JP2001194369A - Screening method of medicine acting on central nervous system, and method for evaluating status under stress - Google Patents
Screening method of medicine acting on central nervous system, and method for evaluating status under stressInfo
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- JP2001194369A JP2001194369A JP2000325071A JP2000325071A JP2001194369A JP 2001194369 A JP2001194369 A JP 2001194369A JP 2000325071 A JP2000325071 A JP 2000325071A JP 2000325071 A JP2000325071 A JP 2000325071A JP 2001194369 A JP2001194369 A JP 2001194369A
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Abstract
Description
【０００１】[0001]
【発明の属する技術分野】本発明は、中枢神経系作用薬
をスクリーニングするための方法及び免疫系に異常のあ
る哺乳動物のストレス状態を評価する方法に関する。The present invention relates to a method for screening a drug acting on the central nervous system and a method for evaluating a stress state of a mammal having an abnormal immune system.
【０００２】[0002]
【従来の技術】従来、動物を用いて中枢神経系に作用す
る化合物をスクリーニングとする方法としては、例え
ば、抗てんかん薬評価のための最大電撃けいれん（ＭＥ
Ｓ）法及びしきい値法（ＰＴＺ法）、抗うつ薬評価のた
めの強制水泳試験、抗不安薬評価のための抗コンフリト
試験、脳機能改善薬評価のための抗ハイポキシア試験、
抗痴呆薬評価のための電気痙攣ショック誘発健忘モデル
及びシクロヘキシミド誘発健忘モデルを用いる方法が挙
げられ、正常動物又は病態モデル動物に薬物を投与する
ことにより各種の中枢神経薬がスクリーニングされてい
る。2. Description of the Related Art Conventionally, as a method for screening a compound acting on the central nervous system using an animal, for example, a maximum electric shock seizure (ME) for evaluating an antiepileptic drug is used.
S) method and threshold method (PTZ method), forced swimming test for antidepressant evaluation, anti-conflit test for anti-anxiety drug evaluation, anti-hypoxia test for evaluation of brain function improving drugs,
Examples include a method using an electroconvulsive shock-induced amnesia model and a cycloheximide-induced amnesia model for evaluating anti-dementia drugs. Various central nervous drugs have been screened by administering the drug to normal animals or diseased model animals.
【０００３】斯かる中枢神経系作用薬をスクリーニング
するための評価系は、殆どが行動観察によるものであ
り、ダブルブラインドを組み合わせて評価しなければな
らないことが多く、また、いずれも客観的に薬物を評価
できるものではなかった。また、上記のように薬理作用
に応じた中枢神経系作用薬のスクリーニング方法は多数
存在するものの、薬理作用に関係なく中枢神経系作用薬
を広くスクリーニングする方法についてはこれまでに知
られていない。一方、ストレス関連生体内物質として、
カテコールアミン類およびそれらの代謝物、ステロイド
ホルモン類等が知られており、これらの濃度を測定し、
ストレス状態の指標とすることができる。しかし、それ
らは共存する測定妨害物質の影響を受けやすいため、必
ずしも満足できるものではない。Most of such evaluation systems for screening for drugs acting on the central nervous system are based on behavioral observation, and often must be evaluated in combination with double blinds. Could not be evaluated. As described above, although there are many screening methods for drugs acting on the central nervous system depending on the pharmacological action, there is no known method for widely screening drugs acting on the central nervous system irrespective of the pharmacological action. On the other hand, as a stress-related biological substance,
Catecholamines and their metabolites, steroid hormones, etc. are known, and their concentrations are measured,
It can be used as an indicator of a stress state. However, they are not always satisfactory because they are susceptible to coexisting measurement interfering substances.
【０００４】[0004]
【発明が解決しようとする課題】本発明は、中枢神経に
作用する薬物を広く且つ簡易にスクリーニングするため
の方法及び免疫系に異常のある哺乳動物のストレス状態
を評価する新たな指標を提供することを目的とする。SUMMARY OF THE INVENTION The present invention provides a method for widely and easily screening for a drug acting on the central nervous system and a new index for evaluating the stress state of a mammal having an abnormal immune system. The purpose is to:
【０００５】[0005]
【課題を解決するための手段】本発明者は，斯かる状況
の下、血中インターロイキン−６（以下、「ＩＬ−６」
という）の濃度と中枢神経作用の相関関係について鋭意
検討した結果、免疫系に異常のあるモデル動物におい
て、ある薬物投与後短時間の間に炎症等の免疫異常に基
づく症状変化とは無関係に血中ＩＬ−６濃度が変化し、
これを指標とすることにより、中枢神経系作用薬を簡易
にスクリーニングできること及び血中ＩＬ−６濃度の変
化がストレス状態の指標となることを見出し、本発明を
完成するに至った。Under such circumstances, the present inventor has proposed that blood interleukin-6 (hereinafter referred to as "IL-6")
As a result of intensive studies on the correlation between the concentration of) and central nervous system activity, in model animals with abnormal immune system Medium IL-6 concentration changes,
By using this as an index, they have found that a drug acting on the central nervous system can be easily screened, and that a change in blood IL-6 concentration can be an index of a stress state, thereby completing the present invention.
【０００６】即ち、本発明は、免疫系に異常のある哺乳
動物に対して被験化合物を投与し、該動物の血中ＩＬ−
６濃度を測定することを特徴とする中枢神経系作用薬の
スクリーニング方法及び免疫系に異常のある哺乳動物の
血中ＩＬ−６濃度を測定することにより免疫系に異常の
ある哺乳動物のストレス状態を評価する方法を提供する
ものである。That is, the present invention relates to a method of administering a test compound to a mammal having an abnormal immune system, and administering the test compound to the blood IL-
6. A screening method for a central nervous system acting drug, comprising measuring the concentration of IL-6, and the stress state of a mammal having an abnormality in the immune system by measuring the blood IL-6 concentration in the mammal having an abnormality in the immune system Is provided.
【０００７】[0007]
【発明の実施の形態】本発明のスクリーニング方法及び
ストレス状態の評価方法は、免疫系に異常のあるモデル
動物を用いて、血中ＩＬ−６濃度を測定することを特徴
とするものである。BEST MODE FOR CARRYING OUT THE INVENTION The screening method and the stress state evaluation method of the present invention are characterized by measuring blood IL-6 concentration using a model animal having an abnormal immune system.
【０００８】ＩＬ−６は、単球、Ｔ細胞、Ｂ細胞、血管
内皮細胞、線維芽細胞、骨芽細胞等種々の細胞から産生
され、Ｂ細胞の抗体産生細胞への分化誘導、肝細胞から
の急性期蛋白質合成誘導、脳神経系細胞の分化誘導、造
血系細胞の増殖分化誘導、破骨細胞の分化誘導等の多様
な生理活性作用を有するサイトカインであるが（実験医
学、７巻、１号、１９８９年）、近年、ＩＬ−６が急性
相蛋白を誘導することが報告され炎症マーカーとして有
用であることが示唆され、更に開心術症例における検討
から、炎症の初期に血中ＩＬ−６が上昇し、ＩＬ−６を
炎症マ−カ−とすることにより初期の炎症を感度よく把
握できる可能性が示唆されている（臨床病理、４２巻、
１１号、１１５１−１１５７頁、１９９４年）。また、
最近では慢性関節リウマチ患者において、笑うことによ
り、血中ＩＬ−６濃度が低下することが報告されている
（ジャ−ナル・オブ・リウマトロジ−（Journal of Rhe
umatology）、２３巻、４号、793−794頁、1996年）。[0008] IL-6 is produced from various cells such as monocytes, T cells, B cells, vascular endothelial cells, fibroblasts, and osteoblasts, and induces differentiation of B cells into antibody-producing cells and hepatocytes. Are various physiologically active cytokines, such as induction of protein synthesis in the acute phase, induction of differentiation of brain nervous cells, induction of proliferation and differentiation of hematopoietic cells, and induction of osteoclast differentiation (Experimental Medicine, Vol. 7, No. 1, In recent years, it has been reported that IL-6 induces an acute phase protein, and it is suggested that IL-6 is useful as an inflammatory marker. In addition, studies in open heart surgery cases show that IL-6 in blood can be expressed in the early stage of inflammation. It has been suggested that the initial inflammation may be detected with high sensitivity by using IL-6 as an inflammatory marker (Clinical Pathology, Vol. 42,
11, 1151-1157, 1994). Also,
Recently, it has been reported that laughing lowers blood IL-6 levels in rheumatoid arthritis patients (Journal of Rheumatology).
umatology), Vol. 23, No. 4, pp. 793-794, 1996).
【０００９】しかし、免疫系に異常のある哺乳動物にお
いて、ＩＬ−６と中枢作用全般について詳細に検討され
た報告はこれまでになく、該哺乳動物の血中ＩＬ−６濃
度が、催眠作用、解熱作用、鎮痛作用、脳機能改善作用
等の作用を有する中枢神経系作用薬によって著しく減少
することは全く予期することができなかった事実であ
る。[0009] However, in a mammal in abnormal immune system than ever reports which are discussed in detail IL-6 and central action in general, the blood IL-6 concentration in the mammalian, hypnotic, It is a fact that the central nervous system acting drug having an action such as antipyretic action, analgesic action, cerebral function improving action, etc. could not be expected to significantly decrease.
【００１０】本スクリーニング方法においては、免疫系
に異常のある哺乳動物が使用される。ここで、「免疫系
に異常のある」とは、炎症等によって免疫系が亢進した
状態をいい、自己免疫疾患を発症した状態が挙げられ
る。自己免疫疾患としては、慢性関節リウマチ等の慢性
炎症疾患が挙げられ、本スクリーニング方法においては
特に関節炎が好ましい。また、当該自己免疫疾患は、特
定の手段により誘発させたものでも、自然発症型の自己
免疫疾患であってもよい。In the present screening method, a mammal having an abnormal immune system is used. Here, “there is an abnormality in the immune system” refers to a state in which the immune system is enhanced by inflammation or the like, and includes a state in which an autoimmune disease has developed. Autoimmune diseases include chronic inflammatory diseases such as rheumatoid arthritis, and in the present screening method, arthritis is particularly preferred. The autoimmune disease may be one induced by a specific means or a spontaneous autoimmune disease.
【００１１】自己免疫疾患の誘導手段としては、アジュ
バントあるいは化学物質を接種する方法、トランスジェ
ニック動物の使用、移植及び遺伝子ノックアウト等が挙
げられる。アジュバントを接種して免疫異常を誘発し、
関節炎を発症させる方法としては、例えば、結核死菌を
流動パラフィンに懸濁し、ラットの尾根部皮内に接種す
る方法（アルツナイミッテル−フォ−シュング／ドラッ
グリサ−チ（Arzneim.-Forsch./Drug Res.）、42巻、2
号、935-944頁、1992年）等が挙げられる。また、自然
発症型の免疫異常を有する動物としては、例えば、MRL
／lprマウス等が挙げられる。Means for inducing an autoimmune disease include a method of inoculating an adjuvant or a chemical substance, use of a transgenic animal, transplantation, and gene knockout. Inoculating an adjuvant to induce immune abnormalities,
As a method of causing arthritis, for example, a method of suspending killed tuberculosis in liquid paraffin and inoculating the suspension into the skin of the ridge of a rat (Arzneim.-Forschung / Drug Research (Arzneim.-Forsch./ Drug Res.), Volume 42, 2
No. pp. 935-944, 1992). In addition, animals having spontaneously occurring immune abnormalities include, for example, MRL
/ Lpr mouse and the like.
【００１２】免疫系に異常のある哺乳動物に更にストレ
ス負荷を与えることにより、血中ＩＬ−６濃度を有効に
上げることができる。斯かるストレス負荷としては、心
理的ストレス及び／または物理的ストレス等の如何なる
方法を用いてもよいが、例えば、強制水泳、強い光や音
の照射、拘束、水浸拘束、過密飼育、電気ショック、低
温及び／または高温等が挙げられる。By further applying a stress load to a mammal having an abnormal immune system, the blood IL-6 concentration can be effectively increased. As such a stress load, any method such as psychological stress and / or physical stress may be used. For example, forced swimming, irradiation of strong light or sound, restraint, water immersion restraint, overcrowding, electric shock, , Low temperature and / or high temperature.
【００１３】上記哺乳動物としては、ヒト以外の哺乳動
物が挙げられ、好ましくは、マウス及びラット等のげっ
歯類が挙げられる。本発明スクリーニング方法において
用いられる好ましいモデル動物としては、関節炎を発症
したヒト以外の哺乳動物、特にマウス及びラット等のげ
っ歯類が挙げられ、更にそれらがストレス負荷された状
態である場合が好ましい。The above mammals include mammals other than humans, and preferably include rodents such as mice and rats. Preferable model animals used in the screening method of the present invention include mammals other than humans that have developed arthritis, particularly rodents such as mice and rats, and more preferably, they are under stress.
【００１４】本発明スクリーニング方法で評価される中
枢系作用薬としては、例えば、麻酔作用、催眠作用、鎮
静作用、解熱作用、鎮痛作用、抗抑欝作用、脳機能改善
作用又は抗（向）精神神経作用等を有する化合物のスク
リーニングに適している。The drugs acting on the central system evaluated by the screening method of the present invention include, for example, an anesthetic action, a hypnotic action, a sedative action, an antipyretic action, an analgesic action, an antidepressant action, a cerebral function improving action or an anti (psychotropic) mind It is suitable for screening a compound having a nerve effect or the like.
【００１５】また、血中のＩＬ−６濃度を測定するに
は、例えば、採取した血液あるいは血液から分離した血
漿又は血清を使用し、全血中、血漿中又は血清中のＩＬ
−６濃度をＲＩＡ法、ＥＩＡ法、ＥＬＩＳＡ法、バイオ
アッセイ法等通常用いられる方法により測定すればよ
い。In order to measure the concentration of IL-6 in blood, for example, collected blood or plasma or serum separated from blood is used, and IL-6 in whole blood, plasma or serum is used.
The -6 concentration may be measured by a commonly used method such as an RIA method, an EIA method, an ELISA method, and a bioassay method.
【００１６】本発明スクリーニング方法は、具体的に
は、（１）免疫系に異常のある哺乳動物に無毒性量の化
合物を経口又は非経口に投与し、（２）ストレスが負荷
されない又は負荷された一定時間後の血液を採取し、
（３）血液中のＩＬ−６濃度を測定することにより実施
できる。特に、炎症を惹起された哺乳動物に被験化合物
を投与し、ストレス負荷により血中ＩＬ−６の誘導が見
られるストレス負荷の至適時間後に血液を採取し、血中
ＩＬ−６濃度を測定することが好ましい。尚、血中のＩ
Ｌ−６濃度の比較は、同一群を用いてもよいし、化合物
投与群及び非投与群で比較してもよい。この様に測定さ
れた血中ＩＬ−６の濃度を被験化合物の投与前後で比較
し、その濃度が低下した場合に、被験化合物が中枢神経
系作用薬であると判断することができる。Specifically, the screening method of the present invention comprises the steps of (1) administering a non-toxic amount of a compound orally or parenterally to a mammal having an abnormal immune system, and (2) applying no or no stress. After a certain period of time,
(3) It can be carried out by measuring the concentration of IL-6 in blood. In particular, a test compound is administered to a mammal in which inflammation has been induced, and blood is collected after the optimal time of the stress load in which induction of the blood IL-6 is observed by the stress load, and the blood IL-6 concentration is measured. Is preferred. In addition, I in blood
The L-6 concentration may be compared in the same group or in the compound administration group and the non-administration group. The blood IL-6 concentration thus measured is compared before and after administration of the test compound, and when the concentration decreases, it can be determined that the test compound is a drug acting on the central nervous system.
【００１７】一方、免疫系に異常のある哺乳動物にスト
レス負荷を与えることにより、血中ＩＬ−６濃度が上昇
することから、免疫系に異常のある哺乳動物の血中ＩＬ
−６濃度を測定することにより、免疫系に異常のある哺
乳動物のストレス状態を測定することができる。On the other hand, when a stress load is applied to a mammal having an abnormal immune system, the blood IL-6 concentration is increased.
By measuring the -6 concentration, the stress state of a mammal having an abnormal immune system can be measured.
【００１８】免疫系に異常のある哺乳動物のストレス状
態を評価するための血中ＩＬ−６濃度の測定法は、上記
した本発明のスクリーニング方法と同様の方法を使用す
ればよい。すなわち、採取した血液あるいは血液から分
離した血漿又は血清を使用し、全血中、血漿中又は血清
中のＩＬ−６濃度をＲＩＡ法、ＥＩＡ法、ＥＬＩＳＡ
法、バイオアッセイ法等通常用いられる方法により測定
すればよい。The method for measuring the blood IL-6 concentration for evaluating the stress state of a mammal having an abnormal immune system may be the same as the above-described screening method of the present invention. That is, using the collected blood or plasma or serum separated from the blood, the IL-6 concentration in whole blood, plasma or serum was determined by the RIA method, EIA method, or ELISA.
It may be measured by a commonly used method such as a method and a bioassay method.
【００１９】ストレス状態の評価は、具体的には、
（１）免疫系に異常のある哺乳動物に無毒性量の中枢神
経系作用薬を経口又は非経口に投与し、（２）一定時間
後の血液を採取し、（３）血液中のＩＬ−６濃度を測定
する、（４）測定されたＩＬ−６の濃度を化合物の投与
前後で比較することにより実施できる。特に、炎症を惹
起された哺乳動物に催眠作用、解熱作用、鎮痛作用又は
脳機能改善作用を有する化合物、とりわけ、催眠作用又
は脳機能改善作用を有する化合物を投与し、至適時間後
に血液を採取し、血中ＩＬ−６濃度を測定し、比較する
ことが好ましい。The evaluation of the stress state is, specifically,
(1) a non-toxic amount of a central nervous system agonist is orally or parenterally administered to a mammal having an abnormal immune system, (2) blood is collected after a certain period of time, and (3) IL- (4) comparing the measured IL-6 concentration before and after administration of the compound. In particular, a compound having a hypnotic, antipyretic, analgesic or cerebral function improving action, particularly a compound having a hypnotic or cerebral function improving action, is administered to a mammal in which inflammation has been induced, and blood is collected after an optimal time. Then, it is preferable to measure and compare the blood IL-6 concentration.
【００２０】[0020]
【実施例】以下、実施例により本発明を具体的に説明す
る。The present invention will be described below in detail with reference to examples.
【００２１】実施例１ 結核死菌（M.tuberculosis H37Ra、ディフコ社製）を流
動パラフィン（メルク社製）に懸濁し(6 mg/mL）、ラッ
ト(ルイス系、6-7週齢、雄)の尾根部皮内に0.1mL接種し
た［アルツナイミッテル−フォーシュング／ドラッグリ
サーチ（Arzneim.-Forsch./Drug Res.,）42巻、2号、93
5-944頁、1992年］。18日後に、プレチスモメーター ウ
ゴ・バシル7150（PLETHYSMO METER Ugo Basile 7150）
で測定した足蹠容積が均等になるように一群5又は6匹に
群分けした。ストレス負荷群は、東大薬作型ストレスケ
ージに入れ、23℃の水槽内に胸部まで浸して、ストレス
負荷した［薬物活性の前臨床的評価法、96-97頁、藤村・
福田編、薬事日報社］。30分後、エーテル麻酔下、採血
し、血清分離剤入スピッツ（セパクリーン、栄研器材）
により血清を分離した。各血清中のＩＬ−６濃度をMH60
・BSF2細胞［ヨーロピアン・ジャーナル・オブ・イムノ
ロジー (Eur. J. Immuol.）、18巻、951-956頁、1988
年]による増殖活性により測定し、既知量のＩＬ−６の
増殖活性をもとに血清中のＩＬ−６量を算出した。尚、
血清中ＩＬ−６は、ストレスを負荷した関節炎ラットの
ＩＬ−６濃度（units/ml）を100％として相対的（T/C：
％）に示した。また、ELISAキットを用いて、血清中の
コルチゾール濃度を測定した。結果を表１に示す。Example 1 M. tuberculosis H37Ra (manufactured by Difco) was suspended in liquid paraffin (manufactured by Merck) (6 mg / mL) and rat (Lewis strain, 6-7 weeks old, male) 0.1 mL was inoculated intradermally into the ridge of the ridge [Arzneim.-Forsch./Drug Res., 42, No. 2, 93
5-944, 1992]. 18 days later, plethysmometer Ugo Basile 7150
The animals were divided into groups of 5 or 6 animals so that the footpad volumes measured in 1) were equal. The stress-loaded group was placed in a medicinal-type stress cage at the University of Tokyo, and immersed in the aquarium at 23 ° C to the chest, and subjected to stress [Preclinical evaluation method of drug activity, pp. 96-97, Fujimura
Fukuda, Pharmaceutical Daily.] 30 minutes later, under anesthesia with ether, blood is collected and spitz containing serum separating agent (Sepaclean, Eiken Instruments)
To separate the serum. The concentration of IL-6 in each serum was adjusted to MH60
BSF2 cells [European Journal of Immunology (Eur. J. Immuol.), 18, 951-956, 1988
The amount of IL-6 in serum was calculated based on the proliferation activity of a known amount of IL-6. still,
Serum IL-6 was relative (T / C: 100%) with the IL-6 concentration (units / ml) of stressed arthritic rats as 100%.
%)Pointing out toungue. The serum cortisol concentration was measured using an ELISA kit. Table 1 shows the results.
【００２２】[0022]
【表１】 [Table 1]
【００２３】上記の結果は、免疫異常状態を起こしたア
ジュバント関節炎ラットに水浸拘束ストレスを負荷する
ことにより血清中ＩＬ−６の上昇と同時にストレスマー
カーとして知られている血清中コルチゾールが上昇して
いることを示している。すなわち、この結果は、免疫系
に異常のある哺乳動物において、血中ＩＬ−６濃度は、
ストレス状態の指標となりうることを示している。[0023] The above results indicate that by applying water immersion restraint stress to an adjuvant arthritis rat having an abnormal immune state, serum IL-6 increases simultaneously with an increase in serum cortisol known as a stress marker. It indicates that That is, this result indicates that in a mammal having an abnormal immune system, the blood IL-6 concentration is
It indicates that it can be an indicator of a stress state.
【００２４】実施例２ 結核死菌（M.tuberculosis H37Ra、ディフコ社製）を流
動パラフィン（メルク社製）に懸濁し(6mg/mL）、ラッ
ト(ルイス系、6−7週齢、雄)の尾根部皮内に0.1mL接種
した［アルツナイミッテル−フォーシュング／ドラッグ
リサーチ（Arzneim.-Forsch./Drug Res.,）42巻、2号、
935-944頁、1992年］。18日後に、プレチスモメータ
ー、ウゴ・バシル7150（PLETHYSMO METER、Ugo Basile
7150）で測定した足蹠容積が均等になるように一群6又
は7匹に群分けした。各被験化合物を投与し、60分後に
東大薬作型ストレスケージに入れ、23℃の水槽内に胸部
まで浸して、ストレス負荷した［薬物活性の前臨床的評
価法、96-97頁、藤村・福田編、薬事日報社］。30分
後、エーテル麻酔下、採血し、血清分離剤入スピッツ
（セパクリーン、栄研器材）により血清を分離した。各
血清中のＩＬ−６濃度をMH60・BSF2細胞[ヨーロピアン
・ジャーナル・オブ・イムノロジー(Eur. J. Immuo
l.）、18巻、951-956頁、1988年］による増殖活性によ
り測定し、既知量のＩＬ−６の増殖活性をもとに血清中
のＩＬ−６量を算出した。尚、血清中ＩＬ−６は、スト
レスを負荷した関節炎ラットのＩＬ−６濃度（units/m
l）を100％として相対的（T/C：％）に示した。被験化
合物としては、催眠作用を有するペントバルビタールナ
トリウム（PE）、シナプス伝達効率改善作用、脳内アセ
チルコリン系神経調節作用及び脳エネルギ−代謝改善作
用を有するアニラセタム（AN）を用い、ペントバルビタ
ールナトリウムは、ストレス負荷直前に投与した。結果
を表２に示す。Example 2 M. tuberculosis H37Ra (manufactured by Difco) was suspended in liquid paraffin (manufactured by Merck) (6 mg / mL), and rat (Lewis strain, 6-7 weeks old, male) was suspended in liquid paraffin. 0.1 mL was inoculated in the ridge skin [Arzneim.-Forsch./Drug Res., 42, No. 2,
935-944, 1992]. 18 days later, plethysmometer, Ugo Basil 7150 (PLETHYSMO METER, Ugo Basile
The groups were divided into groups of 6 or 7 animals so that the footpad volumes measured in 7150) were equal. Each test compound was administered, 60 minutes later, placed in a medicinal stress-type stress cage at the University of Tokyo, and immersed in a 23 ° C. water tank up to the chest, and subjected to stress [Preclinical evaluation method of drug activity, pp. 96-97, Fujimura Fukuda, Pharmaceutical Daily.] Thirty minutes later, blood was collected under ether anesthesia, and serum was separated using Spitz (Sepaclean, Eiken Instruments) containing a serum separating agent. The concentration of IL-6 in each serum was determined using MH60 / BSF2 cells [European Journal of Immunology (Eur. J. Immuo
l.), 18, 951-956, 1988], and the amount of IL-6 in serum was calculated based on the proliferative activity of a known amount of IL-6. The serum IL-6 was measured at the IL-6 concentration (units / m 2) of the stressed arthritic rat.
l) is shown relative (T / C:%) as 100%. As test compounds, pentobarbital sodium (PE) having a hypnotic effect, aniracetam (AN) having a synaptic transmission efficiency improving effect, cerebral acetylcholine neuromodulating effect and brain energy-metabolism improving effect were used. It was administered immediately before stress loading. Table 2 shows the results.
【００２５】[0025]
【表２】 [Table 2]
【００２６】免疫異常状態を起こしたアジュバント関節
炎ラットでは、ストレス無負荷の群でも血清中ＩＬ−６
は検出されたが、ストレス負荷ににより血清中ＩＬ−６
濃度は更に上昇した。これに対し、ペントバルビタール
ナトリウム及びアニラセタムを投与した群では、血清中
ＩＬ−６の低下が見られた。In the adjuvant arthritis rats having an abnormal immune state, serum IL-6 levels were observed even in the unstressed group.
Was detected, but IL-6 in serum was
Concentrations further increased. In contrast, in the group to which pentobarbital sodium and aniracetam were administered, a decrease in serum IL-6 was observed.
【００２７】[0027]
【発明の効果】本発明のスクリーニング方法を用いれ
ば、中枢神経系に作用する薬物を広く且つ簡易にスクリ
ーニングすることができ、新規な中枢神経系作用薬の開
発及び副作用として中枢作用を有する薬物のスクリーニ
ングが可能となる。また、本発明の評価方法は、免疫系
に異常のある哺乳動物のストレス状態を見るための有用
な手段の一つとなる。According to the screening method of the present invention, a drug acting on the central nervous system can be screened widely and easily, and a new drug acting on the central nervous system can be developed and a drug having a central action as a side effect can be obtained. Screening becomes possible. Further, the evaluation method of the present invention is one of useful means for monitoring the stress state of a mammal having an abnormal immune system.
フロントページの続き (51)Int.Cl.7 識別記号 ＦＩ テーマコート゛(参考） Ａ６１Ｐ 25/04 Ａ６１Ｐ 25/04 25/20 25/20 25/28 25/28 Continued on the front page (51) Int.Cl. 7 Identification symbol FI Theme coat II (Reference) A61P 25/04 A61P 25/04 25/20 25/20 25/28 25/28
Claims (5)
験化合物を投与し、該動物の血中インターロイキン−６
濃度を測定することを特徴とする中枢神経系作用薬のス
クリーニング方法。1. A test compound is administered to a mammal having an abnormality in the immune system, and interleukin-6 in the blood of the animal is administered.
A method for screening a drug acting on the central nervous system, comprising measuring the concentration.
用、鎮痛作用又は脳機能改善作用を有するものである請
求項１記載のスクリーニング方法。2. The screening method according to claim 1, wherein the drug acting on the central nervous system has a hypnotic, antipyretic, analgesic or cerebral function improving effect.
のである請求項１または２記載のスクリーニング方法。3. The screening method according to claim 1, wherein the mammal is further subjected to stress.
３のいずれか１項記載のスクリーニング方法。4. The mammal according to claim 1, wherein the mammal is a rodent.
4. The screening method according to claim 3.
ることを特徴とする免疫系に異常のある哺乳動物のスト
レス状態の評価方法。5. A method for evaluating a stress state of a mammal having an abnormal immune system, comprising measuring a blood interleukin-6 concentration.
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| JP2015517813A (en) * | 2012-05-28 | 2015-06-25 | ザ・ロイヤル・インスティテューション・フォア・ザ・アドバンスメント・オブ・ラーニング／マクギル・ユニヴァーシティ | Polypeptides that allow inflammation and uses thereof |
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