JP2007051138A - Ischemic neuropathy medicine - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a medicine for preventing and/or treating neuropathy caused by spinal ischemia. <P>SOLUTION: This medicine for preventing and/or treating neuropathy caused by spinal ischemia comprises a compound represented by formula [I], its salt, or their solvates as an active ingredient, and a use thereof is provided. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a medicament for preventing and / or treating nerve cell damage due to spinal cord ischemia.

  The spinal cord is an elongated columnar organ that descends in the spinal canal following the medulla oblongata in vertebrates and is responsible for nerve conduction between the brain and spinal nerves. The spinal cord is composed of three layers: a central canal, an H-shaped gray matter, and white matter from the center to the outside. Motor nerves are gathered in the front part (anterior horn) of the H-shaped gray matter, and these nerves transmit information from the brain and spinal cord to the muscles, causing movement, and the rear corners ( Sensory nerves gather in the back horn), and these nerves convey sensory information from other parts of the body through the spinal cord to the brain. A number of nerve fiber bundles pass through the surrounding white matter, carrying sensory information from other parts of the body to the brain (upward), and conversely transmitting electrical signals from the brain to the muscles. (Down road). In humans, 31 pairs of spinal nerves play a major role in the transmission of information between the brain and each part of the body, so the spinal cord is a trauma caused by traffic accidents, spinal diseases such as spinal deformities, tumors, and imaginary damage caused by blockage of blood flow. When nerves are damaged by being damaged by blood or the like, movement disorders such as limb paralysis and apraxia and sensory paralysis may occur.

  Since most of the blood flow to the spinal cord, especially the blood flow to the front of the spinal cord, is supplied by the branches of the aorta, the spinal cord becomes ischemic regardless of which branch is blocked . In addition to severe atherosclerosis, aortic detachment, thrombus, and the like, blood flow blockage includes artificial blockage during aortic aneurysm surgery. In recent years, with the aging of society and changes in eating habits, arteriosclerotic diseases are increasing, and the risk of ischemic spinal cord injury tends to increase.

Among arteriosclerotic diseases, aortic aneurysms are usually asymptomatic until rupture, but once they rupture, they present severe symptoms and have fatal effects that require appropriate treatment before rupture. It is. Treatment is performed medically and / or surgically depending on the site, size, patient condition, etc. of the aneurysm, but when a certain size is reached, a surgical procedure is essential. A general surgical procedure is a method (aneurysm repair) in which the aorta is blocked before and after the aortic aneurysm, the aneurysm is removed, and replaced with an artificial blood vessel.
Among the aortic aneurysms, in the repair of the descending thoracic aortic aneurysm or thoracoabdominal aortic aneurysm, it is necessary to block the blood flow in the descending aortic region including an important blood supply site to the spinal cord. Therefore, spinal cord ischemia may be damaged by spinal cord ischemia during or after the operation, and serious complications such as paraplegia and rectal bladder disorder may occur. In fact, it is said that the incidence of paraplegia in the repair of a descending thoracic aortic aneurysm or thoracoabdominal aortic aneurysm is as high as about 10%. Considering milder complications, the complications associated with spinal cord ischemia during aortic aneurysm repair have a serious impact on patients and society.

The mechanism of spinal cord injury during repair of descending thoracic aortic aneurysms and thoracoabdominal aortic aneurysms is thought to be primarily related to direct tissue ischemia. Conventionally, many surgical techniques and pharmacological studies have been made to protect the spinal cord, but this serious complication could not be completely prevented (Non-Patent Documents 1 and 2).
Although the details of neurochemical changes after ischemia remain unclear, the actions of voltage-sensitive sodium channels (VSSC) and voltage-sensitive calcium channels are thought to be related to nerve damage caused by ischemia. (Non-Patent Documents 3 and 4). When VSSC opens, the release of glutamic acid due to ischemia increases, and the overloading of calcium ions into cells is promoted (Non-patent Documents 5 and 6). This is considered to be a major cause of neuronal degeneration after cerebral ischemia (Non-Patent Documents 7 and 8). These findings are consistent with the fact that sodium channel and calcium channel blockers (Na ⁺ / Ca ⁺ channel blockers) show neuroprotective effects in spinal cord and brain ischemia models (non-patent literature). 9, 10). However, since Na ⁺ / Ca ⁺ channel blockers reported so far have serious cardiovascular side effects such as blood pressure lowering and arrhythmia, their clinical application of cerebrospinal neuroprotective action has been limited (Non-patent Document) 11).

As described above, when spinal cord ischemic damage occurs and nerve cells are damaged, there is a high risk of causing paraplegia, rectal bladder injury, etc., so that nerve damage during spinal cord ischemia is prevented and / or treated. Means to do so have been studied. For example, 2-amino-6-trifluoromethoxybenzothiazol (Riluzole ^™ ) is disclosed to be effective in preventing ischemic spinal cord injury caused by aortic cross clamping (Patent Document 1). Moreover, the effect of the pyrazolone derivative in the prevention and / or treatment of the disorder | damage | failure by ischemic spinal cord injury in the spinal cord injury model animal after aneurysm repair is suggested (patent document 2).
However, since ischemic spinal cord injury resulting from accidents and surgery has a great influence on patients and society, there is still a strong demand for the development of a new medicine having an excellent nerve cell protective action in spinal cord ischemia.

International Publication No. 00/66121 JP 2004-67585 A Svensson LG et al., J. Vasc. Surg., 1993; 17: 357-70 Gharagozloo F et al., Chest., 1996; 109: 799-809 Barone FC et al., Stroke. 1995; 26: 1683-90 Lipton P., Physiol Rev. 1999; 79: 1431-568 Koch RA et al., J Neurosci. 1994; 14: 2585-93 Kanai Y et al., Trends Neurosci. 1993; 16: 365-70 Choi DW et al., J Neurosci. 1988; 8: 185-96 Oka M et al., Life Sci. 2000; 67: 2331-43 Gangemi JJ and others, Ann Thorac Surg. 2000; 69: 1744-9 Gemba T et al., J Pharmacol Exp Ther. 1993; 265: 463-7 Tanaka K et al., Brain Res. 2002; 924: 98-108

  The present invention provides a medicament for preventing and / or treating neuropathy caused by spinal cord ischemia, that is, a therapeutic agent for ischemic neuropathy.

で表される化合物、若しくはその塩、又はそれらの溶媒和物を有効成分とする、脊髄虚血による神経細胞障害を予防及び／又は治療するための医薬を提供するものである。
式〔Ｉ〕において、Ｒ¹は、置換されていてもよいアリール又は環を構成する原子の数が５〜１０個の芳香族複素環基を表す。ここで、芳香族複素環基は、単環又は縮合環であってよく、環構成原子として窒素、酸素又は硫黄を一つ以上含む。さらに、アリール又は芳香族複素環基は、ヒドロキシ、ハロゲン、アルキル、ハロアルキル、ヒドロキシアルキル、アラルキル、アルケニル、アルコキシ、ハロアルキルオキシ、アルキルチオ、シクロアルキル、シクロアルキルアルキル、シクロアルキルオキシ、アルキルスルホニル、スルファモイル、アルカノイル、アミノ、モノアルキルアミノ、ジアルキルアミノ、カルボキシ、アルコキシカルボニル、シアノ及びニトロからなる群から選択される同一又は異なった１〜３個の置換基により置換されていてもよい。
Ｒ²は、水素、アルキル、アルケニル、シクロアルキル、シクロアルキルアルキル、ヒドロキシアルキル、ハロアルキル、アルコキシ、アルキルチオ、アミノ、モノアルキルアミノ、ジアルキルアミノ又は置換されていてもよいフェニルを表す。かかるフェニルは、ハロゲン、アルキル及びアルコキシからなる群から選択される同一又は異なった１〜３個の置換基により置換されていてもよい。
Ｒ³、Ｒ⁴は、同一又は異なって、水素若しくは置換されていてもよいアルキル（かかるアルキルは、ヒドロキシ、アルコキシ、アミノ、モノアルキルアミノ及びジアルキルアミノからなる群から選択される同一又は異なった１個又は２個の置換基によって置換されていてもよい。）を表すか、Ｒ³とＲ⁴が隣接するＮと一緒になってＮＲ³Ｒ⁴で４〜８員の環状アミノを表す。かかる環状アミノは、当該窒素のほかに環構成原子として窒素、酸素又は硫黄を有していてもよく、更にアルキル、アルコキシ、ヒドロキシ、オキソ、アミノ、モノアルキルアミノ、ジアルキルアミノ、置換されていてもよいアリール及びピリジルからなる群から選択される同一又は異なった１〜３個の置換基によって置換されていてもよい。
さらに、Ｒ³、Ｒ⁴が結合しているＮは、オキシドを形成していてもよい。
Ａは、炭素数２〜１０のアルキレンを表す。かかるアルキレンは、任意の位置においてアルコキシ、ヒドロキシ及びオキソからなる群から選択される置換基によって置換されていてもよい。
Ｅは、Ｏ又はＳを表す。
Ｗは、単結合、Ｏ、Ｓ又は（ＣＨ₂）ｎ（ＣＨ₂は、アルキルによって置換されていてもよい。ｎは１又は２を表す。）を表す。
Ｘ、Ｙ、Ｚは、同一又は異なってＣＨ、ＣＲ（Ｒはアルキルを表す。）又はＮを表す。但し、Ｘ、Ｙ、Ｚが同時にＣＨ又はＣＲの場合は除く。
Ｇ環は、ピリジン、ピリミジン、1,3,5−トリアジンを表す。
Ｘ、Ｙ、Ｚの一つ乃至三つがＮの場合、その中の一つがオキシドを形成してもよい。
The present inventors have found that compounds known as sodium / calcium channel blockers include compounds useful for the prevention and / or treatment of spinal nerve cell damage during spinal cord ischemia, and complete the present invention. It came to. That is, the present invention relates to the formula [I]:

And a salt thereof, or a solvate thereof as an active ingredient, to provide a medicament for preventing and / or treating nerve cell damage due to spinal cord ischemia.
In the formula [I], R ¹ represents an optionally substituted aryl or an aromatic heterocyclic group having 5 to 10 atoms constituting the ring. Here, the aromatic heterocyclic group may be a monocyclic ring or a condensed ring, and contains one or more of nitrogen, oxygen or sulfur as ring constituent atoms. In addition, aryl or aromatic heterocyclic groups are hydroxy, halogen, alkyl, haloalkyl, hydroxyalkyl, aralkyl, alkenyl, alkoxy, haloalkyloxy, alkylthio, cycloalkyl, cycloalkylalkyl, cycloalkyloxy, alkylsulfonyl, sulfamoyl, alkanoyl , Amino, monoalkylamino, dialkylamino, carboxy, alkoxycarbonyl, cyano and nitro may be substituted by 1 to 3 identical or different substituents.
R ² represents hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkylalkyl, hydroxyalkyl, haloalkyl, alkoxy, alkylthio, amino, monoalkylamino, dialkylamino or optionally substituted phenyl. Such phenyl may be substituted by the same or different 1 to 3 substituents selected from the group consisting of halogen, alkyl and alkoxy.
R ³ and R ⁴ are the same or different and are hydrogen or optionally substituted alkyl (the alkyl is the same or different 1 selected from the group consisting of hydroxy, alkoxy, amino, monoalkylamino and dialkylamino) R ³ and R ⁴ together with the adjacent N represent NR ³ R ⁴ representing a 4- to 8-membered cyclic amino. Such cyclic amino may have nitrogen, oxygen or sulfur as a ring constituent atom in addition to the nitrogen, and may further be alkyl, alkoxy, hydroxy, oxo, amino, monoalkylamino, dialkylamino, or substituted. It may be substituted by 1 to 3 identical or different substituents selected from the group consisting of good aryl and pyridyl.
Furthermore, N to which R ³ and R ⁴ are bonded may form an oxide.
A represents an alkylene having 2 to 10 carbon atoms. Such alkylene may be substituted at any position with a substituent selected from the group consisting of alkoxy, hydroxy and oxo.
E represents O or S.
W represents a single bond, O, S, or (CH ₂ ) n (CH ₂ may be substituted with alkyl. N represents 1 or 2).
X, Y and Z are the same or different and represent CH, CR (R represents alkyl) or N. However, the case where X, Y, and Z are simultaneously CH or CR is excluded.
Ring G represents pyridine, pyrimidine, 1,3,5-triazine.
When one to three of X, Y, and Z are N, one of them may form an oxide.

The compound of the formula [I], which is an active ingredient of the medicament of the present invention, is a known compound and is known to exhibit an excellent inhibitory effect on neuronal necrosis in the acute phase of cerebrovascular disorder (WO / 96/07641) . It is known that excitatory amino acids are involved in neuronal damage, and receptors for such amino acids include receptors other than N-methyl-D-aspartate (NMDA) receptor and NMDA receptor. There is a body (non-NMDA). It has been clarified that the compound [I] according to the present invention does not act on the NMDA receptor (WO / 96/07641).
However, it has not been known at all that Compound [I] has a nerve cell protective action under spinal cord ischemia and is useful as a medicament for prevention and / or treatment of nerve injury caused by ischemic spinal cord injury. .

Examples of the compound of the formula [I] useful as the active ingredient of the medicament of the present invention include the compound groups shown in the following (i) to (iv).
(I) A compound in which NR ³ R ⁴ is a 4- to 8-membered cyclic amino, and A is an alkylene having 4 to 10 carbon atoms. Such a cyclic amino may have oxygen or sulfur as a ring atom, and the cyclic amino has alkyl, alkoxy, hydroxy, oxo, amino, monoalkylamino, dialkylamino, pyridyl or aryl as a substituent. The arylene may be hydroxy, halogen, alkyl, haloalkyl, hydroxyalkyl, aralkyl, alkenyl, alkoxy, haloalkyloxy, alkylthio, cycloalkyl, cycloalkylalkyl, cycloalkyloxy, alkylsulfonyl, sulfamoyl, alkanoyl, amino, mono It may be substituted with 1 to 3 identical or different substituents selected from the group consisting of alkylamino, dialkylamino, carboxy, alkoxycarbonyl, cyano and nitro.
(Ii) In the general formula [I], R ¹ is an aromatic heterocyclic group having 5 to 10 atoms constituting the ring, R ² is hydrogen, and A is 2 to 3 carbon atoms. A compound wherein alkylene is optionally substituted by alkoxy, hydroxy or oxo at any position and E is O. The aromatic heterocyclic group is a monocyclic ring or a condensed ring and contains one or more nitrogen, oxygen or sulfur as ring constituent atoms, and is hydroxy, halogen, alkyl, haloalkyl, hydroxyalkyl, aralkyl, alkenyl, alkoxy, haloalkyloxy 1 or the same or different selected from the group consisting of alkylthio, cycloalkyl, cycloalkylalkyl, cycloalkyloxy, alkylsulfonyl, sulfamoyl, alkanoyl, amino, monoalkylamino, dialkylamino, carboxy, alkoxycarbonyl, cyano and nitro May be substituted by ~ 3 substituents.
(Iii) R ¹ is an aromatic heterocyclic group having 5 to 10 atoms constituting the ring, and the aromatic heterocyclic group is a monocyclic ring or a condensed ring, such as nitrogen, oxygen or Contains one or more sulfur and is hydroxy, halogen, alkyl, haloalkyl, hydroxyalkyl, aralkyl, alkenyl, alkoxy, haloalkyloxy, alkylthio, cycloalkyl, cycloalkylalkyl, cycloalkyloxy, alkylsulfonyl, sulfamoyl, alkanoyl, amino , Monoalkylamino, dialkylamino, carboxy, alkoxycarbonyl, cyano and nitro, which may be substituted by 1 to 3 identical or different substituents, wherein R ² is alkyl, alkenyl, cyclo Alkyl, cycloalkylalkyl, hydroxy Alkyl, haloalkyl, alkoxy, alkylthio, amino, monoalkylamino, dialkylamino or phenyl, wherein the phenyl is substituted by 1 to 3 identical or different substituents selected from the group consisting of halogen, alkyl and alkoxy A compound in which A is alkylene having 2 to 3 carbon atoms, and the alkylene may be substituted with alkoxy, hydroxy or oxo at any position.
(Iv) R ¹ is an aromatic heterocyclic group having 5 to 10 atoms constituting the ring, and the aromatic heterocyclic group is a single ring or condensed ring, and nitrogen, oxygen or One or more sulfur may be contained, and such aromatic heterocyclic groups may be hydroxy, halogen, alkyl, haloalkyl, hydroxyalkyl, aralkyl, alkenyl, alkoxy, haloalkyloxy, alkylthio, cycloalkyl, cycloalkylalkyl, Substituted with 1 to 3 identical or different substituents selected from the group consisting of cycloalkyloxy, alkylsulfonyl, sulfamoyl, alkanoyl, amino, monoalkylamino, dialkylamino, carboxy, alkoxycarbonyl, cyano and nitro. at best, NR ³ R ⁴ is piperazino, the piperazino Dino may be unsubstituted or substituted with alkyl, alkoxy, hydroxy, oxo, amino, monoalkylamino, dialkylamino, pyridyl or aryl, where aryl is hydroxy, halogen, alkyl, haloalkyl, hydroxyalkyl, aralkyl, alkenyl , Alkoxy, haloalkyloxy, alkylthio, cycloalkyl, cycloalkylalkyl, cycloalkyloxy, alkylsulfonyl, sulfamoyl, alkanoyl, amino, monoalkylamino, dialkylamino, carboxy, alkoxycarbonyl, cyano and nitro A compound which may be substituted by 1 to 3 identical or different substituents.

  In the present specification, alkyl includes linear or branched ones having 1 to 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl. , N-pentyl, isopentyl, n-hexyl, and isohexyl. Of these, those having 1 to 4 carbon atoms are preferred.

Examples of alkenyl include those having 2 to 6 carbon atoms such as vinyl, allyl, 3-butenyl, 2-pentenyl, and 4-hexenyl.
Cycloalkyls preferably have 3 to 10 carbon atoms, and examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 1-adamantyl, and 2-adamantyl.

Examples of aryl include phenyl having 6 to 13 carbon atoms, 1-naphthyl, 2-naphthyl and biphenyl. In particular, phenyl is preferred.
Aralkyl has 7 to 13 carbon atoms and an alkyl moiety that is linear or branched, and examples thereof include benzyl, phenethyl, phenylpropyl, phenylbutyl, diphenylmethyl, and naphthylmethyl.
Examples of halogen include chlorine, fluorine, bromine and iodine.

  Alkoxy is preferably a straight chain or branched chain having 1 to 6 carbon atoms, such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, n -Pentyloxy, isopentyloxy, n-hexyloxy, isohexyloxy can be mentioned.

  The alkanoyl is preferably a linear or branched one having 1 to 6 carbon atoms, and examples thereof include acetyl, propanoyl, butanoyl, isobutanoyl, pentanoyl, hexanoyl and 2-methylpentanoyl.

  Alkylthio is preferably a straight chain or branched chain having 1 to 6 carbon atoms, such as methylthio, ethylthio, n-propylthio, isopropylthio, n-butylthio, isobutylthio, sec-butylthio, tert-butylthio, Examples include n-pentylthio, isopentylthio, n-hexylthio, and isohexylthio.

  As the alkylsulfonyl, linear or branched ones having 1 to 6 carbon atoms are preferable, for example, methylsulfonyl, ethylsulfonyl, n-propylsulfonyl, isopropylsulfonyl, n-butylsulfonyl, isobutylsulfonyl, sec-butyl. Examples include sulfonyl, tert-butylsulfonyl, n-pentylsulfonyl, isopentylsulfonyl, n-hexylsulfonyl and isohexylsulfonyl.

  Hydroxyalkyl includes linear or branched ones having 1 to 6 carbon atoms, such as 2-hydroxyethyl, 3-hydroxypropyl, 2-hydroxypropyl, 4-hydroxybutyl, 3-hydroxybutyl, 5- Examples thereof include hydroxypentyl and 6-hydroxyhexyl.

  Examples of haloalkyl include linear or branched ones having 1 to 6 carbon atoms, such as trifluoromethyl, fluoromethyl, 2-bromoethyl, and 3-chloroethyl.

  Examples of monoalkylamino include linear or branched ones having 1 to 6 carbon atoms, such as methylamino, ethylamino, propylamino, butylamino, heptylamino, and hexylamino.

  Examples of the dialkylamino include linear or branched ones having 1 to 6 carbon atoms, such as dimethylamino, diethylamino, dipropylamino, dibutylamino, diheptylamino, and dihexylamino.

  As specific examples of alkoxycarbonyl, linear or branched ones having 2 to 7 carbon atoms are preferable. For example, methoxycarbonyl, ethoxycarbonyl, n-propoxycarbonyl, isopropoxycarbonyl, n-butoxycarbonyl, isobutoxy Examples thereof include carbonyl, sec-butoxycarbonyl, tert-butoxycarbonyl, n-pentyloxycarbonyl, isopentyloxycarbonyl, n-hexyloxycarbonyl and isohexyloxycarbonyl.

  Specific examples of cycloalkyloxy are preferably those having 3 to 10 carbon atoms, such as cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, cycloheptyloxy, cyclooctyloxy, and 2-adamantyloxy. Can do.

  Specific examples of cycloalkylalkyl are preferably those having 4 to 11 carbon atoms, such as cyclopropylmethyl, cyclopropylethyl, cyclobutylmethyl, cyclobutylethyl, cyclopentylmethyl, cyclopentylethyl, cyclopentylpropyl, cyclohexylmethyl, cyclohexyl. Mention may be made of ethyl, cyclohexylpropyl, cycloheptylmethyl and 2-adamantylmethyl.

Examples of the 4- to 8-membered cyclic amino include azetidin-1-yl, pyrrolidin-1-yl, piperidino, hexamethyleneimino, tetrahydropyridino, octahydroazocin-1-yl, piperazin-1-yl, homo Examples include piperazin-1-yl, morpholino and thiomorpholino.
Such cyclic amino substituents include alkyl, alkoxy, hydroxy, oxo, amino, monoalkylamino, dialkylamino, optionally substituted aryl or pyridyl. Examples of the substituent for aryl or pyridyl include those listed as the substituent for R1.

  The aromatic heterocyclic group having 5 to 10 atoms constituting the ring includes one or more arbitrarily selected oxygen, sulfur or nitrogen atoms in the ring, and may be monocyclic or condensed. It may be a ring. For example, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-thienyl, 2-furyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, 3-pyridazinyl, 4-pyridazinyl, 1-isoquinolyl, 4-isoquinolyl, Examples thereof include 2-quinazolinyl and 1-methyl-2-indolyl.

The alkylene represented by A may be linear or branched. As a pharmaceutical for protecting nerves from spinal cord ischemia and preventing and / or treating nerve injury, A is preferably an alkylene having 3 to 6 carbon atoms, more preferably an alkylene having 4 to 6 carbon atoms.
E is preferably O.
W is preferably a single bond.
X, Y, and Z are preferably X = Z = N and Y = CH or Z = N and X = Y = CH. The former is particularly preferable.
R ¹ is preferably halogen-substituted phenyl, particularly fluorophenyl.
R ² is preferably alkyl or haloalkyl, and more preferably alkyl. Particularly preferred is methyl.
As R ³ and R ⁴ , those which form a cyclic amino together with adjacent N together with —NR ³ R ⁴ are preferable, and among them, those containing only one nitrogen atom as a ring-constituting hetero atom are preferable. Piperidino is particularly preferable.

で表される化合物を挙げることができる。
式〔Ｉａ〕において、Ａ^２１は、炭素数４〜６のアルキレンを表す。
Ｅ^２１は、Ｏを表す。
Ｘ^２１＝Ｚ^２１＝ＮでＹ^２１＝ＣＨ又はＸ^２１＝Ｙ^２１＝ＣＨでＺ^２１＝Ｎを表す。
Ｒ^２１は、ハロゲン置換フェニルを表す。
Ｒ^２２は、アルキル又はハロアルキルを表す。
Ｒ^２３、Ｒ^２４としては、隣接するＮと一緒になって−ＮＲ^２３Ｒ^２４で４〜８員の環状アミノを形成するもので、環構成ヘテロ原子として窒素原子１個のみを含むものを表す。
Ｗ^２は単結合を表す。 Preferred compounds include those of the formula [Ia]

The compound represented by these can be mentioned.
In the formula [Ia], A ²¹ represents alkylene having 4 to 6 carbon atoms.
E ²¹ represents O.
X ²¹ = Z ²¹ = N and Y ²¹ = CH or X ²¹ = Y ²¹ = CH and Z ²¹ = N.
R ²¹ represents a halogen-substituted phenyl.
R ²² represents alkyl or haloalkyl.
R ²³ and R ²⁴ together with the adjacent N form a 4- to 8-membered cyclic amino group with —NR ²³ R ²⁴ , and represent one containing only one nitrogen atom as a ring-constituting hetero atom. .
W ² represents a single bond.

As a more preferred compound,
4- (4-fluorophenyl) -2-methyl-6- (4-piperidinobutoxy) pyrimidine,
4- (4-fluorophenyl) -2-methyl-6- (1-methyl-4-piperidinobutoxy) pyrimidine,
4- (4-fluorophenyl) -2-methyl-6- (5-piperidinopentyloxy) pyrimidine,
4- (4-fluorophenyl) -2-methyl-6- (6-piperidinohexyloxy) pyrimidine,
2- (4-fluorophenyl) -4-methyl-6- (4-piperidinobutoxy) pyrimidine, 4- (4-fluorophenyl) -2-methyl-6- (3-piperidinopropoxy) pyridine and Mention may be made of 4- (4-fluorophenyl) -2-methyl-6- (5-piperidinopentyloxy) pyridine and their salts.

The most preferred compounds include 4- (4-fluorophenyl) -2-methyl-6- (5-piperidinopentyloxy) pyridine and salts thereof.
The compound which is an active ingredient of the pharmaceutical of the present invention is preferentially distributed in the synaptosomal membrane fraction of the brain rather than the heart in an experiment using rats, and therefore has an extremely high affinity for brain VSSC compared to cardiac VSSC. Have been confirmed to have excellent properties such as low side effects on cardiac function (Shimizuzu et al., Naunyn Schmiedebergs Arch. Pharmacol. 1997; 355: 601-8).

Examples of the solvate of compound (I) useful as an active ingredient of the medicament of the present invention include hydrates and ethanol solvates.
Examples of the salt of compound [I] include salts of hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrofluoric acid, hydrobromic acid with mineral acids, or acetic acid, tartaric acid, lactic acid, citric acid, fumar Examples include acids, maleic acid, succinic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, naphthalenesulfonic acid, and salts of camphorsulfonic acid with organic acids.
Although compounds having an asymmetric carbon are included in the compound of the formula [I], each optical isomer and any of these racemates are useful for the medicament of the present invention. Moreover, although compound [I] may take a crystal polymorph, these crystal polymorphs are also useful for this invention.
The compound represented by the formula [I], its salt, and solvate that can be used for the medicament of the present invention are produced by known methods, for example, the methods described in WO96 / 07641, WO02 / 20492, and if necessary And can be purified.

The medicament of the present invention comprising a compound represented by the formula [I], a salt thereof or a solvate thereof as an active ingredient protects nerve cells under spinal cord ischemia and prevents cell damage (injury). Or useful for treatment.
Here, “protecting nerve cells under spinal cord ischemia” refers to preventing damage to spinal nerve cells in a situation where the blood flow to the spinal cord is blocked or restricted, and if damaged, Means to promote recovery.
In another aspect, the present invention provides an effective amount of a compound represented by the formula [I], a salt thereof or a solvate thereof, for preventing and / or treating damage to nerve cells due to spinal cord ischemia. The present invention provides a method for preventing and / or treating nerve cell damage due to spinal cord ischemia, comprising the step of administering to a mammal including a human in need thereof.
In still another aspect, the present invention provides the use of a compound of formula [I] or a salt thereof, or a solvate thereof in the manufacture of the above medicament.

  The dosage of the medicament of the present invention is appropriately determined in consideration of the patient's condition such as age and weight, administration route, etc. Usually, the daily dose of the compound of formula [I], which is an active ingredient, to a human adult The amount per dose is 0.1 mg to 1 g / human, preferably 1 to 300 mg / human for oral administration, and 0.01 mg to 100 mg / human for parenteral administration, preferably 0.1 to 30 mg / human. In some cases, doses below or above these dosages may be required. It is desirable to divide the dose into 2-4 times a day.

  In the administration of the medicament of the present invention, the compound of the formula [I] which is an active ingredient or a salt thereof, or a solvate thereof can be used as it is, but generally the active ingredient is pharmaceutically acceptable. It is preferably administered as a pharmaceutical composition contained in a non-toxic and inert carrier. The amount of the active ingredient in the pharmaceutical composition varies depending on the dosage form, but is usually 0.01 to 99.5% by weight, preferably 0.5 to 90% by weight.

  As the carrier, a solid, semi-solid, or liquid diluent, filler, and one or more auxiliary agents for formulation are used. The pharmaceutical composition is desirably administered in dosage unit form. The pharmaceutical composition of the present invention can be administered orally, intra-tissue (eg, intravenous administration), local administration (eg, transdermal administration) or rectally. Intravenous administration or oral administration is preferred, and intravenous administration is particularly preferred. Preparation of the preparation can be carried out by methods known to those skilled in the art, but the method described in WO96 / 07641 can be preferably used.

  Oral administration should be carried out in solid or liquid dosage units such as powders, powders, tablets, dragees, capsules, granules, suspensions, solutions, syrups, drops, sublingual tablets and other dosage forms. Can do.

The powder is manufactured by making the active substance fine. Powders are made by mixing the active substance with appropriate fineness and then mixing it with a similarly finely divided pharmaceutical carrier such as edible carbohydrates such as starch, mannitol and the like. A flavoring agent, a preservative, a dispersing agent, a coloring agent, a fragrance, and the like may be mixed as necessary.
Capsules are produced by first filling a powdered powder, powder or powder as described above into a capsule shell such as a gelatin capsule as described in the section of tablets. The Lubricants and fluidizing agents, such as colloidal silica, talc, magnesium stearate, calcium stearate, solid polyethylene glycol, etc., can be mixed in powder form and then filled. it can. When a capsule is ingested by adding a disintegrant or solubilizer such as carboxymethylcellulose, carboxymethylcellulose calcium, low-substituted hydroxypropylcellulose, croscarmellose sodium, carboxymethyl starch sodium, calcium carbonate, sodium carbonate The effectiveness of the medicine can be improved.

  In addition, a fine powder of the compound of the present invention can be suspended and dispersed in vegetable oil, polyethylene glycol, glycerin, and a surfactant and wrapped in a gelatin sheet to form a soft capsule. Tablets are made by adding excipients to make a powder mixture, granulating or slugging, then adding a disintegrant or lubricant and then tableting. The powder mixture is prepared by mixing an appropriately powdered substance with the above-mentioned diluent or base, and if necessary a binder (for example, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, gelatin, polyvinylpyrrolidone, polyvinyl alcohol, etc.), A dissolution retardant (for example, paraffin), a resorbent (for example, quaternary salt) and an adsorbent (for example, bentonite, kaolin, dicalcium phosphate, etc.) may be used in combination.

  The powder mixture can be first wetted with a binder such as syrup, starch paste, gum arabic, cellulose solution or polymer solution, stirred and mixed, dried and pulverized into granules. Instead of granulating the powder in this way, it is also possible to first crush the tablet and then to crush the resulting incomplete slag into granules. The granules thus produced can be prevented from adhering to each other by adding stearic acid, stearate, talc, mineral oil or the like as a lubricant. The mixture thus lubricated is then tableted. Film coating or sugar coating can be applied to the uncoated tablets thus produced.

  Further, the compound of the present invention may be directly tableted after being mixed with a fluid inert carrier without going through the steps of granulation or slag as described above. A transparent or translucent protective coating made of a shellac hermetic coating, a coating of sugar or polymer material, and a polishing coating made of wax can also be used. Other oral dosage forms such as solutions, syrups, elixirs and the like can also be in dosage unit form so that a given quantity contains a certain amount of a compound of the invention. A syrup is produced by dissolving the compound of the present invention in a suitable flavor aqueous solution, and elixir is produced by using a non-toxic alcoholic carrier. Suspensions are formulated by dispersing the compound in a non-toxic carrier. Solubilizers and emulsifiers (eg, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol esters), preservatives, flavoring agents (eg, peppermint oil, saccharin) and others can also be added as needed. .

  If necessary, dosage unit formulations for oral administration may be microencapsulated. The formulation can also provide extended action time or sustained release by coating or embedding in polymers, waxes, and the like.

Intra-tissue administration is performed by injection, infusion, or continuous infusion into arteries and veins. Injection can be performed by using a liquid dosage unit form for subcutaneous, intramuscular or intravenous injection, for example, a solution or suspension form.
A pharmaceutical formulation for injection, infusion or continuous infusion into an artery or vein is suspended or dissolved in a non-toxic liquid carrier suitable for the purpose of injection, such as an aqueous or oily medium, and then the suspension or Prepared by sterilizing the solution. Non-toxic salts or salt solutions may be added to make the injection solution isotonic. Furthermore, stabilizers, preservatives, emulsifiers and the like can be used in combination.

  Rectal administration involves the compound being soluble or insoluble in low melting water such as polyethylene glycol, cocoa butter, semi-synthetic oils (eg, Witepsol®), higher esters (eg, myristyl palmitate) and those It can be carried out by using a suppository or the like produced by dissolving or suspending in the mixture. Preparations for oral administration are solid or liquid dosage units such as powders, powders, tablets, dragees, capsules, granules, suspensions, solutions, syrups, drops, sublingual tablets and other suitable agents May be in the form of, for example, excipients such as glucose, lactose, D-mannitol, starch, or crystalline cellulose; disintegrants or disintegration aids such as carboxymethylcellulose, starch, or carboxymethylcellulose calcium; hydroxy Binders such as propylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, or gelatin; lubricants such as magnesium stearate or talc; coating agents such as hydroxypropylmethylcellulose, sucrose, polyethylene glycol or titanium oxide; petrolatum, liquid paraffin, polyethylene G Call, gelatin, kaolin, glycerin, purified water, or can be prepared using a base such hard fat.

  The medicament of the present invention is effective for the prevention and / or treatment of cell damage during spinal cord ischemia. That is, when the spinal cord is exposed to ischemia, the medicament of the present invention is used as a therapeutic agent that protects spinal cord neurons to prevent the progress of cell damage and / or restores damaged cells to a normal state. Useful. Therefore, ischemic invasion to the spinal cord can be prevented or reduced by using the medicament of the present invention. Here, “ischemic invasion of the spinal cord” means that the spinal cord is in an ischemic state, the spinal nerve necrosis (necrosis) and apoptosis progress, the stimulation conduction through the spinal cord injury is impaired, and the movement disorder Or a state of sensory disturbance.

  When the medicament of the present invention is used for ischemic spinal cord invasion in aortic aneurysm repair, it can be administered prophylactically. Prophylactic administration can be either oral or parenteral. It can also be administered prophylactically during or before surgery by parenteral administration such as injection, infusion or continuous infusion into arteries and veins. In particular, in the operation of a descending thoracic aortic aneurysm or thoracoabdominal aortic aneurysm, it is preferable to administer multiple times prior to or during the operation, preferably at the time of releasing the aortic block or after the operation. In addition, for the patient who has nerve cell damage due to spinal cord ischemia, the medicament of the present invention can be administered parenterally (intravenously or intraarterially) or orally in order to prevent or reduce the deterioration of symptoms.

  As described above, the medicament of the present invention is effective even before administration during spinal cord ischemia, during the period, and at any time of reopening, so that ischemic invasion to the spinal cord can be predicted. The present invention is not limited to cases (such as in the case of aortic aneurysm repair), and can also be applied to the prevention and / or treatment of neurological disorders during spinal cord ischemia (for example, accidents and strokes) that cannot be predicted. Therefore, the medicament of the present invention is also useful for preventing and / or treating nerve injury due to spinal cord ischemia caused by accidents, seizures, etc., for example, by administering as an emergency treatment, and / or preventing progression of neuropathy. Recovery can be promoted and possible sequelae can be prevented or reduced.

  EXAMPLES The present invention will be described in more detail below with reference to examples, but the present invention is not limited thereto.

Production Example 1 2- (4-Fluorophenyl) -4- (4-piperidinobutoxy) -6-methylpyridine hydrochloride (1) 4- (4-chlorobutoxy) -2- (4-fluorophenyl)- 6-Methylpyridine 2- (4-fluorophenyl) -4-hydroxy-6-methylpyridine (2.5 g), 1-bromo-4-chlorobutane (3.16 g), silver carbonate (1.7 g) and toluene (100 ml) were heated to reflux for 40 hours. The reaction solution was filtered to remove insolubles, the filtrate was concentrated, and the residue was purified by silica gel column chromatography to obtain 1.45 g of the objective compound as white crystals. Melting point 59-61 ° C

(2) 2- (4-Fluorophenyl) -4- (4-piperidinobutoxy) -6-methylpyridine hydrochloride 4- (4-chlorobutoxy) -2- (4- A mixture of 1.45 g of fluorophenyl) -6-methylpyridine, 1.26 g of piperidine and 12 ml of DMF was stirred at 100 ° C. for 1.5 hours. The reaction mixture was cooled, poured into ice water, and extracted with ethyl acetate. The organic layer was washed several times with brine, dried over anhydrous magnesium sulfate, and concentrated. The residue was purified by silica gel column chromatography to obtain 1.2 g of the objective compound as an oil. This was dissolved in methanol, added with 3.5 ml of 1N hydrochloric acid to pH 5, and concentrated. Ether was added to the residue, and the crystals were collected by filtration. The crystals were recrystallized from a mixed solvent of acetonitrile and ether to obtain 1.02 g of the objective compound as white crystals. Melting point 164-166 ° C
Elemental analysis (as C ₂₁ H ₂₇ FN ₂ O · HCl)
Calculated value (%): C: 66.57 H: 7.45 N: 7.39; Actual value (%): C: 66.21 H: 7.45 N: 7.09

The following compounds were synthesized in the same manner as in Production Example 1.
Production Example 2 4- (4-Fluorophenyl) -2-methyl-6- (5-piperidinopentyloxy) pyridine hydrochloride (hereinafter abbreviated as “NS-7”)
Melting point 138-140 ° C
Elemental analysis (as C ₂₂ H ₂₉ FN ₂ O · HCl)
Calculated value (%): C: 67.25 H: 7.70 N: 7.13; Actual value (%): C: 67.00 H: 7.68 N: 6.95

Production Example 3 4- (4-Fluorophenyl) -2-methyl-6- (5-piperidinopentyloxy) pyrimidine (1) 4- (4-fluorophenyl) -6-hydroxy-2-methylpyrimidine 3- (4-Fluorophenyl) -3-oxopropionic acid ethyl ester 8.7 kg, acetamidine hydrochloride 11.7 kg and potassium carbonate 28.6 kg were stirred in methanol 34.5 kg at about 50 ° C. for 5 hours. Insoluble matters were separated from the reaction solution, washed with methanol, water was added to the mother liquor and the washing solution, and neutralized with an 18% aqueous hydrochloric acid solution. After neutralization, the crystals were aged by stirring for 4 hours under reflux, and the precipitated crystals were isolated by centrifugation, washed with water and dried to obtain 7.4 kg of the target compound as white crystals.

(2) 4-chloro-6- (4-fluorophenyl) -2-methylpyridine 13.8 kg of 4- (4-fluorophenyl) -6-hydroxy-2-methylpyrimidine and 11.5 kg of phosphorus oxychloride are refluxed in acetonitrile. And stirred for 4 hours. Water was added to the reaction solution, the precipitated crystals were separated, and the obtained crystals were washed with water and dried to obtain 14.6 kg of the target compound as pale yellow crystals.

(3) 4- (4-Fluorophenyl) -2-methyl-6- (5-piperidinopentyloxy) pyrimidine 4-chloro-6- (4-fluorophenyl) -2-methylpyridine 13.8 kg, 60% 4.8 kg of sodium hydride, 130 kg of cyclohexane, and 10.6 kg of 5-piperidino-1-pentanol were stirred under reflux for 4 hours.

Production Example 4 4- (4-Fluorophenyl) -2-methyl-6- (5-piperidinopentyloxy) pyrimidine 1/2 hydrate The reaction solution obtained in Production Example 3 was cooled, and 55 kg of water was added. In addition, after stirring at room temperature for 1 hour, the aqueous layer was removed. The organic layer was extracted with 3% aqueous hydrochloric acid. The extract was neutralized with 4% aqueous sodium hydroxide solution, and the precipitated crystals were separated. The obtained crystals were treated with activated carbon in 120 kg of acetone, and insolubles were removed by filtration. 85.2 kg of water was added to the filtrate for crystallization. The precipitated crystals were separated and dried to obtain 19.2 kg (yield: 87%) of the target compound as white crystals. Melting point: 62.0-63.5%
Elemental analysis value (as C ₂₁ H ₂₈ FN ₃ O, 1 / 2H ₂ O)
Calculated value (%): C: 68.82 H: 7.98 N: 11.47; actual value (%): C: 68.86 H: 7.98 N: 11.42
¹ H NMR (200 MHz): δ 7.95-8.05 (m, 2H), 7.09-7.21 (m, 2H), 6.83 (s, 1H), 4.38 (t, J = 13.2Hz, 2H), 2.65 (s, 3H), 2.27-2.38 (Complex m, 6H), 1.74-1.88 (m, 2H), 1.43-1.61 (Complex m, 10H)

Production Example 5 NS-7
18.0 kg of 4- (4-fluorophenyl) -2-methyl-6- (5-piperidinopentyloxy) pyrimidine 1/2 hydrate obtained in Production Example 4 was dissolved in 142.2 kg of acetone, and hydrochloric acid was added. 5.0 kg was added and stirred at about 20 ° C. for 1 hour. The precipitated crystals were separated, washed with acetone and dried to obtain 17.4 kg of the target compound as crude crystals. 17.0 kg of this crude crystal was recrystallized from ethanol to obtain 15.4 kg of the target compound (yield: 91%) as white crystals.

Formulation Example 1 Injection According to the following formulation, 1 ml of injection can be prepared by a conventional method.
Formulation: NS-7 1mg, Sodium chloride 9mg, Water for injection

Formulation Example 2 Injection According to the following formulation, 1 ml of injection can be prepared by a conventional method.
Formulation: NS-7 1mg, glucose 48mg, sodium dihydrogen phosphate 1.25mg, sodium monohydrogen phosphate 0.18mg, water for injection

Formulation Example 3 Injection A 1 ml injection can be prepared by a conventional method according to the following formulation.
Formulation: NS-7 1mg, sorbit 48mg, benzyl alcohol 20mg, sodium dihydrogen phosphate 2.5mg, sodium monohydrogen phosphate 0.36mg, water for injection

Formulation Example 4 Tablet According to the following formulation, a 120 mg tablet can be prepared by a conventional method.
Formulation: NS-7 3 mg, lactose 58 mg, corn starch 30 mg, crystalline cellulose 20 mg, hydroxypropyl cellulose 7 mg, magnesium stearate 2 mg

Formulation Example 5 Infusion A 500 ml infusion bag injection for infusion can be prepared by a conventional method according to the following formulation.
Formula: NS-7 25mg, Sodium chloride 4.5g, Water for injection

Formulation Example 6 Continuous Injection A 20 ml prefilled syringe kit can be prepared by a conventional method according to the following formulation.
Formula: NS-7 20mg, Sodium chloride 180mg, Water for injection

Test Example 1 Suppression of nerve cell damage due to rabbit spinal cord ischemia Experimental Method (1) Creation of Rabbit Spinal Cord Ischemia Model Using 26 New Zealand White Rabbits (weight: 2.2-3.0 kg), the aorta below the kidneys is occluded for 20 minutes to spinal cord ischemia model It was created.
Preoperative preparation was performed according to the method described in the literature (Terada et al., J. Thorac. Cardiovasc. Surg. 2001; 122: 979-85). A 24 gauge intravenous catheter was placed in the animal's earlobe vein for drug administration. The animals were anesthetized with sodium pentobarbital (25 mg / kg iv) and allowed to breathe spontaneously. 0.5% lidocaine was administered to the skin resection site as a local anesthetic. A 24 gauge catheter was placed in the left common carotid artery for arterial pressure monitoring. Body temperature was continuously monitored with a rectal probe and maintained at 38.5 ° C. with a warm lamp. After laparotomy, the abdominal aorta below the kidney was exposed. After heparin administration (200 U / kg), the abdominal aorta was occluded for 20 minutes with a snare just distal to the renal artery and just above the aortic bifurcation. Thereafter, the snare was removed and the aortic branch was resumed, and the abdomen was closed.

(2) Drug administration NS-7 (Nippon Shinyaku, available from Japan) obtained in Production Example 2 was dissolved in 0.9% physiological saline to prepare a 2.5 mg / ml solution. The animals were randomly divided into 4 groups, and the above treatment (1) was performed as follows. The aortic occlusion treatment was not performed in the sham operation group (n = 3). In the control group (n = 7), physiological saline (1 ml) was intravenously administered 15 minutes before occlusion of the aorta. Group A (n = 8) received NS-7 (1 mg / kg) intravenously 15 minutes before ischemia. Group B (n = 8) received NS-7 (1 mg / kg) intravenously immediately after reopening.

(3) Extraction of tissue Pentobarbital (200 mg / kg) was administered 48 hours after the operation to cause the animals to die. The spinal cord is quickly collected and the lower thoracic to lower lumbar segments are divided into the following four segments: the lower thoracic spinal cord (LT = T10-T12), the upper lumbar spinal cord (UL = L1-L2), and the middle lumbar spinal cord ( ML = L3-L4) and lower lumbar spinal cord (LL = L5-L6).

(4) Statistical analysis Data were expressed as mean ± standard deviation. Statistical analysis of neurological and histological scores was performed using the Mann-Whitney U test. Physiological data was processed by means of repeated measures analysis of variance. The number of TUNEL positive neurons and the infarct area of the spinal cord were analyzed using a one-way analysis of variance and Dunnett test if there was a significant difference.

2. Experimental results (1) Physiological parameters All animals were alive during the experimental period. Table 1 shows body weight and baseline, rectal temperature and mean arterial pressure 10 minutes after ischemia and 10 minutes after reopening. There was no significant difference in body weight. Rectal temperature and mean arterial pressure were not different at each time point among the 4 groups.

Table 1 Physiological parameters

(2) Neurological evaluation The hindlimb motor function was scored using the Tarlov modified method 24 and 48 hours after surgery. Evaluation of neurological function was performed by an observer who was unaware of the nature of the treatment.
0: No movement, 1: Faint movement, 2: Positioned with assistance, 3: Positioned without assistance, 4: Weak jumping, 5: Normal jumping.
The results are shown in Table 2. The animals in the sham-operated group were normal (Tarlov score: 5) during the observation period. In the control group, severe severe neurological impairment was observed due to aortic occlusion below the kidney for 20 minutes. NS-7 administration (Group A and Group B) showed a recovery of hindlimb motor function 24 and 48 hours after surgery (P <0.01, Group A and Group B vs. control group, both time points). Better Tarlov scores were obtained in animals that received NS-7 at the same time as reopening compared to Group A (Group B), but the difference was not significant.

^※P<0.01 対24時間後の対照群、 ^# P<0.01 対48時間後の対照群
(Mann-Whitney U tests) Table 2 Neurological score at 24 and 48 hours after surgery

^* P <0.01 vs. control group after 24 hours, ^# P <0.01 vs. control group after 48 hours
(Mann-Whitney U tests)

(3) Observation of spinal cord infarct size by TTC staining In order to measure the infarct size of the spinal cord, 2,3,5-triphenyltetrazolinium chloride (TTC) (Sigma Chemical, St. Louis, Missouri, USA) was stained, and the living tissue was examined. Necrotic tissue was stained white with a red brick color. The spinal cord (middle lumbar spinal cord) was quickly removed and 2 mm 2 circular slices were made. Slices were incubated in 1% TTC for 30 minutes at 37 ° C. After fixing with 10% formalin solution, a photograph of the TTC stained slice was taken.
A photograph of TTC staining of the obtained spinal cord (mid lumbar spinal cord) is shown in FIG. In the figure, the living tissue looks red brick and the infarct appears whitish. White matter also appears white. In the sham-operated group (A), no infarction is observed in the gray matter, but in the control group, infarct is observed in almost the entire dorsal horn. Only a minute infarction was observed in the animals (Group A and Group B) administered NS-7 before ischemia (C) and simultaneously with reopening (D).
The area of the infarct was expressed as a percentage of the total gray matter area using National Institute of Health Image Software. The results are shown in FIG. The vertical axis represents the infarct size (%), and from the left are the results of the sham operation group (Sham), the control group (Control), the group A (Group A), and the group B (Group B). * P <0.05 vs. control group.
After 48 hours of spinal cord ischemia, the mean infarct size in the control group was 54.7 ± 19.8%. With NS-7 administration, the infarct size is 14.0 ± 14.3% (P <0.05 vs. control group) when administered before ischemia, and 12.1 ± 12 when administered simultaneously with recanalization. Effectively decreased to 8% (P <0.05).
As described above, a good correlation was observed between the results of TTC staining and the average neurological score after 48 hours.

(4) Histopathological examination The spinal cord was fixed with 10% formalin and embedded in paraffin. Two circular sections (4 μm) were cut out from each segment, stained with hematoxylin and eosin (HE) and photographed. The results are shown in FIG. 3 (mid lumbar spinal cord, 200 ×). From FIG. 3, there is no pathological change in the ventral horn of sham-operated animals (A), and eosin-favorable neurodegeneration (arrow), vacuolation and necrosis are observed in the control group. However, only minute cell damage is observed in animals administered NS-7 before ischemia (C) and simultaneously with reopening (D) (Group A and Group B, respectively).
Researchers who were not informed of the group of animals or neurological findings examined each section.
Histological damage was graded using a semi-quantitative scoring method (Table 3) (Martelli E. J. Vasc. Surg. 2002; 35: 547-53).

グレード化した結果を図４に示す。図中、縦軸は脊髄各部位における病理学的スコアの平均値を表し、左から、下部胸髄（LT）、上部腰髄（UL）、中部腰髄（ML）、下部腰髄（LL）の結果である。＊P<0.05, # P<0.01 対 対照群 (Mann-Whitney U tests.)。 Table 3 Semi-quantitative scoring for pathological assessment of spinal cord invasion

The graded results are shown in FIG. In the figure, the vertical axis represents the mean value of the pathological score in each part of the spinal cord. From the left, the lower thoracic spinal cord (LT), upper lumbar spinal cord (UL), middle lumbar spinal cord (ML), lower lumbar spinal cord (LL) Is the result of * P <0.05, # P <0.01 vs. control group (Mann-Whitney U tests).

  Thus, in the sham-operated group, no signs of spinal cord damage were observed in the HE-stained sections at any part of the spinal cord. In the lower thoracic and upper lumbar areas, the other three groups showed no or very little damage. Although most of the animals in the control group had small histopathological changes, the difference in histopathological scores was not statistically significant. In the middle and lower lumbar spinal cords of the control group, major nerve damage was observed as shown by eosin-favorable neurodegeneration, vacuolation and necrosis (Fig. 3, B). However, in the animals that received NS-7 15 minutes before ischemia (Group A) and the animals that received NS-7 immediately after reopening (Group B), the changes in the spinal cord at these two sites were negligible. Yes, the average histopathological score was significantly lower than the control group (Figure 4).

(5) TUNEL staining Paraffin-embedded sections (middle lumbar spinal cord) using Apop Tag (Intergen Co, New York, New York, USA) according to the manufacturer's recommended method, in situ terminal oxidative transfer (TDT) -mediated dTP -Biotin nick end labeling (TUNEL) staining was performed. In summary, after removing paraffin, the sections were treated with protease and 0.3% hydrogen peroxide. The sections were then incubated for 60 minutes at 37 ° C. in terminal deoxynucleotidyl transfer enzyme. Color was developed with DAB / hydrogen peroxide. Sections were counterstained with methyl green and photographed. The results are shown in FIG. 5 (200 ×). In FIG. 5, A is a sham operation group, B is a control group, C is a group administered NS-7 before ischemia (Group A), and D is a group administered NS-7 simultaneously with reopening (Group B). Represents. Arrows indicate TUNEL positive motor neurons.
Next, the number of TUNEL-stained motor neurons was counted in 3 sections, and the average value was calculated (Sakurai M. J. Thorac. Cardiovasc. Surg. 2000; 120: 1148-57, Lang-Laznski et al. (Lang -Lazdunski L), J. Vasc. Surg. 2003; 38: 564-75). The results are shown in FIG. In FIG. 6, the vertical axis represents the average value of TUNEL positive motoneurons per section. From the left, results of sham operation group (Sham), control group (Control), group A (Group A) and group B (Group B) It is. * P <0.05 vs. control group.

  Cells in which the double strand of DNA is broken, that is, cells that have undergone apoptosis, can be detected by TUNEL staining. Nothing was detected by TUNEL staining in the spinal cord of animals in the sham operation group. In the sample of the control group, many TUNEL positive motor neurons were observed in the ventral gray matter (FIG. 5, B). By administering NS-7 15 minutes before ischemia or simultaneously with reopening, the number of apoptotic cells was greatly reduced compared to the control group (p <0.05, control group, FIG. 6). There was no significant difference in the number of apoptotic cells between the two groups administered NS-7.

  The above experimental results show that the pharmaceutical agent of the present invention has a protective action against both neuronal necrosis and apoptosis during spinal cord ischemia without adverse effects on cardiovascular, etc. It shows that it has an excellent preventive or ameliorating effect. It is also apparent that the medicament of the present invention is effective even if administered at any time point before the blood flow is interrupted and at the same time as the release of the blood block.

  The active ingredient [I] of the medicament of the present invention has a spinal neuroprotective action during spinal cord ischemia. Therefore, the medicament of the present invention can be used for preventing and / or treating nerve cell damage due to spinal cord ischemia.

It is a photograph which shows the typical example of the lumbar spinal cord section which performed TTC dyeing | staining. A is a sham operation group, B is a control group, C is an animal group to which NS-7 was administered before ischemia, and D is an animal group to which NS-7 was administered simultaneously with reopening. Surviving tissue looks red brick and infarct is whitish. White matter also appears white. FIG. 2 is a diagram showing the area of the infarct of the middle lumbar spinal cord based on the photograph of FIG. 1 as a percentage of the total gray matter area using the National Institute of Health image software. *: P <0.05 vs. control group. It is a photograph which shows the result of the HE dyeing | staining of the pathological section (middle lumbar spinal cord, 200x) of a lumbar spinal cord. A is a result of a sham operation group, B is a control group, C is an animal group to which NS-7 was administered before ischemia, and D is a result of an animal group to which NS-7 was administered simultaneously with reopening. Arrows indicate eosin-favorable neurodegeneration. It is a figure which shows the average value of the pathological score in each site | part of a spinal cord. In the figure, LT is the lower thoracic spinal cord; UL is the upper lumbar spinal cord; ML is the middle lumbar spinal cord; LL is the lower lumbar spinal cord. *: P <0.05, #: P <0.01 vs. control group (Mann-Whitney U tests). It is a photograph of a TUNEL stained section (200 ×) of the lumbar spinal cord. A is a result of a sham operation group, B is a control group, C is an animal group to which NS-7 was administered before ischemia, and D is a result of an animal group to which NS-7 was administered simultaneously with reopening. Arrows indicate TUNEL positive motor neurons. It is a figure which shows the spinal cord (middle lumbar spinal cord) TUNEL positive motor neuron number (average of 3 pieces) 48 hours after ischemia. *: P <0.05, vs. control group.

Claims (7)

Formula [I]:

Or a salt thereof, or a solvate thereof as an active ingredient, a medicament for preventing and / or treating neuronal cell damage due to spinal cord ischemia.
In the formula, R ¹ represents an optionally substituted aryl or an aromatic heterocyclic group having 5 to 10 atoms constituting the ring. Here, the aromatic heterocyclic group may be a monocyclic ring or a condensed ring, and contains one or more of nitrogen, oxygen or sulfur as ring constituent atoms. In addition, aryl or aromatic heterocyclic groups are hydroxy, halogen, alkyl, haloalkyl, hydroxyalkyl, aralkyl, alkenyl, alkoxy, haloalkyloxy, alkylthio, cycloalkyl, cycloalkylalkyl, cycloalkyloxy, alkylsulfonyl, sulfamoyl, alkanoyl , Amino, monoalkylamino, dialkylamino, carboxy, alkoxycarbonyl, cyano and nitro may be substituted by 1 to 3 identical or different substituents.
R ² represents hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkylalkyl, hydroxyalkyl, haloalkyl, alkoxy, alkylthio, amino, monoalkylamino, dialkylamino or optionally substituted phenyl. Such phenyl may be substituted by the same or different 1 to 3 substituents selected from the group consisting of halogen, alkyl and alkoxy.
R ³ and R ⁴ are the same or different and are hydrogen or optionally substituted alkyl (the alkyl is the same or different 1 selected from the group consisting of hydroxy, alkoxy, amino, monoalkylamino and dialkylamino) R ³ and R ⁴ together with the adjacent N represent NR ³ R ⁴ representing a 4- to 8-membered cyclic amino. Such cyclic amino may have nitrogen, oxygen or sulfur as a ring constituent atom in addition to the nitrogen, and may further be alkyl, alkoxy, hydroxy, oxo, amino, monoalkylamino, dialkylamino, or substituted. It may be substituted by 1 to 3 identical or different substituents selected from the group consisting of good aryl and pyridyl.
Furthermore, N to which R ³ and R ⁴ are bonded may form an oxide.
A represents an alkylene having 2 to 10 carbon atoms. Such alkylene may be substituted at any position with a substituent selected from the group consisting of alkoxy, hydroxy and oxo.
E represents O or S.
W represents a single bond, O, S, or (CH ₂ ) n (CH ₂ may be substituted with alkyl. N represents 1 or 2).
X, Y and Z are the same or different and represent CH, CR (R represents alkyl) or N. However, the case where X, Y, and Z are simultaneously CH or CR is excluded.
Ring G represents pyridine, pyrimidine, 1,3,5-triazine.
When one to three of X, Y, and Z are N, one of them may form an oxide. R ¹ is halogen-substituted phenyl, R ² is alkyl or haloalkyl, and R ³ and R ⁴ together with the adjacent N are one nitrogen atom as a ring-forming hetero atom represented by —NR ³ R ⁴ 4-membered cyclic amino containing only A, alkylene having 3 to 6 carbon atoms, E being O or S, W being a single bond, X and Z being N, Y being CH The medicament according to claim 1, wherein Z is N and X and Y are CH. NR ³ R ⁴ is piperidino, A is alkylene having 4 to 6 carbon atoms, E is O, W is a single bond, X and Z are N and Y is CH, or Z is N The medicament according to claim 1, wherein X and Y are CH. The active ingredient is 4- (4-fluorophenyl) -2-methyl-6- (4-piperidinobutoxy) pyrimidine,
4- (4-Fluorophenyl) -2-methyl-6- (5-piperidinopentyloxy) pyrimidine, 4- (4-fluorophenyl) -2-methyl-6- (6-piperidinohexyloxy) Pyrimidine,
4- (4-fluorophenyl) -2-methyl-6- (1-methyl-4-piperidinobutoxy) pyrimidine,
2- (4-fluorophenyl) -4-methyl-6- (4-piperidinobutoxy) pyrimidine,
4- (4-Fluorophenyl) -2-methyl-6- (3-piperidinopropoxy) pyridine and 4- (4-fluorophenyl) -2-methyl-6- (5-piperidinopentyloxy) pyridine The pharmaceutical in any one of Claims 1-3 containing the compound or its salt selected from the group which consists of, or its solvate.   An agent for protecting spinal cord nerve cells during spinal cord ischemia, comprising the compound according to any one of claims 1 to 4 or a salt thereof, or a solvate thereof as an active ingredient.   An amount of the compound represented by the formula [I], a salt thereof or a solvate thereof according to any one of claims 1 to 4, in an amount effective for preventing and / or treating nerve cell damage due to spinal cord ischemia. A method for preventing and / or treating nerve cell damage due to spinal cord ischemia, comprising the step of administering to a mammal, including a human being in need thereof. The compound represented by the formula [I] according to any one of claims 1 to 4, or a salt thereof, or a solvation thereof, in the manufacture of a medicament for preventing and / or treating nerve cell damage due to spinal cord ischemia Use of things.

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