Preventive and / or therapeutic agent of senile xerosis

Abstract

To provide a preventive and / or therapeutic agent of senile xerosis based on a new action mechanism.SOLUTION: Provided is a preventive and / or therapeutic agent of senile xerosis which contains a compound having SIRT2 activating action as an active ingredient. Further, also provided is a preventive and / or therapeutic agent which contains a compound having SIRT2 activating action as an active ingredient, and is used for a disease in which reduction in an expression level of claudin-4 causes the disease or forms its pathosis, and an expression level-increasing agent of claudin-4 which contains a compound having SIRT2 activating action as an active ingredient.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a preventive and / or therapeutic agent for senile xeroderma, more specifically, to a preventive and / or therapeutic agent for senile xeroderma in which decreased expression of claudin-4 is observed. [Background technology]

[0002] With the arrival of a super-aging society worldwide, the soaring medical and nursing care costs caused by diseases of the elderly have become a social issue. It is said that more than 70% of elderly people have some kind of skin problem, but the exact number of people with skin diseases is unknown.

[0003] Senile xerosis is not a fatal disease, but it has been reported that itching can cause sleep deprivation and impaired concentration, resulting in a decline in quality of life. Furthermore, xerosis can lead to eczema and inflammation, so appropriate treatment is necessary. Currently, moisturizers, steroids, antihistamines, and other medications are used to treat xerosis, but no fundamental cure exists. Therefore, there is a need to elucidate the molecular mechanisms involved in the onset of xerosis and develop new preventive and therapeutic methods.

[0004] In response to this, Patent Document 1 discloses an agent for improving and preventing senile xeroderma, which is characterized by containing a hydroxyether carboxylate, and Patent Document 2 discloses an agent for treating xeroderma, which contains a DP receptor selective agonist as an active ingredient. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Japanese Patent Application Laid-Open No. 2016-124828 [Patent Document 2] Japanese Patent Application Laid-Open No. 2005-247842 Summary of the Invention [Problem to be solved by the invention]

[0006] However, research and development into senile xerosis is still insufficient, and the development of new types of preventive and / or therapeutic drugs is desired.

[0007] Under these circumstances, the main object of the present invention is to provide a drug for preventing and / or treating senile xerosis based on a new mechanism of action. [Means for solving the problem]

[0008] The present invention provides a drug for preventing and / or treating senile xerosis, which contains as an active ingredient a compound having SIRT2 activating activity. In the present invention, the senile xerosis may be caused by a decrease in the expression level of claudin-4.

[0009] The present invention also provides a preventive and / or therapeutic agent for diseases caused by or resulting in the pathology of a decrease in the expression level of claudin 4, which contains as an active ingredient a compound having SIRT2 activation activity.

[0010] Furthermore, the present invention also provides an agent for increasing the expression level of claudin-4, which comprises as an active ingredient a compound having SIRT2 activation activity. [Effects of the Invention]

[0011] According to the present invention, it is possible to provide a preventive and / or therapeutic agent for senile xerosis based on a new mechanism of action. The effects described here are not necessarily limited to those described herein, and may be any of the effects described in this specification. [Brief explanation of the drawings]

[0012] [Figure 1] FIG. 1 shows the effect of senescence-promoting agents on claudin-4 expression. [Figure 2] FIG. 1 shows the effect of SIRT2 knockdown on claudin-4 expression. [Figure 3]FIG. 1 shows the expression analysis of claudin-4 and SIRT2 in mouse skin tissue. [Figure 4] FIG. 1 shows SIRT2 activity in mouse skin tissue. [Figure 5] FIG. 1 shows the effect of SMT on SIRT2 activity. [Figure 6] FIG. 1 shows the interaction analysis between SIRT2 and SMT-1. [Figure 7] FIG. 1 shows the effects of senescence-promoting agents and SMT-1 on claudin-4 expression. [Figure 8] FIG. 1 shows the ameliorating effect of SMT-1 on the deterioration of intercellular barrier function caused by an aging accelerator. DETAILED DESCRIPTION OF THE INVENTION

[0013] Preferred embodiments for carrying out the present invention will now be described. The embodiment described below is an example of a typical embodiment of the present invention, and the scope of the present invention should not be construed as being narrow.

[0014] 1. Background It has been reported that aging reduces the expression of skin moisturizing factors and reduces moisturizing function, but the impact on intercellular barrier function has not been fully investigated. We found that the expression of claudin-4, which forms the intercellular barrier, decreases in the skin tissue of elderly people. Analysis using human epidermal-derived HaCaT cells showed that decreased expression of claudin-4 reduced the transepidermal electrical resistance (TER), an index of intercellular barrier function, and increased the permeability of a water-soluble low-molecular-weight compound (Lucifer Yellow). Furthermore, the expression of hyaluronan synthase, ceramide synthase, and aquaporin-3, which are involved in maintaining moisturizing function, decreased. The physiological role of claudin-4 in the skin was previously unknown, but our research suggests that claudin-4 is involved in maintaining moisturizing function.

[0015] The mechanism underlying the age-related decline in claudin-4 expression was unknown, but we discovered the involvement of SIRT2. Because no SIRT2-activating drugs had been reported to date, we screened food ingredients and discovered 10-hydroxy-2-decenoic acid (10H2DA) contained in royal jelly. 10H2DA and royal jelly containing 10H2DA have been patented as compounds for improving xeroderma (e.g., JP 10-114652, JP 10-120561, JP 2019-214516). 10H2DA has also been patented as a compound that activates epidermal stem cells and promotes skin turnover (e.g., JP 2023-178803). However, the target molecules and mechanism of action of 10H2DA have not been fully explored. Furthermore, there is no literature suggesting a relationship between SIRT2 and claudin-4.

[0016] SIRT2 is thought to be a new therapeutic target for senile xerosis, but no SIRT2 activating drugs have been developed. Furthermore, the role of SIRT2 in maintaining skin barrier function remains unclear. Therefore, the problems to be solved by the present invention are as follows: (i) Effect of senescence-promoting agents on claudin-4 expression (ii) Relationship between SIRT2 and claudin-4 expression (iii) Effect of aging on SIRT2 expression and activity (iv) Development of compounds with SIRT2 activation activity (v) Effects of SIRT2 activators on intercellular barrier function

[0017] 2. Drugs for the prevention and / or treatment of senile xerosis As mentioned above, the present inventors discovered that the expression level of claudin-4, which forms an intercellular barrier, decreases in the skin tissue of elderly people. Claudins are known to have 27 subtypes and are membrane proteins present at the tight junctions of epithelial and endothelial cells.

[0018] On the other hand, SIRT2 is an enzyme belonging to the Sirtuin family that deacetylates acetylated lysine using NAD+ as a coenzyme. Lysine acetylation is a type of protein post-translational modification that regulates gene expression, energy metabolism, and the like. The inventors have targeted SIRT2 as a therapeutic target and discovered compounds with SIRT2 activating activity, for example, those represented by the following chemical formulas (1) to (7). By applying a compound with SIRT2 activating activity to skin tissue with reduced expression levels of claudin-4, an improvement effect was observed, demonstrating that compounds that activate SIRT2 are effective against senile xerosis.

[0019] [ka]

[0020] [ka]

[0021] The compounds represented by the above chemical formulas (1) to (7) can be prepared by conventionally known synthesis methods. Alternatively, commercially available compounds may be used. The compounds represented by the above chemical formulas (1) to (7) have the advantage that they are low molecular weight compounds and can be produced relatively inexpensively. Therefore, they can also contribute to reducing medical costs.

[0022] In the present invention, it is preferable to use a compound that has a strong SIRT2 activation effect, and among the compounds shown in the above chemical formulas (1) to (7), it is particularly preferable to use the compound shown in the above chemical formula (3), which has a particularly strong SIRT2 activation effect.

[0023] The prophylactic and / or therapeutic drug for senile xeroderma according to the present invention is particularly used for the prevention and / or treatment of senile xeroderma, which is characterized by a decrease in the expression level of claudin-4. In the present invention, "prevention" also includes the meaning of preventing or delaying the onset (including recurrence) of symptoms or diseases in a subject, or reducing the risk of developing symptoms or diseases in a subject. Furthermore, "treatment" also includes the meaning of alleviating (alleviating) symptoms or associated symptoms in a subject, preventing or delaying the worsening of symptoms, etc.

[0024] The prophylactic and / or therapeutic agent for senile xeroderma according to the present invention is preferably used for the prophylaxis and / or treatment of senile xeroderma in which the expression level of claudin-4 is significantly reduced. In the present invention, "significantly" means that the expression level of claudin-4 is significantly reduced compared to a control group.

[0025] The preventive and / or therapeutic agent for senile xerosis according to the present invention may consist solely of a compound having SIRT2 activation activity as an active ingredient, or may be a composition containing the active ingredient together with any other ingredient.

[0026] The preventive and / or therapeutic agent for senile xerosis according to the present invention can be formulated by a conventionally known method. When the agent is formulated, other pharmaceutically acceptable ingredients (e.g., carriers, excipients, disintegrants, buffers, emulsifiers, suspending agents, soothing agents, stabilizers, preservatives, antiseptics, surfactants, lubricants, diluents, coating agents, sugar-coating agents, flavoring agents, emulsifying / solubilizing / dispersing agents, pH adjusters, isotonic agents, solubilizing agents, flavoring agents, coloring agents, solubilizing aids, physiological saline, etc.) may be contained.

[0027] The dosage form of the preventive and / or therapeutic agent for senile xerosis according to the present invention when formulated is not particularly limited, and examples of dosage forms include tablets, powders, fine granules, granules, capsules, syrups, liquids, suspensions, emulsions, jellies, injections, topical preparations, inhalants, nasal drops, eye drops, and suppositories.

[0028] The preventive and / or therapeutic drug for senile xerosis according to the present invention contains an active ingredient in an amount necessary to achieve the effects of the present invention (i.e., a therapeutically effective amount). The effective amount in the preventive and / or therapeutic drug for senile xerosis according to the present invention generally varies depending on the dosage form, but is appropriately set to an effective amount that achieves the desired dosage (for example, within the range of 0.01% by weight to about 100% by weight). The subject is usually a human, but also includes non-human mammals, such as pet animals such as dogs and cats, livestock such as cows, sheep, pigs, and goats, and laboratory animals such as mice, rats, rabbits, guinea pigs, hamsters, and monkeys.

[0029] The prophylactic and / or therapeutic agent for senile xerosis according to the present invention is administered to a subject by oral administration or parenteral administration (e.g., intravenous, intraarterial, subcutaneous, intradermal, intramuscular, or intraperitoneal injection, transdermal, nasal, transmucosal, etc.) depending on its dosage form. These administration routes are not mutually exclusive, and any two or more of them may be used in combination. Furthermore, instead of systemic administration, local administration may be used, and a drug delivery system (DDS) or the like may be utilized.

[0030] 3. Preventive and / or therapeutic drugs for diseases caused by or resulting from decreased expression of claudin 4 The preventive and / or therapeutic agent of the present invention contains a compound having SIRT2 activation activity as an active ingredient, and is used for the purpose of preventing and / or treating diseases caused by or resulting in the pathology of a decrease in the expression level of claudin-4.

[0031] Compounds with SIRT2 activation activity have the effect of increasing the expression (abundance) of claudin-4, and therefore can exert their unique effects on diseases in which a decrease in the expression level of claudin-4 is the cause or pathology of the disease.

[0032] 4. Agents that increase the expression of claudin-4 The agent for increasing the expression level of claudin-4 according to the present invention contains, as at least an active ingredient, a compound having SIRT2 activation activity.

[0033] Claudin-4 may be involved in the onset and progression of senile xerosis and other diseases, making it a potential target for basic research and the development of preventive and / or therapeutic drugs or treatments. Therefore, the binding agent of the present invention is useful, for example, as various tools (e.g., research reagents, diagnostic reagents, etc.) in such research and development. [Example]

[0034] The present invention will be described in more detail below based on examples. The embodiment described below is merely a representative example of the present invention, and the scope of the present invention should not be construed as being narrow.

[0035] <Experimental Example 1> In Experimental Example 1, HaCaT cells were incubated in a medium containing 0, 5, or 10 μM tenovin-1 for 24 hours, and then the protein expression levels of claudin-4 and β-actin were examined by Western blotting.

[0036] The effect of senescence-accelerating agents on claudin-4 expression is shown in Figure 1. The results of Experimental Example 1 showed that when HaCaT cells were treated with the senescence-accelerating agent tenovin-1, the amount of claudin-4 protein decreased in a concentration-dependent manner.

[0037] <Experimental Example 2> In Experimental Example 2, HaCaT cells were transfected with negative siRNA, SIRT1 siRNA, or SIRT2 siRNA. The protein expression levels of claudin-4 and β-actin were examined by Western blotting.

[0038] The effect of SIRT2 knockdown on claudin-4 expression is shown in Figure 2. The results of Experimental Example 2 showed that knocking down SIRT2 expression using siRNA reduced the amount of claudin-4 protein.

[0039] <Experimental Example 3> In Experimental Example 3, skin tissues were removed from young (6-week-old, hereinafter the same) and old (37-week-old, hereinafter the same) mice, and the protein expression levels of claudin-4, SIRT2, and β-actin were examined by Western blotting.

[0040] Figure 3 shows the expression analysis of claudin-4 and SIRT2 in mouse skin tissue. In Experimental Example 3, the expression levels of claudin-4 and SIRT2 in mouse skin tissue were examined, and it was found that the amount of claudin-4 protein significantly decreased with aging.

[0041] <Experimental Example 4> In Experimental Example 4, skin tissues were excised from young and old mice, and SIRT2 activity was examined using the ELISA method.

[0042] Figure 4 shows SIRT2 activity in mouse skin tissue. It was found that SIRT2 activity decreases with age.

[0043] <Experimental Example 5> From the above-mentioned Experimental Examples 1 to 4, it was found that the expression level of claudin-4 protein, which is responsible for the intercellular barrier function, decreases with age, and since SIRT2 is thought to be involved, a search was conducted for SIRT2 activating drugs. In this Experimental Example 5, the structures of 10H2DA, 4-oxo-2DA, and SIRT1 to 5 are shown. The SIRT2 activating effect of SMT was examined using the ELISA method. Activity was expressed as a relative percentage, with the activity of the solvent set at 100%.

[0044] Figure 5 shows the effect of SMT on SIRT2 activity. We investigated the effect of decenoic acid derivatives on SIRT2 activity and found that 4-oxo-2-decenoic acid (4-oxo-2DA) has the same SIRT2 activation activity as 10H2DA. Based on the structures of these compounds, SMT-1 to SMT-5 were synthesized and their SIRT2 activation activity was evaluated. SMT-1 showed the most potent activity. Furthermore, the activation activity of SMT-1 was stronger than that of 4-oxo-2DA and 10H2DA.

[0045] In Experimental Example 6, SIRT2 recombinant protein was adsorbed onto a sensor chip, and then SMT-1 was added to examine the interaction between the two. Kd was approximately 19.5 μM.

[0046] Figure 6 shows the interaction analysis between SIRT2 and SMT-1. When the binding between SMT-1 and SIRT2 was examined using quartz crystal microbalance analysis, the dissociation constant (Kd) was approximately 19.5 μM (Figure 6). Although the affinity was not very strong, direct binding between SMT-1 and SIRT2 was confirmed.

[0047] <Experimental Example 7> In Experiment 7, HaCaT cells were incubated for 24 hours in medium containing vehicle, 10 M tenovin-1, and 10 M SMT-1. The protein expression levels of claudin-4 and β-actin were examined by Western blotting. The cellular localization of claudin-4 (green) was also examined by immunofluorescence staining. Cell nuclei were stained with DAPI (blue).

[0048] Figure 7 shows the effects of senescence-promoting agents and SMT-1 on claudin-4 expression. In HaCaT cells, the decrease in claudin-4 expression caused by tenovin-1 was reversed by co-treatment with SMT-1 (Figure 7). Furthermore, tenovin-1 treatment reduced claudin-4 expression in the cell-adjacent region, and this effect was reversed by co-treatment with SMT-1.

[0049] <Experimental Example 8> In Experiment 8, HaCaT cells were cultured in a transwell to form a monolayer. After 24 hours of incubation in medium containing solvent, 10 μM tenovin-1, or 10 μM SMT-1, electrical resistance was measured using a voltmeter. The permeation rate of the water-soluble fluorescent molecule Lucifer Yellow was also measured, and small molecule permeability was calculated.

[0050] Figure 8 shows the ameliorative effect of SMT-1 on the reduction of intercellular barrier function caused by aging accelerators. Intercellular barrier function was evaluated by measuring electrical resistance and small molecule permeability. As a result, the reduction of electrical resistance and the enhancement of small molecule permeability caused by tenovin-1 were improved by co-treatment with SMT-1.

[0051] <Summary> It has been revealed that SMT-1 (a compound with SIRT2 activation activity) has an improving effect on the decline in intercellular barrier function caused by decreased SIRT2 activity and claudin-4 expression, and it is expected that this will lead to the development of new preventive and / or therapeutic drugs for senile xerosis.

Claims

1. A preventive and / or therapeutic agent for senile xerosis, which comprises as an active ingredient a compound having SIRT2 activation activity.

2. The drug for preventing and / or treating senile xeroderma according to claim 1, wherein the senile xeroderma is caused by a decrease in the expression level of claudin-4.

3. A preventive and / or therapeutic agent for a disease caused by or resulting in the pathology of a decrease in the expression level of claudin 4, comprising a compound having SIRT2 activation activity as an active ingredient.

4. An agent for increasing the expression level of claudin-4, which comprises as an active ingredient a compound having SIRT2 activation activity.

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