Novel use of nonapeptide

Abstract

Provided is a novel use of a peptide comprising the amino acid sequence of SEQ ID NO: 1 or a pharmaceutically acceptable salt thereof. According to the present invention, the composition has the advantage of being capable of effectively treating, preventing, improving and / or inhibiting one or more of obesity or obesity complications.

Description

(19) State Intellectual Property Office (12) Invention Patent Application (10) Application Publication Number (43) Application Publication Date (21) Application Number 202480048869.5 (22) Application Date 2024.08.12 (30) Priority Data 10-2023-0111132 2023.08.24 KR (85) PCT International Application Entering National Phase Date 2026.01.23 (86) PCT International Application Application Data PCT / KR2024 / 012000 2024.08.12 (87) PCT International Application Publication Data WO2025 / 042118 KO 2025.02.27 (71) Applicant: Insoo Bioscience Co., Ltd. Address: South Korea (72) Inventors: Kim Hae-jin, Moon Eun-jung, Lee Young-hoon (74) Patent Agency: Beijing Linda Liu Intellectual Property Agency (General Partnership) 11277 Patent Agent Li Maojia Yan Junping (51) Int.Cl. A61K 38 / 08 (2019.01) A61P 3 / 04 (2006.01) A23L 33 / 18 (2016.01) (54) Invention Title Novel Use of Nonapeptide (57) Abstract This invention provides a novel use of a peptide composed of the amino acid sequence of sequence number 1 or a pharmaceutically acceptable salt thereof. According to the invention, it has the advantage of being able to effectively treat, prevent, improve and / or inhibit one or more of obesity or obesity complications. Claims 1 page Description 11 pages Sequence Listing (electronic publication) Drawings 2 pages CN 121646471 A 2026.03.10 CN 1 21 64 64 71 A 1. A pharmaceutical composition comprising a peptide composed of the amino acid sequence of sequence number 1 or a pharmaceutically acceptable salt thereof as an active ingredient for treating or preventing obesity. 2. A food composition for improving or preventing obesity, comprising a peptide consisting of an amino acid sequence of sequence number 1 or a pharmaceutically acceptable salt thereof as an active ingredient. 3. The food composition according to claim 2, wherein the food composition is a health-functional food composition. 4. A method for treating, preventing, improving or inhibiting obesity, comprising: administering to a non-human animal a peptide consisting of an amino acid sequence of sequence number 1 or a pharmaceutically acceptable salt thereof. Claims 1 / 1 page 2 CN 121646471 A New Use of Nonapeptide Technical Field

[0001] The present invention relates to a new use of a nonapeptide, and more specifically, to a new use of a peptide consisting of an amino acid sequence of sequence number 1 or a pharmaceutically acceptable salt thereof. Background Art

[0002] Overweight and obesity are typical adult metabolic diseases and are also considered diseases (see Powell-Wiley™, Poirier P,Burke LE, Despres JP, Gordon-Larsen P, Lavie CJ, Lear SA, Ndumele CE, Neeland IJ, Sanders P, St-Onge MP. "Obesity and cardiovascular disease: A scientific statement from the American Heart Association." Circulation. 2021 May 25;143(21):e984-e1010). Due to advancements in healthcare and improved living standards, life expectancy has increased. Urban-centric population growth has led to decreased physical activity, resulting in a significant increase in overweight and obesity. Obesity can lead to obesity-related complications, including hypertension, high LDL-C, low HDL-C, or high triglycerides (dyslipidemia), type 2 diabetes, coronary heart disease, stroke, gallbladder disease, osteoarthritis (destruction of articular cartilage and bone), sleep apnea and breathing problems, cancer, depression, anxiety and other mental disorders, body pain and physical dysfunction (see Bluher M. "Obesity: global epidemiology and pathogenesis" Nat Rev Endocrinol. 2019 May;15(5):288-298, Whitlock G, Lewington S, Sherliker P, Clarke R, Emberson J, Halsey J, Qizilbash N, Collins R, Peto R. “Body-mass index and cause-specific mortality in 900,000 adults: collaborative analyses of 57 prospective studies” Lancet. 2009 Mar 28;373(9669):1083-96.).

[0003] Furthermore, studies using mice with induced obesity have confirmed that weight loss through dietary control can normalize glucose metabolism and reverse stroke caused by middle cerebral artery occlusion (see Karampatsi D, Zabala A, Wilhelmsson U, Dekens D,Vercalsteren E, Larsson M, Nystrom T, Pekny M, Patrone C, Darsalia V. “Diet-induced weight loss in obese / diabetic mice normalizes glucose metabolism and promotes functional recovery after stroke” Cardiovasc Diabetol. 2021 Dec 22;20(1):240). Furthermore, clinical trials have demonstrated that weight loss is effective in lowering blood pressure in overweight and hypertensive patients, indicating a direct role in the treatment and prevention of obesity complications (see Reisin E, Abel R, Modan M, Silverberg DS, Eliahou HE, Modan B. “Effect of weight loss without salt restriction on the reduction of blood pressure in overweight hypertensive patients” N Engl J Med. 1978 Jan 5;298(1):1-6.).

[0004] Therefore, there is a need to develop technologies capable of addressing overweight and obesity, as well as obesity complications.

[0005] On one hand, the inventors have previously developed a nonapeptide {a peptide consisting of an amino acid sequence of sequence number 1 (an amino acid sequence consisting of 9 amino acids; YGAGAGAGY) or a pharmaceutically acceptable salt thereof}. It is known that the nonapeptide is effective against pancreatitis and even type 1 diabetes (see International Patent Publication No. WO 2020 / 184901 A1). The inventors have been working to find new uses for this effective nonapeptide. However, when using the nonapeptide to treat type 1 diabetes, some individuals have experienced weight gain, which makes it unpredictable to apply the nonapeptide to obesity requiring weight loss. (See specification 1 / 11 page 3 CN 121646471 A)

[0006] [Prior Art Documents]

[0007] [Patent Documents]

[0008] (Patent Document 1) International Patent Application Publication No. WO 2020 / 184901 A1, 2020.9.17, Description

[0009] [Non-Patent Documents]

[0010] (Non-Patent Document 1) Powell-Wiley™, Poirier P, Burke LE, Despres JP, Gordon-Larsen P, LavieCJ , Lear SA, Believe me CE, Neeland IJ , Sanders P, St‐ Onge MP. "Obesity and Cardiovascular Disease: A Scientific Statement From the American Heart Association." Circulation. 2021 May 25;143(21):e984‐e1010

[0011] (Extracellular Reagent2)Bluher M . "Obesity: global epidemiology and pathogenesis" Nat Rev Endocrinol. 2019 May;15(5):288‐298

[0012] Whitlock G, Lewington S, Sherliker P, Clarke R, Emberson J, Halsey J, Qizilbash N, Collins R, Peto R . “Body‐mass index and cause‐specific mortality in 900 ,000 adults: collaborative analyzes of 57 prospective studies” Lancet. 2009 Mar 28;373(9669):1083‑96

[0013] (Experimental Issue 4)Karampatsi D , Zabala A , Wilhelmsson U , Dekens D , Vercalsteren E, Larsson M, Nystrom T, Pekny M, Patrone C, Darsalia V. 2009 Mar 28; “Diet‐induced weight loss in obese / diabetic mice normalizes glucose metabolism and promotes functional recovery after stroke”Cardiovasc Diabetol. 2021 Dec 22;20 (1):240

[0014] (Exploitation 5)Reisin E , Abel R , Modan M , Silverberg DS, Eliahou HE , Modan B. “Effectof weight loss without salt restriction on the reduction of blood pressure in overweight hypertensive patients” N Engl J Med. 1978 Jan 5; 298(1):1-6 Summary of the Invention

[0015] Technical Problem

[0016] The problem to be solved by the present invention is to provide a novel use of a peptide composed of an amino acid sequence of sequence number 1 or a pharmaceutically acceptable salt thereof.

[0017] The problem of the present invention is not limited to the problem mentioned herein, and other unmentioned problems will also be clearly understood by those skilled in the art through the following description.

[0018] Technical Solution

[0019] The inventors of the present invention have accidentally discovered that a peptide composed of an amino acid sequence of sequence number 1 (YGAGAGAGY) or a pharmaceutically acceptable salt thereof (also known as a nona-peptide; nona represents the number 9) is effective for obesity or obesity complications, thereby completing the present invention.

[0020] In the amino acid sequence, “Y” represents tyrosine (Tyr), and “G” represents glycine (Glycine; Gly), “A” represents alanine (Ala).

[0021] The amino acids constituting the peptide include L-form, D-form, and DL-form, and the amino acids constituting the peptide of the present invention include all of these types. Furthermore, it is obvious that “Y” can be interpreted as including all amino acids called tyrosine or 4-hydroxyphenylalanine.

[0022] The peptide contains variants, including variants that, through natural or artificial variation, alter a portion of the structure of the peptide of the present invention, but do not alter its principal activity.

[0023] Examples of pharmaceutically acceptable salts include, for example, hydrochloride, sulfate, phosphate, acetate, citrate, tartrate, succinate, lactate, maleate, fumarate, oxalate, methanesulfonate, trifluoromethanesulfonate, benzenesulfonate, p-toluenesulfonate, sodium salt, potassium salt, calcium salt, etc.

[0024] This invention provides a peptide composed of the amino acid sequence of sequence number 1 or a pharmaceutically acceptable salt thereof for medicinal and / or food use in treating, preventing, improving, and / or inhibiting one or more selected from obesity or obesity complications. "Treatment" includes the improvement or reduction of symptoms associated with obesity or obesity complications, and "prevention" includes inhibiting the development of the disease from its pre-disease stage.

[0025] As a specific example, this invention provides a peptide for treating or preventing one selected from obesity or obesity complications.The above pharmaceutical composition comprises a peptide consisting of the amino acid sequence of sequence number 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0026] The obesity includes overweight.

[0027] The obesity complications are not limited to obesity-related complications, and may preferably be selected from one or more of the following: hypertension, high low-density lipoprotein cholesterol (LDL-C), low high-density lipoprotein cholesterol (HDL-C) or high triglycerides (dyslipidemia), type 2 diabetes, coronary heart disease, stroke, gallbladder disease, osteoarthritis (destruction of articular cartilage and bone), sleep apnea, breathing problems, cancer, depression, anxiety, mental illness, body pain, physical dysfunction, glucose metabolism disorders, stroke (preferably stroke caused by middle cerebral artery occlusion), and hypertension (preferably hypertension in overweight hypertensive patients). This is because it is well known that obesity (including overweight) and its complications, as described above, can be treated, prevented, improved, and / or suppressed (see Bluher M. "Obesity: global epidemiology and pathogenesis" Nat Rev Endocrinol. 2019 May;15(5):288- 298; Whitlock G , Lewington S, Sherliker P, Clarke R , Emberson J , Halsey J , Qizilbash N, Collins R, Peto R. “Body-mass index and cause-specific mortality in 900,000 adults: collaborative analyses of 57 prospective studies” Lancet. 2009 Mar 28;373(9669):1083-96; Karampatsi D, Zabala A, Wilhelmsson U, Dekens D , Vercalsteren E , Larsson M , Nystrom T , Pekny M , Patrone C, Darsalia V. “Diet‑induced weight loss in obese / diabetic mice normalizes glucose metabolism and promotes functional recovery after stroke” Cardiovasc Diabetol.2021 Dec 22;20(1):240; Reisin E, Abel R, Modan M, Silverberg DS, Eliahou HE, Modan B. “Effect of weight loss without salt restriction on the reduction of blood pressure in overweight hypertensive patients” N Engl J Med . 1978 Jan 5; 298(1):1-6).

[0028] In addition, the composition may further include pharmaceutically acceptable additives, and the composition consists of the peptide or its pharmaceutically acceptable salt and the additives.

[0029] The peptide can be prepared using methods commonly used in the field of peptide synthesis. For example, the peptide can be prepared using known methods such as liquid-phase synthesis or solid-phase synthesis.

[0030] Examples of methods for forming peptide bonds include methods for activating carboxylic acid groups such as acyl azide method, acyl halide method, acyl imidazole method, carbodiimide method, phosphonium method, acid anhydride method, and mixed acid anhydride method; methods for direct dehydration condensation using a condensing agent (see page 3 / 11, CN 121646471 A); redox methods; and methods using Woodward reagent K, etc.

[0031] Before the condensation reaction, unreacted carboxyl groups, amino groups, etc., can be protected, and carboxyl groups participating in the condensation reaction can be activated using methods known in the art.

[0032] Examples of groups for protecting carboxyl groups include ester groups such as methyl, tert-butyl, aryl, pentafluorophenyl, benzyl, p-methoxybenzyl, or methoxyethoxymethyl.

[0033] Examples of groups protecting the amino group include triphenylmethyl carbonyl, aryloxycarbonyl, cyclohexyloxycarbonyl, trichloroethoxycarbonyl, benzyloxycarbonyl, tert-butyloxycarbonyl and / or 9-fluorenemethoxycarbonyl, etc.

[0034] Examples of active forms of the carboxyl group include mixed anhydrides, azides, acyl chlorides or active esters [esters with alcohols (e.g., pentachlorophenol, 2,4-dinitrophenol, cyanoethanol, p-nitrophenol, N-hydroxy-5-norbornene-2,3-dicarboxylic acid imine, N-hydroxysuccinimide, N-hydroxyphthalamide or 1-hydroxybenzotriazole)], etc.

[0035] Solvents that can be used in the condensation reaction for forming peptide bonds can be single solvents or mixtures thereof, such as benzene, toluene, hexane, acetone, nitromethane, cyclohexane, diethyl ether, chloroform, dichloromethane, ethyl acetate, N,N-dimethylformamide, dimethyl sulfoxide, pyridine, dioxane, tetrahydrofuran, water, methanol, or ethanol.

[0036] The reaction temperature can be in the range of approximately -70°C to 100°C, which is commonly used in reactions, and more preferably in the range of -30°C to 30°C.

[0037] The reaction for removing the peptide protecting group varies depending on the type of protecting group, but it can be removed using acidic compounds, basic compounds, or transition metals that do not affect the peptide bond.

[0038] The protecting group can be removed by acid treatment, for example by treatment with hydrogen chloride, hydrogen bromide, hydrogen fluoride, acetic acid, methanesulfonic acid, trifluoromethanesulfonic acid, trifluoroacetic acid, trimethylchlorosilane, or mixtures thereof.

[0039] When carrying out the reaction for removing the protecting group using the acid, the reaction can be promoted by adding anisole, phenol, and thioanisole, etc., as auxiliaries.

[0040] The protecting group can be removed by base treatment, for example by treatment with ammonia, diethylamine, hydrazine, morpholine, N-methylpyrrolidine, piperidine, sodium carbonate, or mixtures thereof.

[0041] In addition, the protecting group can be removed by treatment with transition metals, for example by treatment with zinc, mercury, palladium / hydrogen, etc.

[0042] After the reaction is complete, the peptide can be purified by conventional peptide purification methods, such as extraction, layering, solid-phase precipitation, recrystallization, or column chromatography.

[0043] In addition, the peptide composed of the amino acid sequence number 1 can be converted into a variant or a pharmaceutically acceptable salt thereof by conventional methods.

[0044] The peptide can be synthesized using an automated peptide synthesizer or produced using genetic engineering techniques. For example, a fusion gene encoding a fusion protein composed of a fusion chaperone and the peptide of the present invention can be manufactured by genetic engineering. After the host microorganism is transformed using the fusion gene, it is expressed in the host microorganism as a fusion protein. Then, the peptide of the present invention is cleaved and isolated from the fusion protein using a protease or compound to produce the desired peptide.

[0045] The peptide or a pharmaceutically acceptable salt thereof can be mainly administered by non-oral methods, such as intravenous or subcutaneous injection, intraspinal administration, transdermal administration, nasal administration, or rectal administration. In addition, oral administration is of course possible as appropriate.

[0046] The peptide or its pharmaceutically acceptable salt or composition can be formulated with pharmaceutically acceptable additives to form injections, suppositories, powders, nasal drops, granules, tablets, transdermal patches, etc.

[0047] Pharmaceutically acceptable additives can be applied based on various factors known to those skilled in the art, such as the specific bioactive substance used, its concentration, stability and expected bioavailability; the disease or ailment to be treated; the individual to be treated, age, size and general condition; the route of administration of the composition, such as nasal, oral, ocular, topical, transdermal and intramuscular, etc., but not limited to these. In addition to oral administration, the physiologically active substance is usually administered via...Pharmaceutically acceptable additives used in the medicine include D5W (5% glucose in water), glucose, and aqueous solutions containing physiological saline at a volume of no more than 5%. When injected locally into the lesion site, various injectable hydrogels can be used to enhance efficacy and prolong duration. Additionally, pharmaceutically acceptable additives may also contain preservatives and antioxidants that enhance the stability of the active ingredient. The peptides of the present invention, or their pharmaceutically acceptable salts or compositions, can be formulated using appropriate methods in the relevant art, and reference can be made to well-known literature in the relevant art.

[0048] These peptides can be preserved using physiological saline, can be freeze-dried in ampoules after the addition of mannitol or sorbitol, and can be dissolved in physiological saline or the like when used for administration.

[0049] Furthermore, the present invention provides a food composition for improving or preventing one or more of obesity or obesity complications, the food composition comprising a peptide or its pharmaceutically acceptable salt consisting of the amino acid sequence of sequence number 1 as an active ingredient.

[0050] The food composition can be included in various foods, including beverages, and can be in the form of beverages, chewing gum, tea, health functional foods, etc. The health functional foods can be formulated into dosage forms such as tablets and capsules. The health functional foods refer to foods manufactured (including processed; hereinafter the same) using raw materials or ingredients with functions beneficial to the human body in accordance with the laws related to health functional foods in Korea. "Functionality" refers to the effect of adjusting nutrients for human structure and function or having effects beneficial to health purposes, such as physiological effects.

[0051] In addition to the active ingredient, the food composition can contain other ingredients as necessary ingredients without special restrictions. For example, ordinary beverages can contain various flavoring agents or natural carbohydrates as additional ingredients. Examples of the above-mentioned natural carbohydrates can be monosaccharides, such as glucose, fructose, etc.; disaccharides, such as maltose, sucrose, etc.; and polysaccharides, such as common sugars such as dextrin, cyclodextrin, etc., and sugar alcohols such as xylitol, sorbitol, erythritol, etc. In addition to the flavoring agents mentioned above, natural flavoring agents (such as samaram, stevia extract (e.g., leboside A, glycyrrhizic acid, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. Those skilled in the art can appropriately determine the proportion of the natural carbohydrates. Besides the above-mentioned components, the food composition may contain various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavoring agents, coloring agents and thickeners (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. These components can be used alone or in combination, and the proportions of these additives can be appropriately selected by those skilled in the art.

[0052] Unless otherwise specified, the food composition is equally applicable to the pharmaceutical composition of the present invention, unless it contradicts the matters mentioned in the pharmaceutical composition of the present invention. The term "improvement" is included in "treatment," meaning an improvement in the condition or symptoms. The term "prevention" includes "inhibition," meaning preventing the occurrence of a condition or symptoms.

[0053] The peptide consisting of the amino acid sequence of sequence number 1 or its pharmaceutically acceptable salt can be formulated by adding pharmaceutically or food-acceptable carriers, excipients, or diluents, etc., for details of formulation, as well as well-known literature in the relevant art.

[0054] Therefore, the composition of the present invention may also contain pharmaceutically acceptable or food-acceptable additives, and may consist of the active ingredient and the pharmaceutically acceptable or food-acceptable additives.

[0055] The composition of the present invention may contain 0.01 to 99.99% by weight of the active ingredient.

[0056] The present invention also provides a method for treating, preventing, improving, or inhibiting one or more selected from obesity or obesity complications, as described on page 5 / 11 of the specification, CN 121646471 A, the method comprising administering to an animal (including human animals or animals excluding human animals) a peptide consisting of the amino acid sequence of sequence number 1 or a pharmaceutically acceptable salt thereof.

[0057] The animal may be a mammal.

[0058] The animal may be an animal for which administration of the peptide consisting of the amino acid sequence of sequence number 1 or a pharmaceutically acceptable salt thereof is required.

[0059] The animal may be an animal that suffers from or has the potential to suffer from one or more selected from obesity or obesity complications.

[0060] Furthermore, the administered peptide consisting of the amino acid sequence of sequence number 1 or a pharmaceutically acceptable salt thereof may be an effective amount of the peptide consisting of the amino acid sequence of sequence number 1 or a pharmaceutically acceptable salt thereof.

[0061] Furthermore, the present invention also provides the use of the peptide consisting of the amino acid sequence of sequence number 1 or a pharmaceutically acceptable salt thereof for the preparation of an formulation for improving, inhibiting, treating, and / or preventing one or more selected from obesity or obesity complications. The formulation may be a pharmaceutical preparation and / or a food preparation.

[0062] Unless otherwise stated, the references to the pharmaceutical compositions, food compositions, methods, and uses of the present invention are identical in application unless they contradict each other.

[0063] The dosage of the peptide or its pharmaceutically acceptable salt contained in or used in the compositions of the present invention is, for non-oral administration, 0.0243 mg / day to 4860 mg / day, preferably 0.0486 mg / day to 2430 mg / day. The dosage for oral administration is 5 to 10 times the non-oral dosage. It may also be administered once daily or in multiple divided doses as described above.The dosage can be based on an adult (60 kg), but can naturally vary depending on weight, physical condition, etc. On the other hand, when administering the drug to animals other than humans, the dosage can naturally vary depending on the animal species, which is derived from the well-known conversion method based on body surface area to calculate the human equivalent dose (HED) (see USFDA. Guidance for Industry: Estimating the Maximum Safe Starting Dose in Adult Healthy Volunteer. Rockville, MD: US Food and Drug Administration; 2005.).

[0064] Effects of the Invention

[0065] According to the present invention, it is possible to effectively treat, prevent, improve and / or inhibit one or more of obesity or obesity complications. Brief Description of the Drawings

[0066] Figure 1 is a graph showing the change in weight loss effect over time.

[0067] Figure 2 is a graph showing the weight loss effect of each experimental group. Detailed Description of the Embodiments

[0068] The advantages, features and implementation methods of the present invention will become clearer below with reference to the embodiments described in detail with reference to the accompanying drawings. However, the present invention is not limited to the embodiments disclosed below, but can be implemented in various different forms. The embodiments provided are only for the purpose of fully disclosing the present invention and fully elucidating the scope of the invention to those skilled in the art. The present invention is defined only by the scope of the claims.

[0069] Throughout the specification, "and / or" includes each and more than one combination of all the mentioned constituent elements.

[0070] The terminology used in this specification is only for describing embodiments and is not intended to limit the invention. In this specification, unless otherwise expressly stated, the singular form also includes the plural form. The use of "comprises" and / or "comprising" in the specification does not exclude the presence or addition of more than one other constituent element and / or step mentioned in the specification.

[0071] Hereinafter, the examples have demonstrated the effectiveness of peptides composed of the amino acid sequence of sequence number 1, and even their salts, against obesity and obesity complications using obesity-induced experimental animals.

[0072] The reagents used in the examples are the highest quality commercially available reagents, purchased from Sigma, etc.

[0073] <Example 1> Confirmation of the effects of peptides on improving, treating, inhibiting and / or preventing obesity / obesity complications

[0074] 1-1. Preparation of peptides

[0075] The peptides listed in Table 1 below were prepared using a solid-phase peptide synthesis method. Specifically, Fmoc (9-fluorenemethyloxycarbonyl;The 9-fluorenyl-methoxycarbonyl (FMO) resin was synthesized using a solid-phase method. More specifically, 0.55 mmol / g of solid-phase resin (Wang resin; Sigma-Aldrich), 5 mL of dimethylformamide (DMF), 1.1 mmol of Fmoc-Tyr(tBu)-OH, and 0.55 mmol of O-benzotriazole-N,N,N',N'-tetramethyluronium-hexafluoro-phosphate (HBTU) were added to a completely dry reactor and stirred at room temperature for 2 hours to synthesize Fmoc-Tyr(tBu) resin. The synthesized resin was filtered and washed with dimethylformamide. Add 8 mL of 20% piperidine (dissolved in dimethylformamide) solution to the washed resin and stir at room temperature for 30 minutes to synthesize deprotected Fmoc (fluorenylmethyloxycarbonyl protecting group) Tyr(tBu) resin. Filter the synthesized Tyr(tBu) resin and wash with dimethylformamide.

[0076] Add 5 mL of dimethylformamide, 1.1 mmol of Fmoc-Gly-OH and 0.55 mmol of O-benzotriazole-N,N,N',N'-tetramethyluronium-hexafluoro-phosphate (HBTU) to the washed Tyr(tBu) resin and stir at room temperature for 2 hours to synthesize Fmoc-Gly-Tyr(tBu) resin. Filter the synthesized resin and wash with dimethylformamide. Add 8 mL of a 20% piperidine (dissolved in dimethylformamide) solution to the washed resin and stir for 30 minutes at room temperature to synthesize deprotected Fmoc Gly-Tyr(tBu) resin. Filter the synthesized Gly-Tyr(tBu)- resin and wash with dimethylformamide. Through the process described above, peptide bonds are formed between glycine and tyrosine.

[0077] Then, Fmoc-Ala-OH, Fmoc-Gly-OH, Fmoc-Ala-OH, Fmoc-Gly-OH, Fmoc-Ala-OH, Fmoc-Gly-OH, Fmoc-Tyr(tBu)-OH, repeating the same process as forming the peptide bond between glycine and tyrosine, thereby preparing a peptide-resin compound having the amino acid sequence as described in Table 1.

[0078] Then, 10 mL of a solution of trifluoroacetic acid and water mixed in a 95:5 (v / v) ratio was added, and the mixture was stirred at room temperature for 3 hours. Then, diethyl ether was added to the filtered filtrate to crystallize the solid. The resulting solid was filtered, washed with diethyl ether, dried, and the crude peptide compound having the amino acid sequence described in Table 1 was synthesized.

[0079] The crude peptide compound was purified by reverse-phase high-performance liquid chromatography (RP-HPLC) using a Shimadzu 5 mm Shimpak ODS C18 column (20x250 mm) and lyophilized to obtain the white solid peptide of the example.

[0080] The purified peptides were confirmed by analytical reversed-phase high-performance liquid chromatography (RP-HPLC) using a 5 mm Shimpak ODS C18 column (4.6 x 250 mm), and the molecular weight was confirmed by matrix-assisted laser desorption / ionization (MALDI) mass spectrometer (Axima CFR, Kratos Analytical, Manchester, UK).

[0081] [Table 1] Specification 7 / 11 pages 9 CN 121646471 A

[0082]

[0083] In Table 1, Y represents tyrosine (Tyr), G represents glycine (Gly), and A represents alanine (Ala).

[0084] 1-2. Confirmation of the effect of inhibiting obesity-induced weight gain in animals

[0085] The peptide prepared in 1-1 was confirmed to inhibit obesity-induced weight gain in animals through experiments.

[0086] Six-week-old male C57BL / 6 and db / db (BKS.Cg-m+ / + Leprdb / J, hereinafter referred to as db / db mice) were used as experimental animals (Jackson Laboratory). C57BL / 6 mice are normal mice that do not develop obesity and were used as the normal group in this experiment. db / db mice are induced to be obese through mutations in the leptin receptor.The mice used in this study exhibited overeating, obesity, hyperinsulinemia, and hyperglycemia, with obesity appearing 3 to 4 weeks after birth. Animal housing was conducted after obtaining approval from the Ensol Biosciences Animal Ethics Committee (ES-2023-004). The acclimatization period was one week, with a 12-hour light-dark cycle (lights on at 08:00 and off at 20:00). The temperature was 20±2℃, and the relative humidity was 60-80%. The experimental animals were fed a standard diet.

[0087] After the one-week acclimatization period, all experimental animals were weighed and tested. The peptides prepared in 1-1. (10, 30, 100 mg / kg) were dissolved in PBS and administered orally to db / db mice at a dose of 200 μL using a feeding tube. For the normal group and negative control group mice, PBS (phosphate buffered saline, Welgene) was administered instead of the peptides prepared in 1-1. The administration was twice daily, morning and evening, for five consecutive days each week for four weeks, for a total of 40 administrations. Mice were weighed twice weekly. After the four-week administration period, mice were weighed again for three weeks to observe the persistence of the anti-obesity effect. Five mice were used in all groups, including the normal control group. Figures 1 and 2 show the observed weight changes.

[0088] Figures 1 and 2 are graphs showing the results of the peptides (examples) prepared in Analysis 1-1 on the effect of inhibiting or reducing weight gain. Specifically, Figure 1 shows the weight loss effect over time, and Figure 2 shows the weight loss effect of each experimental group.

[0089] In Figure 1, the x-axis represents time (in weeks) based on the moment when administration of the example peptides began, and the y-axis represents the weight gain (g) compared to before administration of the example peptides. Similarly, in Figure 2, the x-axis represents the experimental groups, and the y-axis represents the weight gain (g) compared to before administration of the example peptides.

[0090] First, as shown in Figure 1, the weight of the normal mice increased steadily over 7 weeks, increasing by 4.1g at the end of peptide administration (week 4) and by 7.1g after 7 weeks. The weight gain rate of the negative control group of db / db mice was faster than that of the normal group, increasing by 10.3g at the end of 4 weeks of administration and by 14.5g after 7 weeks. This is about 2 to 2.5 times the weight gain of the normal group, confirming its applicability as an animal model of obesity. Four weeks after the start of administration, which is the end of administration, the weight gain of the 10, 30, and 100 mg / kg groups was 9.1, 6.5, and 8.8g, respectively, confirming that oral administration of the nonapeptide has the effect of inhibiting weight gain.

[0091] Furthermore, as shown in Figure 2, the weight gain was 13.0, 11.2, and 10.8g, respectively, three weeks after the end of administration in the example. That is, it can be shown that even if the administration of the nonapeptide is stopped, the weight gain will not increase rapidly as in the negative control group.

[0092] Furthermore, Table 2 below shows the results of comparing the weight gain / reduction of each experimental group 3 weeks after the end of reagent administration with the weight gain of the negative control group. The weight gain / reduction rate of the normal group or the administration group of the examples can be calculated using the following Formula 1.

[0093] [Formula 1] Specification 8 / 11 pages 10 CN 121646471 A

[0094] Weight gain / reduction rate (%) = (weight gain of the negative control group - weight gain of the normal group or the administration group of the examples) / weight gain of the negative control group × 100

[0095] [Table 2]

[0096]

[0097] As shown in Table 2, the weight of the normal group showed a reduction rate of 50.6% compared to the negative control group. Furthermore, in the 10, 30, and 100 mg / kg examples, the reduction rates were 10.4%, 22.5%, and 25.4%, respectively, indicating that although the reduction rate was not as high as that of the normal group, it still showed a reduction effect according to volume compared to the negative control group. This result confirms that oral administration of the nonapeptide inhibits weight gain in db / db mice, an obese animal model. Therefore, it is known that the peptide of the present invention has the effects of improving, treating, inhibiting and / or preventing obesity / obesity complications.

[0098] On the one hand, the weight gain that occurred when the nonapeptide was administered to type 1 diabetic animals can be explained as the weight initially reduced due to type 1 diabetes being restored to normal during the treatment of type 1 diabetes with the nonapeptide.

[0099] <Preparation Example 1> Preparation of Powder

[0100] 500 mg of peptide

[0101] 100 mg of lactose hydrate

[0102] 10 mg of talc

[0103] The ingredients were mixed and filled into a sealed bag to prepare a powder.

[0104] <Preparation Example 2> Preparation of Tablets

[0105] 500 mg of peptides

[0106] 100 mg of corn starch

[0107] 100 mg of lactose hydrate

[0108] 2 mg of magnesium stearate

[0109] were prepared by the same method as in Example 1-1. The ingredients were mixed and compressed into tablets according to conventional tablet preparation methods.

[0110] <Preparation Example 3> Preparation of Capsules

[0111] 500 mg of crystalline cellulose

[0112] 3 mg of crystalline cellulose

[0113] 14.8 mg of lactose

[0114] 0.2 mg of magnesium stearate

[0115] were prepared by the same method as in Example 1-1. The ingredients were mixed according to conventional capsule preparation methods and filled into gelatin capsules to form capsules.

[0116] <Preparation Example 4> Preparation of Injection

[0117] 500 mg of the injection was prepared using the same method as in Example 1-1. (Instructions for Use, Page 9 / 11, CN 121646471 A)

[0118] 180 mg of mannitol

[0119] 2793 mg of sterile distilled water for injection

[0120] 26 mg of sodium dihydrogen phosphate

[0121] Prepared according to conventional injection preparation methods, with the contents of the ingredients per ampoule (2 ml).

[0122] <Preparation Example 5> Preparation of Liquid

[0123] Prepared by the same method as in Example 1-1: 500 mg of peptide

[0124] 10 g of isomerized sugar

[0125] 5 g of mannitol

[0126] an appropriate amount of purified water

[0127] an appropriate amount of lemon flavoring

[0128] According to conventional liquid preparation methods, each ingredient was dissolved in purified water, and an appropriate amount of lemon flavoring was added. Then purified water was added to adjust to a total of 100 ml, and the solution was filled into a brown bottle to prepare a liquid.

[0129] <Preparation Example 6> Preparation of Health Functional Food

[0130] 1 mg of peptide

[0131] A suitable amount of vitamin mixture

[0132] 70 μg of vitamin A acetate

[0133] 1.0 mg of vitamin E

[0134] 0.13 mg of vitamin B1

[0135] 0.15 mg of vitamin B2

[0136] 0.5 mg of vitamin B6

[0137] 0.2 μg of vitamin B12

[0138] 10 mg of vitamin C

[0139] 10 μg of biotin

[0140] 1.7 mg of nicotinamide

[0141] 50 μg of folic acid

[0142] 0.5 mg of calcium pantothenate

[0143] A suitable amount of inorganic mixture

[0144] 1.75 mg of ferrous sulfate

[0145] 0.82 mg zinc oxide

[0146] 25.3 mg magnesium carbonate

[0147] 15 mg potassium dihydrogen phosphate

[0148] 55 mg calcium hydrogen phosphate

[0149] 90 mg potassium citrate

[0150] 100 mg calcium carbonate

[0151] 24.8 mg magnesium chloride

[0152] The composition ratio of the vitamin and mineral mixture is a mixture of ingredients relatively suitable for health food in a preferred embodiment, but the mixing ratio can be arbitrarily modified. The ingredients can be mixed according to conventional health food preparation methods, made into granules, and used in the preparation of health functional food compositions according to conventional methods.

[0153] <Preparation Example 7> Instructions for Preparation of Health Beverage 10 / 11 pages 12 CN 121646471 A

[0154] 0.1 mg of peptide

[0155] 15 g of vitamin C

[0156] 100 g of vitamin E (powder)

[0157] 19.75 g of ferrous lactate

[0158] 3.5 g of zinc oxide

[0159] 3.5g of nicotinamide

[0160] 0.2g of vitamin A

[0161] 0.25g of vitamin B1

[0162] 0.3g of vitamin B2

[0163] Appropriate amount of purified water

[0164] The ingredients can be mixed according to conventional health beverage preparation methods, and then stirred and heated at 85°C for about 1 hour. The resulting solution is filtered, placed in a sterilized 2L container, sealed and sterilized, and then refrigerated for storage. It can then be used in the preparation of health functional food (e.g., beverage form) compositions.

[0165] The composition ratio is the mixture of ingredients that are more suitable for the taste of the beverage, according to the preferred embodiment. However, the ratio can also be arbitrarily modified according to the consumer class, consumer country, use, and other regional and ethnic preferences.

[0166] Industrial Applicability

[0167] According to the present invention, it has the effect of effectively treating, preventing, improving and / or inhibiting one or more of obesity and obesity complications. Therefore, the present invention has industrial applicability. Specification 11 / 11 page 13 CN 121646471 A Figure 1 Specification drawing 1 / 2 page 14 CN 121646471 A Figure 2 Specification drawing 2 / 2 page 15 CN 121646471 A Abstract The present invention provides a novel use of a peptide consisting of the amino acid sequence of SEQ ID NO: 1, or a pharmaceutically acceptable salt thereof. According to the present, there is an advantage that one or more selected from obesity or obesity complications can be effectively treated, prevented, improved and / or inhibited.

Claims

1. A pharmaceutical composition for treating or preventing obesity, comprising, as an effective ingredient, a peptide consisting of the amino acid sequence of SEQ ID NO: 1 or a pharmaceutically acceptable salt thereof.

2. A food composition for improving or preventing obesity, comprising, as an effective ingredient, a peptide consisting of the amino acid sequence of SEQ ID NO: 1 or a pharmaceutically acceptable salt thereof.

3. The food composition according to claim 2, wherein the food composition is a health functional food composition.

4. A method of treating, preventing, improving, or inhibiting obesity, comprising the step of administering to a non-human animal a peptide consisting of the amino acid sequence of SEQ ID NO: 1 or a pharmaceutically acceptable salt thereof. ​ ​

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