WO2025143530A1 - Pharmaceutical composition for preventing or treating depression comprising gv1001 - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating depression and, more specifically, to a pharmaceutical composition for preventing or treating depression, comprising a peptide having the amino acid sequence of SEQ ID NO: 1, whereby the composition reduces the concentration of stress hormones in the blood, alleviates various symptoms caused by depression, and is safe for the human body with minimal side effects including adverse reactions.

Description

Pharmaceutical composition for preventing or treating depression comprising GV1001

The present invention relates to a pharmaceutical composition for preventing or treating depression comprising GV1001.

Depression is one of the most common mental illnesses, with approximately 3.5% of the world's population, or 280 million people, experiencing depression in 2020, according to a WHO report.

In Korea, according to the 2021 Mental Health Survey by the National Center for Mental Health under the Ministry of Health and Welfare, the lifetime prevalence of depression was 7.7%. In addition, the socioeconomic loss due to depression is reported to amount to 2.525 trillion won and is on the rise.

Selective serotonin reuptake inhibitors (SSRIs), serotonin-norepinephrine reuptake inhibitors (SNRIs), serotonin-norepinephrine-dopamine reuptake inhibitors (SNDRIs), norepinephrine reuptake inhibitors (NRIs), and norepinephrine-dopamine reuptake inhibitors (NDRIs) are mainly used as antidepressants.

Although selective reuptake inhibitors are known to be relatively safe compared to monoamine oxidase inhibitors (MAOIs) or tetracyclic antidepressants (TeCAs), which are accompanied by serious side effects, it has been reported that they can still be accompanied by many side effects, such as sexual dysfunction, emotional dullness, glaucoma, and increased risk of suicide in children and adolescents.

Therefore, there is a need to develop a method that can treat depression more safely and effectively.

Accordingly, the inventors of the present invention conducted extensive research to develop a method for preventing or treating depression more safely and effectively, and as a result, they confirmed that when GV1001, whose safety has been proven, was administered to a mouse model of depression, various symptoms of depression were improved and the concentration of blood corticosterone was reduced, thereby completing the present invention.

The present invention aims to provide a pharmaceutical composition for preventing or treating depression.

The present invention aims to provide a kit for preventing or treating depression.

The purpose of the present invention is to provide a health functional food for preventing or improving depression.

1. A pharmaceutical composition for preventing or treating depression, comprising a peptide having an amino acid sequence of sequence number 1.

2. A kit for preventing or treating depression, comprising the pharmaceutical composition of the above 1; and an instruction sheet describing a method for preventing or treating depression.

3. A kit according to claim 2 above, wherein the method for preventing or treating depression comprises a step of administering a pharmaceutical composition to a subject who has developed or is at risk of developing depression.

4. A health functional food for preventing or improving depression, comprising a peptide having an amino acid sequence of sequence number 1.

When the pharmaceutical composition of the present invention was administered to a mouse model of depression, not only was the behavioral pattern improved in the behavioral evaluation, but the concentration of blood corticosterone was also reduced. Therefore, the pharmaceutical composition of the present invention is expected to be effective in the treatment, improvement, or prevention of depression.

The pharmaceutical composition and health functional food of the present invention are expected to be useful in preventing, improving or treating depression by including GV1001 as an active ingredient, which has been proven to have excellent safety by being shown to have no cytotoxicity or fatal side effects in numerous clinical trials.

Figure 1 schematically illustrates the schedule of a behavioral assessment experiment to confirm the therapeutic efficacy of GV1001 for depression.

Figure 2 shows the experimental results according to Example 2 (1).

Figure 3 shows the experimental results according to Example 2 (2).

Figure 4 shows the experimental results according to Example 2(3).

Figure 5 shows the experimental results according to Example 2 (4).

Figure 6 schematically illustrates the experimental schedule to confirm the preventive efficacy of GV1001 on depression.

Figure 7 shows the experimental results according to Example 2 (5).

The present invention provides a pharmaceutical composition for preventing or treating depression comprising a peptide having an amino acid sequence of sequence number 1.

In the present invention, the peptide having the amino acid sequence of SEQ ID NO: 1 includes a functional equivalent thereof. The term "functional equivalent" means a peptide exhibiting substantially the same physiological activity as the peptide having the amino acid sequence of SEQ ID NO: 1.

Depression is a mental disorder that is accompanied by symptoms such as low mood and avoidance of activities.

In the present invention, “depression” can be used interchangeably with “depressive disorder” and is used to collectively refer to major depressive disorder and dysthymia.

In the present invention, “prevention” means any act of suppressing or delaying depression.

In the present invention, "treatment" or "improvement" means any act of improving or beneficially altering the symptoms of a subject suspected of having depression or having developed depression. Therefore, a pharmaceutical composition for treating depression can be used interchangeably with "antidepressant" or "antidepressant."

The present applicant confirmed that by administering the pharmaceutical composition of the present invention to a mouse model of depression, the blood corticosterone concentration of the mouse decreased. Corticosterone is the main corticosteroid hormone of rats and mice and is a type of stress hormone, and the main adrenal stress hormone of humans is called cortisol.

In the present invention, stress hormone refers to corticosterone or cortisol.

The pharmaceutical composition of the present invention can prevent, improve or treat depression by reducing stress hormones in the blood of a subject.

The pharmaceutical composition of the present invention can improve various symptoms of depression, such as helplessness, depression, and anxiety.

In the present invention, “subject” means any animal, such as livestock or mice, that has developed or may develop depression, and may be a mammal, including a human.

The pharmaceutical composition of the present invention may be provided as a pharmaceutical composition containing the active ingredient alone or containing one or more pharmaceutically acceptable carriers, excipients or diluents.

The carrier, excipient or diluent that may be included in the pharmaceutical composition of the present invention may be, but is not limited to, lactose, dextrose, sucrose, dextrin, maltodextrin, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil.

The present invention provides a kit for preventing or treating depression, comprising a pharmaceutical composition for preventing or treating depression and instructions describing a method for preventing or treating depression.

In one embodiment, a method for preventing or treating depression may comprise administering a pharmaceutical composition of the invention to a subject suffering from or at risk of developing depression.

In the present invention, “administration” means introducing a predetermined substance to an individual by an appropriate method.

The route of administration of the pharmaceutical composition of the present invention may be oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual or rectal, but is not limited thereto.

In one embodiment, the composition of the present invention may be administered orally or parenterally.

When administering the composition of the present invention parenterally, it is preferable to select an injection method such as external skin application, intraperitoneal injection, intrarectal injection, subcutaneous injection, intravenous injection, intramuscular injection or intrathoracic injection, but is not limited thereto.

The pharmaceutical composition of the present invention may be a solid preparation for oral administration, such as a tablet, pill, powder, granule or capsule.

The pharmaceutical composition of the present invention may be a liquid preparation for oral administration, such as a suspension, a solution, an emulsion or a syrup.

The pharmaceutical composition of the present invention may be a preparation for parenteral administration, such as a sterile aqueous solution, a non-aqueous solution, a suspension, an emulsion, a lyophilized preparation or a suppository.

In one embodiment, a method for preventing or treating depression may comprise administering a pharmaceutically effective amount of a pharmaceutical composition of the present invention to a subject suffering from or at risk of developing depression.

In the present invention, “pharmaceutically effective amount” means an amount sufficient to treat depression at a reasonable benefit/risk ratio applicable to medical treatment.

A pharmaceutically effective amount of the pharmaceutical composition of the present invention can be determined based on factors including the severity of depression, activity of the pharmaceutical composition, sensitivity of the subject or patient to the pharmaceutical composition, time of administration, route of administration and excretion rate, duration of treatment and concomitant medication, and other factors well known in the medical field, and can be appropriately selected by a person skilled in the art.

The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or in multiple doses, which can be readily determined by a person skilled in the art.

The present invention provides a health functional food for preventing or improving depression, comprising a peptide having an amino acid sequence of sequence number 1.

The health functional food of the present invention refers to a food manufactured and/or processed in various forms to provide useful functionality to the human body.

The health functional food of the present invention can be included in various foods or medicines known in the art.

There is no particular limitation on the type of food that can contain the health functional food of the present invention. For example, the health functional food of the present invention can be contained in meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes.

The health functional food of the present invention includes all forms such as functional food, nutritional supplement, health food, and food additives, and these types of food can be manufactured in various forms according to conventional methods known in the art. For example, the health food can be manufactured in the form of a liquid drink and consumed, or can be granulated, encapsulated, spherical tableted (pills, etc.) and powdered and consumed, and can also be manufactured in the form of a powder, capsule, soft capsule, tablet, gum, or adhesive-type liquid composition and consumed. In addition, functional foods include beverages (including alcoholic beverages), fruits and processed foods thereof (e.g., canned fruits, bottled fruits, jams, marmalades, etc.), fish, meat and processed foods thereof (e.g., ham, sausages, corned beef, etc.), breads and noodles (e.g., udon, buckwheat noodles, ramen, spaghetti, macaroni, etc.), fruit juice, various drinks, cookies, taffy, dairy products (e.g., butter, cheese, etc.), edible vegetable oils, margarine, vegetable proteins, retort foods, frozen foods, seasonings, various seasonings (e.g., soybean paste, soy sauce, sauces, etc.).

The health functional food of the present invention may further include ingredients commonly added during food manufacturing, as long as it does not deviate from the ultimate purpose of the present invention, and may further include, for example, proteins, carbohydrates, fats, other nutrients, seasonings, and flavoring agents.

The health functional food of the present invention may additionally contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc.

The health functional food of the present invention may contain fruit pulp for the production of natural fruit juice, fruit juice beverage, and vegetable beverage. These ingredients may be used independently or in combination.

Hereinafter, the present invention will be described in detail by way of examples in order to specifically explain the present invention. However, the following examples are provided only to make it easier to understand the present invention, and the content of the present invention is not limited by the following examples.

Example

1. Synthesis of GV1001

A peptide consisting of 16 amino acids having the structural formula of the following chemical formula 1 selected from human telomerase (hereinafter referred to as GV1001) was synthesized:

[Chemical Formula 1]

GV1001 was synthesized by coupling amino acids one by one from the C-terminus according to the Fmoc solid phase peptide synthesis (SPPS) method using ASP48S (Peptron, Inc., Daejeon, Korea).

All amino acid starting materials used for peptide synthesis were used with the first amino acid at the C-terminus attached to the resin. Examples include:

NH ₂ -Lys(Boc)-2-chloro-Trityl Resin

NH ₂ -Ala-2-chloro-Trityl Resin

NH ₂ -Arg(Pbf)-2-chloro-Trityl Resin

All amino acid starting materials used for peptide synthesis were protected at the N-terminus with Fmoc, and all residues were protected with Trt, Boc, t-Bu (t-butylester), Pbf (2,2,4,6,7-pentamethyl dihydro-benzofuran-5-sulfonyl), etc., which can be removed by acid. For example, as follows:

Fmoc-Ala-OH, Fmoc-Arg(Pbf)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Pro-OH, Fmoc-Leu-OH, Fmoc-Ile-OH, Fmoc-Phe-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Lys(Boc)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Trp(Boc)-OH, Fmoc-Met-OH, Fmoc-Asn(Trt)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Ahx-OH, Trt-Mercaptoacetic acid.

HBTU [2-(1H-Benzotriazole-1-yl)-1,1,3,3-tetamethylaminium hexafluorophosphate] / HOBt [N-Hydroxxybenzotriazole] / NMM [4-Methylmorpholine] were used as coupling reagents. Piperidine in DMF (20% of total DMF) was used for Fmoc removal. Cleavage Cocktail [TFA (trifluoroacetic acid) / TIS (triisopropylsilane) / EDT (ethanedithiol) / H ₂ O = 92.5/2.5/2.5] was used to detach the synthesized peptide from the resin and remove the protecting group of the residue.

The peptide was synthesized by sequentially reacting each amino acid, washing with a solvent, and deprotecting it by repeating the process of using the starting amino acid with an amino acid protecting group attached to the solid support. The synthesized peptide was cleaved from the resin, purified by HPLC, confirmed to be synthesized by MS, and freeze-dried.

The specific synthesis process of GV1001 is as follows:

(1) Coupling: Protected amino acid (8 equivalents) and coupling reagents HBTU (8 equivalents)/HOBt (8 equivalents)/NMM (16 equivalents) were added to NH2 _- Lys(Boc)-2-chloro-Trityl resin, dissolved in DMF, and reacted at room temperature for 2 hours, and washed with DMF, MeOH, and DMF in that order.

(2) Fmoc deprotection: 20% piperidine in DMF was added and reacted twice for 5 minutes at room temperature, then washed with DMF, MeOH, and DMF in that order.

(3) Preparation of basic skeleton: The basic skeleton of the peptide was prepared by repeatedly performing reactions (1) and (2).

(4) Cleavage: The synthesized peptide resin was treated with a cleavage cocktail to separate the peptide from the resin.

(5) Precipitation: Cooling diethyl ether was added to the obtained mixture, and the resulting peptide was precipitated by centrifugation.

(6) Purification and powder production: After purification by prep-HPLC, the molecular weight was confirmed by LC/MS, and the product was frozen to produce a powder.

2. Confirmation of the efficacy of GV1001 in an animal model of depression

Among various depression-inducing methods such as stress, transformation, and nerve damage, the present inventors established an animal model of depression using chronic restraint stress (CRS). To this end, mice were restrained in a 50 mL conical tube for 3 hours every day for 2 weeks so that they could not move freely, thereby inducing stress-induced depression. In order to confirm the therapeutic efficacy of GV1001 on depression symptoms, GV1001 was administered via subcutaneous injection (SC injection) every day for 1 week starting from the 1st week of depression induction, and then 3 types of behavioral assessments were conducted (Fig. 1).

In addition, to confirm the preventive efficacy of GV1001 on depression, GV1001 was administered as a subcutaneous injection every day for 7 days, and CRS was administered for 2 days starting from the 5th day of GV1001 administration to induce depression. Then, blood was collected and the changes in the concentration of stress hormones in the blood were measured (Fig. 6).

(1) Confirmation of the efficacy of GV1001 through tail suspension test (TST)

Normal mice were set as the 'control group', and mice in which depression was induced by applying CRS for 3 hours every day for 2 weeks were set as the 'CRS group'. The group that was subjected to CRS for 2 weeks in the same way as the CRS group, but was administered GV1001 by subcutaneous injection once a day at a dose of 1 mg/kg for 1 week starting from 1 week after the start of CRS was set as the 'CRS+GV1001 group'.

As a result of conducting TST on the three groups, it was confirmed that the immobilization time of the CRS group significantly increased compared to the control group, indicating the development of depression, and the immobilization time of the CRS+GV1001 group significantly decreased compared to the CRS group, confirming that GV1001 has a therapeutic effect on depression (Fig. 2).

(2) Confirmation of the efficacy of GV1001 through sucrose preference test (SPT)

Example 2. (1) After setting the mice into three groups in the same manner, a sucrose preference experiment, a behavioral assessment method for measuring anhedonia, a major symptom of depression, was performed. To this end, the mice were exposed to water containing 1% sucrose for 24 hours and fasted for 24 hours. When the mice were simultaneously exposed to water containing 1% sucrose and regular water without it, the intake of the two types of water was compared to measure the preference for sucrose.

As a result, the sucrose preference of the CRS group decreased compared to the control group, and the sucrose preference of the CRS+GV1001 group increased compared to the CRS group, confirming the therapeutic effect of GV1001 on depression (Fig. 3).

(3) Confirmation of the efficacy of GV1001 through nest building test (NBT)

The following four groups of mice were prepared: a normal mouse group that was not administered anything; a normal mouse group that was administered GV1001; a group of mice in which depression was induced by CRS for 3 hours every day for 2 weeks; and a group in which depression was induced by CRS for 3 hours every day for 2 weeks, but GV1001 was administered by subcutaneous injection once a day at a dose of 1 mg/kg for 1 week starting from 1 week after the start of CRS (hereinafter referred to as the depressed mouse group administered GV1001).

In order to measure the level of anxiety, one of the representative symptoms of depression, a nest-building experiment was conducted on these four groups. To do this, mice were individually placed in cages containing a piece of Nestlet, and the nest-building score was measured on a five-point scale. According to the inherent characteristics of rodents, the greater the anxiety of the animal, the less likely it is to build a nest.

As a result, there was no significant difference in the nest building scores between the normal mouse group and the normal mouse group administered GV1001, but the scores of the mice induced with depression were significantly reduced. In the depressed mouse group administered GV1001, the scores significantly increased compared to the depressed mouse group, confirming the effect of GV1001 on depression (Fig. 4).

(4) Confirmation of the efficacy of GV1001 through Y-maze

Example 2. Four groups of mice were prepared in the same manner as in (3). Based on the natural behavior of rodents preferring to explore a new environment rather than a familiar environment, a Y-maze experiment was performed on these four groups to measure the spatial working memory ability of animals. To this end, the mice were allowed to freely explore the three arms of a Y-shaped maze with three plastic arms arranged at a 120 degree angle, and the number of times they entered an arm that they had not recently visited was measured.

As a result, there was no significant difference in the spontaneous alternation value (%) between the normal mouse group and the normal mouse group administered GV1001, but the value significantly decreased in the depressed mouse group. In the depressed mouse group administered GV1001, the value significantly increased compared to the depressed mouse group, confirming the effect of GV1001 on depression (Fig. 5).

(5) Confirmation of the efficacy of GV1001 through measurement of stress hormone levels

The following four groups of mice were prepared: normal mice that were not administered anything; normal mice administered GV1001; mice in which depression was induced by CRS for 3 hours every day for 2 days; and a group of mice in which depression was induced by CRS for 3 hours every day for 2 days, administered subcutaneously with GV1001 at a dose of 1 mg/kg once a day for 7 days, starting from the 5th day of administration (hereinafter referred to as the group of mice administered GV1001 before induction of depression).

Blood samples were collected from these four groups and blood corticosterone concentrations were measured.

As a result, there was no significant difference in the blood corticosterone concentration between the normal mouse group and the normal mouse group administered GV1001, but the concentration of corticosterone significantly increased in the mouse group in which depression was induced. In the mouse group administered GV1001 before depression was induced, the concentration of blood corticosterone statistically significantly decreased compared to the mouse group in which depression was induced. This confirmed that GV1001 has not only a therapeutic effect on depression but also a preventive effect (Fig. 7).

Claims (4)

A pharmaceutical composition for preventing or treating depression, comprising a peptide having an amino acid sequence of sequence number 1. A pharmaceutical composition according to claim 1; and A kit for the prevention or treatment of depression, comprising instructions describing a method for the prevention or treatment of depression. A kit according to claim 2, wherein the method for preventing or treating depression comprises a step of administering the pharmaceutical composition to a subject who has developed or is at risk of developing depression. A health functional food for preventing or improving depression, comprising a peptide having an amino acid sequence of sequence number 1.

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