KR102272907B1 - Composition for preventing and treating dementia - Google Patents

Abstract

The present invention relates to a use as a therapeutic agent for Alzheimer's disease and dementia through the removal of amyloid beta by concurrent use of mirodenafil, which is one of the phosphodiesterase 5 inhibitors (PDE5 inhibitors), and memantine, which is one of the NMDA receptor antagonists.

Description

Composition for preventing and treating dementia

The present invention relates to amyloid by using mirodenafil, one of the phosphodiesterase 5 inhibitors (PDE5 inhibitors), and memantine, one of the NMDA receptor antagonists, in parallel. It relates to use as a therapeutic agent for Alzheimer's disease and dementia through the elimination of beta.

Dementia is an acquired brain disease with multifaceted pathogenesis caused by various genetic and environmental risk factors, and refers to a clinical disease that suffers from multiple cognitive deficits. The most representative disease causing dementia is Alzheimer's disease, which mainly occurs in the elderly and accounts for about 60-70% of all dementias (Ann Neurol. 1993 May;33(5):494-501.).

Research on amyloid beta protein (Aβ), which is known to be a common cause of hereditary and sporadic Alzheimer's disease, is being actively conducted. Even in normal people, Aβ is produced in small amounts throughout the body. In normal people, Aβ is rapidly decomposed after being made and does not accumulate in the human body, but in Alzheimer's disease patients, Aβ is abnormally produced and accumulated in tissues without being degraded to form senile plaques or the hippocampus or cerebral cortex, which plays an important role in memory and learning It piles up excessively in the same place. Accumulated Aβ triggers an inflammatory response in surrounding cells. As a result, nerve cells are damaged, and even the neural networks that maintain the normal functioning of the brain are damaged. In addition, the accumulated Aβ produces a lot of free radicals that activate the signaling system that kills nerve cells.

Aβ is a part of amyloid precursor protein cleaved by β-secretase. Aβ has various forms depending on the number of amino acids constituting it. In Alzheimer's disease patients, the proportion of Aβ consisting of 40 or 42 amino acids increases rapidly. There are many reports that Aβ induces neuronal cell death when treated in vitro, and that the mechanism of cell death is similar to the type of apoptosis seen in Alzheimer's disease patients. Neuronal damage caused by Aβ1-42 or Aβ1-43 protein has been identified as one of the important causes of Alzheimer-type disease (Yan, SD et al. Nature 1997 Oct 16;389(6652):689-95.) (Haass C, Selkoe DJ. Cell 1993 Dec 17;75(6):1039-42.), Aβ25-35 is known as an important toxic fragment of Aβ1-42 or 43 that induces nerve cell damage (Pike). CJ, Ramezan-Arab N, Cotman CW. J Neurochem 1997 Oct;69(4):1601-11.) (Lorenzo A, Yankner BA. Ann NY Acad Sci 777, 89-95, 1996.)

The most common drugs currently FDA-approved and used to treat dementia include AChEI and N-Methyl-D-aspartate receptor antagonists, as well as antioxidants and nonsteroidal anti-inflammatory drugs (N-Methyl-D-aspartate receptor antagonists). Various drugs such as inflammatory drugs, NSAIDs, anti-inflammatory agents, statins, and hormones are used in combination with them. However, these drugs are only being used for symptom relief and delay and improvement of cognitive function, and a fundamental treatment for dementia has not been developed.

Currently, global pharmaceutical companies are developing drugs that inhibit the action of β-secretase and are making efforts to fundamentally block the production of Aβ. Recently, a lot of research has been conducted on drug companies, universities and research institutes in the United States to isolate antioxidants from medicinal plants for the development of dementia treatments. Among them, Ginkgo biloba L (Ginkgo biloba L (Ginkgo biloba) trees) (Le Bars, PL et al. JAMA. 1997 Oct 22-29;278(16):1327-32.) (Oken, BS et al. Arch Neurol. 1998 Nov;55(11):1409-15. ) and Huperzia serrata Travis (Chinese moss) (Badia, A et al. Bioorg Med Chem. 1998 Apr;6(4):427-40.) (Skolnick, A A. JAMA. 1997 Mar 12;277(10): 776.) is a typical case. In addition, curcuminoids isolated from Curcuma longa L. (turmeric) have also been reported to have therapeutic effects on dementia (Bastianetto, S et al. Eur J Neurosci. 2000 Jun;12(6):1882-90) (Zangara, Andrea. Pharmacol Biochem Behav. 2003 Jun;75(3):675-86.)(Park SY, Kim DS. J Nat Prod. 2002 Sep;65(9):1227-31) Ginseng known to inhibit the toxicity of amyloid metabolites It has been reported that , tetrandrine, and ginkgo leaf extract (Egb761) can increase neuronal survival or slow the aggravation of dementia symptoms in Alzheimer's patients (Bastianetto, S et al. Eur J Neurosci. 2000 Jun; 12 ( 6):1882-90)

On the other hand, there is no satisfactory treatment method for Alzheimer's dementia and the like so far, and an effective anti-dementia agent has not yet been developed.

Republic of Korea Patent Registration No. 10-0358083 Republic of Korea Patent Registration No. 10-2000382 Republic of Korea Patent Publication No. 10-2020-0010224 Republic of Korea Patent Registration No. 10-1605576

Degenerative neurological diseases, including dementia, are caused by the decrease or loss of nerve cell function, resulting in our body's motor control function, cognitive function, perceptive function, and sensory function. In addition to all the functions of the body that can be sensed, it shows abnormalities in a wide variety of functions, including the autonomic nervous system, which is self-regulating in a state that we do not perceive.

Until now, the cause of dementia is unknown, so fundamental treatment is impossible, and the five commercially available drugs can only relieve symptoms in some diseases, and these drugs do not fundamentally change the progression of dementia. Later tolerance and serious drug side effects have emerged, further limiting symptom improvement in dementia patients.

The present invention has been devised to solve the above-mentioned problems, and an object of the present invention is to inhibit the growth and differentiation of nerve cells and to degenerate learning and memory in parallel with mirodenafil and memantine against the toxic protein Aβ To provide a composition capable of treating dementia by inducing a decrease in intracellular Aβ by reducing the expression of toxic protein by use, thereby increasing the protection of neurons and synaptic plasticity.

The present invention relates to a phosphodiesterase 5 inhibitor; And NMDA receptor antagonist (NMDA receptor antagonist); relates to a composition for preventing and treating dementia containing as an active ingredient.

Phosphodiesterase 5 inhibitor of the present invention is mirodenafil (mirodenafil), sildenafil (sildenafil), vardenafil (vardenafil), tadalafil (tadalafil), udenafil (udenafil), da santafil (dasantafil), avanafil (avanafil); and at least one selected from the group consisting of pharmaceutically acceptable salts, solvates and hydrates thereof.

The pharmaceutically acceptable salt refers to a formulation of a compound that does not cause serious irritation to the organism to which the compound is administered and does not impair the biological activity and properties of the compound. The pharmaceutically acceptable salts are prepared by conventional methods well known in the art using pharmaceutically acceptable, substantially non-toxic organic and inorganic acids. The acid may be an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid or phosphoric acid; Sulfonic acids such as methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, tartaric acid, formic acid, citric acid, acetic acid, trichloroacetic acid, trifluoroacetic acid, capric acid, isobutanoic acid, malonic acid, succinic acid, phthalic acid, gluconic acid, benzoic acid , organic acids such as lactic acid, fumaric acid, maleic acid, salicylic acid, and the like. In addition, ammonium salts can be obtained by reacting the compounds of the present invention with a base; alkali metal salts such as sodium or potassium salts; salts such as alkaline earth metal salts such as calcium or magnesium salts; salts of organic bases such as dicyclohexylamine, N-methyl-D-glucamine, and tris(hydroxymethyl)methylamine; and amino acid salts such as arginine and lysine.

According to one embodiment of the present invention, the pharmaceutically acceptable salt may include mirodenafil hydrochloride, sildenafil citrate, or vardenafil hydrochloride.

The hydrate is a compound of the present invention containing a stoichiometric or non-stoichiometric amount of water bound by a non-covalent intermolecular force, or a compound thereof means the salt of

The solvate means a compound of the present invention or a salt thereof that contains a stoichiometric or non-stoichiometric amount of a solvent bound by non-covalent intermolecular forces. Preferred solvents therefor are those that are volatile, non-toxic and/or suitable for administration to humans.

NMDA receptor antagonist of the present invention, memantine (memantine), amantadine (amantadine), ketamine (ketamine), traxoprodil (traxoprodil), lanicemine, rislenemdaz; and at least one selected from the group consisting of pharmaceutically acceptable salts, solvates and hydrates thereof.

More preferably, the phosphodiesterase 5 inhibitor of the present invention is mirodenafil and the NMDA receptor antagonist is memantine and its pharmaceutically acceptable salts, solvates and hydrates. It is characterized in that it is at least one selected from the group consisting of.

Here, the mirodenafil and memantine may be co-administered in a concentration ratio of 1: 0.2 to 5.

The pharmaceutical composition of the present invention may be administered orally or parenterally.

According to one embodiment of the present invention, the pharmaceutical composition of the present invention is administered orally to a subject or non-oral administration to a site other than the head. That is, the composition of the present invention may exhibit the intended effect in the present invention even when not directly administered to the brain tissue, the body tissue surrounding the brain tissue (eg, the scalp), and a region adjacent thereto. In one specific example, the non-oral administration is subcutaneous administration, intravenous administration, intraperitoneal injection, transdermal administration or intramuscular administration, and in another specific example, subcutaneous administration, intravenous administration or intramuscular administration.

Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are commonly used in formulation, and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methyl hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium stearate, and mineral oil; it's not going to be The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like, in addition to the above components. Suitable pharmaceutically acceptable carriers and agents are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention is prepared in unit dosage form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily carried out by a person of ordinary skill in the art to which the present invention pertains. or may be prepared by incorporation into a multi-dose container. In this case, the formulation may be in the form of a solution, suspension, or emulsion in oil or an aqueous medium, or may be in the form of an extract, powder, granule, tablet, film or capsule, and may additionally include a dispersant or stabilizer.

The present invention relates to a dementia inhibitory effect through reduction of amyloid beta by using memantine, which is one of phosphodiesterase 5 inhibitor and NMDA receptor antagonist, in combination. The results of the invention can be widely used as a preventive and therapeutic pharmaceutical use as an agent for treating dementia.

1 is an experimental result for the decrease in intracellular Aβ according to the combination treatment of mirodenafil (mirodenafil, AR1001) and memantine according to an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, these examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.

[Experimental Example 1] Cell culture

The SH-SY5Y human neuroblastoma cell line used in the experiment was purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA) and was supplemented with 10% fetal bovine serum (FBS; Australian Orgin, HyClone, Logan, UT, USA) and 1% penicillin. Using DMEM/F12 Complete Medium (HyClone) containing /streptomycin (P/S;HyClone) at 37°C, 5% CO ₂ conditions, in a CO ₂ incubator (311-TIF, Thermo Fisher Scientific Forma, MA, USA) cultured.

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

To evaluate cytotoxicity, 2 × 104 cells/well were dispensed into a 96-well plate, and neuronal cell death, neuronal inflammatory response, neurotransmitter and synaptic plasticity-related protein expression changes, and acetylcholinesterase (AChE) activity were confirmed. In order to do this, 2×105 cells were aliquoted into a T-25 flask. For cell fixation and stabilization, use DMEM/F12 Complete Medium (HyClone) containing 10% FBS (HyClone) and 1% P/S (HyClone) under a CO ₂ incubator (Thermo Fisher _{) at 37°C and 5% CO 2 conditions.} Scientific Forma) for 24 hours. 24 hours after cell dispensing, the cell culture medium was removed for neuron-like differentiation, 1% FBS (HyClone), 1% P/S (HyClone), 10 μM all-trans-retinoic acid (RA; Sigma-Aldrich, St (Louis, MO, USA) was replaced with DMEM/F12 differentiation medium. On the third day of differentiation, the medium was replaced with a new DMEM/F12 medium for differentiation. On the 6th day of differentiation, the untreated control group was replaced with new DMEM/F12 differentiation medium, and the sample treated group was replaced by adding new DMEM/F12 differentiation medium under various conditions.

[Experimental Example 3] Amyloid β(Aβ)1-42 Formation and Treatment

To form Aβ1-42 oligomer, human Aβ1-42 (Abcam, Cambridge, MA, USA) was added to DMEM/F12 Complete Medium (HyClone) containing 1% FBS (HyClone) and 1% P/S (HyClone) 10 It was added so as to become μM, and left in a CO ₂ incubator (Thermo Fisher Scientific Forma) for 3 hours at _{37° C., 5% CO 2 condition to form Aβ1-42 oligomer.}

To check the BDNF change, the existing cell culture medium was removed from the RA-differentiated SH-SY5Y neuron-like cells and replaced with DMEM/F12 Complete Medium (HyClone) containing Aβ1-42 oligomer (1 μM), 37°C, 5% Aβ1-42 oligomer-induced cell damage was induced by incubation for 72 hours in a CO ₂ incubator (Thermo Fisher Scientific Forma) under CO _{2 conditions.}

After removing the culture solution after 72 hours Aβ1-42 oligomer (1 or 10 μM) DMEM / F12 Complete Medium CO 2 (HyClone) , either alone or under the mirodenafil and memantine in combination treatment 37 ℃ 5% CO ₂ conditions including After incubation for 24 hours in an incubator (Thermo Fisher Scientific Forma), subsequent experiments were performed.

[Examples 1-3]

In Table 1 below, Example 1 is a combination treatment of mirodenafil 0.1 μM and memantine 0.1 μM, Example 2 is a combination treatment of mirodenafil 0.5 μM and memantine 0.1 μM, and Example 3 is labyrinthine It is a combination treatment of 0.5 µM pil and 0.5 µM memantine.

For reference, AR1001 in Table 1 below means mirodenafil.

[Comparative Examples 1-4]

In Table 1 below, Comparative Examples 1 and 2 were treated with mirodenafil 0.1 μM and 0.5 μM alone, and Comparative Examples 3 and 4 were treated with memantine 0.1 μM and 0.5 μM alone.

In conclusion, 0.1 μM of mirodenafil and 0.5 μM of memantine were treated in combination, and the Aβ reduction rate of Example 1 was 15.97%; Aβ reduction rate 13.15% of Example 2 treated with mirodenafil 0.5 μM and memantine 0.1 μM; The 23.18% reduction rate of Aβ in Example 3, in which mirodenafil 0.5 μM and memantine 0.5 μM were combined, was significantly higher than the sum of reduction rates A and B when mirodenafil and memantine alone were treated, indicating that the additive effect was recognized. could confirm that

That is, the Aβ reduction rates of Comparative Examples 1 and 2 treated alone with mirodenafil 0.1 μM and 0.5 μM were A1 and A2, respectively, and the Aβ reduction rates of Comparative Examples 3 and 4 treated with memantine 0.1 μM and 0.5 μM alone When B1 and B2, respectively, the Aβ reduction rate of 15.97% of Example 1 is significantly higher than 'A1+B2 = -0.18%', and the Aβ reduction rate of Example 2 15.97% is significantly higher than 'A2+B1 = -1.63%' It can be seen that the Aβ reduction rate of 15.97% of Example 3 is significantly higher than 'A2+B2 = -1.72%'.

The present invention described above is merely exemplary, and those of ordinary skill in the art to which the present invention pertains will appreciate that various modifications and equivalent other embodiments are possible therefrom. Therefore, it will be well understood that the present invention is not limited to the form mentioned in the above detailed description. Therefore, the true technical protection scope of the present invention should be determined by the technical spirit of the appended claims. It is also to be understood that the present invention includes all modifications, equivalents and substitutions falling within the spirit and scope of the invention as defined by the appended claims.

Claims (5)

phosphodiesterase 5 inhibitors; and
NMDA receptor antagonist; contains as an active ingredient,
The phosphodiesterase 5 inhibitor is at least one selected from the group consisting of mirodenafil and pharmaceutically acceptable salts, solvates and hydrates thereof;
The NMDA receptor antagonist (NMDA receptor antagonist) is a composition for preventing and treating dementia, characterized in that at least one selected from the group consisting of memantine and pharmaceutically acceptable salts, solvates and hydrates thereof.
delete delete delete According to claim 1,
The phosphodiesterase 5 inhibitor and the NMDA receptor antagonist are co-administered in a molar ratio of 1: 0.2 to 5 to have a synergistic effect on the removal of amyloid beta (Aβ). A composition for preventing and treating dementia.

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