield) as a white solid carboxylic acid.

^{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, IH, ArH), 7.78 (d, IH, J = 2.4 Hz, ArH), 8.04 1H, J = 2.4 Hz, ArH).

Example 4 Synthesis of 5,7-dichloro-4- [3- (4-aminophenylsulfanyl) propylamino] -quinoline-

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

The reaction was carried out in the same manner as in step 1 of Example 1 to give the title compound.

(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). Purification by flash column chromatography (30% EtOAc in hexanes) after workup provided pure sulphide 2o as the title compound (1.11 g, 82% yield) as a white solid.

_{^{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 = 7.2Hz, 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 = 8.6Hz , 7.48 (d, 2H, J = 8.4 Hz, ArH), 7.01 (d, 2H, J = 8.6 Hz, ArH), 7.28 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-

The reaction was carried out in the same manner as in step 3 of Example 3 with the methyl ester (1.0 g, 1.66 mmol) prepared in this Example (Step 2) to give 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 = 7.0Hz, 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), 9.38 (br s, 1H, NH).

Example 5 Synthesis of 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 reaction was carried out in the same manner as in step 1 of Example 1 to give the title compound.

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

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

_{^{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, 2OCH 2), 4.02 (s, 3H, OCH 3), 7.30 (d, 2H, J = (D, 2H, ArH), 7.46 (d, 2H, J = 8.0 Hz, ArH), 7.48 = 2.2 Hz, ArH).

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

The reaction was carried out in the same manner as in step 3 of Example 3 with the methyl ester prepared in this Example (Step 2) (1.0 g, 1.66 mmol) to give the title compound as a white solid (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 = 2.3Hz, ArH), 8.01 (d, 1H, J = 2.3 Hz, ArH);

The 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, 2OCH 2),} 7.43 (s, 1H, ArH), 7.92 (d, 1H, J = 2.4Hz, ArH), 8.37 (d, 1H, J = 2.4Hz, ArH), 9.18 (br t, 1H, J = 7.6 Hz, NH).

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

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

5,7-Dichloro-4- [3- (toluene-3-sulfonyl) propylamino] -quinoline-

^{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-

^{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 , ≪ / RTI > 1H, ArH), 7.75 (m, 3H, ArH).

Ⅰ. In vitro screening for binding activity

1) Preparation of synaptic membrane

Male Sprague-Dawaur (300-400 g) was obtained from the laboratory animal laboratory of the Korea Research Institute of Chemical Technology. Experimental animals were exposed to water and standard water in an air-conditioned room (22 + 1 C (relative humidity, 60 + 5%)) with slight darkening (brightness at 8 am) during the 4-10 day preparation period prior to use. They were fed with laboratory food. The synaptic membrane for receptor binding studies is a modified method of Foster and Fagg [Eur. J. Pharmacol. 133, 291 (1987)] and Murphy et al. [Br. J. Pharmacol. 95, 932 (1988). To summarize, the male Sprague-Dawaur rat was killed, the hippocampus of the brain cortex and brain were chopped with a scalpel and homogenized 5 times with 10 volumes of 0.32 M sucrose using a Teflon-fiber homogenizer. The mixture was centrifuged for 10 minutes at 1000 x g for 10 minutes with a Beckman J2 / 21 centrifuge (roto: JA20), and the supernatant was collected and centrifuged at 20,000 x g for 20 minutes. The supernatant was removed and homogenized with a pellet homogenizer (setting 5, 30 sec).

After incubating for 30 minutes at 4 ° C, the membrane suspension was centrifuged at 39,800 x g for 25 minutes with a Beckman L8-M ultracentrifuge. The pellet was left overnight at -70 < 0 > C. The next day, the pellet was dissolved at room temperature for 10 minutes and resuspended in 20 volumes of 50 mM Tris-acetone (pH 7.1, 4 DEG C) containing 0.04% Triton X-100, incubated at 37 DEG C for 20 minutes, Lt; RTI ID = 0.0 > 3900 x g. ≪ / RTI > The pellet was centrifuged as above and washed 3 times 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 the protein concentration was measured using Bio-Rad agent (Bradford, 1976). The resuspension buffer solution 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 the [ ³ H] glycine saturate binding assay, the synaptic membrane (100 μg of membrane protein) was incubated with a final volume of 0.5 ml containing 50 mM Tris-acetate buffer solution pH 7.1, 5- 500 nM [ ³ H] glycine The reaction was carried out in a borosilicated glass tube containing the mixture at a temperature of 4 ° C for 30 minutes. For drug potential measurements, the synaptic membrane (100 μg of membrane protein) was incubated in a reaction mixture containing 50 mM Tris-acetate buffer at pH 7.1, 50 nM [ ³ H] glycine and various concentrations of the experimental compound as described above .

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

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

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

Compound No. Inhibition at 100 [mu] M% 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 convulsions experiment

Based on the method of Chugai, a dose-effect curve was obtained in which 40-160 ng of NMDA per mouse was administered iv. Lt; RTI ID = 0.0 > convulsions. ≪ / RTI > A 28-gauge needle attached to a 50-μl syringe was inserted through the 1-mm right brachymal bone on the sagittal suture and the test solution was inserted using a manipulator. Insertion volume was 5 μl per mouse. The spasm that occurred in response to NMDA occurred within 5 minutes and was characterized by 1) rough running or jumping, 2) myoclonic seizures (causing seizures and separation), and 3) (clonic seizures, loss of sense of direction and repeated movement of the limbs). The animals were i.c.v. 10 min after injection and observed seizure rate when seizure occurred. To investigate the NMDA-antagonistic properties of the experimental drug, the following procedure was performed. Experimental drug (infusion volume, 1 ml / 100 g) was administered ip. After 15 minutes of injection, mice were injected with 160 ng of NMDA at 5 μl per mouse. i.c.v. Experimental drug and NMDA solution were injected simultaneously for administration. Ten rats were used for the same dose. The dose was increased until the antagonistic NMDA reached a limit (eg, solubility, mortality) that could induce convulsions or could no longer be tested. NMDA was dissolved in a 0.9% sodium chloride solution, and AP5,5,7-dichloroquinolenic acid and 7-chloroquinolenic acid were dissolved in a minimum amount of 0.1 N sodium hydroxide to which 0.9% physiological saline had been added.

2) Permanent ligation of Middle Celebral Atery (MCA)

The middle cerebral artery ligation was performed by the method of 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% halothane and maintained at 1.5% halothane oxide: oxygen (70:30) using a halothane evaporator. After laying the animal on its side, the left temporalis muscles were cut and the anatomic coagulant was used to partially separate the angular arc. Small craniofacial resection was performed using micro drills cooled with cold saline in the front of the mandibular nerve under the surgical microscope. The innervation of the olfactory nerve and the middle cerebral artery were seen through the thin dura of the brain. The middle cerebral artery near the lenticulostriate artery (LSA) of the lens nucleus was exposed and ligated using a mini clip. The middle cerebral artery and the lens nucleus artery behind the mini clip were corroded using anodic coagulant. During surgery, the temperature of the right temporalis muscles and rectum was maintained at about 37 ° C using a heating pad. The contraction of the temporalis muscle was reversed and sutured. The animal was placed in a cage and kept warm overnight using a heating lamp. Twenty-four hours after ligation of the middle cerebral artery, the animal's neck was quickly removed, the brain was quickly removed, immersed in cold salt, and sliced at 2 mm intervals from the frotal pole. The slices were freshly prepared in saline and cultured in a 2% TTC solution preheated to 37 < 0 > C. The stiffened slices were fixed in 4% formalin solution at 37 ° C for 30 minutes. The infarct area of each slice was measured with an image analyzer using a stereoscopic microscope. The infarct size in the left hemisphere was measured by subtracting the left hemisphere infarct size from the right hemisphere infarct size to exclude the effect of edema formation.

The 4- (terminally substituted-alkylamino) -quinoline-2-carboxylic acid derivatives of the present invention are potent and unique antagonists acting on the strychnine nonsensitive glycine binding site in the NMDA receptor complex, It is well penetrated 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 the damage of the central nervous system caused by anemia such as heart attack, hypoglycemia, ischemia, arrest of heart beat, trauma, or hypoxia.

The compounds of the present invention are also useful as prophylactic and therapeutic agents for chronic neurodegenerative diseases including epilepsy, Alzheimer's disease, Huntington's disease and Parkinson's disease as well as antispasmodics, analgesics, antidepressants, antianxiety agents and antipsychotic agents have.

Claims (17)

(Terminally substituted-alkylamino) -quinoline-2-carboxylic acid derivatives, the tautomeric isomers thereof, the pharmaceutically acceptable salts thereof, and the prodrugs thereof, Formula 1 In Formula 1, n is selected from 0 to 10; Nu is an optionally substituted or unsubstituted arylthio, alkylphosphonate, arylsulfonyl, thiourea, non-aromatic thio, heterocyclic thio, arylphosphonate, heterocycle, alkyl, aryl An alkylthio group, an alkylsulfonyl group, an alkylsulfonyl group, an arylsulfonyl group, an arylsulfonyl group, an arylsulfonyl group, an arylsulfonyl group, an arylsulfonyl group, an arylsulfonyl group, Thiocarbamate, amidine, guanidine, imidate, thioimidate, phosphorylamide, sulfonamide or amine. 4. The compound according to claim 1, wherein the appropriately substituted arylthio is a substituent of the formula (3): wherein R < 1 > Salts and prodrugs (3) Wherein R 'is selected from the group consisting of halogen, alkyl, aryl, alkylamino, arylamino, alkoxy, aryloxy, fused heterocycle, guanidine, imidate, phosphorylamide, sulfonamide, urea, Furnace or carbocycle. The method of claim 2 wherein, R 'is a m-CH _3, or 4, characterized in that p-NH ₂ (terminal-substituted-alkyl-amino) -quinoline-2-carboxylic acid derivative, its tautomeric isomer, pharmaceutically acceptable Possible salts and prodrugs thereof 4. A compound according to claim 1, wherein the non-aromatic thio is an alkylthio, aralkylthio, carbocyclic thio or a substituted bicyclic thio, or a 4- (terminally substituted-alkylamino) -quinoline- Its tautomer, its pharmaceutically acceptable salt and its prodrug The positive resist composition according to claim 1, wherein the alkyl group is an alkyl group having _{1 to 20} carbon atoms, The aryl group is an aryl group having _{6 to 12} carbon atoms, The heterocyclic group is selected from the group consisting of a C ₄ -C ₇ heterocycloalkyl, a C ₃ -C ₇ heterocycloalkyl (C ₁ -C ₆ ) alkyl, a heteroaryl and a heteroaryl (C ₁ -C ₆ ) - (terminally substituted-alkylamino) -quinoline-2-carboxylic acid derivatives, their tautomeric isomers, their pharmaceutically acceptable salts and their prodrugs 6. A compound according to claim 5, wherein the C ₁ -C ₂₀ alkyl group is methyl, ethyl, propyl, isopropyl, butyl, sec-butyl and tert- The aryl group having from _{6 to 14} carbon atoms includes 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, benzoxazolyl, imidazolyl, tetrazolyl, triazolyl, pyridyl, pyrimidinyl and phthalimidyl groups (Terminally substituted-alkylamino) -quinoline-2-carboxylic acid derivatives, their tautomeric isomers, their pharmaceutically acceptable salts and prodrugs thereof, The method of claim 1 wherein, n is selected from 0 to 3, Nu is R 'is arylthio of m-CH ₃ or p-NH ₂ of the formula 3; Alkyl phosphonates; Arylsulfonyl; (Terminally substituted-alkylamino) -quinoline-2-carboxylic acid derivatives, their tautomeric isomers, their pharmaceutically acceptable salts and prodrugs The compound of claim 1, wherein the compound is 5,7-dichloro-4- [3- (3-phenylthioureido) propylamino] -quinoline- 5,7-dichloro-4- [3- (4-methoxyphenylthioureido) propylamino] -quinoline- 5,7-dichloro-4- [3- (m-tolylsulfanyl) propylamino] -quinoline- 5,7-dichloro-4- [3- (4-aminophenylsulfanyl) propylamino] -quinoline- 5,7-dichloro-4- [3- (diethoxyphosphoryl) propylamino] -quinoline-2-carboxylic acid, 5,7-dichloro-4- [3- (toluene-3-sulfinyl) propylamino] -quinoline- 5,7-Dichloro-4- [3- (toluene-3-sulfonyl) propylamino] -quinoline- (Terminally substituted-alkylamino) -quinoline-2-carboxylic acid derivatives, their tautomeric isomers, their pharmaceutically acceptable salts and prodrugs thereof, A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier Use of a compound of claim 1, a pharmaceutically acceptable salt thereof or a prodrug thereof as an antagonist of an excitatory amino acid to an NMDA receptor Use of the compound of claim 1, a pharmaceutically acceptable salt thereof or a prodrug thereof for the reduction of central nervous system damage caused as a result of anemia such as stroke, hypoglycemia, ischemia, Use of the compound of claim 1, a pharmaceutically acceptable salt thereof or a prodrug thereof for the prophylactic and therapeutic treatment of neurodegenerative diseases Use of the compound of claim 1, a pharmaceutically acceptable salt thereof or a prodrug thereof for the treatment of epilepsy, stroke, Alzheimer's disease, Huntington's disease and Parkinson's disease Use of the compound of claim 1, a pharmaceutically acceptable salt thereof or a prodrug thereof as an anticonvulsant, analgesic, antidepressant, antianxiety, and antipsychotic agent 1) a step of alkylating the compound of formula 31 with 1,3-dibromopropane to obtain the bromide compound of formula 32 (first step); 2) a step of treating the compound of the formula 32 obtained in the first step with sulfuric acid to remove the N-tosyl group (-NTs), followed by replacement of the bromine with NaN ₃ to obtain an azido compound of the formula 33 (second step); 3) catalytic hydrogenation of the compound of formula 33 obtained in the second step and reaction with an appropriate thiocyanate to obtain the thiourea compound of formula 34 (step 3); And 4) a step of basic hydrolysis of the compound of the structural formula 34 prepared in the third step to obtain the compound of the formula 1 (the fourth step) Wherein Nu in the general formula (1) is at least one selected from the group consisting of thiourea, carbamate, amide, urea, acylurea, sulfonylurea, acylthiourea, sulfonylthiourea, thiocarbamate, amidine, guanidine, imidate, thio (Terminal substituted-alkylamino) -quinoline-2-carboxylic acid derivative of the present invention applied in the case of imidate, phosphorylamide, sulfonamide or amine, etc. Scheme 1 1) treating the compound of formula 32 with an arylthiol (HSAr) compound to prepare the sulfide compound of formula 36 or treating with a triethyl phosphite [P (OEt) ₃ ] to give the phosphonate of formula 37 Stage 1); And 2) a step of removing the N-tosyl group (-NTs) by subjecting the compound of the structural formula 36 or 37 obtained in the first step to sulfuric acid and performing basic hydrolysis to obtain the compound of the formula (1) of the present invention (second step) (Terminal substituted-alkylamino) -quinoline-2-carboxylic acid derivative of the present invention which is applied when Nu of formula (1) is an appropriately substituted arylthio or alkylphosphonate. Scheme 2 In the above Reaction Scheme 2, X is the same as Nu in Formula (1). The method according to claim 16, wherein, in the case where Nu in formula (1) is sulfide, in addition to the second step, the target compound is reacted with m-chloroperbenzoic acid (m- (Terminal substituted-alkylamino) -quinoline-2-carboxylic acid derivative of the present invention which is characterized in that a sulfinyl compound of formula Scheme 3