JP2010024198A - Pharmaceutical for treating and preventing type 1 diabetes comprising dopamine d2-like receptor agonist as effective component, and screening method - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a pharmaceutical effective against type 1 diabetes and an efficient screening method for the pharmaceutical. <P>SOLUTION: The pharmaceutical for treating and preventing type 1 diabetes is characterized in that its effective component is a dopamine D2-like receptor agonist. The screening method for the pharmaceutical is characterized in that it uses bondability to a dopamine D2-like receptor or activation action to a dopamine D2-like receptor as the indicator. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a medicament for type 1 diabetes and a screening method for the medicament.

  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. 20066.7: 929-936

  An object of the present invention is to solve the above-mentioned conventional problems, and to provide an effective medicine for type 1 diabetes, among other immune-related diseases, and an efficient screening method for the medicine.

  As a result of intensive studies to solve the above problems, the present inventor has an excellent effect on type 1 diabetes by using a dopamine D2-like receptor agonist such as pramipexole hydrochloride hydrate. I found out that I can do it. Furthermore, as a result of the analysis of the mechanism of action of the dopamine D2-like receptor agonist, the present inventor has found that the dopamine D2-like receptor agonist decreases IL-17 production in T cells or cAMP in antigen-presenting cells. It has been found that it has an action of reducing the concentration, and the present invention has been completed.

  This invention is based on the said knowledge by this inventor, and as a means for solving the said subject, it is as follows.

  <1> A medicament for treating or preventing type 1 diabetes, 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 pramipexole hydrochloride hydrate.

  <4> The method for screening a medicine according to any one of <1> to <3>, wherein binding to a dopamine D2-like receptor is used as an index.

  <5> The method for screening a pharmaceutical according to any one of <1> to <3>, wherein the screening is performed using an activity action on a dopamine D2-like receptor as an index.

  According to the present invention, conventional problems can be solved, and an effective medicine for type 1 diabetes and an efficient screening method for the medicine can be provided.

(Medicine)
The medicament of the present invention is a medicament that can be used for the treatment or prevention of type 1 diabetes, and contains a dopamine D2-like receptor agonist as an active ingredient, and further contains other ingredients as necessary.

<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.

<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 exhibit excellent effects 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 And a method using the active action on the body as an index (second screening method).

<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 in antigen-presenting cells. Or the method etc. which investigate the production of IL-17 in a T cell are mentioned. The various changes can be examined using, for example, a conventionally known method.

  For example, in the presence of the test substance, compared to the absence of the test substance, the intracellular cAMP concentration is decreased, the intracellular DA synthesis amount is decreased, the intracellular DA storage amount is decreased, or IL-17 production is performed. In the case where a decrease is observed, 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: Effect of dopamine D2-like receptor agonist in a spontaneous mouse model of type 1 diabetes)
As a prophylactic administration to NOD mice, which is a spontaneous model of type 1 diabetes, “bi-ciflor” (generic name; pramipexole hydrochloride hydrate; Nippon Boehringer Ingelheim Co., Ltd.) twice a week from 4 weeks of age ) Was administered orally. The drug was dissolved in phosphate buffered saline (PBS) and administered at an amount of 0.1 mg / kg / day. As a control group, phosphate buffered saline (PBS) was orally administered. Thereafter, urine sugar was monitored at 17 weeks of age, and the incidence of diabetes in each group (n = 20) was compared. As a result, the incidence of diabetes in the control group was 25% (5 out of 20 animals), whereas the incidence in the biciflor administered group was 0% (0 out of 20 animals) (p = 0.023). From this fact, it has been clarified that biciflor is effective in the onset of diabetes in type 1 diabetes model mice.

(Example 2: Relationship between action of dopamine D2-like receptor agonist and expression of IL-17 in T cells of human patients)
Biciflor is orally administered to a patient with extrapyramidal symptoms (1 patient) (0.5 mg / day, 1 tablet once a day), 3 points on day 0, 7 and 150 Then, peripheral blood mononuclear cells (PMBC) were collected, stimulated with an antibody (anti-CD3 + anti-CD28), and cytokine production was measured (FIG. 1). As a result, although the production amount of IL-5 was at the same level, the IL-17 / IFNγ ratio was significantly reduced (p <0.01) on the 150th day. That is, IL-17 productivity was reduced in T cells of patients. Among the T cells, Th1 cells produce IFNγ, Th2 cells produce IL-5, and Th17 cells produce IL-17, indicating that the IL-17 / IFNγ ratio was reduced in Th1 in patients. The / Th2 / Th17 balance is deflected to Th1, and Th17 is suppressed.

(Example 3: Effect of ergot and non-ergot dopamine D2-like receptor agonists on intracellular cAMP concentration)
THP-1 cells cultured in 10% FCS / RPMI were stimulated with PMA (50 ng / ml) for 48 hours. Then, the stimulated cells were collected, centrifuged (1200 rpm, 5 min), the supernatant was removed, and 30 mg / ml ergot alkaloid derivative kabasal and 500 pg / ml non-ergot alkaloid derivative were used as dopamine D2-like receptor agonists. Was added. Sixty minutes later, it was pretreated with 1 mM IBMX (PDE inhibitor) for 10 minutes and stimulated with 1 μM dopamine for 10 minutes. As a control, forskolin (positive control) was used. Thereafter, THP-1 cells were thawed with a cell lysis buffer (Molecular Devices), and then CatPoint Cyclic-AMP Fluorescent Assay Kit (Bulk kit) (Molecular Devices, Inc. was used with MP). The results were analyzed with statistical analysis software (statview). As a result, as shown in FIG. 2, when dopamine (−) was pretreated with RECIP, the cAMP concentration was significantly reduced, but even when dopamine was stimulated, the cAMP concentration was reduced. On the other hand, when pre-treated with cabasal, the cAMP concentration was significantly increased in dopamine (−), but the cAMP concentration was maintained even when dopamine was stimulated.

  The medicament of the present invention is useful as an agent for treating or preventing type 1 diabetes, and the screening method of the present invention is useful as a method for efficiently screening the medicament.

FIG. 1 is a graph showing the effect of a dopamine D2-like receptor agonist (biciflor) on IL-17 production in patient T cells. FIG. 2 is a graph showing the effect of a dopamine D2-like receptor agonist (Kabasar, Requip) on intracellular cAMP concentration.

Claims (5)

  A medicament for treating or preventing type 1 diabetes, 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 system.   The medicament according to claim 2, wherein the dopamine D2-like receptor agonist is pramipexole hydrochloride hydrate.   The method for screening a medicine according to any one of claims 1 to 3, wherein binding to a dopamine D2-like receptor is used as an index.   The method of screening for a medicine according to any one of claims 1 to 3, wherein an activity of the dopamine D2-like receptor is used as an index.

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