JPH01500513A - Antagonist of specific excitatory amino acid neurotransmitter receptors - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Background of the invention Industrial applications The present invention provides antagonism to specific excitatory amino acid (EEA) neurotransmitter receptors. A new and powerful anticonvulsant, antiepileptic, analgesic and Related to sensory enhancers. In particular, the present invention provides ω-[2-(phosphonoalkylene phenyl-2-aminoalkanoic acid, pharmaceutically possible salts and derivatives thereof The present invention relates to bodies and methods of synthesizing them.

Conventional technology L-glutamate and L-albaragate were initially thought to be involved in brain metabolism. At present, molecular pharmacology, biochemistry, and electrophysiology are being studied. There is sufficient evidence that these amino acids are thought to be neuroexcitatory transmitters. There is.

[D.R.Curtis, A1. Duggar, D, Fe1ix, C, ^ , Rjohnston, ^, K.

Tebecis, J.C., Watkins, Brain Res, 41:283 -301 (1972) since the first analysis of the neuroexcitatory effects of core amino acids. For a long time, all compounds of this type (GeI'' both as agonists and as antagonists) 1) 98 K l: f’“′”t 60) T: is “WI W (D It had been pulled down. Different effects of FAA or different FAA compounds The discovery of relatively selective antagonists of the action has dispelled this assumption. It is now believed that there are multiple FAA recognition sites in the vertebrate nervous system. It is. [J.C. Watkins.

R, H, Evans, Ann, Rev, Pharmacol, Toxicol, 21:165-204 (1981)] These were treated with basic agonists or antagonists. 1, L-glutamate (Glu), conformationally restricted Gl u analogue, activated by quisqualic acid (Quis), diethyl glutamate A receptor that is selectively antagonized by esters.

2. Synthetic analogue of L-aspartate (Asp), N-methyl-D-aspara Gic acid (NMDA), isoxazole neurotoxin, ibotenic acid (Ibo), bilino dicarboxylic acid neurotoxin, quinolinic acid (Quin), and possibly (A, s A receptor that is reactive to p itself. These receptors are D(-)-2-amino- 5-phosphonobencunic acid (AP 5), D(-)-2-amino-7-phos It is antagonized by honohebutanoic acid (AP?), a divalent cation Mg′+.

3. Pyrrolidine pre-excitatory, activated by kainic acid (KA), but especially Receptors for which no antagonists have yet been identified.

4, t, -(+)-2-amino-4-phosphonobutyric acid (LAP4) receptors that are antagonized. Originally identified as an antagonist of FAA by electrophysiological means LAP4 is an unknown endogenous trigger at lateral perforant pathway synapses in the hippocampus. Inhibits the response to stimulants. It is highly unlikely that this neurotransmitter is Glu. Most recently, evidence suggests that the N-inhibitory dipeptide, N-acetylaspartyl Routine glutamic acid is believed to perform this function. [J.M. H, ff-French-mullen, K, J, Koller, R.

Zaczek, Li Hori, J. T., Coyle, D. O., Carpent er, Proc, Nat, ^cad.

Sci, 1jSA, 82. 3897-4001 (1985)] EAAs are may act through one or more of these receptors and have effects on the CNS. It has been implicated in the etiology of various pathological conditions that affect humans. For this reason, KA [K, Biziere.

J, T, S] evin, R, Zaczek, J, C, Colins, J , T. Coyle Advances in Pharmacology and Tberapeutics (H, Yoshida, Y, Hagihara , S.

Ebashi (ed.), Pergamon, New York, pp. 27 1-276 (1,9g2)], NMDA [R, Zaczek, J, Co 11ins, J, T, Coyle, Neurosci, Letts24: 181-186 (+98’l): l and endogenous excitatory amino acid Q u i n [R, Schwarcz, W, 0. Whetsell, R.M., Mango , 5science 219::06-318 (1983) used to produce symptoms similar to epilepsy and other convulsive disorders in humans. and the anatomical and neurochemical events caused by these compounds in animals. The lesions and defects are those of Huntington's disease [-J, Coyle, R, 5ch varcz.

Nature 263:244-246 (1976)] and patients who died from epilepsy. These characteristics are similar to those found in human brains after death. By administration of kainic acid , the internal hardness of A m m o n, which is an abnormality frequently seen in temporal lobe epilepsy. It is possible to create lesions with a linear structure similar to that of a scar. In this temporal lobe epilepsy experimental system, Studies have shown that endogenous EAAs are involved in this disease, especially in the case of existing antiepileptic drugs. It has been shown to be resistant to drugs. [J.J., Nadler, B.Y. Perry.

C, W, Cotman, Nature 271:676-677 (197g) EAAs are used not only for Huntington's disease and epilepsy, but also for Alzheimer's disease. er disease [^, C, Foster, J, F, Colins, R, Set+v arcz, NeurophallIac, 2Crook, L, Gersh on edition) Medium Mark Power As5oc, New Camarin, C.

nn, pp. 247-230 (1981)], poststroke causing cerebral ischemia. neuronal death and other factors, [R, P, Simon, J, H, Swan, T, Griffiths, B, S, Meldrum, 5science, 22 6.850-852. (1984); S, Rothman, J, Neur oscience 4:1g84-1891 (1894) and hereditary olive It has been suggested that it may also be a cause of pontocerebellar atrophy.

EAA activity at specific brain receptors in vitro and in vivo; Excitotoxic lesions caused by FAA in animals, the aforementioned neurodegenerative diseases There is a conceptual link between the pathogenesis of endogenous excitability and It is logical to try to antagonize excitotoxic neurotransmitters. brain There are specific endogenous substances that act on the KA receptors that have not yet been discovered. Therefore, it is important to develop antagonists against exogenous excitotoxic substances such as KA. Both are valid. α-amino-ω-phosphonoalkylenylcarboxylic acids (most Also strong and selective (-)-2-amino-7-phosphonohebutanoic acid, D(-) With the advent of potent and selective EAA antagonists exemplified by AP7, etc. , which triggered developments toward pharmacological intervention of FAA's actions at receptors. Ta.

Exogenous excitotoxic substance, IBO1 Endogenous excitotoxic substance Quin (KA is different) [^ ,C,Foster,G,E,Fagg,Brain Res,Rev, 7:103-184 (1984); A, C, Foster, J, F, Co1 1ins, R, Scbwarcz, Neuropharmac, 22:1331 -1341 (1983); R, Scbwarcz, J, F, Colins, D.

A. Parks, Neurosci, Letts 33:85-90 (19 82)] and AP7 (i, c.

ν, and i, v,) not only inhibit the neurotoxic and convulsive effects of NMDA but also activity in protecting against audiogenic seizures induced in genetically susceptible mice. I'm getting sex. [M, J, Croucher, J, F, Colins, B, S, Meldrum, 5science illusion 6+899-901 (1982) ]1, v, AP7 suppresses photogenic muscle clonics in baboons [B, S, Meldr um, M, J, Croucher, G, Badman, J, F, Co11 ins, Neurosi, Letts 39:101-104 (1983) ],? Increases the current threshold for electrical shock-induced seizures in mice and rodents Prevents chemically induced seizures in the neck. [S, J, Czuczvar, G. Meldrum, Eur, J. Phar++ac.

: 335-338 (1982) :] Very recently, AP7 (in the hippocampus) has become Significantly reduces and eliminates ischemic brain damage in carotid artery occlusion model of stroke in the neck It has been reported that [R, P, Simon, J.] 1. Swan.

T, Grirriths, B, S, IIIeldrum, 5science 226+850-852 (1984)], another less powerful FAA counterpart. The anti-Glu drug σ-D-glutamylglycine was administered exogenously under conditions of oxygen depletion. and protects against degeneration of cultured rat hippocampal neurons while inhibiting Asp toxicity. It has been shown that [S, Rotbman, J, Neuroscience ±:1884-1891 (1984) 3 Recently, kainic acid and quiscal Acid receptor antagonists have also been shown to have anticonvulsant activity. [M, J.

Croucher, B.S., Meldrum, ^, Y., Jones, J.C. , Yatkins, Brain Res.

377:111-114 (1984)] Finally, and importantly, excitability Amino acids, especially glutamate A , Greenamyre, J.B., Penny, B., Young, C.D.” mato, S, P, H4cks, l.

5choulson, 5science 227:1496-1498 (198 5)] and the onset of age-related neurodegenerative diseases such as late movement abnormalities [J, Y, 01n ey, Excitotoxins (lf, Fuxe, R, Roberts, There is some circumstantial evidence of involvement in It is.

Summary of the invention The present invention is a potent and selective excitatory amino acid neurotransmitter receptor having the general structural formula: It is a condition antagonist.

R3 and R1 may be the same group or different groups, and may be hydrogen, lower alkyl, halogen, -CH =CH-CHMaCH-, amino, nitro, trifluoromethyl, cyano group selected from among the group. n and m are 0. I, any of 2.3, and pharmaceutically Contains possible salts and derivatives of this substance.

Detailed description of the invention The structures and chemical formulas of the novel compounds of the present invention include various excitatory amino acid neurotransmitters. This was obtained as a result of detailed research on the antagonistic effect on the receptors. be.

These receptors are classified according to their basic agonists or antagonists as follows.

! , L-glutamate (Glu), a conformationally restricted Glu analogue, KISS Activated by calic acid (Quis) and activated by glutamic acid diethyl ester. A receptor that is selectively antagonized by

2. Synthetic analogue of L-aspartate (Asp), N-methyl-D-aspara Gic acid (NMDA), isoxazole neurotoxin, ibotenic acid (Ibo), pyridine Dicarboxylic acid neurotoxin, quinolinic acid (Quin), and possibly <Asp itself receptors that are reactive to These receptors are D(-)-2-amino-5-phosphorus Honopencunic acid (AP5), D(-)-2-amino-7-phosphonohebutanoic acid (AP 7), divalent cation Mg! Antagonized by +.

3. It is activated by pyrrolidine neuroexcitotoxin, kainic acid (KA), A receptor for which no specific antagonist has yet been identified.

4, antagonized by L(+)-2-amino-4-phosphonobutyric acid (LAP4) receptor. Originally identified as antagonists of FAA by electrophysiological means Therefore, LAP4 acts as an unknown endogenous excitatory substance in the lateral perforant pathway synapses of the hippocampus. inhibits the reaction to There is little possibility that this neurotransmitter is Glu. , recent evidence suggests that the N-11fl-toxic dipeptide, N-acetylaspartyl- It is believed that glutamic acid fulfills this function. [J, M, H, ff-French-mullen, Kj, Koller, R, Zacze K, Li Hori, J, T, Coyle, D, O, Carpenter, Pr. oc, Nat, Acad, Sci, (tlsA) 肚, 3897-4001 (1 985)] The structure of the new compound has a high affinity for one of the receptors and It has no affinity for some of them, making it selective for the compounds. It is a powerful antagonist. In other words, this allows multiple EAA receptors to be activated. It is possible to selectively antagonize only one EA receptor in one tissue. Become.

The present invention has higher affinity and selectivity, so the new compound has fewer side effects. It has become.

For example, 3-[2-(2-phosphonoethyl)phenyl)-2-aminopropanoic acid such as 3-[2-(2-phosphonomethyl)phenyl-2-aminopropanoic acid The high affinity and selectivity of the compound has been demonstrated in receptor binding experiments in mice. demonstrated by its ability to protect against pentylentitrazol (PTZ)-induced seizures. Ru.

The novel compounds of the present invention can be easily prepared by the following synthetic route.

Route 1 Example in route 1! and hydrogen bromide in a sealed test tube to introduce the compound of 4. By using an inchroman reaction with a solution of acid and acetic acid, the required intermediate o- (2-bromoethyl)-benzyl bromide is obtained in high yield. (^nders on, E, L,; H.

11iman, F, G, J, Chem, Soc, 1950.103 7) By reacting this compound with triethyl phosphorous acid, the compound of Example 1 was obtained. The compound is obtained with a yield of 70%. The compound of Example 2, ethyl 4-[2-(diethyl) phosphono-methyl)-phenyl] ~2-acetamido-2-carboethoxyb The bromophosphonate described in Example 1 was converted into diethyl acetamide malone. It was produced by reacting with the sodium salt of 6N) Addition in ICI2 The compound of Example 3 is obtained by carrying out water splitting. Intermediate o -(2-but Lomoethyl) benzyl bromide is added to diethyl acetamide malonate' separately from the above. The compound of Example 4 is obtained by reacting with the sodium salt of . This compound The compound of Example 5 was obtained in a yield of 75% when the compound was reacted with triethylphosphorous acid. It will be done. The compound of Example 6 was obtained by hydrolysis in 6N HC (! It will be done.

Route 2 In route 2, commercially available α,α゛-dibromo〇-xylene triethyl phosphorous acid The compound of Example 7 is obtained by reacting with. This intermediate is diethyl amine Reaction with the sodium salt of cetamide malonate gives the compound of Example 8. . The compound of Example 9 is obtained by hydrolysis in GN HCQ.

Route 3 Example 12 Example 15 In route 3, the intermediate 0-<3-bromopropyl)benzyl bromide is synthesized. It is necessary to [Rieche, person; Gross, H, Chew- thing Chloromethyl 3-buenyl fluorophyl ether is obtained in high yield. CS Frdeldel-Crafts cyclization reaction using AQCb in 2- Penzoxebine is obtained.

When this compound is reacted with a solution of hydrobromic acid-acetic acid in a sealed test tube, the necessary common The intermediate 0-(3-bromopropyl)benzyl bromide is obtained. this compound Example I by reacting with the sodium salt of diethylacetamide malonate A compound of O is obtained. By reacting this compound with triethyl phosphite, The phosphonomalonate compound of Example 1.1 is obtained. eN　Hydrated in HCQ The compound of Example 12 is obtained by decomposition. Intermediate〇-(3-bro Mopropyl) benzyl bromide is reacted with triethyl phosphorous acid separately from the above and the compound of Example 13 are obtained. This compound is diethyl acetamide malone-1 The compound of Example 14 is obtained by reacting with the sodium salt of . GN The compound of Example 14 is obtained by hydrolysis in HCQ.

Methods for preparing compounds for administration as pharmaceutical formulations are subject to the expertise of experts in pharmaceutical technology. There are probably many methods that are well known among people. This new compound is especially suitable for acid salts. , i.e. HCl2 salt, sulfate, phosphate, nitrate, metasulfonate, tartaric acid salts, or base salts and other pharmaceutically possible salts and compounds. Let's become.

In parenteral administration of the novel compounds and compositions of the invention, antioxidants, buffers, It will be formulated as an aqueous injectable solution containing bacteriostatic agents and the like. for instant injection Solutions include sterile pills, granules, etc. containing diluents, dispersing surfactants, binders, lubricants, etc. It will be prepared from tablets.

For oral administration, fine powders or granules of the compound are combined with a diluent and a dispersing surfactant. Formulated together, single-dose capsules in water or syrup or dry or in cachets or, when a suspending agent is included, in a non-aqueous suspension. I'll have to make it. The compound may also be prepared in tablets with optional binders and lubricants. or as a suspension in water, oil, water/oil pore agents. may contain flavorings, preservatives, suspending agents, thickeners, and emulsifiers. Ru. Granules or tablets for oral administration may be coated and pharmaceutical technology Other pharmaceutically possible drugs and dosage forms that are well known to the medical profession are: It may be used.

The following examples are illustrative of the compounds of the present invention; This does not mean that it is limited to.

Example Manufacturing example Example! Round bottom flask for diethyl 2-(2-bromoethyl)penzyl phosphonate distillation Medium, 2-(2-bromoethyl)bennorbromide (15, OL 54m mo l) and triethyl phosphite (9, Oy, 54mj01) on an oil bath. The mixture was heated at 00° C. with stirring.

After the ethyl bromide has distilled (1 hour), the residual volatile by-products and triester Chill phosphite was removed by vacuum distillation. The remaining viscous liquid is used as an eluent. Purified by silica gel column chromatography using ethyl acetate (+ + 1) did.

The fractions were combined and concentrated under reduced pressure to obtain a yellow oil (12.5 g, yield 70%). Ta.

IR (neat): 2987, 1249.1170.1064.964.802 cm”’HN M R(CD CIh>δ: 1.2 (t, 6B) 3.0-4.35 (m, 10tD 7.2 (s, 4B) Example 2- Ethyl 4-[2-(diethylphosphonomethyl)phenyl-2-acetamide -2-carboethquin butanoate sodium (1,03y. 44.8m mo Solid diethyl acetamidomalo in ethanol (50gawa 1) dissolved in A small amount of phosphoric acid (9.72 L, 44°8 mmol) was added. This solution is passed through a nitrogen gas stream. The mixture was stirred for 2 hours, then refluxed. After cooling to room temperature, the solvent was removed under reduced pressure and a yellow-brown color was obtained. A solid was obtained. This solid was dried under reduced pressure for about 2 hours.

Diethyl acetamidomalonate sodium salt in dry toluene (50ml) 2-(2-bromoethyl) suspended in toluene (25ml) ) benzyl phosphonate (is, og, 44.8 imol) was added dropwise.

This solution was refluxed with stirring in a nitrogen gas stream for 36 hours. The solution was cooled to room temperature Afterwards, the solid precipitate was removed by filtration and washed with toluene (20ml). Tor The ene solutions were combined and concentrated under reduced pressure to give a viscous oil. This oil is eluent It was purified by silica gel column chromatography using ethyl acetate. distillate The mixture was collected and concentrated under reduced pressure to obtain a transparent, viscous oil (7.2 g, yield 34%). Ta.

IR (neat): 1745.1675(C=O)am-''HNMR(CDCρ,)δ: +, o -6 (m, 12H) 1.8-3.2 (complex tn, 9B) 3.7-4.6 (m, 8H) 64-7.4 (+n, 5) 1) C□l-[,,NO,P・0.58! Theoretical elemental analysis value as O: C154, 99: H, 7, 34 + N, 2.92 Experimental value: C, 54, 75: H, 7 , 37; N, 2.97 Example 3 Ethyl 4-[2-phosphonomethylphenyl]-2-aminobutyrate 4-C2-C di Ethylphosphotumedel)-phenyl]-2-acetamido-2-carboethquin Butyric acid (2,5iF, 5.3mn+ol) in 6N hydrochloric acid (50n+1) The mixture was stirred vigorously and refluxed for a period of time. After cooling to room temperature, the reaction mixture was concentrated under reduced pressure to form an oil. I got it. This oil was washed three times with water (50 ml) and dissolved in 95% ethanol. , a small excess of propylene oxide was added. The precipitated acid was collected by filtration, and water-containing ethanol was collected. A white solid (1,26 iJ, yield 77%) was obtained by recrystallization from a filtrate. Melting point 24 7-249℃.

,IR(KBr): 1725.1620cm-’ 'HNMR(D*O)δ: 2.0-3.4 (6H, unresolved) 3.9-4.3 (m, l [+] 7.4 (s, 48) Elemental analysis theoretical value as Cr + H + a N Os P・0.5HtO: C, 46, 62, H, 6, Q5. N, 4.94 Experimental value: C, 46,73, H, 6, 04, N, 4.79 Example 4 Ethyl 3-[2-(2-bromoethyl)phenyl-2-ace)amide-2 -Sodium carboetquin propionate (0.41y, +8m mol) Solid diethylacetamidomalonic acid was dissolved in dry ethanol (100 mC). (3,99,18 mmol) was added in small amounts. This mixed solution was heated with nitrogen gas for 2 hours. Stir and reflux for 2 hours, then cool to 0-10°C. Then, 2-(2- Bromoethyl)benzyl bromide was added in one portion.

The reaction solution was stirred at 0-10°C for 2 hours and then at room temperature for 24 hours. precipitated The inorganic salts were filtered and discarded. The solvent was removed under reduced pressure to give a golden oil.

This oil is chromatographed on a reverse phase column (C-18), methanol - Eluted with water (1:1). The fractions were collected and concentrated under reduced pressure to give a white solid (5, 69, yield 75%).

Melting point: 86.0-86.5°C.

T R (nujo+): 1785, 1637 cm” (C=0)’HNMRCCDCQ3>δ: 1.2 (t, 6H) 1.9 (s, 3H) 2.8-3.5 Cm, 4H) 3J (5,28) 4.2 (q, 4H) 6.8 (s, IH) 7.2 (m, 4H) Theoretical elemental analysis value as C+sH*-N OsBr, C, 52.1g, old 5. 84, N, 3.38 experimental value; c, 52.26, H, 5,86, N, 3.34 Example 5 Ethyl 3-[2-(2-diethylphosphonoethyl)phenyltoco-2-aceto Amido-2-carboethoxy-propionic acid 3-[2-(2-diethylphosphono ethyl)phenyl]-2-acetamido-2-carboethoxypropionic acid (0 ,59,1,2 to mol) in triethylphosphite I (IOmQ), The mixture was stirred and refluxed for 4 hours.

Excess triethyl phosphite and volatile by-products were removed under reduced pressure. residual viscosity The thick oil was eluted with column chromatography (C-18゜methanol:water (4:l)). ) and then preparative 14PLC (C-18, methanol:water (7:3) A clear viscous oil (0.48 L, 86% yield) was obtained.

IR (Nujol): +745.9,1676.5 cm-I (C=0)'HNMR (CD Cl2 3) δ: 1.1-1.5 (m, I 28) t, 8-3.1 (complex m, ?H) 3.7 (s, 2H) 3.8-4.4 (m, 8H) 6.6 (s, 1M) 7.0-7.3 (m, 4H) CttH,s4N Elemental analysis theoretical value as OsF; c, 56.04, H, 7 ,27,N,2.97Experimental value;c,55.91　,H,7,31,N,2.84 Example 6 3-[2-(2-phosphonoethyl)phenyl coke-aminopropionic acid Ethyl 3-[2-(2-(diethylphosphonoethyl)-phenyl]-2-acetyl Toamide 2-carboethoxyprobionic acid (7゜9g, 16.8mmol ) was vigorously stirred and refluxed in 6N hydrochloric acid (4 (1 ml)) for 14 hours. Cooled to room temperature. After that, the reaction mixture was concentrated under reduced pressure to obtain an oily substance. Add this oil to water (25IIIQ) Wash 4 times with dripped.

The precipitated impure acid was filtered off and recrystallized from aqueous ethanol to form a white solid (4,1y , yield 90%). Melting point 241-243°C.

IR (Nujol): 1715cm-' (C=0) 'l-I NMR (D, O) δ: 1.5-2.2 (m, 2H) 2.6-3.3 (m, 4H) 3.9-4.2 (t, IH) 7.2 (m, 4H) CzH+aN Elemental analysis theoretical value as OsP: C, 48,35; H, 5,9 0; N, 5.13 Experimental value: C, 48, 69, H, 6, +6; N, 4.9 5 Example 7 In a round bottom flask for distillation, α, α-nobromo O-xy 1 non (20°Q9.75 ．． 8m mof) and triethyl phosphite (12.6g.

75.8 mmol) was heated at 70-75° C. with stirring. ethyl bromide After the distillation is complete (approximately 2-3 hours), the remaining volatile by-products and triethylphosph The nitrite was removed by vacuum distillation. The remaining oil was chromatographed on silica gel. The product was purified by using ethyl acetate as an eluent. The fractions are collected and concentrated under reduced pressure to give a yellow color. A colored oil was obtained.

IR (as is): 1249; 1164.8. 1038.8; 966.8; 794.5cm- ’ 'HN MR (CD C123) δ: 1.0-1.4 (m, 6H) 3.6-4.2 (m, 4H) Example 8 Ethyl 3-[2-(diethylphosphonomethyl)phenyl-2-acetamide Sodium 2-carboethoxypropionate (0,439,18,8m mo 1) in dry ethanol (50 ml), solid diethyl acetamide Ronic acid (4.09 g, 1.8.8 mmol) was added in small portions. After this mixture The mixture was refluxed with stirring in a fluorine stream for 2 hours, and then cooled to room temperature.

Then 2-(diethylphosphonomethyl)ben dissolved in ethanol (40 ml) Zilb(7?ide) (6.05g, 18.8mmol) was added dropwise and the mixture was Stir for hours.

The precipitated salts were removed by filtration and the solvent was concentrated under reduced pressure to give a viscous oil. . This oil was analyzed using silica gel column chromatography using ethyl acetate as an eluent. and purified.

The fractions were combined and concentrated under reduced pressure to give the product as a clear oil (6°679, yield 7. 9%).

IR (neat): 1745.9, 1676.5, 1501.6.1375.6゜1247.1,1 649.1,969.4 am-''HNMR (CDC123) δ: ■, 0-1.4 (m, 12H) 2.0 (s, 3H) 2.9 (s, IH) 3.3 (s.IH) 3.8-4.4 (m, l0H) 5.8-7.4 (m, 5H) Elemental analysis theoretical value as C2+H3tN P Os・0.5 HtO: C, 54 ,06:8.6.92:N,3.00Experimental value:C,53,73:H,7,11: N, 3.27 Example 9 Ethyl 3-[2-phosphonomethylphenyl-2-aminopropionate 3-'2 -(diethylphosphonomethyl)-phenyl]-2-ace)amide-2-carbo Ethoxypropionic acid (4.5g, 9°8@mol) in 6N hydrochloric acid (50ml) The mixture was refluxed vigorously with stirring for 12 hours. After cooling to room temperature, the reaction mixture was placed under reduced pressure. Concentration gave an oil. This oil was washed 3 times with water (50 ml) and contained 95% water. Dissolved in ethanol and added excess propylene oxide. The precipitated acid It was filtered and recrystallized from aqueous ethanol to give a white solid product (1.6 g, yield 63 %) was obtained. Melting point 259-261"C0 IR (Nujol): 1722; 1625; 1128; 1049 cm-''HNMR (DtO) δ: 2.8-3.7 (unresolved, 4H) 4.2 (m, IH) 7.73 (s, 4H) Elemental analysis value as C1eH+4NOsP-0,25HtO Theoretical value: C, 45, 55; H, 5, 54; N, 5.32 Experimental value: C, 45, 57, H2S, 55. N , 5.38 Example 10 Ethyl 3-[2-(3-bromopropyl)phenyl-2-acetamide-2 -Sodium carboethoxy-3-propionate (0.61g, 27m +5o A small amount of 86 g (27 s mol) of dry ethanol in which 1) was dissolved was added. This mixture The mixture was refluxed with stirring in a nitrogen stream for 2 hours. Then, in an ice water bath for 0-1H. Cooled to ℃. Thereafter, 2-(3-bromine) dissolved in dry ethanol (40 ml) was added. Mopropyl) benzyl bromide (8.0 g, 27 nmol) was rapidly added. Ta. The reaction mixture was stirred at 0-10<0>C for 2 hours, then at room temperature for 24 hours. precipitation The resulting inorganic salts were filtered, washed with ethanol (20 ml) and discarded. Ame the solvent and removed under reduced pressure to give an orange oil.

This oil was purified by silica gel column chromatography using hexane-acetic acid ester as an eluent. Purified using chill (3:I). The distillate was concentrated under reduced pressure to obtain an oil. Ta. It solidifies on standing (9.5 g, yield 82%), melting point 73-74.5°C.

iR (Nujol): 1745.1648 cm-' (C=O)'HNMR (D, O) δ: 1.3 (t, 6H) 1.9-2.4 (m, 5H) 2.7(t, 2H) 3.4 (t, 2H) 6.6 (s, IH) 7.0-7.3 (m, 4H) Elemental analysis value theoretical value as C+sHtaN OsBr: C, 53, 28; H , 6, 12; N, 3.27; Br.

18.66 Actual value: C, 53,33; H2S, 13; N, 3.23: Br.

18.67 Example 11 Ethyl 3-r, 2-(3-diethylphosphonopropyl)phenylcou 2- Ethyl acetamido-2-carboethoxy-propionate 3-[2-(3-bropropionate) mopropyl) phenylcou 2-acetamido-2-carboethoxypropionic acid (7.5 g, 17.5 mmol) was added to triethyl phosphite (20 m 1) was refluxed with stirring for 6 hours. Excess triethyl phosphite and volatile By-products were removed by vacuum distillation. The remaining viscous oil is used as an eluent. Purification was performed by silica gel column chromatography using ethyl acid. heat the distillate The mixture was concentrated under reduced pressure to obtain a viscous yellow oil (4.2 g, yield 49%). .

IR (Neat): 1746, +680c+n-' (C=O)'HNMR (CDC123) δ : 1.1-2.1 (complex m, 19H) 2.4-2.7 (t, 2H ) 3.6 (5,2H) 3.8-4.3 (m, 8H) 6.55 (s, IH) 6.9-7.2 (m, 4l-1) Example 12 3-[2-(3-phosphonopropyl)phenyl-io2-aminopropionic acid Ethyl 3-[2-(diethylphosphonopropyl)phenylcou-2-acetamide Do-2-carbetoquine propionic acid (3.6 g.

6.9 mmol) was refluxed vigorously in 6N hydrochloric acid (25 ml) with stirring for 12 hours. Ta. After cooling to room temperature, the reaction mixture was concentrated under reduced pressure to obtain an oil. oily substance Washed three times with water (25 ml), dissolved in 95% aqueous ethanol (25 ml), Propylene oxide was added dropwise. The precipitated acid was filtered and re-purified from aqueous ethanol. Crystallization gave a white solid (0.76 g, yield 38%). Melting point: 95℃ or higher (decomposition) point).

IR (Nujol): 1717.6cm-' (C=O) 'HNMR(D,O)δ: 1.1-2.0 (complex m, 4H) 2.6-3.1 (m, 4H ) 3.35-3.65 (m, IH) 7.3 (m, 4H) Theoretical elemental analysis value as C, tH, 5NO5P -H, O ・C, 47, 21:H , 6, 60: N, 4.58 Experimental value: C, 47, 47, H, 6, 72, N, 4.5 3 Example 13 Round bottom flask for diethyl 2-(3-bromopropyl)penzyl phosphonate distillation In the co, 2-(3-bromopropyl)benzyl bromide (11.36 g, 38 ．． 9mmol) and triethyl phosphite (6.46g, 38.9mmol) immediately after distillation. 9 nmol) was heated on an oil bath at 100-110°C with stirring. ethyl bro When the amide has finished distilling (approximately 2 hours), the remaining volatile by-products and triethyl phosphorus Phi) was removed by vacuum distillation. The remaining oil is extracted with Hexazy as an eluent. Equipped with silica gel column chromatography using ethyl acetate (+, :I) and purified. The fractions were combined and concentrated under reduced pressure to yield a clear oily product (11.2 g, yield). 83%).

IR (neat) 2985.1496.1450.1391S 1252.1162.1104. 967.843.802.758cm-''HN MR (CD Cl2s) δ :1.2 (t, 6H) 1.8-2.3 (m, 2H) 2.7-3.55 (m, 4H) 3, 8-4, 2 (m, 21-1) 7.1-7.4 (m, 4H) Example 14 Ethyl 5-[2-(diethylphosphonomethyl)phenyl]-2-acetamide Sodium -2-carboethoxybencuinate (0.48g, 21nmol) Solid diethylacetamidomalon in dry ethanol (50n+l) dissolved in Acid (4.56 g, 21 Illmol) was added in small portions. Add this solution to nitrogen The mixture was refluxed with stirring in a stream of air for 2 hours. After cooling to room temperature, the solvent was removed under reduced pressure. A tan solid was obtained. This solid was dried under reduced pressure for about 2 hours. diethyl acetate The sodium salt of toamidomalonic acid was suspended in dry toluene (25 ml) and added to the dry toluene (25 ml). Diethyl 2-(3-bromopropyl)penzyl dissolved in toluene (25wl) Phosphonate (9.0 g, 21 mmol) was added dropwise. Add this solution to nitrogen The mixture was refluxed with stirring in a stream of air for 20 hours. After cooling to room temperature, filter the precipitated solid and washed with toluene. The toluene solutions were combined and concentrated under reduced pressure to give a dark black color. of oil was obtained.

This oil was subjected to silica gel column chromatography using ethyl acetate as the eluent. It was purified by filtration. The fractions were collected and concentrated under reduced pressure to give a yellow viscous oil (4.2 g, yield 42%). It solidifies on standing and has a melting point of 76-79°C.

IR (neat) +745.9, I 680 cra-' (C=O)' HNMR (CD CC5 ) 6: 1.0-1.4 (m, I 28) 2.0 (s, 3H) 2.2-3.3 (m, 6H) 3.7-4.4 (m, 8H) 6.8 (5, IH) 7.0-7.3 (m, 4H) Ct3H3@N Elemental analysis theoretical value as OsP: C, 56,90, H, 7,4 8: N, 2.89 Experimental value: C, 56, 27, H, 7, 51; N, 2.86 Example l5 5-[2-phosphonomethylphenyl-2-aminopentanoate ethyl 5-[2 -diethylphosphonomethyl)phenyl]-2-acetamido 2-carboethoxy Cypentanoic acid (3.8 g, 7.8 mol) in 6N hydrochloric acid (25+a1.) The mixture was vigorously refluxed with stirring for an hour. After cooling to room temperature, the reaction mixture was concentrated under reduced pressure. It was condensed to give an oil.

This oil was washed three times with water (25 ml) and then washed with 95% aqueous ethanol (25 ml). m1. ), and propylene oxide was added dropwise.

The precipitated impure acid was filtered and recrystallized from aqueous ethanol to produce a white solid product ( 1.7 g, yield 76%) was obtained. Melting point: 152°C or higher (decomposition point).

IR (nujo+) 1717.6 cva-' (C=O)'HN MR (D to) δ: 1.6-1.9 (broad, 4H) 2.8-3.3 (m, 4H) 3.4 5 (m, IH) 7.3-7.7 (m, 4H) C+t)t+llN　Elemental analysis theoretical value as OSP・0.5HtO: C, 4 8,65; H, 6,46: N, 4.73 Experimental value: C, 48,56; H, 6,46 ;N, 4.72 Example 16 In vitro receptor binding test example standard in vitro ligand binding technique Using the method, the compound shown in Example 3.5.9.12.15 was synthesized into various excitatory amino acids. The ability of acid ligands to inhibit specific binding to rat brain membranes was investigated. Special The compound ['H]kainic acid ['H]KA, RS-α-amino-3-hydroxy C-5-methyl-4-isoxazolepropionic acid [”H]AMPA, [”H] DL(+)2-74/-7-E, 2. Specific binding of ho/heptanoic acid ['H]AP7 It was evaluated for its ability to inhibit

The method is as follows. Rat forebrain with Enna and 5nyder (Mol.

Pharmacol, 13.422-453.1977) Repeatedly resuspend the final pellet in freshly prepared buffer for the test. 20vol; w/v) further centrifugation 3 times (45,000g; 10 minutes , 4°C). [For the 3HcoAP4 test, the tissue immediately after preparation was used. that For other tests, tissues were stored frozen (-40°C) until use. exam All cells were cultured in triplicate.

The sediment was transferred to 111 Protosol (New England Nuclear , Boston, MA) and 6 ml of Enconofluor (N ew England Nuclear, Boston.

Radioactivity was measured by conventional liquid scintillation counting after adding MA). .

[3H]AP4 (specific radioactivity (S, A,) = 26.I Ci/mmol.

New Er+gIand Nuclear, Boston9MA) specific Binding was performed using HEPES KO) (buffer (0.05M: pH 7.1)). tcher and investigated. Incubation was carried out at 37°C for 45 minutes (2+sl), followed by centrifugation ( 45,000 g: I 0 min, 4° C.) to terminate the reaction.

The supernatant was discarded, and only the surface of the precipitate was quickly washed twice with 3.5 ml of water-cooled buffer.

The final concentration of ligand in the test was 50 nM to clarify non-specific binding. Monoglutamate (10-M) was used.

['H] AMPA (S, A, = 25.6 Ci/s mol, liew E The specific binding of ngland Nuclear) Using tsHcL buffer (0,05M, pH 6,9; 23°C), Murphy et al. method (Soc, Neurosci, Abs, Dan, 109.1985) Therefore, we investigated. Tissues were co-treated with Triton-X-100 (0.05%; V/V). After pre-incubation for 30 minutes (37°C), incubation was performed at 4°C for 60 minutes (2III). . The supernatant was discarded, and the surface of the sediment was quickly washed twice with 3.5 ml of water-cooled buffer. . The final concentration of Sugant in the test was +6 nM, to clarify non-specific binding. Rice monoglutamate (10-M) was used.

['H] AP7 (S, A, = 58.4 Ci/m mol, New Eng The specific binding of landNuclear) was achieved using Tris citrate buffer (0,0 5M; pH 7,5: 23°C) by Ferkany and Coyle. Reported method (Life Sci, 33.1295-1305.1983) The investigation was conducted according to the following. After pre-incubating the tissue (30 minutes at 37°C), incubate at 37°C for 90 minutes. culture (2111) and centrifugation (45,000 g: I 0 min, 4°C). The reaction was terminated. Discard the supernatant and rinse the pellet twice with 3.5 ml of water-cooled buffer. Only the surface was cleaned. The final concentration of ligand in the test was 500 μM, with non-specific Monoglutamate (10-M) was used to clarify the binding.

[3H] KA (S, A, == 6 0 Ci 7m mol, New En The specific binding of Grand Nuclear) was confirmed using Tris HCL41 buffer (0 , 05M: pH 7, /1.23°C) using the method of London and Coyle ( Mo1. Prepared according to Pharmacol ■, 492-505.1979). I investigated. Culture Jl (2 ml) was incubated at 4°C for 90 minutes, and centrifuged (45,000 g; The reaction was terminated by 0 minutes (4°C). Discard the supernatant and transfer the precipitate to water-cooled buffer. Only the surface was quickly washed twice with 3.5 ml. Final concentration of ligand in the test The concentration was 5 nM and monoglutamate (10-M) was added to clarify non-specific binding. was used.

The results are shown in Table 1. Compound 3.5 when tested at a final concentration of 100 μM ．． 6.9.12.15 shows the specific binding of [3H]KA or [”H]AMPA. It inhibited less than 20%. Compound 9 was similarly ['H, The specific binding of FA, P4 and [3H]AP7 could not be inhibited either. Compound 3.6. 12.15 inhibits the specific binding of [311coAP4 and ['H]AP7 in a concentration-dependent manner. The order of strength in each test was 15>3>12n6. Compound 3.1 2.15 has the ability to inhibit the asymmetric binding of ['H]AP4 and ['HiAP7 -amino-ω-phosphonoic acid, DL(±)AP7, but the same level as that of compound 6. The latter were 3 to 10 times less capable in this regard. Compound 6 was also tested in two tests. There is a clear difference in the specific binding of [H]AP7 than ['H]CoAP4. It had a stronger ability to inhibit

Table 1 The compound of the example inhibits the specific binding of [3H] excitatory amino acids to rat brain membranes. Ability to [3H] 8P4 [H]AP? [31(]Kainate C’) I ]AMPAXV1.03 2.29 >>100 >>100III 6.8 6.1 >>100 >>100Xll 9.5 N, T, >>100 >> 100Vl 16.7 52.5 >>100 >>100IX >>100 >>100)>100 >>100V >>too >>100 >>100 N, T.

AP7 5.1 6.8 >>100 >>100 Method is as described in the text It is. The values in the table are determined by triplicate measurements using 8 different drug concentrations, each with a minimum of 3 This is the average value measured twice. Where the value is >>100 μM, the values were used for the test. Less than 20% of specifically bound ligand was found even at the highest concentration of drug. This shows that.

Example 17 Maximal electroshock seizure (MES’) protective compounds 3.5.6.9.12.1 5, and the control compound DL(Sat) P7 to seizures induced by maximum electric shock. The anticonvulsant activity was evaluated.

For the test, CF-1 male mice (20-259; Charles Ri Attach electrodes to the ears of vers) and apply a current of 0.5 mA for 0.2 seconds to induce a seizure. caused it. Anticonvulsant activity is demonstrated by loss of extensor muscle segment in seizures; It was defined as when the extension of the hind limbs did not make an angle of 90" or more with respect to the body plane. Above The percentage of mice in which hindlimb extension was not observed was calculated and used as data.

Drugs were dissolved in a solution of propylene glycol and distilled water (5:95; V/V). . For i, c, and v administration, the drug was administered in a final volume of 5 μg to the 15th part of the study. . For i, p administration, the drug should be administered at a dose of 12,5 mQlkg 30 minutes before the test. gave.

The results are shown in Table 2. The control compound AP7 had a dose-dependent effect on MES-induced seizures as expected. ED b. Calculated values are IC, V, and i, p, injection site. 84 μg (n = 8), +27rb9/kflcn = 1 6). Example compound 3.5.6.12.15 and control compound E When administered at a molar dose equivalent to or twice the s o value, both No effect on MES-induced convulsions was observed depending on the route of administration. Example 9 The compound showed some protective activity against MES-induced seizures, and the EDso value was I 5u9Ci, c, v,), E D fs value is 500 ta9/kg (ip, ) was calculated. The compound of Example 9 caused significant ataxia in animals at high doses. It was impossible to test because of the possibility of rubbing.

Table 2 The compounds of the examples showed maximum electric shock or bench response in CF-1 male mice. Ability to antagonize seizures induced by lentitrazol Example　E　D　s.

(ug) (II1g/kg) (ug) (mg/kg) AP7 g, 3 1 27 1.8 199Vl > 100 N, T, 3.2 >> 350D (16 500* 24 >> 250V >> 100 N, T, >> 28 350 H [IN, T, >>250 >>6 >>500 size >>23 >>155 >>5 >>300XV >>22 >>150 >>s >>300 The method is As stated in the text. ED in the table. Values are 6-8 for each concentration for each drug. It was calculated by drawing a dose-response curve at a minimum of 5'a degree using 10 animals.

E.D. Where the value is 〉〉, there is no protection even at the highest concentration of the drug used in the test. This indicates that less than 20% of the animals were seen.

*Highest dose tested; ED□ **Highest dose tested; 50% of animals protected against seizures; 7 Three of the animals died before the end of the observation period.

Example 18 pendylenetetrazol-induced seizure (PTZ) protective compound 3.5.6.9.12, I5 and control compound DL, (out) AP7 with pendylenetetrazol Anticonvulsant activity against induced seizures (PTZ) was evaluated.

For the test, PTZ was dissolved in saline (0.9%: w/v) and CF-1 male Mouse (Charles Rivers; 20-259) 85 m97 9 15 minutes (i, c, v,) or 30 minutes (i, p,) after test compound administration at dose was administered. Mice were observed for 0 minutes after PTZ administration to see if they had seizures or not. was recorded. Data were expressed as the percentage of animals that developed seizures.

In the test, the example compound 5.6.12.15 and the control compound were propylene. Example compounds 3 and 9 were dissolved in a solution of recall and distilled water (5:95; V/V). was dissolved in 0.2M bicarbonate ion. Drugs are for i, c, v, and i, p. Each was administered in a volume of 9 at 5 μQ or 1.2, 5 mQ/.

The results are shown in Table 2. Example compound 3.12.15 was used as a control compound for PTZ-induced seizures. ED, which suppresses. i, c, v, or in an amount equal to or twice the value No activity was observed even after i and p administration. Intermediates in the synthetic route of Example 6 Compound Example 5 has an f, p of 350 to 9/9, and some degree of post-injection It showed anti-seizure activity (4 out of 7 animals). Example 5 was administered at a high dose (500 mg/kg ), 40% of the test animals died, so seizure-protective activity could not be recorded. It was.

Example 6 has +, c, v, and the PTZ-induced convulsion suppressing effect after administration is comparable to that of the control compound. It was degree. (Table 2) However, after administration of i, p, doses up to 350 mg/kg This seizure system did not significantly protect the animals at any dose.

Example 9 also prevented mice from seizures induced by PTZ when administered intraventricularly. The EDso value is 1.0 times that of the control compound and 6 times that of Example 6. Met. Similar to Example 6, Example 9 also showed almost no anticonvulsant activity upon intraperitoneal injection. I couldn't stand it at all.

Compounds 6 and 9 exhibited strong anti-inflammatory effects in the PTZ-induced seizure system when administered i, c, v. Although it is a convulsant drug, it has protective activity only in this seizure system and not in the MES-induced seizure system. It differs from the control compound in that it has the selectivity of showing .

41 bundles 71τ100- 1=-next℃htouna1general formula1 Excitatory amino acids, their inducers or their salts.

I. Display of incidents International application number PCT US/871004442, title of the invention Antagonist of specific excitatory amino acid neurotransmitter receptor 3, corrector Relationship to the incident: Patent applicant Address Pax, Millersville, Maryland 21108, United States Bar coat 407 Name: Rezotarsky, Janutz Wakurou (and 2 others) )4. Agent 6. Subject of amendment: Description, translation of claims, power of attorney 7. Contents of amendment: Description , international search report of the translation of the claims

Claims (1)

[Claims] 1. A general expression such as ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ [In the formula, R1 and R2 are either the same or different, and a hydrogen atom, a lower Alkyl group, halogen atom, butadiene-CH=CH-CH=CH- group, amino group, nitro group, trifluoromethyl group or cyano group. . n and m represent integers of 0, 1, 2, or 3. ] Neurotransmitters represented by Potent specific excitatory amino acids, derivatives thereof, that are receptor antagonists Or its salt. 2. As described in claim 1, R1 and R2 are butadiene-CH=CH-C Neurotransmitter receptor antagonists with H=CH- group, n=1 and m=2 A potent specific excitatory amino acid, its inducer or its salt that is a stimulant. 3. Among the claims described in claim 1, 4-[2-phosphonomethyl-phenyl]-2 -A substance that is amino-butyric acid. 4. Among the claims described in claim 1, 3-[2-(2-diethyl-phosphonoethyl)-phenyl]-2-acetamide -2-carboethoxypropionic acid substance 5. Of what is stated in claim 1 , 3-[2-(2-phosphonoethyl)-phenyl]-2-aminopropionic acid. substance 6. Among the claims described in claim 1, A substance that is 3-[2-phosphonomethylphenyl]-2-amino-propionic acid 7. Among the claims described in claim 1, 3-[2-(3-phosphonopropyl)-phenyl]-2-aminopropionic acid a certain substance 8. Among the claims described in claim 1, A substance that is 5-[2-phosphonomethylphenyl]-2-amino-pentanoic acid 9. Among the claims 1 to 7, pivaloyloxymethyl ester a certain substance 10. Among those described in claim 8, pivaloyloxymethyl carbamate material 11. Compatible with pharmaceutically acceptable carriers and/or diluents Contains one or more substances that effectively alleviate the pain described in item 1 of the scope of the request. Pharmaceutical composition (product) for pain relief 12. Parenteral, nasal, oral, rectal or administration of the above-mentioned substances according to claims 1 to 10 in a combined manner; Treatment of pain relief in animals in need thereof, including 13. Compatible with pharmaceutically acceptable carriers and/or diluents Scope of Claim Containing an effective amount of one or more substances listed in item 1. Pharmaceutical compositions (products) used for the treatment of epilepsy or epilepsy 14. Parenteral, oral, nasal, rectal or Administering the above-mentioned substances according to claim 1 or 12 in a combined manner. 15. Treatments used to treat convulsions or epilepsy, including pharmaceutical Claim 1 in coexistence with acceptable carriers and/or diluents. Sensation-promoting, containing a sensation-promoting amount of one or more of the substances mentioned. Pharmaceutical composition (thing) 16. Parenteral, oral, nasal, rectal or Administering the above-mentioned substances according to claim 1 or 14 in a combined manner. Treatments that promote sensations that include