Composition for preventing and treating dementia by combining a PDE5 inhibitor and a glucocorticoid receptor antagonist

Abstract

The present invention relates to the removal of amyloid-β by the combination of mirodenafil, a phosphodiesterase 5 inhibitor (PDE5 inhibitor), and CORT-108297, a glucocorticoid receptor (GR) antagonist, for the treatment of Alzheimer's disease or dementia.

Description

[Technical field]

[0001] The present invention relates to the treatment of Alzheimer's disease and dementia via the removal of amyloid beta by the co-administration of mirodenafil, a phosphodiesterase 5 inhibitor (PDE5 inhibitor), and CORT-108297, a glucocorticoid receptor (GR) antagonist. [Background technology]

[0002] Dementia is an acquired brain disease with a multifaceted etiology caused by various genetic and environmental risk factors, and refers to a clinical disease suffering from multiple cognitive impairments. The most common cause of dementia is Alzheimer's disease, which occurs mainly in the elderly and is reported to account for approximately 60-70% of all dementia cases (Non-Patent Document 1).

[0003] Amyloid beta protein (Aβ) is known to be a common cause of hereditary and sporadic Alzheimer's disease and is actively studied. Aβ is produced in small amounts throughout the human body, even in normal individuals. In normal individuals, Aβ is quickly broken down and does not accumulate in the human body, but in Alzheimer's disease patients, Aβ is produced in abnormally large amounts and accumulates in tissues without being broken down, resulting in the formation of senile plaques or excessive accumulation in the hippocampus and cerebral cortex, which play important roles in memory and learning. Accumulated Aβ induces an inflammatory response in surrounding cells. As a result, nerve cells are damaged and the neural circuits that maintain normal brain function are gradually impaired. In addition, accumulated Aβ generates large amounts of free radicals that activate signaling systems that kill nerve cells.

[0004] Aβ is a part of the amyloid precursor protein that is cleaved by β-secretase. There are several forms of Aβ, depending on the number of amino acids that compose it. In patients with Alzheimer's disease, the ratio of Aβ consisting of 40 or 42 amino acids increases dramatically. There are many reports that treatment of in vitro cultured nerve cells with Aβ induces nerve cell death, and the mechanism of cell death is similar to the type of apoptosis seen in Alzheimer's disease patients. Damage to nerve cells by Aβ1-42 or Aβ1-43 proteins has been identified as one of the important causes of Alzheimer's disease (Non-Patent Document 2) (Non-Patent Document 3), and Aβ25-35 is known to be an important toxic fragment of Aβ1-42 or 43 that causes damage to nerve cells (Non-Patent Document 4) (Non-Patent Document 5).

[0005] The most common FDA-approved drugs currently used to treat dementia include AChEI and N-methyl-D-aspartate receptor antagonists, antioxidants, nonsteroidal anti-inflammatory drugs (NSAIDs), anti-inflammatory drugs, statins, hormones, etc. Various drugs such as preparations are used interchangeably with these. However, these drugs are only used to alleviate symptoms, slow down and improve cognitive function, and no fundamental treatment for dementia has been developed.

[0006] Typical AChEIs include donepezil, rivastigmine, and galantamine, which have a symptomatic effect by temporarily increasing the concentration of the neurotransmitter acetylcholine. These drugs are also prescribed for mild to moderate Alzheimer's disease, vascular dementia, Parkinson's disease dementia, stroke, or subcortical ischemic vascular disease (Non-Patent Document 6).

[0007] Currently, pharmaceutical companies around the world are trying to develop drugs that inhibit the action of β-secretase and block the production of Aβ at the source. In recent years, pharmaceutical companies, universities, and research institutes in the United States have been actively researching the isolation of components that exhibit antioxidant effects from medicinal plants in order to develop dementia treatments. Among these, Ginkgo biloba L (Ginkgo biloba) (Non-Patent Document 7) (Non-Patent Document 8) and Huperzia serrata Travis (Huperzia serrata Travis) (Non-Patent Document 9) (Non-Patent Document 10) are typical examples. In addition, curcuminoids isolated from Curcuma longa L. (Turmeric) have also been reported to have therapeutic effects on dementia (Non-Patent Document 11) (Non-Patent Document 12) (Non-Patent Document 13). Ginseng, tetrandrine, and ginkgo extract (EGB761), which are known to inhibit the toxicity of amyloid metabolites, have been reported to increase the survival of nerve cells in patients with Alzheimer's disease and slow the rate of deterioration of dementia symptoms (Non-Patent Document 14).

[0008] On the other hand, there is no satisfactory treatment for Alzheimer's disease, and no effective anti-dementia drugs have been developed. [Prior art documents] [Non-patent literature]

[0009] [Non-Patent Document 1] Ann Neurol.1993 May;33(5):494-501 [Non-Patent Document 2] Yan, S, D, et al. Nature 1997 Oct 16;389(6652):689-95. [Non-Patent Document 3] Haass C, Selkoe DJ.Cell 1993 Dec 17;75(6):1039-42. [Non-Patent Document 4] Pike CJ, Ramezan-Arab N, Cotman CW.J Neurochem 1997 Oct;69(4):1601-11.

Non-Patent Document 5

Non-Patent Document 6

Non-Patent Document 7

Non-Patent Document 8

Non-Patent Document 9

Non-Patent Document 10

Non-Patent Document 11

Non-Patent Document 12

Non-Patent Document 13

Non-Patent Document 14

Summary of the Invention

Problems to be Solved by the Invention

[0010] Degenerative neurological diseases, including dementia, are caused by a decline or loss of nerve cell function, causing abnormalities in a variety of functions, including all bodily functions that we can sense, such as motor control, cognitive function, perception, and sensory function, as well as the autonomic nervous system, which regulates the body without our awareness.

[0011] To date, the cause of dementia has not been identified, so there is no fundamental treatment available. The five drugs available on the market only alleviate some of the symptoms of the disease. These drugs only have the effect of terminating symptoms, but cannot fundamentally change the progression of dementia.

[0012] The present invention provides a method for treating dementia by reducing amyloid-β, which is the direct cause of neuronal cell death, and protecting neuronal cells by using mirodenafil alone or in combination with CORT-108297, which is being developed as a therapeutic agent for post-traumatic stress disorder (PTDS). [Means for solving the problem]

[0013] The present invention relates to a composition for preventing and treating dementia, which contains as active ingredients a phosphodiesterase 5 inhibitor and a glucocorticoid receptor (GR) antagonist.

[0014] In the present invention, dementia includes Alzheimer's disease, AIDS-induced dementia, dementia with Lewy bodies, frontotemporal dementia, multi-infarct dementia, semantic and vascular dementia, Huntington's disease, Parkinson's disease, and amyotrophic lateral sclerosis.

[0015] The phosphodiesterase 5 inhibitor of the present invention includes at least one selected from the group consisting of mirodenafil, sildenafil, vardenafil, tadalafil, udenafil, dasantafil, avanafil, and pharma- ceutically acceptable salts, solvates, and hydrates thereof.

[0016] Pharmaceutically acceptable salts refer to preparations of compounds that do not cause significant irritation to the organism to which the compound is administered and do not impair the biological activity and physical properties of the compound. Pharmaceutically acceptable salts are prepared by conventional methods well known in the art using substantially non-toxic pharma- ceutically acceptable organic and inorganic acids. The acids include inorganic acids such as hydrochloric acid, bromic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, sulfonic acids (e.g., p-toluenesulfonic acid), tartaric acid, formic acid, citric acid, acetic acid, trichloroacetic acid, trifluoroacetic acid, capric acid, isobutanoic acid, and organic acids such as malonic acid, succinic acid, phthalic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid, salicylic acid, and the like. Furthermore, by reacting the compound of the present invention with a base such as an ammonium salt, an alkali metal salt such as a sodium or potassium salt, a salt such as a calcium or magnesium salt, an alkaline earth metal salt, or a salt of an organic base such as dicyclohexylamine, N-methyl-D-glucamine, or tris(hydroxymethyl)methylamine, salts of amino acids such as arginine and lysine can be produced.

[0017] According to one embodiment of the present invention, the pharma- ceutically acceptable salt can be exemplified by mirodenafil hydrochloride, sildenafil citrate, vardenafil hydrochloride, and the like.

[0018] Hydrate refers to a compound of the present invention or a salt thereof that contains a stoichiometric or non-stoichiometric amount of water bound by non-covalent intermolecular forces.

[0019] Solvate refers to a compound of the present invention or a salt thereof that contains a stoichiometric or non-stoichiometric amount of solvent bound by non-covalent intermolecular forces. Preferred solvents are volatile, non-toxic, and / or suitable for administration to humans.

[0020] At least one selected from the group consisting of glucocorticoid receptor (GR) antagonists), CORT-108297, prednisolone, dexamethasone, mifepristone, ciclesonide, budesonide, cortisone, flunisolide; and pharma- ceutically acceptable salts, solvates, and hydrates thereof.

[0021] More preferably, the phosphodiesterase 5 inhibitor of the present invention is at least one selected from the group consisting of mirodenafil and its pharma- ceutically acceptable salts, solvates, and hydrates, and the glucocorticoid receptor (GR) antagonist is at least one selected from the group consisting of CORT-108297 and its pharma- ceutically acceptable salts, solvates, and hydrates.

[0022] CORT-108297 of the present invention has the following structure: (R)-(4a-ethoxymethyl-1-(4-fluorophenyl)-6-(4-trifluoromethyl-benzenesulfonyl)-4,4a,5,6,7,8-hexahydro-1H,1,2,6-triaza-cyclopenta[b]naphthalene.

[0023] [ka]

[0024] The pharmaceutical compositions of the present invention can be administered orally or parenterally.

[0025] According to one embodiment of the present invention, the pharmaceutical composition of the present invention is orally administered to a subject or parenterally administered to a site other than the head.That is, the composition of the present invention can show the intended effect in the present invention even if it is not directly administered to brain tissue, body tissue surrounding brain tissue (e.g., scalp), and adjacent areas.In one specific example, parenteral administration is subcutaneous administration, intravenous administration, abdominal injection, transdermal administration, or intramuscular administration, and in another specific example, it is subcutaneous administration, intravenous administration, or intramuscular administration.

[0026] The pharmaceutically acceptable carriers contained in the pharmaceutical composition of the present invention are commonly used in the formulation of lactose, glucose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginic acid, gelatin, calcium silicate, crystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil, but are not limited thereto. In addition to the above-mentioned components, the pharmaceutical composition of the present invention can further include lubricating oils, wetting agents, sweeteners, flavorings, emulsifiers, suspensions, preservatives, etc. Suitable pharmacologically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

[0027] The pharmaceutical composition of the present invention can be prepared in a unit dose form or in a multi-volume container using a pharma- ceutically acceptable carrier and / or excipient according to a method that can be easily carried out by a person skilled in the art to which the present invention belongs. In this case, the preparation can be in the form of a solution, suspension, or emulsion, in an oil medium or an aqueous medium, or in the form of a tablet, powder, granule, tablet, film, or capsule, and can further include a dispersant or stabilizer. Effect of the Invention

[0028] The present invention relates to the effect of suppressing dementia by reducing amyloid β through the combined use of a phosphodiesterase 5 inhibitor and a glucocorticoid receptor antagonist. [Brief description of the drawings]

[0029] [Figure 1] FIG. 1 shows experimental results of intracellular Aβ reduction by combined treatment with mirodenafil (AR1001) and CORT-108297, according to one embodiment of the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0030] The present invention will be described in more detail below with reference to the following embodiments. However, these embodiments are merely for the purpose of explaining the present invention, and the scope of the present invention is not limited by these embodiments.

[0031] [Experimental Example 1] Cell culture

[0032] The SH-SY5Y human neuroblastoma cell line used in the experiments was obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA) and incubated with 10% fetal bovine serum (FBS; Australia; HyClone, Logan, UT, USA) and 1% penicillin / streptomycin (P / S; HyClone) at 37°C in a CO2 incubator (311-TIF, Thermo Fisher Scientific Forma, MA, USA) in 5% CO2.

[0033] [Experimental Example 2] Neuron-like differentiation of SH-SY5Y cells using all-trans-retinoic acid (RA)

[0034] To determine the change in the amount of amyloid beta, * ? *5 The cells were dispensed into T-25 flasks.

[0035] For cell attachment and stabilization, the cells were incubated in DMEM / F12 complete medium (HyClone) containing 10% FBS (HyClone) and 1% P / S (HyClone) for 24 hours at 37°C and 5% CO in a CO incubator (Thermo Fisher Scientific Forma).

[0036] Twenty-four hours after cell dispensing, the cell culture medium was removed for neuronal-like differentiation and replaced with DMEM / F12 differentiation medium containing 1% FBS (HyClone), 1% P / S (HyClone), and 10 μM all-trans-retinoic acid (RA; Sigma-Aldrich, St. Louis, MO, USA).

[0037] On the third day of differentiation, the medium was replaced with fresh DMEM / F12 differentiation medium. On the sixth day of differentiation, the medium of the untreated control group was replaced with fresh DMEM / F12 differentiation medium, and the medium of the sample-treated group was replaced under various conditions by adding fresh DMEM / F12 differentiation medium under various conditions.

[0038] [Experimental Example 3] Amyloid β (Aβ) 1-42 formation and processing

[0039] Human Aβ1-42 (Abcam, Cambridge, MA, USA) was added up to 10 μM to DMEM / F12 complete medium (HyClone) containing 1% FBS (HyClone) and 1% P / S (HyClone) to form Aβ1-42 oligomers, and the mixture was left in a CO2 incubator (Thermo Fisher Scientific Forma) at 37°C and 5% CO2 for 3 hours to form Aβ1-42 oligomers.

[0040] To confirm Aβ1-42 changes, the existing cell culture medium was removed from RA-differentiated SH-SY5Y neuron-like cells and replaced with DMEM / F12 complete medium (HyClone) containing Aβ1-42 oligomers (10 μM) and incubated in a CO2 incubator (Thermo Fisher Scientific Forma) at 37°C and 5% CO2 for 72 h to induce Aβ1-42 oligomer-induced cell damage.

[0041] After 72 hours, the medium was removed, and the cells were treated with DMEM / F12 complete medium (HyClone) containing 10 μM Aβ1-42 oligomers alone or a combination of mirodenafil and CORT-108297, and incubated in a CO2 incubator (Thermo Fisher Scientific Forma) at 37°C and 5% CO2 for 24 hours before moving on to the next experiment.

[0042] [Example 4] Amyloid β42 human ELISA (enzyme-linked immunosorbent assay) measurement results

[0043] To measure the amount of Aβ42 (pg / mL) in cells, cells were harvested and treated with cell lysis buffer.

[0044] After centrifugation at 14,000 rpm for 10 minutes at 4°C, the supernatant was transferred and the protein was collected. The amount of protein was quantified using Pierce™ BCA Protein Assay Kit (Thermo Fisher Scientific). The amount of Aβ42 in the cells was then measured using a human Aβ42 ELISA kit (Invitrogen).

[0045] [Table 1]

[0046] In summary, the Aβ reduction rate in the group treated with 0.1 μM mirodenafil and 0.3 μM CORT-108297 was 7.06%; the Aβ reduction rate in the group treated with 0.1 μM mirodenafil and 0.6 μM CORT-108297 was 9.52%; the Aβ reduction rate in the group treated with 0.1 μM mirodenafil and 1.2 μM CORT-108297 was 12.32%; The Aβ reduction rate in the 1.8 μM combination treatment group was 14.95%; the Aβ reduction rate in the 0.1 μM mirodenafil and 2.4 μM CORT-108297 combination treatment group was 20.00%, all of which were higher than the combined reduction rates of A and B from treatment with mirodenafil or CORT-108297, respectively, confirming greater than additive and synergistic effects.

[0047] The present invention described above is merely an example, and those skilled in the art to which the present invention belongs will fully recognize that various modifications and other equivalent embodiments are possible from the present invention. Therefore, it will be well understood that the present invention is not limited to the forms mentioned in the above detailed description. Therefore, the true scope of technical protection of the present invention should be determined by the technical ideas of the appended claims. Furthermore, the present invention should be understood to include all modifications, equivalents, and alternatives within the spirit and scope of the present invention defined by the appended claims.

Claims

1. A composition for preventing or treating dementia, the composition comprising: a phosphodiesterase 5 inhibitor; and a glucocorticoid receptor antagonist as an active ingredient, The phosphodiesterase 5 inhibitor is at least one selected from the group consisting of mirodenafil, its pharmaceutically acceptable salts, solvates or hydrates; the glucocorticoid receptor antagonist is selected from the group consisting of CORT-108297, a pharmaceutically acceptable salt, solvate, or hydrate thereof; A composition for preventing or treating dementia, wherein at least one of the phosphodiesterase 5 inhibitor and the glucocorticoid receptor antagonist are co-administered at a molar concentration ratio of 1:3 to 1:24, and wherein the composition has a synergistic effect on the removal of amyloid beta (Aβ).

2. A composition for the prevention or treatment of dementia as described in claim 1, wherein the prevention or treatment of dementia includes the prevention or treatment of Alzheimer's disease, AIDS-induced dementia, dementia with Lewy bodies, frontotemporal dementia, multi-infarct dementia, semantic dementia, vascular dementia, Huntington's disease, Parkinson's disease, and amyotrophic lateral sclerosis.

3. The composition for preventing or treating dementia according to claim 1, further comprising a pharmaceutically acceptable carrier selected from the group consisting of lactose, glucose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginic acid, gelatin, calcium silicate, crystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil.

4. A composition for preventing or treating dementia as described in claim 3, further comprising at least one selected from a lubricant, a humectant, a sweetener, a flavoring, an emulsifier, a suspension or a preservative.

5. A composition for preventing or treating dementia as described in claim 1, which is administered orally or parenterally.

6. A composition for preventing or treating dementia as described in claim 1, which is used by subcutaneous administration, intravenous administration, abdominal injection, transdermal administration, or intramuscular administration.

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