Prophylactic or therapeutic agent for myelofibrosis

Abstract

The present invention aims to provide an agent for the prevention or treatment of myelofibrosis. [Solution] Dopamine, dopamine salts, dopamine precursors, dopamine degradation inhibitors, and / or dopamine receptor agonists are useful as active ingredients in agents for the prevention or treatment of myelofibrosis.

Description

Technical Field

[0001] The present invention relates to a prophylactic or therapeutic agent for myelofibrosis.

Background Art

[0002] Myeloproliferative neoplasms (classified as a type of hematological cancer in current classification) based on genetic mutations in hematopoietic stem cells show a significant increase in the number of onset cases with advancing age, starting at around 50 years old. It is known that about half of the genetic abnormalities responsible for this are the JAK2V617F mutation (Non-Patent Document 1).

[0003] Myeloproliferative neoplasms are chronic neoplasms that progress over several years to several decades, and are characterized by blood cell proliferation (and further anemia) and myelofibrosis. Among these, regarding blood cell proliferation, there are already treatment options including antitumor agents that have demonstrated effects. On the other hand, with respect to myelofibrosis, although there are drugs with stated efficacy, in clinical practice, the reality is that there is no treatment that provides a clearly perceptible effect in clinical settings.

Prior Art Documents

Non-Patent Documents

[0004]

Non-Patent Document 1

Summary of Invention

Problem to be Solved by the Invention

[0005] The reason why there are virtually no effective options for myelofibrosis is considered to be that theoretical drug discovery based on the onset mechanism of myelofibrosis in myeloproliferative neoplasms has not been carried out. If the factor that essentially contributes to the onset of myelofibrosis in myeloproliferative neoplasms is clarified, theoretical drug discovery based on the onset mechanism of myelofibrosis can be expected by using a drug having an action that targets that factor.

[0006] Therefore, the object of the present invention is to provide an agent for the prevention or treatment of myelofibrosis. [Means for solving the problem]

[0007] The inventors of this invention, using their own method to measure biofactors in bone marrow, identified dopamine as a fundamentally deficient biofactor (a disease-suppressing factor) in the bone marrow of patients who developed myelofibrosis in myeloproliferative neoplasms. Furthermore, they found that supplementing dopamine could strongly suppress the development of myelofibrosis. This invention was completed by further investigation based on these findings.

[0008] In other words, the present invention provides inventions in the following embodiments. Item 1. A prophylactic or therapeutic agent for myelofibrosis containing dopamine, dopamine salts, dopamine precursors, dopamine degradation inhibitors, and / or dopamine receptor agonists as active ingredients. Item 2. The prophylactic or therapeutic agent according to Item 1, wherein the active ingredient is a dopamine receptor agonist. Item 3. The prophylactic or therapeutic agent according to item 1 or 2, wherein the myelofibrosis is myelofibrosis in a myeloproliferative neoplasm. Item 4. A prophylactic or therapeutic agent according to any of items 1 to 3, applicable to subjects with the JAK2V617F mutation, JAK2 exon12 mutation, CALR mutation, or c-Mpl mutation. [Effects of the Invention]

[0009] According to the present invention, an agent for the prevention or treatment of myelofibrosis is provided. [Brief explanation of the drawing]

[0010] [Figure 1] The results of measuring dopamine levels in the bone marrow of wild-type (WT) mice and JAK2V617F Tg mice are shown. [Figure 2A] This is a study design (culture system construction and DA addition timing) to confirm the effect of dopamine (DA) on suppressing myelofibrosis using bone marrow disease organoids. [Figure 2B] The images show the silver-stained cell layer at the bottom of a culture dish (top panel) and its digitally processed image (bottom panel). [Figure 2C] Figure 2B shows the quantitative results of the fibrosis area in the digital image. ***p<0.001, ns: not significant. [Figure 3A] This is a study design (construction of the transplant model and timing of DA administration) to confirm the effect of dopamine (DA) on suppressing myelofibrosis using the bone marrow transplant model JAK2V617F Tg. [Figure 3B] The images show silver staining (top panel) and its digitally processed image (bottom panel) after 4 weeks of DA administration. [Figure 3C] Figure 3B shows the quantitative results of the fibrosis area in the digital image. [Modes for carrying out the invention]

[0011] The present invention provides a preventive or therapeutic agent for myelofibrosis, characterized by containing dopamine, dopamine salts, dopamine precursors, dopamine degradation inhibitors, and / or dopamine receptor agonists as active ingredients.

[0012] Active ingredients Dopamine is a factor that suppresses the development of myelofibrosis. Therefore, the myelofibrosis preventive or therapeutic agent of the present invention can contain as an active ingredient a component that can replenish dopamine in the body and / or inhibit the breakdown of dopamine in the body. In other words, the myelofibrosis preventive or therapeutic agent of the present invention contains dopamine, dopamine salts, dopamine precursors, dopamine breakdown inhibitors, and / or dopamine receptor agonists as active ingredients.

[0013] Parkinson's disease is known as a disease in which dopamine acts as a disease-suppressing factor. Therefore, as the active ingredient of the myelofibrosis preventive or therapeutic agent of the present invention, existing drugs used for the purpose of supplementing dopamine in the body and / or inhibiting the breakdown of dopamine in the body for Parkinson's disease can be developed (drug repositioning) as preventive or therapeutic agents for myelofibrosis.

[0014] Dopamine is 4-(2-aminoethyl)benzene-1,2-diol. The form of the dopamine salt is not particularly limited as long as it is a pharmaceutically acceptable salt. For example, the salt includes an acid addition salt, preferably a hydrochloride.

[0015] The dopamine precursor is known as a substance that is converted into dopamine in the body, and specific examples thereof include levodopa. Levodopa may be a combination preparation with a decarboxylase (a component that inhibits decarboxylation of levodopa), preferably carbidopa.

[0016] Examples of the dopamine degradation inhibitor include monoamine oxidase B inhibitors (MAO-B inhibitors).

[0017] Examples of the dopamine receptor agonist include rotigotine, bromocriptine, pergolide, cabergoline, and ropinirole.

[0018] The above active ingredients may be used alone or in a combination of two or more. Among the above active ingredients, a dopamine receptor agonist is preferred, and rotigotine is more preferred.

[0019] The blending amount of the active ingredient contained in the prophylactic or therapeutic agent for myelofibrosis of the present invention is not particularly limited, and may be appropriately set according to the formulation form of the prophylactic or therapeutic agent for myelofibrosis, and examples thereof include 40 to 100% by weight.

[0020] Other ingredients The myelofibrosis preventive or therapeutic agent of the present invention may, in addition to the above-mentioned active ingredient, further contain one or more additives and bases depending on the formulation form and application, to the extent that they do not impair the effects of the present invention, or may not contain them. Such additives and bases, whether present or absent, are not particularly limited to the extent that they are pharmaceutically acceptable, but examples include excipients, binders, disintegrants, lubricants, isotonic agents, plasticizers, dispersants, emulsifiers, solubilizers, wetting agents, stabilizers, suspending agents, adhesives, gelling agents, coating agents, glossing agents, water, oils and fats, waxes, hydrocarbons, fatty acids, higher alcohols, esters, water-soluble polymers, surfactants, metal soaps, lower alcohols, polyhydric alcohols, pH adjusters, buffers, antioxidants, UV inhibitors, preservatives, flavoring agents, fragrances, powders, thickeners, colorants, chelating agents, sweeteners, and the like. The amounts of these additives and bases will be determined as appropriate depending on the type of additives and bases used, the formulation form and application of the myelofibrosis preventive or therapeutic agent, etc.

[0021] Furthermore, the myelofibrosis preventive or therapeutic agent of the present invention may contain, in addition to the above-mentioned active ingredients, one or more other nutritional and pharmacological components, to the extent that they do not impair the effects of the present invention, or they may not contain any other components. Such nutritional and pharmacological components, whether present or absent, are not particularly limited as long as they are pharmaceutically acceptable, but examples include antacids, stomachic agents, digestive agents, intestinal regulators, antispasmodics, mucosal repair agents, anti-inflammatory agents, astringents, antiemetics, antitussives, expectorants, anti-inflammatory enzymes, sedatives and hypnotics, antihistamines, caffeines, cardiotonic and diuretic agents, antibacterial agents, vasoconstrictors, vasodilators, local anesthetics, herbal extracts, vitamins, menthols, etc. The content of these components may be appropriately determined depending on the type of component used.

[0022] Formulation The form and properties of the myelofibrosis preventive or therapeutic agent of the present invention are not particularly limited, as long as it contains the above-mentioned active ingredient.

[0023] The administration form of the cell proliferation inhibitor of the present invention includes both oral and parenteral administration forms. Therefore, the myelofibrosis preventive or therapeutic agent of the present invention can be prepared as an oral preparation, injection, intravenous infusion, nasal spray, topical patch, etc.

[0024] Furthermore, the properties of the myelofibrosis preventive or therapeutic agent of the present invention may be liquid or solid. Examples of liquids include liquid preparations, beverages, emulsions, suspensions, alcoholic preparations, syrups, elixirs, soft extracts, etc., while examples of solids include gels, tablets, pills, powders, granules, capsules (including hard and soft capsules), lozenges, chewable tablets, etc. The myelofibrosis preventive or therapeutic agent of the present invention may also be in a sustained-release dosage form of the active ingredient.

[0025] Manufacturing method The method for producing the myelofibrosis preventive or therapeutic agent of the present invention can be carried out by using the above-mentioned active ingredient and other ingredients as needed, following conventionally known formulation procedures according to various forms and properties.

[0026] Purpose The myelofibrosis preventive or therapeutic agent of the present invention is used for the purpose of preventing or treating myelofibrosis. In a preferred embodiment, the myelofibrosis preventive or therapeutic agent of the present invention is used for the purpose of preventing or treating myelofibrosis in myeloproliferative neoplasms. In a preferred embodiment, the myelofibrosis preventive or therapeutic agent of the present invention is applied to subjects having the JAK2V617F mutation, the JAK2 exon12 mutation, the CALR mutation, or the c-Mpl mutation, more preferably to subjects having the JAK2V617F mutation. Furthermore, subjects having these mutations are also suitable as targets when the agent is used for the purpose of preventing myelofibrosis.

[0027] dose The dosage of the myelofibrosis preventive or therapeutic agent of the present invention is such that the amount of the active ingredient in humans is, for example, 0.1 to 10 mg / kg. [Examples]

[0028] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples.

[0029] [Test Example 1] We prepared JAK2V617F transgenic mice (hereinafter also referred to as "JAK2 Tg mice"). JAK2 Tg mice are genetically modified mice in which the JAK2V617F gene mutation (mouse JAK2 human type V617F) is inserted into cells throughout the body, and are widely used as a disease research model that well expresses the symptoms of human myeloproliferative neoplasms (hemocytosis and myelofibrosis).

[0030] As wild-type (normal) mice, we prepared mice of strain C57BL / 6, aged 8-10 weeks. These mice were created by the inventors by crossing JAK2V617F Tg mice with wild-type mice.

[0031] Femurs were collected from both JAK2 Tg mice and wild-type mice, and each femur was processed as follows: Using a syringe with a needle containing methanol (-30 to -20°C), one end of the femur was punctured, and 1 mL of methanol was injected into the bone marrow tissue in the medullary cavity of the femur. Along with the methanol injection, the injection pressure pushed the bone marrow from the opposite end of the femur into a sample tube. The bone marrow cells collected in the sample tube were then ultrasonically disrupted using a Bioruptor BR-II in a bath of cooled water (4-6°C). Ultrasonography was performed for a total of 100 seconds (conditions: Time ON 10 sec / OFF 10 sec: 10 cycles, power high). Afterward, centrifugation was performed, and the supernatant was obtained as a sample for quantification. This method for preparing quantitative samples is a preparation method uniquely developed by the inventor, designed to avoid the influence of fluctuations in biofactors caused by stimulation during tissue acquisition, and to ensure that the content of biofactors in the quantitative sample reflects the content of those biofactors present in the tissue.

[0032] After storing the quantitative samples at -80°C for 1 to 2 months, the amount of dopamine (amount of dopamine obtained from one femur) was measured by HPLC-ECD.

[0033] Figure 1 shows the results of dopamine level measurements in the quantitative sample. As shown in Figure 1, it was found that dopamine levels were sufficient in wild-type mice, but nearly depleted in JAK2 Tg mice.

[0034] [Test Example 2] A study was conducted to confirm the effect of dopamine (DA) on suppressing myelofibrosis using bone marrow disease organoids. The study design (culture system construction and DA addition timing) is shown in Figure 2A.

[0035] As shown in Figure 2A, disease organoids were created using bone marrow from JAK2 Tg mice, and disease bone marrow hematopoiesis was reproduced through culture, thereby establishing a long-term bone marrow culture system for JAK2 mutant mouse bone marrow. 100 μM dopamine was added weekly for three weeks, and fibrosis was quantified by silver staining of adherent cells in the culture (reproducing the bone marrow hematopoietic environment). The results are shown in Figures 2B and 2C.

[0036] As shown in Figures 2B and 2C, the control group without dopamine (vehicle administration group) developed severe fibrosis, but fibrosis was almost completely suppressed in the dopamine-supplemented group.

[0037] [Test Example 3] As a bone marrow transplantation model, JAK2V617F Tg, bone marrow from JAK2 Tg mice was transplanted into irradiated wild-type mice to create chimeric mice in which only hematopoietic cells possessed the JAK2V617F gene mutation. This bone marrow transplantation model, JAK2V617F Tg, is an in vivo model that more closely replicates a condition similar to that of a human disease.

[0038] A study was conducted to confirm the effect of dopamine (DA) on suppressing myelofibrosis using the bone marrow transplant model JAK2V617F Tg. The study design (construction of the transplant model and timing of DA administration) is shown in Figure 3A.

[0039] Dopamine 50 mg / kg (equivalent to a 60 kg human dose: 4 mg / kg) was administered intraperitoneally daily to bone marrow transplant model JAK2V617F Tg for 4 weeks starting 1 week after transplantation, and myelofibrosis was confirmed. The results are shown in Figures 3B and 3C. In Figure 3C, the bar represents the median.

[0040] As shown in Figure 3B, the control group that did not receive dopamine (vehicle administration group) developed severe myelofibrosis, but the dopamine administration group was found to have a strong suppression of myelofibrosis.

[0041] [summary] The results from Test Examples 1-3 showed that dopamine, which is depleted in the bone marrow of myeloproliferative neoplasm models, has a potent myelofibrosis-suppressing effect both in vitro and in vivo.

Claims

1. A prophylactic or therapeutic agent for myelofibrosis, containing dopamine, dopamine salts, dopamine precursors, dopamine degradation inhibitors, and / or dopamine receptor agonists as active ingredients.

2. The preventive or therapeutic agent according to claim 1, wherein the active ingredient is a dopamine receptor agonist.

3. The preventive or therapeutic agent according to claim 1, wherein the myelofibrosis is myelofibrosis in a myeloproliferative neoplasm.

4. The preventive or therapeutic agent according to claim 1, applicable to subjects having the JAK2V617F mutation, JAK2 exon12 mutation, CALR mutation, or c-Mpl mutation.

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