JP6457687B1 - Pharmaceutical composition for inhibiting PDE5 and food composition for inhibiting PDE5 - Google Patents

Abstract

A pharmaceutical composition for inhibiting PDE5 and a food composition for inhibiting PDE5 are provided. The present invention relates to a pharmaceutical composition for inhibiting PDE5 and a food composition for inhibiting PDE5 comprising pyroglutamic acid as an active ingredient. [Selection figure] None

Description

The present invention relates to a pharmaceutical composition and a food composition for inhibiting phosphodiesterase (PDE) 5.

  Pyroglutamic acid is an amino acid in which a glutamic acid carboxyl group and an amino group undergo an intramolecular condensation reaction to form a lactam.

  Conventionally, as shown in Patent Document 1 below, it is known that pyrrolidone carboxylic acid corresponding to pyroglutamic acid has an action of promoting the production of nitric oxide (NO: Nitric Oxid Syntase). It has been.

  That is, the following cited document 1 discloses a NOS production promoter comprising pyroglutamic acid (pyrrolidone carboxylic acid) as an active ingredient, and the agent is cNOS among two types of NOS, iNOS and cNOS. It is disclosed that NO is moderately produced by promoting only the production of NO.

JP 2002-220335 A

  As described above, Patent Document 1 discloses that a NOS production promoter containing pyroglutamic acid as an active ingredient promotes only the production of cNOS. In addition, “as in the case where cNOS is produced” is disclosed. When the amount of NO is small, it is considered to carry various protective actions on the living body, that is, vasodilatory action, blood circulation improvement,... Erection (reproduction), ... through cyclic GMP (cGMP). (Patent Document 1: Paragraph 0003).

  However, this only exemplifies what can be considered as an action of cGMP, and the above-mentioned Patent Document 1 describes an action in which a NOS production promoter containing pyroglutamic acid as an active ingredient promotes only the production of cNOS, and thus moderate It only discloses the action of producing NO.

  Actually, in the example of the above-mentioned Patent Document 1, only the amount of NO produced, the amount of cNOS expressed, and the eyelid improvement effect (vasodilatory effect, blood circulation improvement) are disclosed, and how much the amount of NO produced is cGMP. There is no disclosure or suggestion about whether the bioprotective action through the body is effective, which of the bioprotective actions is effective, particularly about the improvement of male dysfunction.

  The present invention focuses on the improvement of male dysfunction, and focuses on the action of inhibiting the action of PDE5 that reversely degrades cGMP that can be generated as a result of NOS production.

  Here, the difference in action mechanism between NOS production promotion and PDE5 inhibition will be described with reference to FIG. As shown in [1] of FIG. 1, the NOS production promoter of Patent Document 1 promotes the enzyme activity or expression level of “cNOS” as NOS, and as shown in [2] of FIG. Alternatively, the increased amount of “cNOS” promotes the production of NO using “arginine” as a substrate.

  The produced NO activates “guanylate cyclase” as shown in [3] of FIG. 1, and the activated “guanylate cyclase” is obtained as shown in [4] of FIG. It is known to promote the production of “cGMP” using “GTP (guanosine triphosphate)” as a substrate.

  On the other hand, as shown in [A] of FIG. 1, “PDE5 inhibitory action” inhibits the action of “PDE5”, that is, the action of decomposing “cGMP” into “GMP” to effectively relax smooth muscle. To promote blood flow into the penis and finally prevent or treat male dysfunction.

As described above, “NOS production promoting action” indirectly acts on “cGMP”, and there are insufficient substrates such as “arginine” and “GTP” required in the process, or “cGMP” As a result, when smooth muscle relaxation is suppressed by the action of “PDE5”, smooth muscle relaxation is not effectively performed. In addition, as disclosed in Patent Document 1 above, “If a large amount of NO is generated, it reacts with active oxygen to attack exogenous microorganisms and cancer cells, while causing inflammation and tissue damage. "It has been known.
On the other hand, “PDE5 inhibitory action” acts directly on “cGMP” to effectively relax smooth muscles.

  That is, the “NOS production promoting action” ultimately promotes the production of “cGMP”, and the “PDE5 inhibitory action” ultimately aims to suppress the degradation of “cGMP”. However, it is common to have an action related to “cGMP”.

  However, in the first place, PDE5 is unevenly distributed in the penile cavernous body and lung tissue, and the PDE5 inhibitory action is naturally used for the local part where PDE5 is unevenly distributed, particularly the penile corpus cavernosum. The production action is an action on the whole body, both of which are clearly distinguishable in the end use.

  As a result of intensive studies, the present inventors have found that pyroglutamic acid has an action of directly inhibiting the action of PDE5, and have completed the present invention.

That is, the present invention relates to P DE 5 for inhibiting pharmaceutical compositions ing contains pyroglutamic acid as an active ingredient or PDE5 inhibitory food composition.

The pharmaceutical composition for inhibiting PDE5 and the food composition for inhibiting PDE5 of the present invention can contribute to the prevention and / or treatment of a state in which cGMP is excessively degraded by the action of PDE5.

It is explanatory drawing which shows the difference between NOS production promotion effect and PDE5 inhibition effect.

The present invention relates to a pharmaceutical composition for inhibiting PDE5 and a food composition for inhibiting PDE5 containing pyroglutamic acid as an active ingredient.

  Pyroglutamic acid is an amino acid in which a carboxyl group and an amino group of glutamic acid undergo an intramolecular condensation reaction to form a lactam, and there are D-form, L-form and racemate. Preferably, D-form pyroglutamic acid is used alone or L-form pyroglutamic acid is used alone. This is because, as will be described later in Examples, PDE5 inhibition can be achieved with higher efficiency by using D-form pyroglutamic acid or L-form pyroglutamic acid alone.

As described above, PDE5 is an enzyme that hydrolyzes cGMP and is ubiquitous in the vascular smooth muscle of the cavernous corpus cavernosum. A pharmaceutical composition for inhibiting PDE5 is known to be effective as a therapeutic agent for male dysfunction (ED) by suppressing the degradation of cGMP at a site where PDE5 is unevenly distributed and increasing the local blood flow.

The present invention relates to a pharmaceutical composition for inhibiting PDE5 for the prevention and / or treatment of a condition mediated by PDE5, comprising pyroglutamic acid as an active ingredient.

  The state mediated by PDE5 may be a state involving PDE5, and particularly male dysfunction.

  The pharmaceutical composition according to the present invention can be used as a pharmaceutical or a quasi drug. These drugs or quasi drugs can be administered orally, for example, in the form of powders, tablets, coated tablets, dragees, hard or soft gelatin capsules, solutions, emulsions, or suspensions. Others, eg, rectally in the form of suppositories; eg, in the form of injections or infusions; eg, locally or transdermally in the form of ointments, creams, gels, or solutions; It can also be administered topically to the eye in the form of eyewash; parenterally. Oral administration is preferred.

  When the pharmaceutical composition of the present invention is used in the form of tablets, granules, or capsules, tableting processing aids, granule processing aids, capsule processing aids and the like can be used.

  The tableting processing aid is not particularly limited as long as it does not impair the effects of the present invention, and is any granulated sugar, super white sugar, powdered sugar, reduced maltose starch syrup, lactose, glucose, pullulan, erythritol, starch, dextrin, etc. Surfactant, powder such as sugar, crystalline cellulose, gum arabic, okara, etc., dietary fiber such as corn protein, calcium phosphate, food extract, food dry powder, natural fruit juice powder, sucrose fatty acid ester Fats and oils such as fats and oils, glycerin and fatty acid esters, or various sweeteners used in chewable tablets (eating tablets), various acidulants and various flavorings, shellac as a coating material, corn protein, yeast cell wall, starch , Reduced maltose starch syrup, sugarless sugar coating, maltitol, g Serine, sorbitol, HPMC, HPC and the like are exemplified.

  The granule processing aid is not particularly limited as long as the effects of the present invention are not impaired, and any sugar such as granulated sugar, super white sugar, powdered sugar, reduced maltose starch syrup, lactose, glucose, pullulan, erythritol, starch, dextrin, etc. Examples thereof include dietary fibers such as crystalline cellulose and gum arabic, food calcium such as corn protein and calcium phosphate, food extracts, food dry powders, and natural fruit juice powders.

  The capsule processing aid is not particularly limited as long as the effects of the present invention are not impaired. Granulated sugar, fine white sugar, powdered sugar, reduced maltose starch syrup powder, lactose, glucose for preparing hard capsule type capsules , Pullulan, erythritol, starch, dextrin and other sugars, crystalline cellulose, dietary fiber such as gum arabic, food calcium such as corn protein and calcium phosphate, food extracts, food dried powders, natural fruit juice powder, etc., soft capsule type For example, content viscosity modifiers such as food fats and oils, beeswax, glycerin fatty acid esters, and the like for preparing capsules are exemplified.

  Tablets can usually be prepared using a tableting machine, but the tablets are blended with seasoning ingredients to make a chewable tablet, or the tablet surface is coated using an automatic coating machine, spray granulator or hand pan. Or you may. Various types of granulators such as a spray granulator type, a kneading (extruding) type, or a high-speed agitation granulator type can be used for forming the granule. For preparation of capsules, capsule filling machines (hard type and soft type) can be used by mixing capsule aids.

  Moreover, this invention relates to the food composition for PDE5 inhibition for prevention and / or improvement of the state mediated by PDE5 which contains pyroglutamic acid as an active ingredient.

  The food composition for inhibiting PDE5 of the present invention can be used as food, beverage or animal feed, for example, as health food, functional food, food for specified health use, beauty food, or dietary supplement (supplement). . These foods and animal feeds may be in the form of, for example, drinking water such as tea and juice; and ice cream, jelly, candy, chocolate, chewing gum and the like. Moreover, the form of a liquid agent, a powder agent, a granule, a capsule, or a tablet may be sufficient.

  In addition, the food composition for inhibiting PDE5 of the present invention is particularly recommended for those who are concerned about male dysfunction.

Engagement Ru P DE 5 inhibitor pharmaceutical composition for the present invention, intake of PDE5 inhibition food composition is not particularly limited, dosage form, and the age of the taker or ingestion animals such user or patient, weight, and condition It can be selected as appropriate according to the conditions. For example, it is preferable to take orally about 0.01 mg to about 100 g, preferably about 0.2 mg to about 5 g per day, as the amount of the active ingredient per 60 kg of the body weight of the intake person or intake animal.
The intake period can be arbitrarily determined according to the age and symptoms of the intake person or animal.

  EXAMPLES Next, although an Example demonstrates this invention further in detail, this invention is not limited to a following example, unless the summary is exceeded.

Phosphodiesterase assay In order to evaluate the PDE5 inhibitory activity of pyroglutamic acid, a PDE5 inhibition assay by fluorescence polarization was performed. For the assay, an IMAP ™ assay kit (manufactured by Molecular Devices, catalog number R8127) was used. All evaluations were performed by double testing. The assay was based on high affinity binding of phosphate by metal coordination complexes immobilized on nanoparticles. The IMAP binding reagent forms a complex with the phosphate group of the nucleotide monophosphate generated from cGMP by phosphodiesterase. Upon detection by fluorescence polarization detection, binding causes a change in the molecular motion rate of the phosphate-containing molecule, resulting in an increase in the polarization of the fluorescence labeled on the substrate.

(Preparation of reagents)
1) Substrate solution Fluorescein-labeled cGMP (100 nM) was prepared in IMAP reaction buffer containing 2.5 mM Tris-HCl, 2 mM MgCl ₂ , 0.01% NaN ₃ , 0.02% BSA.
2) Enzyme solution Human PDE5 catalytic domain expressed and purified in insect cells using baculovirus was prepared in IMAP reaction buffer.
3) IMAP binding solution An IMAP binding reagent was prepared in IMAP reaction buffer.

(Preparation of test compound solution)
The test compound was dissolved in dimethyl sulfoxide.
(Test method and measurement method)
1) A test compound solution, a substrate solution, and an enzyme solution were added to a 96-well black plate at 10 μL per well (the final concentration of dimethyl sulfoxide was 3%).
2) Incubated for 40 minutes at 37 ° C in the dark
3) 60 μL of IMAP binding solution was added per well.
4) The fluorescence polarization was measured at 485 nm / 530 nm ex / em in each well with a multi-label reader (Victor 3 manufactured by Perkin Elma Co., Ltd.).
5) In place of the test compound, dimethyl sulfoxide was added, and the other operations were performed as described above, and the result was used as a control.
6) The IMAP reaction buffer was added instead of the enzyme solution, and the other operations were performed in the same manner as described above, and the result was used as the background.

(Calculation formula of PDE5 inhibition rate)
The PDE5 damage rate (%) was calculated by the following formula. The result was an average of double tests.
PDE5 inhibition rate (%) = {1− (fluorescence polarization value of test compound−background fluorescence polarization value) / (control fluorescence polarization value−background fluorescence polarization value)} × 100

(IC50 value calculation)
Data analysis was performed using statistical analysis software GraphPad Prism. The 50% inhibitory concentration (IC50) value was determined from a dose response curve in which enzyme activity was measured in the presence of a compound concentration of 0.001 to 100 mg / mL and then the compound concentration was plotted against the fluorescence polarization value.

(Evaluation results)
The IC50 of L-pyroglutamic acid was 100 μg / mL.
The IC50 of D-pyroglutamic acid was 116 μg / mL.
The inhibition rate of L-pyroglutamic acid at 2.0 mg / mL was 72.5%.
The harm rate of D-pyroglutamic acid at 2.0 mg / mL was 79.0%.
The inhibition rate of DL-pyroglutamic acid at 2.0 mg / mL was 66.9%.

(Verification of assay validity)
With a sildenafil known as a PDE5 inhibitor as the reference test compound, the IC50 value of 8.59 nM obtained by carrying out the same operation as the compound of the present application was within an acceptable range, indicating the validity of the assay.

  L-pyroglutamic acid, D-pyroglutamic acid and DL-pyroglutamic acid were all found to have an excellent PDE5 inhibitory action. However, from the evaluation results, it was found that it is desirable to contain L-pyroglutamic acid or D-pyroglutamic acid alone as an active ingredient.

Claims (3)

A pharmaceutical composition for inhibiting PDE5 comprising pyroglutamic acid as an active ingredient (excluding those used as a pharmaceutical composition for promoting in vivo nitric oxide synthase production) . A food composition for inhibiting PDE5 comprising pyroglutamic acid as an active ingredient (except for a food composition for promoting production of nitric oxide synthase in vivo) . The food composition for inhibiting PDE5 according to claim 2, which is labeled "Used for prevention or improvement of ED".

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