Lipid secretion promoter and prophylactic / therapeutic drug for eye disease caused by decreased lipid secretion

Abstract

To provide a lipid secretion promoter and a prophylactic / therapeutic agent for eye diseases caused by lipid secretion reduction.SOLUTION: This lipid secretion-promoting agent and the preventing and treating agent for eye diseases caused by the lowering of lipid secretion contain cyclo-trans-4-L-hydroxyprolyl-L-serine as an active ingredient.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a lipid secretion promoter and a drug for the prevention and treatment of eye diseases caused by decreased lipid secretion. [Background technology]

[0002] Cyclo-trans-4-L-hydroxyprolyl-L-serine is a hydroxyproline derivative known to have cell proliferation and cell protection effects (Patent Document 1), to be effective in the prevention and treatment of allergic diseases and inflammatory diseases (Patent Documents 2 and 3), and to be effective in the prevention and treatment of dry eye (Patent Document 4). [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Patent No. 3969831 [Patent Document 2] Patent No. 4253161 [Patent Document 3] Patent No. 4601118 [Patent Document 4] Patent No. 5782638 Summary of the Invention [Problem to be solved by the invention]

[0004] An object of the present invention is to provide a novel lipid secretion promoter and a novel drug for the prevention and treatment of eye diseases caused by decreased lipid secretion. [Means for solving the problem]

[0005] The present inventors have found that cyclo-trans-4-L-hydroxyprolyl-L-serine (hereinafter also referred to as Compound 1) promotes lipid secretion. The lipid secretion promoting effect of Compound 1 is completely different from the known effects of Compound 1, and the fact that Compound 1 has such an effect was an unexpected result.

[0006] The present invention relates to, for example, the following [1] and [2].

[0007] [1] A lipid secretion promoter containing cyclo-trans-4-L-hydroxyprolyl-L-serine as its active ingredient. [2] A preventive and therapeutic drug for eye diseases caused by decreased lipid secretion, containing cyclo-trans-4-L-hydroxyprolyl-L-serine as its active ingredient. [Effects of the Invention]

[0008] According to the present invention, a novel lipid secretion promoter and a preventive and therapeutic drug for eye diseases caused by decreased lipid secretion are provided. [Brief explanation of the drawings]

[0009] [Figure 1] FIG. 1 is a graph showing LipidTOX staining intensity in an established human meibomian gland epithelial cell line in the presence and absence of Compound 1. DETAILED DESCRIPTION OF THE INVENTION

[0010] Preferred embodiments of the present invention will be described in detail below, but the present invention is not limited to these embodiments.

[0011] The present invention provides a lipid secretion promoter containing, as an active ingredient, Compound 1. By administering such a lipid secretion promoter to a subject, it is possible to promote lipid secretion in the subject, particularly in the meibomian gland cells of the subject.

[0012] In this specification, Compound 1 is a compound represented by the following chemical formula (1): Compound 1 may be in the form of a free form or may form a pharmaceutically acceptable salt. [ka]

[0013] The method for producing Compound 1 is not particularly limited, but it can be obtained by the methods described in Patent Documents 1 and 3, for example.

[0014] The content of Compound 1 in the lipid secretion promoter according to this embodiment, which contains Compound 1 as an active ingredient, varies depending on the form of the preparation, but in the case of eye drops, for example, it can be 0.1 to 2.5% by mass.

[0015] The lipid secretion promoter containing Compound 1 according to this embodiment as an active ingredient can be used by mixing it with appropriate pharmaceutically acceptable additives and other pharmaceutical ingredients required for pharmaceutical purposes, such as carriers, excipients, pH adjusters, and diluents.

[0016] The form of the lipid secretion promoter is not particularly limited, and it is preferably prepared into a pharmaceutical preparation in the form of eye drops, oral medication, or injection. The content of Compound 1 in the preparation can be appropriately adjusted by those skilled in the art, and the preparation method of the preparation is not particularly limited, and known methods can be used.

[0017] The effective dosage and administration schedule of such lipid secretion promoters can be appropriately determined by those skilled in the art depending on the administration method, the condition of the disease, the weight and age of the patient, and the like.

[0018] The content of compound 1 in a preventive and therapeutic drug for eye diseases caused by decreased lipid secretion, which contains compound 1 according to this embodiment as an active ingredient, varies depending on the form of the preparation. For example, in the case of eye drops, the content can be 0.1 to 2.5% by mass.

[0019] The prophylactic and therapeutic drug for eye diseases containing Compound 1 according to this embodiment as an active ingredient can be used by mixing it with appropriate pharmaceutically acceptable additives and other pharmaceutical ingredients, such as carriers, excipients, pH adjusters, and diluents.

[0020] The form of the above-mentioned drug for preventing or treating eye diseases is not particularly limited, and it is preferably prepared as a pharmaceutical preparation in the form of eye drops, oral medication, or injection. The content of Compound 1 in the preparation can be appropriately adjusted by those skilled in the art, and the preparation method of the preparation is not particularly limited, and known methods can be used.

[0021] The effective dosage and administration schedule of such preventive and therapeutic drugs for eye diseases can be appropriately determined by those skilled in the art depending on the administration method, the condition of the disease, the weight and age of the patient, and the like.

[0022] Examples of eye diseases caused by decreased lipid secretion include dry eye caused by meibomian gland dysfunction and dry eye caused by decreased lipid secretion, including decreased lipid secretion due to aging. [Example]

[0023] The present invention will be explained in more detail below by way of examples, but the present invention is not limited to these examples.

[0024] (Analysis of lipid content in meibomian gland epithelial cells) To examine the effect of Compound 1 on lipid secretion, an in vitro experiment was carried out using an immortalized human meibomian gland epithelial cell line (IHMGEC) as follows.

[0025] HMGECs were cultured in a T75 flask using growth medium (Keratinocyte SFM supplemented with bovine pituitary extract (BPE), epidermal growth factor (rEGF), and 1% penicillin-streptomycin) in a humidified CO2 incubator (37°C, 5% CO2, humidified atmosphere). The growth medium was changed every 2–3 days. When the cells reached 70–80% confluence, they were detached, centrifuged (180 × g, room temperature, 5 min), and suspended in fresh growth medium. After cell counting, the cells were seeded at 3,500 cells / 0.1 mL / well in a 96-well black plate and cultured in a CO2 incubator. When the cells reached 75% confluence, the growth medium was replaced with medium containing 10 μM (Example 1) or 100 μM (Example 2) of Compound 1 (test medium: DMEM / F-12 supplemented with 10 ng / mL rEGF / 1% penicillin-streptomycin) and incubated in a CO2 incubator for 3 days. The growth medium was replaced with medium without Compound 1 and incubated in a CO2 incubator for 3 days. This group served as a control group (comparison). The test medium was then removed, the cells were washed with DPBS (Dulbecco's phosphate-buffered saline), and 100 μL of 4% PFA (paraformaldehyde) supplemented with 1 / 1000 volume of Hoechst 33342 (a nuclear staining dye) was added. The cells were then incubated at room temperature for 30 minutes in the dark to fix them. Next, the wells were washed three times with DPBS, and then the DPBS was replaced with 100 μL of LipidTOX™ green neutral lipid stain solution (prepared by adding 1 / 200 volume of LipidTOX™ green neutral lipid stain to DPBS) and incubated for 30 minutes at room temperature in the dark. After staining, images were taken using an Operetta CLS (Revvity). 25 fields in the center of each well were photographed using a 20x objective lens. After image acquisition, the LipidTOX staining intensity (area stained with LipidTOX™ green neutral lipid stain in the field / total number of nuclei) was calculated.The difference in LipidTOX™ staining intensity between the control group and the test substance-treated group was tested using Dunnett's t-test. A two-tailed test was performed, and a P value of <0.05 was considered significant.

[0026] 1 is a graph showing the LipidTOX staining intensity in an established human meibomian gland epithelial cell line in the presence and absence of Compound 1. For the test substance treatment group (Example 1) containing Compound 1 at a concentration of 10 μM, the LipidTOX staining intensity value tended to increase compared to the control group (Comparative Example) on day 3 of Compound 1 treatment, indicating a trend toward an increase in lipid content. For the test substance treatment group (Example 2) containing Compound 1 at a concentration of 100 μM, the LipidTOX staining intensity value significantly increased compared to the control group (Comparative Example) on day 3 of Compound 1 treatment, indicating a significant increase in lipid content.

Claims

1. A lipid secretion promoter containing cyclo-trans-4-L-hydroxyprolyl-L-serine as an active ingredient.

2. A preventive and therapeutic drug for eye diseases caused by decreased lipid secretion, which contains cyclo-trans-4-L-hydroxyprolyl-L-serine as an active ingredient.

Images