New application 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

TECHNICAL FIELD

[0001] The present application relates to a new use of a nonapeptide, more particularly, to a new use of a peptide consisting of the amino acid sequence of SEQ ID NO: 1 or a pharmaceutically acceptable salt thereof. BACKGROUND

[0002] Overweight and obesity are classic adult metabolic diseases and are also considered a disease (see Powell-Wiley TM, 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 medical advances and improvements in living standards, leading to an increase in average life expectancy, urban-centered population growth, resulting in a decrease in the amount of exercise, causing overweight and obesity are increasing dramatically. Obesity can lead to obesity complications, as obesity complications have high blood pressure, 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 joint cartilage and bone), sleep apnea and respiratory problems, cancer, depression, anxiety and other mental disorders, physical pain, and physical dysfunction, etc. (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] Further, it was confirmed in a study using obese mice that weight loss by dietary control can normalize glucose metabolism and restore 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). In addition, it was confirmed by a clinical trial that weight loss has the efficacy of reducing blood pressure in overweight hypertensive patients, which can indicate that weight loss plays 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 a technology capable of coping with overweight or even obesity and obesity complications.

[0005] On the one hand, the present inventors have developed a nonapeptide {a peptide consisting of the amino acid sequence of SEQ ID NO: 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 for pancreatitis or even type 1 diabetes (see International Patent Publication No. WO 2020 / 184901 A1). The present inventors have been working to find new uses for this effective nonapeptide. However, when the nonapeptide is used to treat type 1 diabetes, there are individuals who have weight gain, which makes it impossible to predict the application of the nonapeptide to obesity that requires weight loss.

[0006]

Prior Art Documents

[0007] [PATENT LITERATURE]

[0008] (Patent Literature 1) International Patent Application Publication No. WO 2020 / 184901 Al, September 17, 2020, Specification

[0009] [NON-PATENT LITERATURE]

[0010] (Non-Patent Literature 1) Powell-Wiley TM, 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

[0011] (Non-Patent Literature 2) Bluher M. "Obesity: global epidemiology and pathogenesis" Nat Rev Endocrinol. 2019 May; 15(5): 288-298

[0012] (Non-Patent Literature 3) 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

[0013] (NPL 4) 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

[0014] (NPL 5) 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 SUMMARY

[0015] PROBLEM TO BE SOLVED BY THE INVENTION

[0016] The present invention intends to solve the problem of providing a new use of a peptide consisting of the amino acid sequence of SEQ ID NO: 1 or a pharmaceutically acceptable salt thereof.

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

[0018] TECHNICAL SOLUTION

[0019] The present inventors have unexpectedly found that a peptide consisting of the amino acid sequence of SEQ ID NO: 1 (YGAGAGAGY) or a pharmaceutically acceptable salt thereof (also called nona-peptide; nona means the number 9) is effective for obesity or obesity complications, thereby completing the present invention.

[0020] In the above-mentioned amino acid sequence, "Y" means Tyrosine (Tyr), "G" means Glycine (Gly), and "A" means Alanine (Ala).

[0021] L-form, D-form, DL-form are present in the amino acids constituting the peptide, and the amino acids constituting the peptide of the present application include all of these types. Furthermore, it is obvious that the "Y" can be interpreted to include all of the amino acids called Tyrosine or 4-hydroxyphenylalanine.

[0022] The peptide includes a variant, including a variant that changes a part of the structure of the peptide of the present application by natural mutation or artificial mutation, but does not change its main activity.

[0023] The pharmaceutically acceptable salt can be exemplified by, for example, hydrochloride, sulfate, phosphate, acetate, citrate, tartrate, succinate, lactate, maleate, fumarate, oxalate, methanesulfonate, trifluoromethanesulfonate, benzenesulfonate, p-toluenesulfonate, sodium salt, potassium salt, calcium salt, and the like.

[0024] The present application provides a peptide consisting of the amino acid sequence of SEQ ID NO: 1 or a pharmaceutically acceptable salt thereof for medical and / or food use for treating, preventing, ameliorating, and / or inhibiting one or more selected from obesity or complications of obesity. "Treatment" is a meaning including amelioration, alleviation, and the like of symptoms associated with obesity or complications of obesity, and "prevention" is a meaning including inhibition of progression from a pre-disease stage to a disease.

[0025] As a specific example, the present application provides a pharmaceutical composition for treating or preventing one or more selected from obesity or complications of obesity, the pharmaceutical composition comprising a peptide consisting of the amino acid sequence of SEQ ID NO: 1 or a pharmaceutically acceptable salt thereof as an effective 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 and hypertensive patients).This is because it is well known that obesity (including overweight) and its complications can be treated, prevented, improved, and / or suppressed as described above (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, DekensD, Vercalsteren E, Larsson M, Nystrom T, Pekny M, Patrone C, Darsalia). V. "Diet-induced weight loss in obese / diabetic mice normalizes glucosemetabolism 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 a pharmaceutically acceptable additive, the composition comprising the peptide or a pharmaceutically acceptable salt thereof and the additive.

[0029] The peptide can be prepared using methods commonly used in the field of peptide synthesis. For example, peptides 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 dehydration condensation using condensing agents; redox methods; and methods using Woodward reagent K.

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

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

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

[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)].

[0035] Solvents that can be used in condensation reactions to form 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 peptide protecting groups varies depending on the type of protecting group, but it can be done using acidic compounds, basic compounds, or transition metals that do not affect the peptide bond and can remove the protecting group.

[0038] Protecting groups can be removed by acid treatment, such as 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 to remove the protecting group using the acid, the reaction can be promoted by adding anisole, phenol and thioanisole as auxiliaries.

[0040] Protecting groups can be removed by treatment with bases, such as ammonia, diethylamine, hydrazine, morpholine, N-methylpyrrolidine, piperidine, sodium carbonate, or mixtures of these bases.

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

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

[0043] In addition, peptides consisting of amino acids of sequence number 1 can be converted into variants or pharmaceutically acceptable salts of them using conventional methods.

[0044] The peptides can be synthesized using an automated peptide synthesizer or produced using genetic engineering techniques. For example, a fusion gene encoding a fusion protein consisting of a fusion partner and the peptides of the present invention can be manufactured through genetic engineering. After transforming a host microorganism using this fusion gene, the fusion protein is expressed in the host microorganism. Then, the peptides of the present invention are cleaved and isolated from the fusion protein using a protease or compound, thereby producing the desired peptides.

[0045] The peptide or its pharmaceutically acceptable salt can be administered primarily by non-oral methods, such as intravenous or subcutaneous injection, intraspinal administration, transdermal administration, nasal administration, or rectal administration. Additionally, oral administration may be considered as appropriate.

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

[0047] Pharmaceutically acceptable additives can be applied based on various factors well 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, their age, size, and general condition; and the route of administration of the composition, such as nasal, oral, ocular, topical, transdermal, and intramuscular, but not limited to these. Pharmaceutically acceptable additives commonly used in the administration of physiologically active substances, other than oral administration, include D5W (5% glucose in water), glucose, and aqueous solutions containing physiological saline at a volume of up to 5%. When injected topically at the lesion site, various injectable hydrogels can be used to enhance efficacy and prolong duration. Furthermore, 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 methods appropriate in the relevant art, as can be found in well-known literature in the relevant field.

[0048] These peptides can be preserved using physiological saline, freeze-dried in ampoules after the addition of mannitol or sorbitol, and dissolved in physiological saline or similar solutions when intended 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 a pharmaceutically acceptable salt thereof consisting of an amino acid sequence of sequence number 1 as an active ingredient.

[0050] The food composition can be incorporated into various foods, including beverages, and can take 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) using raw materials or ingredients with functions beneficial to the human body, in accordance with Korean laws concerning health functional foods. "Functionality" refers to the adjustment of nutrients to the structure and function of the human body or having beneficial effects for health purposes, such as physiological effects.

[0051] In addition to the active ingredient, the food composition may contain other ingredients as essential components without particular limitation. For example, a common beverage may contain various flavoring agents or natural carbohydrates as additional ingredients. Examples of the aforementioned natural carbohydrates include monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; and polysaccharides such as common sugars like dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. In addition to the aforementioned flavoring agents, natural flavoring agents (such as sematriene and stevia extracts (e.g., leucodilin A, glycyrrhizic acid, etc.)) and synthetic flavoring agents (such as saccharin and aspartame) can be advantageously used. Those skilled in the art can appropriately determine the proportion of the natural carbohydrates. In addition to 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, and carbonating agents used in carbonated beverages. These components may be used alone or in combination, and the proportions of these additives may be appropriately selected by those skilled in the art.

[0052] Unless otherwise specified, the food compositions described herein are equally applicable to the pharmaceutical compositions of the present invention, unless they contradict any provisions mentioned in the pharmaceutical compositions 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 or its pharmaceutically acceptable salt, consisting of the amino acid sequence of sequence number 1, can be formulated by adding pharmaceutically or food-acceptable carriers, excipients, or diluents, etc. For information on formulation, please refer to well-known literature in the relevant field.

[0054] Therefore, the compositions 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 compositions of the present invention may contain 0.01 to 99.99% by weight of the said active ingredient.

[0056] The present invention also provides a method for treating, preventing, improving or inhibiting one or more of obesity or obesity complications, the method comprising administering to an animal (including human animals or animals excluding humans) a peptide consisting of an amino acid sequence of sequence number 1 or a pharmaceutically acceptable salt thereof.

[0057] The animal in question can be a mammal.

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

[0059] The animal may be an animal that has or is likely to have one or more of the following: obesity or obesity-related 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 a peptide comprising the amino acid sequence of sequence number 1 or a pharmaceutically acceptable salt thereof for the preparation of a 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 and applicable 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 can also be administered once daily or in multiple divided doses. The dosage described above can be based on an adult (60 kg), but naturally can vary depending on weight, physical condition, etc. On the other hand, when administered to animals other than humans, the dosage can naturally vary depending on the animal species, as determined by calculating the human equivalent dose (HED) using a well-known conversion method based on body surface area (see USFDA. Guidance for Industry: Estimating the Maximum Safe Starting Dose in Adult Healthy Volunteer. Rockville, MD: US Food and Drug Administration; 2005.).

[0064] Invention Effects

[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. Attached Figure Description

[0066] Figure 1 It is a graph showing how the weight loss effect changes over time.

[0067] Figure 2 This is a chart showing the weight loss effect of each experimental group. Detailed Implementation

[0068] Below, with appendix Figure 1 The advantages, features, and implementation methods of the present invention will become clearer with reference to the embodiments described in detail below. However, the present invention is not limited to the embodiments disclosed below, but can be implemented in various different forms. These embodiments are provided only to fully disclose the present invention and to fully illustrate 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 constituent elements mentioned.

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

[0071] In the following examples, it has been demonstrated using obesity-inducing experimental animals that the peptide composed of the amino acid sequence number 1, and even its salts, are effective against obesity and obesity-related complications.

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

[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 were prepared using a solid-phase peptide synthesis method. Specifically, the synthesis was carried out using a solid-phase method utilizing the chemical properties of Fmoc (9-fluorenyl-methoxycarbonyl). 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'-tetramethyl-uronium-hexafluoro-phosphate (HBTU) to washed Tyr(tBu) resin. 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 20% piperidine (dissolved in dimethylformamide) solution to the washed resin and stir at room temperature for 30 minutes to synthesize deprotected Fmoc-containing Gly-Tyr(tBu) resin. Filter the synthesized Gly-Tyr(tBu)-resin and wash with dimethylformamide. Through the process described above, a peptide bond is 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 were used sequentially to repeat the same process as forming the peptide bond between glycine and tyrosine, thereby preparing a peptide-resin compound having the amino acid sequence 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. Diethyl ether was then 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 HPLC (RP-HPLC) using a Shimadzu 5mm Shimpak ODS C18 column (20x250mm) and then 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

[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] Experiments confirmed that the peptides prepared in section 1-1 inhibited obesity-induced weight gain in animals.

[0086] Six-week-old male C57BL / 6 and db / db (BKS.Cg-m + / + Leprdb / J, hereinafter referred to as db / db mice) (Jackson Lab) were used as experimental animals. C57BL / 6 mice are normal mice that do not develop obesity and were used as the control group in this experiment. db / db mice are induced to be obese through a mutation in the leptin receptor and are commonly used in obesity research. These mice 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 - lights 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 a one-week acclimatization period, all experimental animals were weighed and tested. The peptides prepared in section 1-1 (10, 30, and 100 mg / kg) were dissolved in PBS and administered orally to db / db mice at a dose of 200 μL via a feeding tube. For the normal control group and negative control group, PBS (phosphate buffered saline, Welgene) was administered instead of the peptides prepared in section 1-1. Administration was twice daily (morning and evening), five days a week for four weeks, for a total of 40 administrations, with weighing 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, in the experiment. Figure 1 and Figure 2 The observed weight changes are shown in the figure.

[0088] Figure 1 and Figure 2 This is a graph showing the results of the effects of the peptides prepared in Analysis 1-1 (Example) on inhibiting or reducing weight gain. Specifically, Figure 1 It is a graph showing how weight loss results change over time. Figure 2 This is a chart showing the weight loss effect of each experimental group.

[0089] Figure 1 The x-axis represents time (in weeks) based on the moment when the peptide of the example was first administered, and the y-axis represents the weight gain (g) compared to before the administration of the peptide of the example was first administered. Furthermore, Figure 2 The x-axis represents the experimental group, and the y-axis represents the weight gain (g) compared to before the administration of the peptide in the example.

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

[0091] In addition, such as Figure 2 As shown, in the example, at week 7 after 3 weeks following the end of administration, the weight increases were 13.0, 11.2, and 10.8 g, respectively. This demonstrates that even after discontinuing administration of the nonapeptide, there is no rapid increase in body weight, as seen in the negative control group.

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

[0093] [Formula 1]

[0094] Weight gain / loss rate (%) = (Weight gain in negative control group - Weight gain in normal group or the treatment group in the example) / Weight gain in negative control group × 100

[0095] Table 2

[0096]

[0097] As shown in Table 2, the normal group showed a 50.6% reduction in body weight 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 lower than that of the normal group, it still showed a reduction efficacy on a volume basis compared to the negative control group. These results confirm that oral administration of the nonapeptide inhibits body weight gain in db / db mice, an obese animal model. Therefore, it is evident that the peptide of the present invention has effects in improving, treating, inhibiting, and / or preventing obesity / obesity complications.

[0098] On the one hand, the weight gain that occurred when nonapeptides were administered to animals with type 1 diabetes can be explained as the weight initially lost due to type 1 diabetes returning to normal during the treatment of type 1 diabetes with nonapeptides.

[0099] <Preparation Example 1> Preparation of Powder

[0100] Prepare 500 mg of peptide using the same method as in Example 1-1.

[0101] 100mg lactose hydrate

[0102] 10mg talc

[0103] The ingredients are mixed and filled into sealed bags to form a powder.

[0104] <Preparation Example 2> Tablet Preparation

[0105] Prepare 500 mg of peptide using the same method as in Example 1-1.

[0106] 100mg corn starch

[0107] 100mg lactose hydrate

[0108] 2mg magnesium stearate

[0109] The ingredients are mixed and compressed into tablets using conventional tablet preparation methods.

[0110] <Preparation Example 3> Preparation of Capsules

[0111] Prepare 500 mg using the same method as in Example 1-1.

[0112] 3mg of crystalline cellulose

[0113] 14.8mg lactose

[0114] 0.2 mg magnesium stearate

[0115] The ingredients are mixed according to conventional capsule preparation methods and filled into gelatin capsules to form capsules.

[0116] <Preparation Example 4> Preparation of Injectables

[0117] Prepare 500 mg using the same method as in Example 1-1.

[0118] 180mg mannitol

[0119] 2793mg of sterile distilled water for injection

[0120] 26mg sodium dihydrogen phosphate

[0121] Prepared according to conventional injection preparation methods, with the stated ingredient content per ampoule (2 ml).

[0122] <Preparation Example 5> Preparation of Liquids

[0123] Prepare 500 mg of peptide using the same method as in Example 1-1.

[0124] 10g of isomerized sugar

[0125] 5g of mannitol

[0126] An appropriate amount of purified water

[0127] A suitable amount of lemon flavoring

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

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

[0130] Prepare 1 mg of peptide using the same method as in Example 1-1.

[0131] Appropriate amount of vitamin mixture

[0132] 70ug of Vitamin A Acetate

[0133] 1.0mg Vitamin E

[0134] 0.13mg of vitamin B1

[0135] 0.15mg of vitamin B2

[0136] 0.5mg of vitamin B6

[0137] 0.2ug of vitamin B12

[0138] 10mg of Vitamin C

[0139] 10ug of biotin

[0140] 1.7mg nicotinamide

[0141] 50ug of folic acid

[0142] 0.5mg calcium pantothenate

[0143] Appropriate amount of inorganic mixture

[0144] 1.75 mg ferrous sulfate

[0145] 0.82 mg of zinc oxide

[0146] 25.3 mg magnesium carbonate

[0147] 15mg potassium dihydrogen phosphate

[0148] 55mg of calcium hydrogen phosphate

[0149] 90mg potassium citrate

[0150] 100mg of calcium carbonate

[0151] 24.8 mg magnesium chloride

[0152] The composition ratio of the vitamin and mineral mixture is a mixture of ingredients that are relatively suitable for health food in a preferred embodiment. However, this 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> Preparation of Health Beverage

[0154] Prepare 0.1 mg of peptide using the same method as in Example 1-1.

[0155] 15g of Vitamin C

[0156] 100g of Vitamin E (powder)

[0157] 19.75g of ferrous lactate

[0158] 3.5g of zinc oxide

[0159] 3.5g of niacinamide

[0160] 0.2g of Vitamin A

[0161] 0.25g of vitamin B1

[0162] 0.3g of vitamin B2

[0163] An appropriate amount of purified water

[0164] The ingredients can be mixed according to conventional health beverage preparation methods, 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, refrigerated, and then used in the preparation of health functional food (e.g., beverage form) compositions.

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

[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 is industrially usable.

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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