Pharmaceutical composition comprising gv1001 for use in the prevention or treatment of depression

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating depression, and more particularly, to a pharmaceutical composition for preventing or treating depression by including a peptide having an amino acid sequence of SEQ ID NO: 1, reducing the concentration of stress hormones in blood, improving various symptoms caused by depression, being safe for human body, and having few side effects including adverse reactions.

Description

Technical Field

[0001] This invention relates to a pharmaceutical composition comprising GV1001 for the prevention or treatment of depression. Background Technology

[0002] Depression is one of the most common mental illnesses. According to a report by the World Health Organization (WHO), as of 2020, approximately 3.5% of the world's population, or about 280 million people, had experienced depression.

[0003] In South Korea, according to the 2021 Mental Health Facts Survey conducted by the National Center for Mental Health under the Ministry of Health and Welfare, the lifetime prevalence of depression is 7.7%. Furthermore, the socioeconomic losses caused by depression amount to 2.0525 trillion won, and this figure is reportedly on the rise.

[0004] As treatments for depression, 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 the main types of selective reuptake inhibitors used.

[0005] It is well known that selective reuptake inhibitors are relatively safer than monoamine oxidase inhibitors (MAOIs) or tetracyclic antidepressants (TeCAs) which have serious side effects, but they are still reported to have many side effects such as sexual dysfunction, emotional apathy, glaucoma, and increased risk of suicide in children and adolescents.

[0006] Therefore, there is a need to develop a safer method that can effectively treat depression.

[0007] To this end, the inventors conducted in-depth research to develop a safer method that can effectively prevent or treat depression. As a result, they confirmed that when the safety-verified GV1001 was administered to a mouse model of depression, various symptoms caused by depression were improved and the concentration of corticosterone in the blood was reduced, thus completing the present invention. Summary of the Invention

[0008] Problems to be solved

[0009] The present invention aims to provide a pharmaceutical composition for the prevention or treatment of depression.

[0010] The present invention aims to provide a kit for the prevention or treatment of depression.

[0011] The present invention aims to provide a health food for the prevention or improvement of depression.

[0012] Problem Solving Methods

[0013] 1. A pharmaceutical composition for the prevention or treatment of depression, comprising a peptide having an amino acid sequence having sequence number 1.

[0014] 2. A kit for the prevention or treatment of depression, comprising: the pharmaceutical composition of 1 above; and instructions for use, the instructions for use describing methods for the prevention or treatment of depression.

[0015] 3. The kit according to 2 above, wherein the prevention or treatment of depression includes the step of administering a pharmaceutical composition to an individual who has depression or is at risk of developing depression.

[0016] 4. A health food for the prevention or improvement of depression, comprising a peptide having an amino acid sequence having sequence number 1.

[0017] Invention Effects

[0018] The results of administering the pharmaceutical composition of the present invention to a mouse model of depression showed not only improvements in behavioral patterns in behavioral assessments, but also a reduction in blood corticosterone concentrations. Therefore, the pharmaceutical composition of the present invention shows promise for treating, improving, or preventing depression.

[0019] The pharmaceutical compositions and health functional foods of the present invention contain GV1001 as an active ingredient, which is expected to be used to prevent, improve or treat depression. GV1001 has been shown to have no cytotoxicity or fatal side effects in multiple clinical trials and has been proven to have excellent safety. Attached Figure Description

[0020] Figure 1This is a brief outline of the behavioral assessment trial schedule used to confirm the therapeutic efficacy of GV1001 for depression.

[0021] Figure 2 The experimental results according to Example 2(1) are shown.

[0022] Figure 3 The experimental results according to Example 2(2) are shown.

[0023] Figure 4 The experimental results according to Example 2(3) are shown.

[0024] Figure 5 The experimental results according to Example 2(4) are shown.

[0025] Figure 6 This outlines the experimental schedule used to confirm the preventive efficacy of GV1001 against depression.

[0026] Figure 7 The experimental results according to Example 2 (5) are shown. Detailed Implementation

[0027] The present invention provides a pharmaceutical composition for the prevention or treatment of depression, comprising a peptide having an amino acid sequence having sequence number 1.

[0028] In this invention, a peptide having the amino acid sequence number 1 includes its functional equivalent. "Functional equivalent" refers to a peptide that has substantially the same physiological activity as the peptide having the amino acid sequence number 1.

[0029] Depression is a mental disorder characterized by symptoms such as depressed mood and avoidance of activities.

[0030] In this invention, "depression" can be used interchangeably with "depressive disorder" and is used as a general term for severe depressive disorder and dysthymia disorder.

[0031] In this invention, "prevention" refers to any behavior that inhibits or delays depression.

[0032] In this invention, "treatment" or "improvement" refers to any action that improves or beneficially alters the symptoms of an individual suspected of having or already suffering from depression. Therefore, pharmaceutical compositions for treating depression may be used interchangeably with "depression treatment agent" or "antidepressant."

[0033] The applicant confirms that administering the pharmaceutical composition of this invention to a mouse model of depression reduces the concentration of corticosterone in the mouse blood. Corticosterone, as the main corticosteroid hormone in rats and mice, belongs to the stress hormones category, while the main adrenal stress hormone in humans is called cortisol.

[0034] In this invention, stress hormones refer to corticosterone or cortisol.

[0035] The pharmaceutical compositions of the present invention can prevent, improve or treat depression by reducing stress hormones in an individual's blood.

[0036] The pharmaceutical composition of the present invention can improve various symptoms caused by depression, such as weakness, depression, and anxiety.

[0037] In this invention, "individual" refers to any domestic animal or mouse that has or may have depression, such as mammals including humans.

[0038] The pharmaceutical compositions of the present invention may independently contain an active ingredient or be provided as pharmaceutical compositions containing one or more pharmaceutically acceptable carriers, excipients or diluents.

[0039] The carrier, excipient, or diluent that may be included in the pharmaceutical composition of the present invention may be lactose, glucose, sucrose, dextrin, maltodextrin, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylparaben, propylparaben, talc, magnesium stearate, or mineral oil, but is not limited thereto.

[0040] The present invention provides a kit for the prevention or treatment of depression, comprising a pharmaceutical composition for the prevention or treatment of depression and an instruction manual describing a method for the prevention or treatment of depression.

[0041] In one embodiment, a method for preventing or treating depression may include the step of administering the pharmaceutical composition of the present invention to an individual who already suffers from depression or is at risk of suffering from depression.

[0042] In this invention, "administration" refers to introducing a specific substance into an individual through appropriate methods.

[0043] The pharmaceutical compositions of the present invention can be administered via oral cavity, intravenous vein, intramuscular vein, intraarterial vein, intramedullary vein, intracardiac vein, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, local, sublingual, or rectal routes, but are not limited thereto.

[0044] In one embodiment, the composition of the present invention can be administered orally or non-orally.

[0045] When the compositions of the present invention are administered non-orally, it is preferred to use topical skin injection, intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection, or intrapleural injection, but not limited thereto.

[0046] The pharmaceutical compositions of the present invention can be solid dosage forms for oral administration, such as tablets, pills, powders, granules or capsules.

[0047] The pharmaceutical compositions of the present invention can be liquid formulations for oral administration, such as suspensions, oral liquids, emulsions, or syrups.

[0048] The pharmaceutical compositions of the present invention can be formulations for non-oral administration, such as sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, or suppositories.

[0049] In one embodiment, a method for preventing or treating depression may include administering a pharmaceutically effective amount of the pharmaceutical composition of the present invention to an individual who already suffers from depression or is at risk of suffering from depression.

[0050] In this invention, "pharmaceuticalally effective dose" refers to a dose that is suitable for medical treatment and adequately treats depression with a reasonable benefit / risk ratio.

[0051] The pharmaceutically effective dosage of the pharmaceutical composition of the present invention can be determined by factors including the severity of depression, the activity of the pharmaceutical composition, the sensitivity of the individual or patient to the pharmaceutical composition, the time of administration, the route of administration and the excretion rate, the duration of treatment, concomitant medications and other factors known in the medical field, and can be appropriately selected by those skilled in the art.

[0052] The pharmaceutical compositions of the present invention can be administered as a single therapeutic agent or in combination with other therapeutic agents, can be administered sequentially or simultaneously with existing therapeutic agents, and can be administered once or multiple times, as can be readily determined by those skilled in the art.

[0053] This invention provides a health food for preventing or improving depression, which contains a peptide having an amino acid sequence with sequence number 1.

[0054] The health functional food of this invention refers to food manufactured and / or processed in various forms to provide functional benefits to the human body.

[0055] The health-functional food of the present invention can be included in various foods or medicines known in the art.

[0056] There are no special limitations on the types of foods that contain the health-functional foods of the present invention. For example, the health-functional foods of the present invention can be contained in meat, sausage, bread, chocolate, candy, snacks, biscuits, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea, beverage preparations, alcoholic beverages, and vitamin complexes, etc.

[0057] The health-functional foods of the present invention include all forms such as functional foods, nutritional supplements, health foods, and food additives. These types of foods can be prepared in various forms according to conventional methods known in the art. For example, as health foods, they can be prepared as liquid beverages for consumption, or as granules, capsules, spherical tablets (pills, etc.) and powders for ingestion. They can also be prepared as powders, capsules, soft capsules, tablets, chewing gum, or viscous liquid compositions for ingestion. In addition, functional foods include beverages (including alcoholic beverages), fruits and their processed foods (such as canned fruit, bottled fruit, jam, orange marmalade, etc.), fish, meat and their processed foods (such as ham, sausage, corned beef, etc.), bread and noodles (such as udon noodles, buckwheat noodles, ramen, pasta, macaroni, etc.), fruit juices, various beverages, cookies, malt syrup, dairy products (such as butter, cheese, etc.), edible vegetable oils, margarine, vegetable protein, high-temperature sterilized foods, frozen foods, decoctions, and various seasonings (such as bean paste, soy sauce, sauces, etc.).

[0058] The health functional food of the present invention may also include ingredients that are usually added during food production, without departing from the ultimate purpose of the present invention. For example, it may also include proteins, carbohydrates, fats, other nutrients, seasonings and spices.

[0059] The health-functional food of the present invention may further contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, and carbonating agents used in carbonated beverages, etc.

[0060] The health-functional foods of this invention may contain fruit pulp used in the production of natural fruit juices, fruit juice beverages, and vegetable beverages. These ingredients may be used alone or in combination.

[0061] The following detailed description of embodiments illustrates the present invention. However, these embodiments are merely illustrative and are not intended to aid in a better understanding of the invention; the scope of the invention is not limited to these embodiments.

[0062] Example

[0063] 1. Synthesis of GV1001

[0064] Synthesize a 16-amino acid peptide (hereinafter GV1001) selected from human telomerase and having the following chemical formula 1:

[0065]

[0066] [Chemical Formula 1]

[0067] GV1001 was synthesized by coupling amino acids one by one from the C-terminus using ASP48S (Peptron, Inc., Daejeon, South Korea) according to the Fmoc solid phase peptide synthesis (SPPS) method.

[0068] All amino acid raw materials used for peptide synthesis are those in which the first amino acid at the C-terminus is attached to the resin. Examples include:

[0069] NH2-Lys(Boc)-2-chlorotrityl resin

[0070] NH2-Ala-2-chloro-Trityl Resin

[0071] NH2-Arg(Pbf)-2-chlorotrityl resin

[0072] All amino acid starting materials used in peptide synthesis are protected at the N-terminus by Fmoc, and the residues are protected by Tlt, Boc, tert-butyl ester (t-Bu), Pbf (2,2,4,6,7-pentamethyl dihydro-benzofuran-5-sulfonyl), etc., which are removed under all acids. Examples are as follows:

[0073] 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, F moc-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).

[0074] HBTU[2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethylaminium hexafluorophosphate] / HOBt[N-hydroxybenzotriazole] / NMM[4-methylmorpholine] was used as the coupling reagent. A solution of piperidine in 20% DMF was used for Fmoc removal. The synthesized peptides were isolated from the resin using a cleavage cocktail [TFA (trifluoroacetic acid) / TIS (triisopropylsilane) / EDT (ethanedithiol) / H2O = 92.5 / 2.5 / 2.5 / 2.5] and the protecting groups of the residues were removed.

[0075] By utilizing the state of starting amino acids with protecting groups bound to a solid support, each amino acid is reacted sequentially, followed by deprotection after solvent washing. This process is repeated to synthesize peptides. After cleaving the synthesized peptides from the resin, they are purified by high-performance liquid chromatography (HPLC), and the synthesis is confirmed by mass spectrometry (MS). Finally, they are lyophilized.

[0076] The specific synthesis process of GV1001 is as follows:

[0077] (1) Coupling: The amino acid (8 equivalents) protected in NH2-Lys(Boc)-2-chlorotriphenylmethyl resin and the coupling reagent HBTU (8 equivalents) / HOBt (8 equivalents) / NMM (16 equivalents) were dissolved in DMF and added. The mixture was reacted at room temperature for 2 hours and washed with DMF, MeOH and DMF in sequence.

[0078] (2) Fmoc deprotection: Add piperidine in 20% DMF solution, react twice at room temperature for 5 minutes, and wash with DMF, MeOH and DMF in sequence.

[0079] (3) Preparation of the basic skeleton: Repeat the reactions in steps (1) and (2) to prepare the basic skeleton of the peptide.

[0080] (4) Cleavage: The peptide chain is treated with resin after synthesis to cleave the mixture, thereby separating the peptide from the resin.

[0081] (5) Precipitation: After adding cooled diethyl ether to the resulting mixture, the precipitated peptides are separated by centrifugation.

[0082] (6) Purification and powder preparation: After purification by preparative high performance liquid chromatography (Prep-HPLC), the molecular weight was confirmed by liquid chromatography-mass spectrometry (LC / MS), and the powder was prepared by freezing.

[0083] 2. Confirmation of the efficacy of GV1001 in an animal model of depression

[0084] In this invention, the inventors established an animal model of depression using chronic restraint stress (CRS) among various methods for inducing depression, including stress, transgenics, and nerve damage. Mice were placed in 50 mL conical tubes and restrained for 3 hours daily for 2 weeks to restrict their free movement, thereby inducing stress-induced depression. To confirm the therapeutic efficacy of GV1001 on depressive symptoms, GV1001 was administered daily via subcutaneous injection (SC injection) starting in the first week after induction of depression. After one week, three types of behavioral assessments were performed. Figure 1 ).

[0085] In addition, to confirm the preventive efficacy of GV1001 against depression, GV1001 was administered subcutaneously daily for 7 days. Starting on the 5th day of GV1001 administration, depression was induced by administering CRS for two days. Blood samples were then collected to detect changes in the concentration of stress hormones in the blood. Figure 6).

[0086] (1) Confirmation of the efficacy of GV1001 through tail suspension test (TST)

[0087] Normal mice were designated as the "control group," and mice were given CRS for 3 hours daily for 2 weeks. Mice that developed depression were designated as the "CRS group." The mice were given CRS for two weeks in the same manner as the CRS group, but one week after the start of CRS, they were given GV1001 once daily via subcutaneous injection at a dose of 1 mg / kg for one week. This group was designated as the "CRS+GV1001 group."

[0088] The results of TST studies on three groups confirmed that, compared with the control group, the immobilization time was significantly prolonged in the CRS group, leading to depression. Conversely, compared with the CRS group, the immobilization time was significantly shortened in the CRS+GV1001 group, confirming the therapeutic effect of GV1001 on depression. Figure 2 ).

[0089] (2) Confirmation of the efficacy of GV1001 through the sucrose preference test (SPT)

[0090] Example 2: Similar to (1), mice were divided into three groups and subjected to a sucrose preference test, a behavioral assessment method used to measure the main symptom of depression—anhedonia. For this purpose, mice were exposed to water containing 1% sucrose for 24 hours and then fasted for 24 hours. The intake of the two types of water was compared when mice were simultaneously exposed to water containing 1% sucrose and plain water without sucrose, thus measuring their preference for sucrose.

[0091] The results showed that, compared with the control group, the CRS group had a decreased sucrose preference, while the CRS+GV1001 group had an increased sucrose preference compared with the CRS group, thus confirming the therapeutic effect of GV1001 on depression. Figure 3 ).

[0092] (3) Confirmation of the efficacy of GV1001 through the nest building test (NBT).

[0093] The following four groups of mice were prepared: normal mice without drug administration; normal mice administered GV1001; mice subjected to CRS for 3 hours daily for 2 weeks to induce depression; and mice subjected to CRS for 3 hours daily for 2 weeks to induce depression, and then administered GV1001 once daily via subcutaneous injection at a dose of 1 mg / kg for 1 week after the start of CRS (hereinafter referred to as the GV1001-administered depressed mouse group).

[0094] Nesting experiments were conducted on these four groups of mice to assess anxiety levels, a representative symptom of depression. Each mouse was placed individually in a cage containing a piece of compressed nesting material, and its nesting score was assessed using a five-point rating scale. Based on inherent characteristics of rodents, higher anxiety levels correlated with a lower likelihood of nesting.

[0095] The results showed no significant difference in nesting scores between the normal mouse group and the GV1001-treated normal mouse group, but the nesting scores of mice with induced depression were significantly reduced. In the GV1001-treated depressed mouse group, the nesting scores were significantly higher than those in the induced depression group, thus confirming the effect of GV1001 on depression. Figure 4 ).

[0096] (4) Confirmation of the efficacy of GV1001 through the Y-maze test

[0097] Similar to Example 2.(3), four groups of mice were prepared. Based on the natural behavior of rodents, which tend to explore new environments rather than familiar ones, a Y-maze experiment was conducted on these four groups of mice to measure their spatial working memory. For this purpose, three plastic arms (Arm) were arranged in a Y-shaped maze at an angle of 120 degrees, and the mice were allowed to freely explore the three plastic arms. The number of times the mice entered an arm that had not been visited recently was measured.

[0098] The results showed no significant difference in the spontaneous alternation rate (%) between the normal mouse group and the normal mouse group treated with GV1001, while the rate was significantly lower in the mouse group with induced depression. In the GV1001-treated depressed mouse group, the rate was significantly higher compared to the mouse group with induced depression, thus confirming the effect of GV1001 on depression. Figure 5 ).

[0099] (5) The efficacy of GV1001 was confirmed by measuring stress hormone levels.

[0100] The following four groups of mice were prepared: a normal mouse group that did not receive the drug; a normal mouse group that received GV1001; a mouse group that received CRS for 3 hours daily for 2 days to induce depression; and a mouse group that received GV1001 once daily via subcutaneous injection at a dose of 1 mg / kg for 7 days, but received CRS for 3 hours daily for 2 days starting from day 5 of administration to induce depression (hereinafter referred to as the mouse group that received GV1001 before inducing depression).

[0101] Blood samples were collected from these four groups of mice, and the concentration of corticosterone in the blood was measured.

[0102] The results showed no significant difference in blood corticosterone concentration between the normal mouse group and the normal mouse group treated with GV1001. However, in the mouse group treated with induced depression, blood corticosterone concentration was significantly increased. In the mouse group treated with GV1001 before induction of depression, blood corticosterone concentration was statistically significantly lower than in the mouse group treated with induced depression. This confirms that GV1001 not only has a therapeutic effect on depression but also a preventive effect. Figure 7 ).

Claims

1. A pharmaceutical composition for the prevention or treatment of depression, comprising a peptide having an amino acid sequence having sequence number 1.

2. A kit for the prevention or treatment of depression, comprising: The pharmaceutical composition according to claim 1; and The instruction manual describes methods for preventing or treating depression.

3. The kit according to claim 2, wherein, The prevention or treatment of depression includes the step of administering the pharmaceutical composition to an individual who already has depression or is at risk of developing depression.

4. A health food for the prevention or improvement of depression, comprising a peptide having an amino acid sequence having sequence number 1.

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