JP2010111581A - Medicine containing dopamine d2-like receptor agonist as active ingredient and screening method - Google Patents

Abstract

Disclosed is an effective drug for a disease (for example, multiple sclerosis, etc.) caused by an excessive reaction of helper T cells with Th2 or Th17, and an efficient screening method for the drug.
A drug for treating or preventing a disease caused by an excessive reaction of Th2 or Th17, which contains a dopamine D2-like receptor agonist as an active ingredient, and binding to a dopamine D2-like receptor and / or dopamine D2-like A screening method for the above-mentioned pharmaceuticals, which uses an activity action on a receptor as an index. As the pharmaceutical, at least one of pramipexole hydrochloride hydrate and ropinirole hydrochloride is particularly preferable.
[Selection figure] None

Description

  The present invention relates to a drug for a disease caused by an excessive reaction of Th2 or Th17 such as multiple sclerosis, and a screening method for the drug.

  Helper T cells that play a central role in acquired immunity are Th1 (type 1 helper T cell) that promotes cellular immunity and Th2 (type 2 helper T cell) that promotes humoral immunity due to differences in cytokines produced. And classified. Dendritic cells (DC) are antigen-presenting cells that play an important role in the early stage of immune response. Recently, functional differences such as promoting Th1 induction (DC1) or Th2 induction (DC2). It became clear that there was a subset with. Several attempts have been made to correct the biased Th1 / Th2 balance by artificially controlling the induction of Th1 and Th2 from naive T cells via DC using this, and reported successful cases. (Non-Patent Document 1).

  Conventionally, abnormalities in Th1 / Th2 balance are considered to be involved in the development of various immune-related diseases. For example, the balance bias toward Th1 cells is caused by rheumatoid arthritis, which is a chronic inflammatory disease, and organ-specific autoimmune diseases (eg, multiple sclerosis, type 1 diabetes, inflammatory bowel disease, glomerulonephritis, hepatitis, liver Disorders, autoimmune hemolytic anemia, leukopenia, thrombocytopenia, demyelinating disease, Hashimoto's thyroiditis, pernicious anemia, psoriasis), and balance bias to Th2 cells It is thought to be involved in sex diseases and many systemic autoimmune diseases. On the other hand, in recent years, Th17, a new Th sub-fraction, has been reported by a plurality of papers. These cells are involved in exacerbating autoimmune inflammation by producing IL-17 exclusively. In particular, the above-mentioned multiple sclerosis is considered to have a strong suspicion due to this bias toward Th17 (Non-patent Document 2). Th17 acts in a mutually inhibitory manner with Th1 and in a mutually potentiating manner with Th2.

  Many methods for adjusting the abnormal Th1 / Th2 balance as desired have been proposed as a method for effectively treating or preventing various immune-related diseases as described above. Specifically, for example, for example, TCCR (T cell cytokine receptor) Method for treating Th1-mediated disease by administering a polypeptide antagonist (Patent Document 1); Excess Th1 cell-mediated immune response caused by autoimmune disease such as multiple sclerosis by using xanthophyll A method of suppressing (Patent Document 2); a pharmaceutical composition containing a specific benzhydryl derivative for treating or preventing an organ-specific autoimmune disease or the like (Patent Document 3);

  However, various factors are involved in the acquired immune system in a complicated manner, and it is still beyond the scope of speculation about which immune-related disease is involved in Th1 bias, Th2 bias, or Th17 bias. it is conceivable that. Therefore, at present, it is still desired to obtain more accurate knowledge about the relationship between Th1 / Th2 / Th17 bias and various diseases, and to develop more effective medicines for various diseases. .

Special table 2003-512824 gazette Special table 2003-510353 gazette JP 2003-3000881 A Morita Y et al .; Dendritic cells genetically engineered to express IL-4 inhibitory collagen collagen-induced arthritis. J Clin Invest. 2001 May; 107 (10): 1275-84. Battern, M et al .; Interleukin 27 limit autoimmune encephalomyloidis by pressing the development of interleukin 17-production T cells. Nat Immunol. 2006. 7: 929-936.

  An object of the present invention is to solve the conventional problems and achieve the following objects. That is, an object of the present invention is to provide an effective medicine for diseases caused by excessive reaction of Th2 or Th17 (for example, multiple sclerosis etc.) and an efficient screening method for the medicine.

  In recent years, the present inventors have found that a disease such as multiple sclerosis, which has been conventionally considered to be caused by an excessive reaction of Th1, is actually a disease caused by an excessive reaction of Th2 or Th17. It has been found that the disease such as the multiple sclerosis can be effectively treated or prevented by positively inducing (see Japanese Patent Application No. 2006-211881). Therefore, as a result of further intensive studies related to this, the present inventors have found that the use of a dopamine D2-like receptor agonist such as pramipexole hydrochloride hydrate, ropinirole hydrochloride and the like, as described above, Th2 Alternatively, the inventors have newly found that an excellent clinical effect can be exerted on diseases caused by excessive reaction of Th17 (for example, multiple sclerosis etc.), and the present invention has been completed.

The present invention is based on the above findings by the present inventors, and means for solving the above problems are as follows. That is,
<1> A medicament for treating or preventing a disease caused by an excessive reaction of Th2 or Th17, comprising a dopamine D2-like receptor agonist as an active ingredient.
<2> The medicament according to <1>, wherein the dopamine D2-like receptor agonist is non-ergot system.
<3> The medicament according to <2>, wherein the dopamine D2-like receptor agonist is at least one of pramipexole hydrochloride hydrate and ropinirole hydrochloride.
<4> The group resulting from Th2 or Th17 overreaction comprises multiple sclerosis, rheumatoid arthritis, autoimmune myositis, bronchial asthma, atopic dermatitis, chronic GVHD, type 1 diabetes, and glomerulonephritis The medicament according to any one of <1> to <3>, which is at least one selected from the above.
<5> A method for screening a pharmaceutical according to any one of <1> to <4>, wherein binding to a dopamine D2-like receptor is used as an index.
<6> The method for screening a pharmaceutical according to any one of <1> to <4>, wherein the activity is an activity on a dopamine D2-like receptor as an index.

  According to the present invention, various conventional problems can be solved, and an effective drug for diseases caused by excessive reaction of Th2 or Th17 (for example, multiple sclerosis) and an efficient screening method for the drug are provided. can do.

(Medicine)
The medicament of the present invention is a medicament that can be used for the treatment or prevention of a disease caused by an excessive reaction of Th2 or Th17, contains a dopamine D2-like receptor agonist as an active ingredient, and, if necessary, other Contains ingredients.

<Dopamine D2-like receptor agonist>
The “dopamine receptor” has five subtypes from D1 to D5, and is generally known to have a function of sending a signal in the cell coupled with the G protein. D1 and D5 are known as “dopamine D1-like receptors” and are coupled to Gs proteins that increase adenyl cyclase activity and increase cAMP concentration. On the other hand, it is known that D2 to D4 are conjugated with a Gi protein that suppresses adenyl cyclase activity as a “dopamine D2-like receptor”. Therefore, as the “dopamine D2-like receptor agonist”, a substance having a function of activating the action of at least one of subtypes D2 to D4 of the dopamine receptor can be used.

That is, the dopamine D2-like receptor agonist is not particularly limited and can be appropriately selected according to the purpose. For example, a known dopamine D2-like receptor agonist (dopamine D2-like receptor agonist) is used. Alternatively, a substance that is evaluated to have a binding ability to a dopamine D2-like receptor and / or an activity effect on a dopamine D2-like receptor by the screening method of the present invention described later may be used.
Specific examples of the dopamine D2-like receptor agonist include, for example, pramipexole hydrochloride hydrate, ropinirole hydrochloride, talipexol hydrochloride, non-ergot dopamine D2-like receptor agonists such as rotigotine, cabergoline, bromocriptine mesylate, etc. Although ergot-type dopamine D2-like receptor agonists are mentioned, among these, non-ergot-type dopamine D2-like receptor agonists such as pramipexole hydrochloride hydrate and ropinirole hydrochloride are particularly preferable. In addition, the pramipexole hydrochloride hydrate is from Nippon Boehringer Ingelheim Co., Ltd. under the trade name “Bi-Siflor”, the ropinirole hydrochloride is from Glaxo Smith Kline Co., Ltd. under the trade name “Requip”, The brand name “Domin” from Nippon Boehringer Ingelheim Co., Ltd., the rotigotine from Schwarzwama as the brand name “New Pro”, and the cabergoline from Pfizer Co., Ltd. under the brand name “Kabasar”, the bromocriptine mesylate Can be obtained from Novartis Pharma Co., Ltd. under the trade name “Perlodel”.

  The content of the dopamine D2-like receptor agonist in the medicament is not particularly limited and can be appropriately selected according to the purpose. The medicament may be the dopamine D2-like receptor agonist itself. .

<Other ingredients>
There is no restriction | limiting in particular as said other component, In the range which does not impair the effect of this invention, it can select suitably according to the objective, For example, a pharmacologically acceptable carrier etc. are mentioned. The carrier is not particularly limited and may be appropriately selected depending on, for example, the pharmaceutical dosage form described below. Moreover, there is no restriction | limiting in particular also as content of the said other component in the said pharmaceutical, According to the objective, it can select suitably.

<Dosage form, production>
The pharmaceutical dosage form is not particularly limited, and can be appropriately selected according to a desired administration method. For example, oral solid preparations (tablets, coated tablets, granules, powders, capsules, etc.), Oral solutions (internal solutions, syrups, elixirs, etc.), injections (solutions, suspensions, solid preparations for erection, etc.), suppositories, ointments, patches, gels, creams, external powders, sprays And inhalable powders.

Examples of the oral solid preparation include, for example, excipients and further additives such as binders, disintegrants, lubricants, colorants, flavoring / flavoring agents, etc., to the dopamine D2-like receptor agonist. In addition, it can be produced by a conventional method.
Examples of the excipient include lactose, sucrose, sodium chloride, glucose, starch, calcium carbonate, kaolin, microcrystalline cellulose, and silicic acid. Examples of the binder include water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, hydroxypropylcellulose, hydroxypropyl starch, methylcellulose, ethylcellulose, shellac, calcium phosphate, polyvinylpyrrolidone and the like. It is done. Examples of the disintegrant include dry starch, sodium alginate, agar powder, sodium hydrogen carbonate, calcium carbonate, sodium lauryl sulfate, stearic acid monoglyceride, and lactose. Examples of the lubricant include purified talc, stearate, borax, and polyethylene glycol. Examples of the colorant include titanium oxide and iron oxide. Examples of the flavoring / flavoring agent include sucrose, orange peel, citric acid, tartaric acid and the like.

The oral solution can be produced by a conventional method, for example, by adding additives such as a taste-masking / flavoring agent, a buffering agent and a stabilizer to the dopamine D2-like receptor agonist.
Examples of the flavoring / flavoring agent include sucrose, orange peel, citric acid, tartaric acid and the like. Examples of the buffer include sodium citrate. Examples of the stabilizer include tragacanth, gum arabic, and gelatin.

As the injection, for example, a pH regulator, a buffer, a stabilizer, a tonicity agent, a local anesthetic, or the like is added to the dopamine D2-like receptor agonist, and then subcutaneously or intramuscularly by a conventional method. Injectables for intravenous use can be produced.
Examples of the pH adjusting agent and the buffering agent include sodium citrate, sodium acetate, sodium phosphate and the like. Examples of the stabilizer include sodium pyrosulfite, EDTA, thioglycolic acid, thiolactic acid, and the like. Examples of the isotonic agent include sodium chloride and glucose. Examples of the local anesthetic include procaine hydrochloride and lidocaine hydrochloride.

  Examples of the suppository include, for example, known dopamine D2-like receptor agonists, known carriers for suppository preparations such as polyethylene glycol, lanolin, cocoa butter, fatty acid triglyceride, and Tween (registered trademark) as necessary. After adding the surfactant, etc., it can be produced by a conventional method.

The ointment can be produced, for example, by mixing a known base, stabilizer, wetting agent, preservative and the like with the dopamine D2-like receptor agonist and mixing them by a conventional method.
Examples of the base include liquid paraffin, white petrolatum, white beeswax, octyldodecyl alcohol, paraffin and the like. Examples of the preservative include methyl paraoxybenzoate, ethyl paraoxybenzoate, propyl paraoxybenzoate, and the like.

  As the patch, for example, a cream, gel or paste as the ointment can be applied to a known support by a conventional method. Examples of the support include woven fabric, nonwoven fabric, soft vinyl chloride, polyethylene, polyurethane and other films made of cotton, suf, and chemical fibers, and foam sheets.

<Target disease>
There is no restriction | limiting in particular as said medical target disease, According to the objective, it can select suitably from the diseases resulting from the excessive reaction of Th2 or Th17. Here, Th2 or Th17 excessive reaction means that the balance of Th1 (type 1 helper T cell), Th2 (type 2 helper T cell), and Th17 (type 17 helper T cell) in the helper T cell is abnormal. This refers to a state biased to Th2 or Th17. As diseases caused by these excessive reactions, for example, allergic diseases and many systemic autoimmune diseases are conventionally known. Further, in recent years, the present inventors have shown that a disease such as multiple sclerosis, which was conventionally thought to be caused by a Th1 overreaction, is actually a disease caused by a Th2 or Th17 overreaction (special feature). Application 2006-211881). Therefore, as the disease caused by the excessive reaction of Th2 or Th17, in addition to allergic diseases and many systemic autoimmune diseases conventionally known as diseases caused by the excessive reaction of Th2 or Th17, Can also include diseases such as multiple sclerosis and rheumatoid arthritis, which have been attributed to Th1 overreaction.
Among these, multiple sclerosis, rheumatoid arthritis, autoimmune myositis, bronchial asthma, atopic dermatitis, chronic GVHD, type 1 diabetes and glomerulonephritis are particularly suitable as the target diseases of the medicine.

<Administration>
There is no restriction | limiting in particular as an administration object of the said pharmaceutical, According to the objective, it can select suitably, For example, a human, a mouse | mouth, a rat, a cow, a pig, a monkey etc. are mentioned.
Moreover, there is no restriction | limiting in particular as the administration method of the said pharmaceutical, According to the dosage form etc. of the said pharmaceutical, it can select suitably, For example, oral administration, administration by injection, etc. are mentioned.
The dosage of the drug is not particularly limited and can be appropriately selected according to the age, weight, sex, symptom, etc. of the patient to be administered, and is an active ingredient per day for a human adult. The amount of the dopamine D2-like receptor agonist is considered to be preferably about 0.1 to 30 mg. Further, the frequency of administration of the medicament is not particularly limited and can be appropriately selected according to the purpose. For example, the daily dose may be administered once a day, Multiple doses may be administered.

  Further, the administration timing of the medicament is not particularly limited and can be appropriately selected according to the purpose. For example, the medicament may be administered prophylactically before the onset of the disease or treated after the onset of the disease. May also be administered. The medicament can exert an excellent effect on a disease caused by the excessive reaction of Th2 or Th17 in both administration before onset and administration after onset.

(Screening method)
The screening method of the present invention is a method for screening the pharmaceutical agent of the present invention. For example, a method using the binding to dopamine D2-like receptor as an index (first screening method) and dopamine D2-like receptor Examples include a method (second screening method) using an active action on the body as an index.

<First screening method (indicating binding)>
The first screening method is not particularly limited as long as it is a method using the binding to the dopamine D2-like receptor as an index, and can be appropriately selected according to the purpose. For example, (a) a test substance Evaluating the ability to bind to the dopamine D2-like receptor, and (b) selecting the test substance evaluated to have the ability to bind to the dopamine D2-like receptor in step (a). And the like.
In addition, there is no restriction | limiting in particular as said test substance, For example, it can select suitably from the said drug candidate substance according to the objective.

-(A) Evaluation process-
The method for evaluating the ability of the test substance to bind to the dopamine D2-like receptor in the evaluation step is not particularly limited and can be appropriately selected depending on the purpose. For example, a dopamine D2-like receptor protein is selected from Examples include a method based on a binding assay between the expressed cell line and the test substance.
In addition, for example, when a result that there is a difference in the degree of binding of the test substance to two types of cell lines that differ only in the expression of the dopamine D2-like receptor, the test substance is the dopamine D2 It can be evaluated that it has a binding ability to a like receptor.
-(B) Selection step-
In the selection step, the test substance evaluated as having the ability to bind to the dopamine D2-like receptor in the step (a) is selected.

<Second screening method (indicating activity action)>
The second screening method is not particularly limited as long as it is a method using the activity of the dopamine D2-like receptor as an index, and can be appropriately selected according to the purpose. For example, (a ′) test A step of evaluating an active action of a substance on a dopamine D2-like receptor, and (b ′) selecting the test substance evaluated to have an active action on the dopamine D2-like receptor in the step (a ′). And the like.

-(A ') Evaluation process-
The method for evaluating the active action of the test substance on the dopamine D2-like receptor in the evaluation step is not particularly limited and can be appropriately selected depending on the purpose. For example, in the presence of the test substance And a method for examining changes in intracellular cAMP concentration, intracellular dopamine (DA) synthesis amount, intracellular dopamine (DA) storage amount, and the like. The various changes can be examined using, for example, a conventionally known method.
For example, in the presence of the test substance, the intracellular cAMP concentration decreases, the intracellular DA synthesis amount decreases, and / or the intracellular DA storage amount decreases compared to the absence of the test substance. When the result is obtained, it can be evaluated that the test substance has an active action on the dopamine D2-like receptor.
-(B ') Selection step-
In the selection step, the test substance evaluated to have an activity on the dopamine D2-like receptor in the step (a ′) is selected.

  As the screening method, only one of the first screening and the second screening may be performed, or both may be performed, but the medicine can be efficiently selected. It is preferable to do both. In this case, the medicine can be selected more efficiently by performing the first screening and the second screening in this order.

  Examples of the present invention will be described below, but the present invention is not limited to these examples.

(Example 1: Clinical effect of dopamine D2-like receptor agonist in EAE model mouse (1))
A dopamine D2-like receptor agonist was administered to mice in which experimental autoimmune encephalomyelitis (EAE), which is a model of multiple sclerosis, was induced, and its clinical effect (prophylactic effect) was examined.

<Method>
Six weeks old SJL / J mice (female) were injected subcutaneously with PLP139-151 (100 μg) + CFA (100 μg) (day 0) to initiate EAE induction. From day 3, the following various dopamine D2-like receptor agonists were orally administered every other day to 4 mice in each group, and EAE clinical scores were observed. Various dopamine D2-like receptor agonists were dissolved in phosphate buffered saline (PBS) and administered at the following doses. In addition, only PBS was administered to the control group.
As for various dopamine D2-like receptor agonists, “Cabasar” (generic name: cabergoline) is from Pfizer Inc. and “Perlodel” (generic name: bromocriptine mesylate) is from Novartis Pharma Co., Ltd. The general name; pramipexole hydrochloride hydrate) obtained from Nippon Boehringer Ingelheim Co., Ltd. was used.
[Experimental group]
Group A: Control (PBS) group B Group: Cabasar 0.3 mg / kg / day (generic name; cabergoline) Group C: Parrodel 0.2 mg / kg / day (generic name: bromocriptine mesylate) group D group: Bi・ Siflor 0.1 mg / kg / day (generic name; pramipexole hydrochloride hydrate) group [EAE clinical score]
1; reduced tail tension 2; hindlimb paraplegia 3; hindlimb paralysis 4; limb paralysis 5; drowning or death

<Result>
The results are shown in FIG. In the bi-ciflor-administered group, a particularly significant decrease in EAE clinical score was observed compared to the control (cont.) Group. That is, in the bi-ciflor administration group, the EAE clinical score did not change from the beginning of EAE induction to a score of 0, indicating that the administration of bi-ciflor could completely prevent the onset of EAE. (Fig. 1).

(Example 2: Clinical effect of dopamine D2-like receptor agonist in EAE model mouse (2))
A dopamine D2-like receptor agonist was administered to mice in which EAE was induced, and its clinical effect (therapeutic effect) was examined.

<Method>
Six weeks old SJL / J mice (female) were injected subcutaneously with PLP139-151 (100 μg) + CFA (100 μg) to initiate EAE induction. Four mice that developed EAE were divided into 2 groups of 2 mice each, which were used as a control group and a requip group. From the 24th day after the start of EAE induction, the requip group was orally administered with requip (generic name; ropinirole hydrochloride), which is a dopamine D2-like receptor agonist, every other day, and the EAE clinical score as in Example 1 was obtained. Observed. Lekip (generic name; ropinirole hydrochloride) was obtained from GlaxoSmithKline, dissolved in phosphate buffered saline (PBS) and administered at a dose of 0.1 mg / kg / day. In addition, only PBS was administered to the control group.

<Result>
The results are shown in FIG. In the requip administration group, a significant decrease in EAE score was observed from the control group on the 10th day after the start of oral administration.

  From the above, it was suggested that a dopamine D2-like receptor agonist can be an excellent drug for diseases caused by excessive reaction of Th2 or Th17 including multiple sclerosis. Among these, bi-ciflor (generic name: pramipexole hydrochloride hydrate) and requip (generic name: ropinirole hydrochloride), which are non-ergot systems, were found to have particularly remarkable clinical effects.

  In addition, the present inventors have previously found that multiple sclerosis is a disease caused by an excessive reaction of Th2 or Th17. Therefore, for example, by actively inducing Th1 differentiation, the above-mentioned disease can be effectively treated. It was found that it can be treated or prevented (see Japanese Patent Application No. 2006-211881). Also in the present invention, dopamine D2-like receptors in dendritic cells are first administered by administration of a dopamine D2-like receptor agonist such as bi-ciflor (generic name; pramipexole hydrochloride hydrate), requip (generic name; ropinirole hydrochloride). As a result, Th1 differentiation of a naive T cell that has received a signal from a dendritic cell is induced, thereby presuming a mechanism of action that biases the Th1 / Th2 / Th17 balance to Th1. Since the expression of D2 and D3 receptors in dendritic cells is at a low level, it is possible that dopamine D2-like receptor agonists directly act on naive T cells and memory T cells to induce Th1 bias. The dopamine D2-like receptor agonist is expected to exert a wide clinical effect not only on multiple sclerosis but also on diseases caused by excessive reaction of Th2 or Th17.

  The medicament of the present invention is a disease caused by an excessive reaction of Th2 or Th17 (for example, multiple sclerosis, rheumatoid arthritis, autoimmune myositis, bronchial asthma, atopic dermatitis, chronic GVHD, type 1 diabetes, glomerulonephritis And the like, and the screening method of the present invention is useful as a method for efficiently screening the aforementioned medicaments.

FIG. 1 is a graph showing the clinical effect (prophylactic effect) on EAE of dopamine D2-like receptor agonists (Kabasar, Parrodel, Bi-Siflor). (The vertical axis represents the average value of the EAE clinical score of each group, and the horizontal axis represents the number of days after the start of EAE induction.) FIG. 2 is a graph showing the clinical effect (therapeutic effect) of a dopamine D2-like receptor agonist (Requip) on EAE. (The vertical axis represents the average value of the EAE clinical score of each group, and the horizontal axis represents the number of days after the start of oral administration.)

Claims (6)

  A medicament for treating or preventing a disease caused by an excessive reaction of Th2 or Th17, comprising a dopamine D2-like receptor agonist as an active ingredient.   The medicament according to claim 1, wherein the dopamine D2-like receptor agonist is non-ergot.   The medicament according to claim 2, wherein the dopamine D2-like receptor agonist is at least one of pramipexole hydrochloride hydrate and ropinirole hydrochloride.   The disease resulting from an excessive reaction of Th2 or Th17 is selected from the group consisting of multiple sclerosis, rheumatoid arthritis, autoimmune myositis, bronchial asthma, atopic dermatitis, chronic GVHD, type 1 diabetes, and glomerulonephritis The medicament according to any one of claims 1 to 3, which is at least one of the following.   The method for screening a medicine according to any one of claims 1 to 4, wherein binding to a dopamine D2-like receptor is used as an index.   The method for screening a medicine according to any one of claims 1 to 4, wherein an active action on a dopamine D2-like receptor is used as an index.