Pharmaceutical composition for preventing or treating obesity, containing GLP analogue as active ingredient

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating obesity, said pharmaceutical composition containing a GLP (Glucagon Like Peptide) analogue as an active ingredient. The GLP analogue of the present invention has been confirmed to be an analogue of GLP-1 and GLP-2, which has been confirmed to have a long in-vivo activity duration due to an increase in the half-life thereof, thereby having an excellent effect in the prevention or treatment of metabolic diseases. Specifically, the peptide disclosed by the invention not only can reduce the weight and fat of an individual, reduce blood sugar, total cholesterol in blood, high-density lipoprotein, low-density lipoprotein and fat factor level in the blood of neutral fat, but also can reduce ASL and AST levels of the liver, the weight of the liver, the degree of hepatic fatty disease and the level of the neutral fat of the liver. From this point, the peptide according to the present invention can be used as an excellent preventive or therapeutic agent for metabolic diseases.

Description

Technical Field

[0001] This invention relates to a pharmaceutical composition for the prevention or treatment of obesity, comprising a GLP analogue as an active ingredient.

[0002] This application claims priority based on Korean Patent Application No. 10-2023-0085302, filed on June 30, 2023, and Korean Patent Application No. 10-2024-0084869, filed on June 27, 2024, the contents of which are disclosed in the description and drawings of these applications are incorporated herein by reference. Background Technology

[0003] Obesity, a chronic disease of modern people, has become one of the most serious health problems worldwide and was recognized as a disease by the World Health Organization (WHO) in 1996. With multiple factors such as industrialization, rising income levels leading to Westernized dietary habits and lifestyles, increased consumption of processed foods, and changes in lifestyle, obesity continues to increase.

[0004] Obesity is a metabolic disorder caused by excessive body fat. While it is not a problem in itself, it should not be taken lightly because it can lead to social barriers and secondary complications such as hyperlipidemia, hypertension, arteriosclerosis, diabetes, fatty liver, and cardiovascular disease caused by excessive fat accumulation.

[0005] Therefore, in 2013, the WHO listed obesity as a new infectious disease of the 21st century. Since then, as people’s understanding of health has deepened, the world has declared a war on obesity. Multifaceted research on the development of obesity treatments is underway, and the market size of obesity treatments has gradually increased.

[0006] The U.S. Food and Drug Administration (FDA) approved sibutramine in 1997 and orlistat in 1999, both of which are obesity treatments that can be used for more than three months. European countries, including the UK, approved rimonabant in 2006. However, rimonabant was withdrawn from the market in 2009 due to psychogenic side effects such as suicide, and sibutramine formulations were also withdrawn in 2010 due to controversy surrounding cardiovascular side effects. Following these controversies, obesity treatments faced a period of stagnation.

[0007] On the other hand, appetite-regulating hormones, such as glucagon-like peptide (GLP), are hormones secreted by the small intestine after eating. They regulate appetite by enhancing satiety and induce the pancreas to secrete insulin to regulate blood sugar. Therefore, although various GLP analogs are currently being studied for therapeutic purposes such as diabetes, there are still few reports on GLP analogs with good sustained efficacy in treating obesity. Summary of the Invention

[0008] Technical issues One object of the present invention is to provide a peptide represented by the following Formula I: HX2EGX5FTX8DX 10 X 11 X 12 X 13 X 14 X 15 X 16 X 17 X 18 X 19 X 20 X 21 FIX 24 X 25 LX 27 X 28 X 29 X 30 X 31 X 32 X 33 X 34 X35X 36 X 37 X 38 X 39 X 40 X 41 X 42 X 43 [Formula I] In equation I, X2 is either Gly or Aib; X5 is Thr or Ser; X8 is either Ser or Asp; X 10 For Leu or Val; X 11 For Ser or Asn; X 12 For Thr or Ser; X 13 For Tyr or Ile; X 14 For Leu or Met; X 15For Asp or Glu; X 16 It is any one of the choices from the group consisting of Ala, Gly, and Asn; X 17 For Leu or Gln; X 18 and X 19 For Ala or His; X 20 For Arg or Cys; X 21 For Asp or Glu; X 24 For Asn or Lys; X 25 For Trp or Thr; X 27 For Ile or Val; X 28 For Gln or Lys; X 29 For Thr or Gly; X 30 For Lys or Arg; X 31 For Ile or missing; X 32 Thr or missing; X 33 For ASP or missing; X 34 To X 43 For missing or Lys.

[0009] Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of metabolic diseases, incorporating the peptide as an active ingredient.

[0010] Another object of the present invention is to provide a kit for the prevention or treatment of metabolic diseases, comprising a composition containing the peptide as an active ingredient and an instruction manual.

[0011] Another object of the present invention is to provide a food composition containing the peptide as an active ingredient for the prevention or improvement of metabolic diseases.

[0012] Another object of the present invention is to provide a cosmetic composition for preventing or improving obesity, incorporating the peptide as an active ingredient.

[0013] However, the technical problem to be solved by the present invention is not limited to the problems mentioned above. For other technical problems not mentioned, those skilled in the art to which the present invention pertains can fully understand them through the following description.

[0014] Methods for solving technical problems This invention provides a peptide represented by the following formula I: HX2EGX5FTX8DX 10 X 11 X 12 X 13 X 14 X 15 X 16 X 17 X 18 X 19 X 20 X 21 FIX 24 X 25 LX 27 X 28 X 29 X 30 X 31 X 32 X 33 X 34 X35X 36 X 37 X 38 X 39 X 40 X 41 X 42 X 43 [Formula I] In equation I, X2 is either Gly or Aib; X5 is Thr or Ser; X8 is either Ser or Asp; X 10 For Leu or Val; X 11 For Ser or Asn; X 12 For Thr or Ser; X 13 For Tyr or Ile; X 14 For Leu or Met; X 15 For Asp or Glu; X 16 It is any one of the choices from the group consisting of Ala, Gly, and Asn; X 17 For Leu or Gln; X 18 and X 19 For Ala or His; X 20 For Arg or Cys; X 21 For Asp or Glu; X 24 For Asn or Lys; X 25 For Trp or Thr; X 27 For Ile or Val; X 28 For Gln or Lys; X 29 For Thr or Gly; X 30 For Lys or Arg; X 31 For Ile or missing; X 32 Thr or missing; X 33 For ASP or missing; X 34 To X 43 For missing or Lys.

[0015] In one embodiment of the present invention, X5 is Thr, X8, and X. 11 For Ser, X 12 For Thr, X 13 For Tyr, X 14 For Leu, X 15 For Asp, X 16 For Ala, X 17 For Leu, X 19 For Ala, X 21 For Asp, X 25 For Trp, X 27 For Ile, X 28 For Gln, X 29 For Thr, X 30 For Lys, X 31 For Ile, X 32 For Thr, X 33 For Asp, X 34 To X 43 or X 36 To X 43 It can be missing, but is not limited to this.

[0016] In one embodiment of the present invention, X 10 For Leu, X18 For Ala, X 34 and X 35 It can be Lys, but is not limited to this.

[0017] In one embodiment of the present invention, when the X 20 When it is Cys, PEG (polyethylene glycol) can be combined on Cys, but it is not limited to this.

[0018] In one embodiment of the invention, from the X 24 and X 35 To X 43 Any one or more amino acids selected from the group can be linked to a fatty acid, but is not limited to this.

[0019] In one embodiment of the present invention, the fatty acid may be C. 16 To C 20 However, it is not limited to this.

[0020] In one embodiment of the present invention, the fatty acid may be any one selected from the group consisting of palmitic acid, octadecanedioic acid, eicosanedioic acid, margaricacid, heptadecanedioic acid, hexadecanedioic acid, stearic acid, nonadecylic acid, nonadecanedioic acid, and eicosanedioic acid, but is not limited thereto.

[0021] In one embodiment of the invention, the peptide may comprise any one selected from the group consisting of amino acid sequences represented by sequence numbers 1 to 9, but is not limited thereto.

[0022] The present invention provides a pharmaceutical composition for the prevention or treatment of metabolic diseases, incorporating the peptide as an active ingredient.

[0023] In one embodiment of the present invention, the metabolic disease may be any one or more selected from the group consisting of obesity, diabetes, non-alcoholic fatty liver disease, hepatic steatosis, dyslipidemia, arteriosclerosis, insulin resistance syndrome and complications thereof, but is not limited thereto.

[0024] In one embodiment of the present invention, the peptide may be a persistent form, but is not limited thereto.

[0025] In one embodiment of the invention, the composition may be administered by any method selected from the group consisting of subcutaneous administration, intravenous administration and oral administration, but is not limited thereto.

[0026] In one embodiment of the present invention, the composition can activate GLP-1 and GLP-2, but is not limited thereto.

[0027] In one embodiment of the invention, the composition has any one or more features selected from the group consisting of, but not limited to: a) Reduce individual weight and fat; b) Lower blood sugar, total cholesterol, high-density lipoprotein, low-density lipoprotein, and triglycerides; c) Reduce liver ASL and AST levels, liver weight, degree of liver steatosis, and liver neutral fat.

[0028] The present invention provides a kit for the prevention or treatment of metabolic diseases, comprising a composition containing the peptide as an active ingredient and instructions.

[0029] The present invention provides a food composition for preventing or improving metabolic diseases by including the peptide as an active ingredient.

[0030] In one embodiment of the present invention, the food is a health functional food, but is not limited thereto.

[0031] The present invention provides a cosmetic composition for preventing or improving obesity, incorporating the peptide as an active ingredient.

[0032] In addition, the present invention provides a method for preventing, improving or treating metabolic diseases, comprising the step of administering a pharmaceutically effective amount of the peptide or a composition containing it as an active ingredient to an individual who requires the peptide.

[0033] In addition, the present invention provides the use of the peptide or a composition containing it as an active ingredient for the prevention, improvement or treatment of metabolic diseases.

[0034] In addition, the present invention provides the use of the peptide or a composition containing it as an active ingredient in the preparation of a formulation for the prevention, improvement or treatment of metabolic diseases.

[0035] Invention Effects Using GLP (glucagon-like peptide) analogs as active ingredients, pharmaceutical compositions for the prevention or treatment of obesity, as analogs of GLP-1 and GLP-2, have demonstrated excellent efficacy in the prevention or treatment of metabolic diseases due to the extended duration of activity in vivo caused by the increased half-life. Specifically, they not only reduce individual weight and fat, and lower blood glucose, total cholesterol, high-density lipoprotein, low-density lipoprotein, and triglyceride levels, but also reduce liver ASL and AST levels, liver weight, degree of hepatic steatosis, and hepatic triglyceride levels. Therefore, the peptides of this invention can be effectively utilized as excellent agents for the prevention or treatment of metabolic diseases. Attached Figure Description

[0036] Figure 1 This is a graph showing the excellent GLP-1 activity of HU020K2FA (with the highest point as the baseline, in order: Exendin-4, HU020, liraglutide, HU020K2FA, and smegglutide).

[0037] Figure 2 This is a graph showing the effect of HU020K2FA on reducing body weight and feed intake in an obese mouse model (Comparison of mouse body weight changes: based on the highest point, in order, HU020, control group, liraglutide, HU020K2FA=semaglutide) (Comparison of body weight loss rate compared with day 0: based on the highest point, in order, HU020, control group, liraglutide, HU020K2FA, semaglutide) (Comparison of cumulative feed intake: based on the highest point, in order, control group, HU020, liraglutide, HU020K2FA, semaglutide).

[0038] Figure 3 These are the results of biochemical experiments when HU020K2FA was applied to an obese mouse model.

[0039] Figure 4 The results of the mouse pharmacokinetic experiment are curves showing the results of subcutaneous administration of HU020K2FA and HU020M1 to HU020M6 with peptide 0.2 mg / kg mouse (based on the highest point, in order: semaglutide, HU020M4, HU020M1, HU020M5, HU020M2, HU020M6, HU020K2FA and HU020M3).

[0040] Figure 5This comparison examines the effects of HU020K2FA, HU020M4, and HU020M5 on body weight changes and cumulative feed intake in obese mouse models. (Comparison of mouse body weight changes: based on the highest point, the order is: control group (Vehicle) (DIO mice, QD), control group (Vehicle) (Normal mice)). (mice), QD), HU020M5 (10 nmol / kg, QD), HU020M4 (10 nmol / kg, QD), HU020K2FA (45 nmol / kg, QD), Smegglutide (10 nmol / kg, QD) (Comparison of cumulative feed intake: based on the highest point, in order are control group (Vehicle) (healthy mice, QD), control group (Vehicle) (DIO mice, QD), HU020M5 (10 nmol / kg, QD), HU020M4 (10 nmol / kg, QD), HU020K2FA (45 nmol / kg, QD), Smegglutide (10 nmol / kg, QD)).

[0041] Figure 6 and Figure 7 The images and bar charts are DEXA images taken after HU020K2FA, HU020M4 and HU020M5 were administered to obese mouse models (from left to right: control group (Vehicle) (healthy mice, QD), control group (Vehicle) (DIO mice, QD), smegglutide (10 nmol / kg, QD), HU020K2FA (45 nmol / kg, QD), HU020M4 (10 nmol / kg, QD), HU020M5 (10 nmol / kg, QD)).

[0042] Figure 8 The results show the fat content of various tissues when HU020K2FA, HU020M4 and HU020M5 were administered to an obese mouse model (from left to right: control group (Vehicle) (healthy mice, QD), control group (Vehicle) (DIO mice, QD), smegglutide (10 nmol / kg, QD), HU020K2FA (45 nmol / kg, QD), HU020M4 (10 nmol / kg, QD), HU020M5 (10 nmol / kg, QD)).

[0043] Figure 9The results show the analysis of plasma levels of obesity-related factors when HU020K2FA, HU020M4, and HU020M5 were administered to an obese mouse model (from left to right: control group (Vehicle) (healthy mice, QD), control group (Vehicle) (DIO mice, QD), smegglutinin (10 nmol / kg, QD), HU020K2FA (45 nmol / kg, QD), HU020M4 (10 nmol / kg, QD), HU020M5 (10 nmol / kg, QD)).

[0044] Figure 10 The results show the analysis of the liver injury index AST / ALT values ​​when HU020K2FA, HU020M4 and HU020M5 were administered to an obese mouse model (from left to right: control group (Vehicle) (healthy mice, QD), control group (Vehicle) (DIO mice, QD), smegglutinin (10 nmol / kg, QD), HU020K2FA (45 nmol / kg, QD), HU020M4 (10 nmol / kg, QD), HU020M5 (10 nmol / kg, QD)).

[0045] Figure 11 and Figure 12 This is a bar chart and photograph showing the results of liver weight and hepatic steatosis examination when HU020K2FA, HU020M4 and HU020M5 were administered to an obese mouse model (from left to right: control group (Vehicle) (healthy mice, QD), control group (Vehicle) (DIO mice, QD), smegglutinin (10 nmol / kg, QD), HU020K2FA (45 nmol / kg, QD), HU020M4 (10 nmol / kg, QD), HU020M5 (10 nmol / kg, QD)).

[0046] Figure 13 The results show the liver neutral fat content measured when HU020K2FA, HU020M4 and HU020M5 were administered to an obese mouse model (from left to right: control group (Vehicle) (healthy mice, QD), control group (Vehicle) (DIO mice, QD), smegglutinin (10 nmol / kg, QD), HU020K2FA (45 nmol / kg, QD), HU020M4 (10 nmol / kg, QD), HU020M5 (10 nmol / kg, QD)).

[0047] Figure 14 and Figure 15The bar chart and photographs show the results of oil red O staining analysis when HU020K2FA, HU020M4 and HU020M5 were administered to an obese mouse model. (From left to right: control group (Vehicle) (healthy mice, QD), control group (Vehicle) (DIO mice, QD), smegglutinin (10 nmol / kg, QD), HU020K2FA (45 nmol / kg, QD), HU020M4 (10 nmol / kg, QD), HU020M5 (10 nmol / kg, QD)).

[0048] Figure 16 This comparison examines the effects of HU020M4 administration to obese mouse models on weight changes and cumulative feed intake (weight change comparison: from left to right: control group (healthy mice, Q2D), control group (DIO mice, Q2D), smegglutinin (10 nmol / kg, Q2D), smegglutinin (25 nmol / kg, Q2D), HU020M4 (10 nmol / kg, Q2D), HU020M4 (25 nmol / kg, Q2D)). (Mouse weight loss rate comparison: based on the highest point, in order: control group (Vehicle) (healthy mice, Q2D), control group (Vehicle) (DIO mice ... (O mice, Q2D), Smegglutinin (10 nmol / kg, Q2D), Smegglutinin (25 nmol / kg, Q2D), HU020M4 (10 nmol / kg, Q2D), HU020M4 (25 nmol / kg, Q2D) (Comparison of cumulative feed intake: based on the highest point, the order is control group (healthy mice, Q2D), control group (DIO mice, Q2D), Smegglutinin (10 nmol / kg, Q2D), HU020M4 (10 nmol / kg, Q2D), Smegglutinin (25 nmol / kg, Q2D), HU020M4 (25 nmol / kg, Q2D)).

[0049] Figure 17 The results show the plasma levels of obesity-related factors when HU020M4 was administered to an obese mouse model every 2 days (from left to right: control group (Vehicle) (healthy mice, Q2D), control group (Vehicle) (DIO mice, Q2D), smegglutinin (10 nmol / kg, Q2D), smegglutinin (25 nmol / kg, Q2D), HU020M4 (10 nmol / kg, Q2D), HU020M4 (25 nmol / kg, Q2D)).

[0050] Figure 18The results show the AST / ALT ratio, a liver injury indicator, when HU020M4 was administered to an obese mouse model every 2 days (from left to right: control group (Vehicle) (healthy mice, Q2D), control group (Vehicle) (DIO mice, Q2D), smegglutinin (10 nmol / kg, Q2D), smegglutinin (25 nmol / kg, Q2D), HU020M4 (10 nmol / kg, Q2D), HU020M4 (25 nmol / kg, Q2D)). Detailed Implementation

[0051] In this invention, "GLP analogue" can refer to a variant, i.e., a peptide or compound, containing a sequence similar to any one or more of GLP, GLP-1, or GLP-2. GLP, GLP-1, and GLP-2 all possess characteristics that prevent their use on their own. In particular, GLP-1 itself has insufficient activity; the two truncated naturally occurring peptides, GLP-1(7-37)OH and GLP-1(7-36)NH2, are rapidly eliminated from the body, resulting in a very short half-life. Therefore, the therapeutic usefulness of GLP-1 peptide intervention has been limited. Specifically, dipeptidyl peptidase-3 (DPP-3), generated endogenously, removes the N-terminal histidine (7) and alanine (8) residues of GLP-1, thereby inactivating the GLP-1 peptide. This is reportedly the main reason for the short half-life in vivo. In other words, the short half-life (t1 / 2 < 2 minutes) of GLP-1 produced by the breakdown of DPP-3 hinders the application of GLP-1 in the treatment of insulin-resistant type 2 diabetes.

[0052] Currently, representative GLP-1 analogs, namely exenatide, lixisenatide, and liraglutide, are marketed under the brand names Byetta, Lismin, and NovoMix, respectively. They are administered once or twice daily via subcutaneous injection into the abdomen, thigh, or upper arm. These commercially available subcutaneous GLP-1 analogs have the following drawbacks: they are difficult to store, require daily disinfection of the injection site, and the injection process is complex, leading to inconvenience. They may also cause rashes and swelling at the injection site.

[0053] Conversely, oral administration eliminates the inconvenience of storing and taking subcutaneous injections for diabetic patients who require continuous treatment, thereby improving their quality of life and reducing health management costs.

[0054] Therefore, in order to utilize the excellent activity of GLP and its analogues, it is necessary to improve the half-life, storage method, and dosing cycle. To solve this problem, the inventors of this invention have conducted repeated research and developed and completed a sustained-release GLP analogue.

[0055] Therefore, the present invention provides a peptide represented by the following formula I: HX2EGX5FTX8DX 10 X 11 X 12 X 13 X 14 X 15 X 16 X 17 X 18 X 19 X 20 X 21 FIX 24 X 25 LX 27 X 28 X 29 X 30 X 31 X 32 X 33 X 34 X35X 36 X 37 X 38 X 39 X 40 X 41 X 42 X 43 [Formula I] In equation I, X2 is either Gly or Aib; X5 is Thr or Ser; X8 is either Ser or Asp; X 10 For Leu or Val; X 11 For Ser or Asn; X 12 For Thr or Ser; X 13 For Tyr or Ile; X 14 For Leu or Met; X 15 For Asp or Glu; X 16 It is any one of the choices from the group consisting of Ala, Gly, and Asn; X 17 For Leu or Gln; X18 and X 19 For Ala or His; X 20 For Arg or Cys; X 21 For Asp or Glu; X 24 For Asn or Lys; X 25 For Trp or Thr; X 27 For Ile or Val; X 28 For Gln or Lys; X 29 For Thr or Gly; X 30 For Lys or Arg; X 31 For Ile or missing; X 32 Thr or missing; X 33 For ASP or missing; X 34 To X 43 For missing or Lys.

[0056] In Formula I of the present invention, the GLP-1 and GLP-2 are activated simultaneously, and the nine sequences necessary for GLP-1 activity are immobilized.

[0057] In Formula I, X, the GLP analogue of the present invention 34 To X 43 It can be missing or Lys. In this case, it depends on X. 35 To X 43 The half-life of the GLP analogue of the present invention may vary depending on the number of Lys in the nine amino acids. For example, in one embodiment of the present invention, the GLP analogue may be X. 34 and X 35 For Lys, X 36 To X 43 It is missing, or it could be X. 34 To X 43 The presence of more than one Lys molecule increases the half-life compared to the case where all Lys molecules are missing, thus prolonging the duration of activity in the body and resulting in excellent obesity treatment effects, but this is not the only possibility.

[0058] Therefore, the present invention can provide a peptide represented by the following Formula II, depending on the number of Lys at the C-terminus: HX2EGX5FTX8DX 10 X 11 X 12 X 13 X 14 X 15 X 16 X 17 X 18 X 19 X 20 X 21 FIX 24 X 25 LX 27 X 28 X 29 X 30 X 31 X 32 X 33 X 34 X 35 [Formula II]. In one embodiment of the present invention, X5 is Thr, X8, and X. 11 For Ser, X 12 For Thr, X 13 For Tyr, X 14 For Leu, X 15 For Asp, X 16 For Ala, X 17 For Leu, X 19 For Ala, X 21 For Asp, X 25 For Trp, X 27 For Ile, X 28 For Gln, X 29 For Thr, X 30 For Lys, X 31 For Ile, X 32 For Thr, X 33 For Asp, X 34 To X 43 or X 36 To X 43 It can be omitted, but is not limited to this. In the case where the peptide of the present invention is represented by Formula II, X 34 To X 35 It may be omitted, but is not limited to this. This is the sequence commonly contained in the peptides of the present invention, excluding the amino acids essential for GLP-1 and GLP-2 activity.

[0059] Therefore, the present invention can provide a peptide represented by the following Formula III: HX2EGTFTSDX 10 STYLDALX 18 AX 20DFIX 24 WLIQTKITDX 34 X 35 [Formula III]. Therefore, the peptides of the present invention can be represented by the above formulas I to III.

[0060] In one embodiment of the present invention, it can be X 10 For Leu, X 18 For Ala, X 34 and X 35 It is Lys, but not limited to this.

[0061] In the peptide sequence of the present invention, the amino acids at positions 2, 10, 18, 20, 24, 34, and 35, based on the N-terminus, can be amino acids that the inventors of the present invention have identified, based on research using the amino acid sequences of GLP-1 and GLP-2, as having particularly excellent therapeutic activity for metabolic diseases. Furthermore, specific experiments have confirmed that the amino acids, PEG binding, fatty acid binding, etc., of the present invention at these positions exhibit superior efficacy compared to existing GLP and GLP analogues.

[0062] The 35 basic sequences of the peptide HU020K2FA, represented by sequence number 1 of the present invention, can be composed of the following: 1) 7 amino acid sequences present only in GLP-1; 2) 14 amino acids present only in GLP-2; 3) 10 amino acids common to GLP-1 and GLP-2; and 4 amino acids with self-mutation. Specifically, among the 16 GLP-1 amino acids, 9 amino acids are the same as GLP-1 / 2, therefore there are 7 amino acids present only in GLP-1 and 24 amino acids the same as GLP-2. That is, HU020K2FA may exhibit a significantly high sequence homology with GLP-2, but is not limited thereto. In addition, HU020K2FA of the present invention may exhibit approximately 57% sequence homology with the GLP-1 amino acid sequence, and the above amino acids may represent a portion of the sequence identical to the GLP-1 amino acid sequence, but is not limited thereto. Therefore, the HU020K2FA of the present invention may exhibit approximately 70% sequence homology with the GLP-2 amino acid sequence. The following amino acids may represent a portion of the same sequence as the GLP-2 amino acid sequence, but are not limited thereto.

[0063] In this invention, the terms "protein," "polypeptide," or "peptide" are used in harmony, for example, referring to a polymer of amino acid residues typically found in proteins in their natural state.

[0064] The sustained-release GLP analog of the present invention, as a GLP-based analog, is characterized by an increased half-life, resulting in excellent duration of activity in vivo. Three methods can be used to prepare this sustained-release GLP analog.

[0065] One approach is to conjugate a fatty acid with Lys, which is an amino acid containing a GLP analog at its C-terminus. In the case of conjugated fatty acids, albumin binding affinity increases, thus extending the half-life; the effect may vary depending on the number of carbon atoms in the fatty acid and the type of fatty acid.

[0066] In one embodiment of the invention, from the X 24 and X 35 To X 43 Any one or more amino acids selected from the group can be linked to a fatty acid, but is not limited to this. In this case, as mentioned above, the amino acid linking the fatty acid can be Lys.

[0067] In one embodiment of the present invention, the fatty acid may be C 16 To C 20 However, it is not limited to this.

[0068] In one embodiment of the present invention, the fatty acid may be any one selected from the group consisting of palmitic acid, octadecanedioic acid, eicosanedioic acid, margaricacid, heptadecanedioic acid, hexadecanedioic acid, stearic acid, nonadecylic acid, nonadecanedioic acid, and eicosanedioic acid, but is not limited thereto.

[0069] In this invention, C 16 To C 20 Fatty acids can be C 16 C 18 and C 20The fatty acids, preferably palmitic acid, octadecanoic acid, or eicosanoic acid, are not limited thereto. In one embodiment of the invention, it was confirmed that the activities of GLP-1 and GLP-2 increased with fatty acid conjugation, and that changes in the type or length of the fatty acids affected the persistence or anti-obesity effect. Although characteristics related to the type or length of the fatty acids were confirmed, specific data confirmed that, regardless of the type of fatty acid, the anti-obesity-related activities of GLP-1 and GLP-2 were equivalent to or greater than those in the absence of fatty acids (Example 5, etc.).

[0070] As a second method to increase the half-life of the GLP analogues of the present invention, PEG binding can be used.

[0071] In this context, conjugating PEG at sites that do not affect activity is important to increase half-life and stability while maintaining high levels of obesity treatment efficacy. In one embodiment of the invention, when the X... 20 When it is Cys, PEG (polyethylene glycol) can be bound to Cys, but it is not limited to this.

[0072] In this case, PEG and X 20 The binding method is unrestricted. For example, it can be bound via a linker or directly; in this case, binding refers to all binding methods, including covalent and non-covalent bonds. Binding can also be achieved through methods commonly used in this industry, such as direct covalent bonding using well-known gene recombination technologies.

[0073] The third method involves replacing an amino acid at a specific position with Aib (a non-coded amino acid, alpha-amino isobutyric acid) to prepare the sustained-release GLP analog of this invention. In this case, an increased half-life can be achieved by preventing peptide degradation dependent on DPP-IV enzymes.

[0074] The sustained GLP analogues of the present invention may replace the second amino acid with Aib, but are not limited thereto.

[0075] In one embodiment of the present invention, the peptide may comprise any one of the amino acid sequences selected from the group consisting of sequences represented by serial numbers 1 to 9, but is not limited thereto. Furthermore, the peptide of the present invention may be a sequence composed of any one of the amino acid sequences selected from the group consisting of sequences represented by serial numbers 1 to 9, but is not limited thereto.

[0076] The present invention provides a pharmaceutical composition comprising the peptide as an active ingredient for the prevention or treatment of metabolic diseases.

[0077] In one embodiment of the present invention, the metabolic disease may be any one or more selected from the group consisting of obesity, diabetes, non-alcoholic fatty liver disease, hepatic steatosis, dyslipidemia, arteriosclerosis, insulin resistance syndrome and complications thereof, but is not limited thereto.

[0078] In one embodiment of the present invention, the peptide may be a persistent form, but is not limited thereto.

[0079] In this invention, "sustained" can refer to a form that increases biostability, prevents peptide degradation by various enzymes from prolonging the half-life, increases the residual time in the blood, or increases the duration of activity and bioavailability, but is not limited to these. In this invention, it was confirmed that when the peptide of this invention is administered once daily, it not only reduces the level of obesity factors in the blood containing blood glucose, but also reduces body weight and fat mass, and produces excellent effects related to various fat-inhibiting factors in the liver. Furthermore, even when administered once every two days, the effects were still confirmed to be very excellent; in this case, even compared with the control group, semaglutide, excellent effects were confirmed at a smaller dosage.

[0080] In one embodiment of the invention, the composition may be administered by any method selected from the group consisting of subcutaneous administration, intravenous administration and oral administration, but is not limited thereto.

[0081] The GLP analogs of the present invention exhibit sustained-release characteristics using the three methods described above. Compared with existing GLP analog weight loss or blood sugar lowering agents, the longer dosing cycle improves the convenience of administration, and not only reduces the burden on the body, but also alleviates the financial burden.

[0082] In one embodiment of the invention, the composition can activate GLP-1 and GLP-2, but is not limited thereto. In particular, the GLP-1 activity of the peptide of the present invention has been confirmed to be particularly excellent, and the GLP-2 activity has also been confirmed to be significant.

[0083] In one embodiment of the invention, the composition is characterized by, but is not limited to, any one or more of the following: a) Reduce individual weight and fat; b) Lowering blood sugar, total cholesterol, high-density lipoprotein (HDL), low-density lipoprotein (LDL), and triglycerides; and c) Reduce liver ASL and AST levels, liver weight, degree of liver steatosis, and liver neutral fat.

[0084] The "pharmaceutical composition" of the present invention may also include suitable carriers, excipients, and diluents commonly used in the preparation of pharmaceutical compositions. For example, the excipient may be one or more selected from the group consisting of diluents, binders, disintegrants, lubricants, adsorbents, humectants, film-coating substances, and controlled-release additives.

[0085] The pharmaceutical compositions of the present invention can be formulated into various dosage forms according to conventional methods, including but not limited to: powders, granules, sustained-release granules, enteric-coated granules, liquids, eye drops, elixirs, emulsions, suspensions, liniments, lozenges, aromatics, lemonade, tablets, sustained-release tablets, enteric-coated tablets, sublingual tablets, hard capsules, soft capsules, sustained-release capsules, enteric-coated capsules, pills, tinctures, decoctions, dry extracts, fluid extracts, injections, capsules, infusion solutions, plasters, lotions, mud dressings, sprays, inhalers, patches, sterile injection solutions, or aerosols, etc. Furthermore, the topical agents may be in the form of creams, gels, patches, sprays, ointments, plasters, lotions, liniments, mud dressings, or pastes.

[0086] The carrier, excipients, and diluents that may be included in the pharmaceutical composition of the present invention may be: lactose, glucose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, povidone, water, methylparaben, propylparaben, talc, magnesium stearate, and mineral oil.

[0087] In the formulation process, conventional fillers, compatibilizers, binders, wetting agents, disintegrants, surfactants and other diluents or excipients are usually used for formulation.

[0088] The following additives may be used as additives in the tablets, powders, granules, capsules, pills, and lozenges according to the present invention: corn starch, potato starch, wheat starch, lactose, white sugar, glucose, fructose, D-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, calcium dihydrogen phosphate, calcium sulfate, sodium chloride, sodium bicarbonate, refined lanolin, microcrystalline cellulose, dextrin, sodium alginate, methylcellulose, sodium carboxymethyl cellulose, kaolin, urea, colloidal silica, hydroxypropyl starch, hydroxypropyl methylcellulose (HPMC) 1928, HPMC 2208, HPMC 2906, HPMC 2910. Excipients include propylene glycol, casein, calcium lactate, and Pranic acid; excipients include gelatin, gum arabic, ethanol, agar powder, cellulose acetate phthalate, carboxymethyl cellulose, calcium carboxymethyl cellulose, glucose, purified water, sodium casein, glycerin, stearic acid, sodium carboxymethyl cellulose, sodium methyl cellulose, methyl cellulose, microcrystalline cellulose, dextrin, hydroxycellulose, hydroxypropyl starch, hydroxymethyl cellulose, refined shellac, starch paste, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, polyvinyl alcohol, and povidone; binders that can be used include hydroxypropyl methyl cellulose, corn starch, agar powder, methyl cellulose, bentonite, hydroxypropyl starch, sodium carboxymethyl cellulose, sodium alginate, calcium carboxymethyl cellulose, calcium citrate, sodium lauryl sulfate, anhydrous silicate, 1-hydroxypropyl cellulose, dextran, ion exchange resin, polyvinyl acetate, formaldehyde-treated casein and gelatin, alginate, amyl starch, and guar gum. Disintegrants include: gum, sodium bicarbonate, povidone, calcium phosphate, gelled starch, gum arabic, amylopectin, pectin, sodium polyphosphate, ethyl cellulose, white sugar, magnesium aluminum silicate, D-sorbitol solution, and hard anhydrous silicate; and lubricants including: calcium stearate, magnesium stearate, stearic acid, hydrogenated vegetable oil, talc, pine nuts, kaolin, petrolatum, sodium stearate, cocoa butter, sodium salicylate, magnesium salicylate, polyethylene glycol 4000, PEG 6000, liquid paraffin, hydrogenated soybean oil, aluminum stearate, zinc stearate, sodium lauryl sulfate, magnesium oxide, polyethylene glycol (Macrogol), synthetic aluminum silicate, anhydrous silicate, higher fatty acids, higher alcohols, silicone oil, liquid paraffin, polyethylene glycol fatty acid ethers, starch, sodium chloride, sodium acetate, sodium oleate, dl-leucine, and hard anhydrous silicate.

[0089] The additives for the liquid formulation according to the present invention can be water, dilute hydrochloric acid, dilute sulfuric acid, sodium citrate, sucrose monostearate, polyoxyethylene sorbitan fatty acid esters (polysorbates), polyoxyethylene monoalkyl ethers, lanolin ethers, lanolin esters, acetic acid, hydrochloric acid, ammonia, ammonium carbonate, potassium hydroxide, sodium hydroxide, proline, polyvinylpyrrolidone, ethyl cellulose, sodium carboxymethyl cellulose, etc.

[0090] The syrups according to the present invention can use a solution of white sugar, other sugars or sweeteners, and may also use flavoring agents, coloring agents, preservatives, stabilizers, suspending agents, emulsifiers, thickeners, etc. as needed.

[0091] The emulsions according to the present invention can use purified water, and emulsifiers, preservatives, stabilizers, fragrances, etc. can be used as needed.

[0092] The suspending agent according to the present invention may use acacia gum, tragacanth gum, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, sodium alginate, hydroxypropyl methylcellulose, HPMC 1828, HPMC 2906, HPMC 2910 and other suspending agents, and may use surfactants, preservatives, stabilizers, colorants and fragrances as needed.

[0093] The injectable formulation according to the present invention comprises distilled water for injection, 0.9% sodium chloride injection, Ringer's solution, glucose injection, glucose + sodium chloride injection, polyethylene glycol (PEG), lactated Ringer's solution, ethanol, propylene glycol, non-volatile oils—sesame oil, cottonseed oil, peanut oil, soybean oil, corn oil, ethyl oleate, isopropyl myristate, benzyl benzoate, and other solvents; sodium benzoate, sodium salicylate, sodium acetate, urea, ethyl carbamate, monoethyl acetanilide, and buprofen. Cosolvents such as tazodine, propylene glycol, Tween compounds, nicotinamide, hexamethylenetetramine, and dimethylacetamide; buffers such as weak acids and their salts (acetic acid and sodium acetate), weak bases and their salts (ammonia and ammonium acetate), organic compounds, proteins, albumin, peptone, and gums; tensioning agents such as sodium chloride; stabilizers such as sodium bisulfite (NaHSO3), carbon dioxide gas, sodium metabisulfite (Na2S2O5), sodium sulfite (Na2SO3), nitrogen (N2), and ethylenediaminetetraacetic acid; antioxidants such as 0.1% sodium dithionite, sodium formaldehyde sulfoxylate, thiourea, disodium ethylenediaminetetraacetate, and sodium acetone bisulfite; analgesics such as benzyl alcohol, chlorobutanol, procaine hydrochloride, glucose, and calcium gluconate; and suspending agents such as sodium carboxymethyl cellulose, sodium alginate, Tween 80, and aluminum monostearate.

[0094] The suppositories according to the present invention may use the following bases: cocoa butter, lanolin, Witdroprol, polyethylene glycol, glycerin gelatin, methylcellulose, carboxymethylcellulose, a mixture of stearic acid and oleic acid, subanal, cottonseed oil, peanut oil, palm oil, cocoa butter + cholesterol, lecithin, Lanette wax, glyceryl monostearate, Tween or Span, Imhausen, Monolen (propylene monostearate), glycerin, Adeps solidus, Buytyrum Tego-G, Cebes Pharma 16, Hexalide Base 95, Cotomar, Hydrokote SP, S-70-XXA, S-70-XX75 (S-70-XX95), Hydrokote 25, Hydrokote 711, Idropostal, Massa Suppositories include estrarium (A, AS, B, C, D, E, I, T), Massa-MF, Masopol, Masopol-15, Neo-Suppositories-N, Paramonte-B, Supocillo (OSI, OSIX, A, B, C, D, H, L), suppository matrix type IV (AB, B, A, BC, BBG, E, BGF, C, D, 299), Supocillo (N, Es), Wickebe (W, R, S, M, Fs), and Tegister triglyceride matrix (TG-95, MA, 57), etc.

[0095] Solid dosage forms for oral administration include tablets, pills, powders, granules, capsules, etc. These solid dosage forms are formulated by mixing the extract with at least one or more excipients (e.g., starch, calcium carbonate, sucrose or lactose, gelatin, etc.). In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used.

[0096] Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups. Besides commonly used simple diluents such as water and liquid paraffin, they also contain various excipients, such as wetting agents, sweeteners, flavoring agents, and preservatives. Preparations for non-oral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate can be used as non-aqueous solvents and suspensions.

[0097] The pharmaceutical compositions according to the invention are administered at a pharmaceutically effective dose. In this invention, a "pharmaceutically effective dose" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment. The effective dose level can be determined based on factors including the type and severity of the patient's disease, drug activity, drug sensitivity, timing of administration, route of administration and excretion rate, treatment duration, concomitant drugs, and other factors known in the medical field.

[0098] The pharmaceutical compositions according to the present invention can be administered as a single therapeutic agent or in combination with other therapeutic agents, sequentially or simultaneously with existing therapeutic agents, and can be administered once or multiple times. It is important to comprehensively consider all the above factors to achieve maximum efficacy with the minimum dose while avoiding side effects, which can be easily determined by those skilled in the art to which this invention pertains.

[0099] The pharmaceutical compositions of the present invention can be administered to an individual via a variety of routes. All routes of administration are predictable, such as oral, subcutaneous, intraperitoneal, intravenous, intramuscular, perispinal (intradural) injection, sublingual, buccal, rectal, vaginal, ocular, ear, nasal, inhalation, oral or nasal spray, skin, and transdermal administration.

[0100] The pharmaceutical compositions of the present invention depend on the type of drug as the active ingredient, as well as various related factors such as the disease to be treated, the route of administration, the patient's age, sex, weight, and disease severity. Specifically, the effective dose of the compositions according to the present invention may vary depending on the patient's age, sex, and weight, typically ranging from 0.001 to 150 mg / kg body weight daily or every other day, preferably from 0.01 to 100 mg / kg body weight, or may be administered in 1 to 3 divided doses daily. However, this dose can be increased or decreased based on factors such as the route of administration, disease severity, sex, weight, and age; therefore, the stated dosage does not limit the scope of the present invention in any way.

[0101] In this invention, the term "individual" refers to a person who needs treatment for a disease. More specifically, it refers to mammals such as humans or non-human primates, mice, rats, dogs, cats, horses, and cattle.

[0102] In this invention, "administration" means providing an individual with the prescribed composition of the invention in any suitable manner. In this invention, "prevention" means all actions that inhibit or delay the onset of a target disease; "treatment" means all actions that improve or beneficially enhance the symptoms of a target disease and its associated metabolic abnormalities by administering the pharmaceutical composition according to the invention; and "improvement" means all actions that alleviate parameters related to the target disease (e.g., the severity of symptoms) by administering the composition according to the invention.

[0103] This invention provides a kit for the prevention or treatment of metabolic diseases, comprising a composition with the peptide as the active ingredient and instructions for use. In this invention, the instructions may specify, but are not limited to, an increased dosing or administration period with increasing half-life.

[0104] In addition to the above-described components, the "kit" of the present invention may also include other components, devices, and substances typically required for storage, management, efficacy enhancement, and application. Furthermore, all components of the kit can be used more than once without limitation on the number of uses, the order of use of each substance is unrestricted, and the application of each substance can be performed simultaneously or in stages.

[0105] In addition to the formulation and instructions, the kit of the present invention may also include a container. The container can be used to package the above-mentioned components and also serves a storage and securing function. For example, the container may be made of materials such as bottles, tubs, sachets, envelopes, tubes, ampoules, etc., and these containers may be partially or wholly made of plastic, glass, paper, foil, wax, etc. The container may be equipped with a fully or partially removable cap, which may initially be part of the container or attached to the container by mechanical, adhesive, or other means. Furthermore, the container may be equipped with a stopper that allows access to the contents through an injection needle. The kit may include outer packaging, which may contain instructions for the use of the components, but is not limited thereto.

[0106] The present invention provides a food composition containing the peptide as an active ingredient for the prevention or improvement of metabolic diseases.

[0107] In one embodiment of the present invention, the food may be a health functional food, but is not limited thereto.

[0108] In this invention, "food" refers to a natural or processed product containing one or more nutrients. Preferably, it refers to an article that has undergone a certain degree of processing to become ready for direct consumption, and generally includes health functional foods, beverages, food additives, and beverage additives.

[0109] In this invention, "functional food" and "food for special health use (FosHU)" are synonymous, referring to foods that, through targeted processing, can efficiently exert biological regulatory functions in addition to nutritional supply, possessing high medical and therapeutic effects. These foods can be formulated as tablets, capsules, pills, granules, powders, liquids, flakes, pastes, syrups, gels, jellies, bars, or films. The term "functional" here refers to achieving beneficial effects on human structure and function, such as regulating nutrients or physiological functions for health purposes. In this invention, the food may include oral beauty foods and oral beauty and health functional foods.

[0110] In this invention, there are no particular limitations on the types of health functional foods. Specifically, examples of foods to which the compositions of this invention can be added include dairy products including ice cream, various soups, beverages, teas, powders, alcoholic drinks, and vitamin complexes, especially food series that add value to the conventional functions of the food in order to act and manifest for a specific purpose, as well as various foods designed to fully stimulate the biological defense rhythm regulation, disease prevention and rehabilitation, and other in vivo regulatory functions of the food components in the body.

[0111] In this invention, "food additives" refers to substances used in the manufacture, processing, or preservation of food by addition, mixing, impregnation, or other methods. Similar to functional foods, long-term intake of such substances must be harmless to the human body.

[0112] When the compositions of the present invention are used as food additives, the food additives can be added directly to the compositions of the present invention, or they can be used with other foods or food ingredients, and can be used appropriately in the usual manner.

[0113] The amount of active ingredients mixed can be appropriately adjusted according to the purpose of use (prevention, health or treatment). Generally, when manufacturing food or beverages, the compositions of the present invention can be used at an addition amount of less than 15% or less than 10% of the weight of the raw materials. However, in cases of long-term intake for health and hygiene purposes or aimed at regulating health, the dosage may be lower than the above range; and since there are no safety concerns, the dosage of the active ingredients may also be higher than the above range.

[0114] In this invention, the composition may contain various food additives permitted by food science, and may also contain appropriate carriers, excipients and diluents commonly used in food manufacturing.

[0115] In addition to the above-mentioned components, the compositions of the present invention may also 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, alcohols, and carbonating agents for carbonated beverages. Furthermore, the compositions of the present invention may contain fruit pulp components used in the manufacture of natural fruit juices, fruit juice beverages, and vegetable beverages. These components may be used alone or in combination. While the proportion of such additives is not a critical factor, it is generally selected within the range of 0.01-0.20 parts by weight, based on 100 parts by weight of the composition of the present invention. However, this is not a limitation, and the optimal or arbitrary amount may be used depending on the type and function of the product being applied.

[0116] In this invention, there are no particular limitations on the types of food products to which the above-mentioned substances may be added. Examples of food products to which the above-mentioned substances may be added include: meat, sausage, bread, chocolate, candy, snacks, biscuits, pizza, ramen and other noodles, chewing gum, dairy products including ice cream, various soups, beverages, teas, powdered drinks, alcoholic beverages, and vitamin complexes, and may cover all health functional foods in the conventional sense, but are not limited thereto.

[0117] Furthermore, the compositions according to the present invention can also be added to health beverages and, like conventional beverages, contain various flavoring agents or additional ingredients such as natural carbohydrates. The natural carbohydrates include: monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As sweeteners, natural sweeteners such as sematriol and stevia extract, or synthetic sweeteners such as saccharin and aspartame, can be used. The proportion of the natural carbohydrates is typically about 0.01-0.20 g, or about 0.04-0.10 g, per 100 mL of the composition of the present invention, but is not limited thereto. Simultaneously, the commonly used amounts in the industry can be adopted, as well as the maximum range for enhancing the efficacy of the composition of the present invention, and also the optimal, arbitrary amounts considering the synergistic effects with other substances added together.

[0118] The present invention provides a cosmetic composition containing the peptide as an active ingredient for preventing or improving obesity.

[0119] In this invention, the concept of "cosmetic material" may cover all items used to beautify the appearance of the human body, but is not limited thereto, and its meaning may include the broadest definition used in the art.

[0120] The cosmetic compositions of the present invention can be used in cosmetics or as cosmetic additives. When used as cosmetic additives, the compositions of the present invention can be used to manufacture cosmetic compositions for keeping hands or feet clean. Examples include, but are not limited to, soaps (solid soaps, liquid soaps, foaming soaps, shower gels, hand soaps, etc.), facial foams, shampoos (hair shampoos, dry shampoo, etc.).

[0121] The cosmetic compositions of the present invention can be prepared using any dosage form conventional in the industry, such as toners (skin lotions), lotions, softening lotions, skin care lotions, astringents, solutions, suspensions, emulsions, creams, powders, gels, face creams, hand creams, hand sanitizers, lotions, moisturizing lotions, nourishing lotions, body lotions, shower gels, pressed powders, soaps, facial cleansers containing surfactants, oils, liquid foundations, cream foundations, wax-based foundations, sprays, and sheet products, but are not limited thereto. More specifically, they can be prepared into moisturizing lotions, nourishing lotions, nourishing creams, massage creams, moisturizing creams, serums, nourishing essences, eye creams, cleansing creams, cleansing foams, makeup removers, care films, sprays, or loose powders, etc.

[0122] Compositions of this type of dosage form can be prepared according to conventional methods in the art. The amount of the added ingredients, such as the humectant, can be easily selected by those skilled in the art without impairing the purpose and effect of the invention.

[0123] In this invention, the composition can be prepared into a conventional emulsifying or soluble formulation. Emulsifying cosmetics include nourishing toners, creams, serums, etc.; soluble cosmetics include moisturizing toners. More specifically, the compositions of this invention can be in the following forms: solutions, gels, solid or paste-like anhydrous products, emulsions formed by dispersing an oil phase in an aqueous phase, suspensions, emulsions, microemulsions, microcapsules, masks, microparticles or ionic (liposome) and nonionic vesicle dispersions, creams, toners, lotions, powders, ointments, sprays, pastes, protective films, cleansers, soaps, surfactant-containing cleaning products, oils, liquid foundations, bath powders, cream foundations, waxy bases, foundations, or concealer sticks, etc. Furthermore, they can also be prepared into foam forms or aerosol compositions containing compressed propellants.

[0124] When the dosage form of the composition is an ointment, cream, or gel, its carrier component may include animal oil, vegetable oil, waxes, paraffin, starch, tragacanth gum, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc.

[0125] When the composition is in the form of a powder or spray, its carrier component may include lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, etc. In particular, when the formulation is a spray, it may also contain propellants such as chlorofluorocarbons, propane / butane, and dimethyl ether.

[0126] When the dosage form of the composition is a solution or emulsion, its carrier component may include solvents, solubilizers, emulsifiers, etc. Specifically, it may include water, ethanol, isopropanol, diethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butanediol oil, glycerol fatty acid esters, polyethylene glycol, sorbitan fatty acid esters, etc.

[0127] When the dosage form of the composition is a suspension, its carrier components may include: liquid diluents such as water, ethanol, and propylene glycol; suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitan ester, and polyoxyethylene dehydrated sorbitan ester; microcrystalline cellulose, aluminum aluminate hydroxide, bentonite, agar, and tragacanth gum.

[0128] When the formulation of the composition is a cleaning product containing surfactants, its carrier component may include: fatty alcohol sulfate salts, fatty alcohol polyoxyethylene ether sulfate salts, sulfosuccinate monoester salts, hydroxyethyl sulfonate, imidazoline derivatives, methyl taurate, acyl sarcosine salts, fatty acid amide ether sulfate salts, alkyl amide betaine, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, ethoxylated glycerol fatty acid esters, etc.

[0129] Furthermore, the additional ingredients that can be added are not limited to the examples above. Any of the above ingredients can be used in combination as long as they do not impair the purpose and effect of the present invention. However, the amount added relative to the total weight is preferably 0.01-10% by weight, more preferably 0.01-5% by weight.

[0130] In addition to the ingredients disclosed in this specification, the cosmetic composition may also contain additional functional additives and ingredients commonly found in cosmetic compositions. Furthermore, it may also contain conventionally used purified water, thickeners, preservatives, stabilizers, solubilizers, surfactants, carriers, fragrances, or combinations thereof.

[0131] Examples of carriers include alcohols, oils, surfactants, fatty acids, silicone oils, wetting agents, humectants, viscosity modifiers, emulsions, stabilizers, ultraviolet scattering agents, ultraviolet absorbers, color developers, and fragrances. Since compounds / compositions that can be used as alcohols, oils, surfactants, fatty acids, silicone oils, wetting agents, humectants, viscosity modifiers, emulsions, stabilizers, ultraviolet scattering agents, ultraviolet absorbers, color developers, and fragrances are well known in the art, those skilled in the art can select suitable substances / compositions for use. In addition, sunscreens, antioxidants (butylated hydroxyanisole, propyl gallate, ascorbic acid, tocopheryl acetate, butylated hydroxytoluene, etc.), preservatives (methylparaben, butylparaben, propylparaben, phenoxyethanol, imidazolidinyl urea, chlorphenesin, etc.), colorants, pH adjusters (triethanolamine, citric acid, malic acid, sodium malate, fumaric acid, sodium fumarate, succinic acid, sodium succinate, sodium hydroxide, sodium monohydrogen phosphate, etc.), moisturizers (glycerin, sorbitol, propylene glycol, butylene glycol, hexanediol, diglyceride, betaine, glyceryl polyether-26, methyl gluceth-20, etc.), lubricants, and other ingredients may be added to the cosmetic composition as needed.

[0132] In this invention, the composition may further comprise one or more known ingredients with skin-improving effects, and may further comprise one or more substances conventionally used in cosmetic compositions. Specifically, it may also contain adjuvants conventionally used in the fields of cosmetics or dermatology, such as: lipids, organic solvents, solvents, thickeners and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, metal ion blocking agents and chelating agents, preservatives, vitamins, blocking agents, humectants, essential oils, dyes, pigments, hydrophilic or lipophilic active ingredients, lipovesicles, or any other ingredients conventionally used in cosmetics. Furthermore, the ingredients may be added in amounts commonly used in the field of dermatology.

[0133] In this invention, the composition may further comprise an ingredient selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high molecular weight peptides, high molecular weight polysaccharides, and sphingolipids.

[0134] In addition to the essential components mentioned above, other ingredients commonly used in cosmetics may be added as needed in the cosmetic composition of the present invention, such as: oil components, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, bactericides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, fragrances, blood circulation promoters, cooling agents, antiperspirants, purified water, etc. Specific Implementation Preferred embodiments are provided below to facilitate understanding of the present invention. However, the following embodiments are only for the purpose of helping to better understand the present invention and are not intended to limit the scope of the present invention.

[0136] [Example] Example 1. Preparation of HU020K2FA, a sustained-release GLP analog for the treatment of metabolic diseases. GLP analogs for the prevention or treatment of metabolic diseases were prepared. Specifically, they were designed to contain 17 amino acids identical to the sequence of GLP-1 and 23 amino acids identical to the sequence of GLP-2, with Lys and C-terminus added. 16 The fatty acid is palmitic acid, which is used to prepare HU020K2FA, whose sequence is shown in Table 1 below.

[0137] Table 1

[0138] Example 2. Confirmation of the excellent GLP-1 activity of HU020K2FA The GLP-1 activity of the sustained-release peptide HU020K2FA synthesized in Example 1 was analyzed. As an analytical method, a cell-based assay was performed to verify the activity of the novel GLP-1 peptide. The cell line used was GLP-1R / CHO cells (DiscoverX, product number: 95-0062C2). The induced increase in cAMP was analyzed by reacting the experimental material in GLP-1R / CHO cells. cAMP concentration was determined using a cAMP assay kit (DiscoverX, 90-0075-LM10). GLP-1 was compared with EC using Exendin-4 (DiscoverX, 92-1115, purity >95%) as a standard. 50 Cell lysis induced by increased cAMP was performed, and the increase in intracellular cAMP was quantified using ELISA to confirm the activity of each test substance relative to the control group. To ensure the accuracy of the activity results, it was preferred to perform at least three replicates; therefore, four replicates were performed for each peptide, and the average value was derived. Comparisons were then made using GLP-1 Control (Exendin-4), HU020 (non-persistent), liraglutide, and semaglutide.

[0139] The results are HU020K2FA and HU020 EC 50 The comparison results are shown in Table 2 below.

[0140]

Table 2

[0141] Specifically, the cAMP activities are presented in the following order: liraglutide (32.6 pM) ≤ semaglutide (31.1 pM) < HU020K2FA (15.1 pM) < Exendin-4 (9.6 pM) < HU020 (4.8 pM). That is, it is confirmed that the GLP-1 activity level of HU020K2FA is higher, about twice that of the positive control group, namely liraglutide and semaglutide. Based on the fact that the control agonist, Exendin-4, which binds to GLP1R overexpressing CHO cells, is mainly used for actual blood glucose regulation, and liraglutide and semaglutide are significantly superior in terms of weight loss effect, the results of this experiment show that the in vitro receptor binding activity of HU020K2FA is more than twice that of liraglutide and semaglutide. This suggests that HU020K2FA exhibits a more excellent weight loss effect than Exendin-4, liraglutide and semaglutide.

[0142] Example 3. Confirmation of the excellent obesity treatment effect of HU020K2FA The obesity treatment effect test was carried out on the sustained GLP analog prepared in Example 1. Specifically, in order to confirm the obesity treatment effect, an obese animal model was made, and after a certain amount of the substance was administered to the model, the changes in body weight, food intake and biochemical detection were carried out. First, to make a diet-induced obese mouse model, C57BL / 6 DIO mice were fed a high-fat diet for 12 weeks, and their body weight increased by 130% compared with the control group (DIO model). Subsequently, grouping was carried out at specific time points, and after confirming the average body weight of the DIO model (46.2 ± 2.2), the mice were then administered drugs for 4 weeks. During this period, the body weight and food intake were measured daily.

[0143] The experimental results (Table 3 and Figure 2 ) show that HU020K2FA exhibits higher weight loss activity than liraglutide and is confirmed to have the same level of weight loss effect as semaglutide. On the other hand, even though the same amount of HU020 as HU020K2FA was administered, not only did its body weight increase instead, but an increase in the cumulative feed intake was also observed.

[0144]

Table 3

[0145] In addition, after the experiment, blood and serum were collected from each mouse model, and biochemical tests were performed on blood glucose, total cholesterol, high-density lipoprotein (HDL), low-density lipoprotein (LDL), and triglycerides.

[0146] Experimental results (Table 4 and) Figure 3 The results showed that among all experimental groups, HU020K2FA had the most significant blood glucose reduction effect, with its efficacy being significantly superior to or similar to that of the practically used drugs liraglutide and smegglutide. Furthermore, compared to the control group, HU020K2FA also demonstrated superior efficacy in reducing total cholesterol and LDL. Moreover, under HU020 treatment, blood glucose, cholesterol, high-density lipoprotein, low-density lipoprotein, and triglycerides were all significantly higher than in the normal control group, confirming its poor efficacy in treating obesity in obese animal models.

[0147] Table 4

[0148] Based on the above results, it can be seen that the sustained GLP analogue of the present invention, namely HU020K2FA, is not only superior to liraglutide and smegglutide in terms of in vitro GLP-1 cAMP activity, but also shows significantly better weight loss, cumulative feed intake and other biochemical test results in an in vivo obese mouse model.

[0149] In particular, although the cAMP activity of the non-persistent GLP analog HU020 of this invention is superior to that of the control group, HU020K2FA, liraglutide, and smegglutide, its efficacy in in vivo experiments is poor, while HU020K2FA exhibits more prominent activity in treating obesity. This comparative result confirms that in vivo persistence is a key factor in enhancing the therapeutic effect of peptides for obesity.

[0150] Therefore, HU020K2FA of the present invention, as one of the optimized sustained GLP analogs, can produce obesity treatment effects that are comparable to or even better than those of GLP analogs, and can be effectively used as a novel sustained obesity treatment agent.

[0151] Example 4. Preparation of additional sustained-release GLP analogues for obesity treatment Based on the sequence of the GLP analog prepared in Example 1, various substances were additionally prepared (Table 5) by amino acid substitution, addition of lysine, fatty acid conjugation (C16, C18, C20), or PEG conjugation. In this case, C16 was palmitic acid, C18 was octadecanedioic acid, and C20 was eicosanedioic acid. Furthermore, the iso-Glu (gamma-glutamyl) linker used in liraglutide was employed when conjugating fatty acids and lysine. Additionally, for HU020M6, a polyethylene glycol (PEG) linker was conjugated at the 20th amino acid position to enhance peptide solubility and in vivo half-life. The conjugated PEG is a 3.4 kDa maleimide-modified PEG, a thiol / SH reactive PEG compound; the peptide is designed to allow PEG to selectively bind to the thiol (SH) group on the 20th cysteine ​​residue.

[0152] Table 5

[0153] Example 5. Confirmation of the excellent GLP-1 and GLP-2 activity of the added GLP analog. The GLP-1 and GLP-2 activities of the GLP analogs with serial numbers 3 to 9, namely HU020M1 to HU020M7, prepared in Example 4 were further analyzed. The analytical methods were the same as in Example 2.

[0154] The results are shown in Table 6 below.

[0155] Table 6

[0156] Specifically, compared with HU020K2FA (serial number 1), the GLP-1 and GLP-2 activities were compared and found to be: except for HU020M2 and HU020M5 (C20 fatty acid conjugation), all maintained GLP-1 activity. Furthermore, serial number 9 (HU020M7), with the fatty acid conjugation site changed to position 24, not only showed equivalent or higher GLP-1 activity, but also further enhanced GLP-2 activity.

[0157] This confirms that even with the same amino acid sequence, the type and conjugation site of the associated conjugated fatty acids can lead to differences in in vitro activity.

[0158] The above results indicate that the peptides prepared according to sequence numbers 1 to 9 exhibit excellent GLP-1 and GLP-2 activities.

[0159] Example 6. PK animal experiments with additional GLP analogs Mouse pharmacokinetic (PK) experiments were conducted on serial number 1 prepared in Example 1 and serial numbers 3 to 6 prepared in Example 4. Specifically, after a single subcutaneous administration of semaglutide and the candidate substance to mice, the concentration of the drug in plasma was determined by LC-MS / MS, and pharmacokinetic parameters (PK parameters) were calculated. Seven-week-old male C57BL / 6N mice were used as experimental animals. The experimental groups received HU020K2FA and HU020M1 to HU020M6, while the control group received semaglutide. All experimental and control groups received a single subcutaneous administration of 0.2 mg / 5 mL / kg to the dorsal region of a total of 12 mice using a 1 mL syringe equipped with a 26-gauge needle. Following drug administration, approximately 0.04-0.05 mL of blood was collected from each mouse via the orbital venous plexus at predetermined time points (four mice were cross-sampled). Blood collection time points were 1, 2, 4, 8, 24, 32, 48, 56, 72, 96, and 120 hours. All plasma samples were analyzed using a self-developed LC-MS / MS method. Pharmacokinetic analysis of the drug in the blood was performed using a non-compartmental model method, and pharmacokinetic parameters were analyzed using Phoenix WinNonlin 64 (version 8.3.5.340).

[0160] The results are shown in Table 7 below.

[0161] Table 7

[0162] Specifically (Table 7 and) Figure 4 The original C16 fatty acid conjugated HU020K2FA and HU020M3, in which the glycine at position 2 is replaced by Aib, showed a similar half-life to smegglutinin. However, the new candidates HU020M1, HU020M2, HU020M4, and HU020M5, which are conjugated with C18 and C20 fatty acids, showed a half-life that is about 1.5 to 2.0 times longer than that of smegglutinin.

[0163] These results indicate that peptides with equivalent or higher half-life characteristics based on sequence numbers 1 to 9 can enhance the therapeutic effect on metabolic diseases.

[0164] Example 7. Confirmation of the excellent anti-obesity effect of the added GLP analogue The anti-obesity effects of sequence number 1 prepared in Example 1 and sequences 6 and 7 prepared in Example 4 were analyzed. HU020M4 and HU020M5, which possess GLP-1 activity and have a longer half-life than semaglutide, were selected for comparison with the control substance semaglutide. Furthermore, the experimental model used was a DIO (Diet Induced Obesity) mouse model induced by a 3-month high-fat diet.

[0165] Example 7-1. Comparison of weight loss effect and cumulative feed intake To analyze the anti-obesity effect, a comparative analysis of the weight loss effect and cumulative feed intake was first conducted. The experimental method used was the same as in Example 3.

[0166] The results are confirmed in Table 8 below. Figure 5 As shown.

[0167] Table 8

[0168] Specifically, when administered once daily (QD) for 4 weeks, the weight loss effects of smegglutinin, HU020K2FA, HU020M4, and HU020M5 were almost identical, and the cumulative feed intake also showed a similar trend to the weight loss effect. This effect is predicted by the GLP-1 activity of the peptides of this invention and the increased half-life.

[0169] Example 7-2. DEXA imaging results confirm the fat reduction effect. To analyze the anti-obesity effect, DEXA (Body Composition Analysis using Dual-Energy X-ray Absorptiometry) analysis was performed. The DEXA imaging system (VETDXA iNSiGHT) measured bone mineral density, body fat, and lean body mass in the experimental animals using dual X-ray transmission. After administration of the test substance, DEXA measurements of fat, muscle mass, etc., were performed on each individual. Based on this, the fat reduction effect was analyzed by categorizing body weight into BMC and lean / fat, thus providing a more objective analysis of the anti-obesity effect according to Example 7-1.

[0170] The results are confirmed in Table 9 below. Figure 6 and 7 As shown.

[0171] Table 9

[0172] Specifically, measurements showed that the fat and lean weight of smegglutide, HU020K2FA, HU020M4, and HU020M5 were almost at the same level. In particular, this effect was still observed even when the dosage of HU020M4 and HU020M5 was the same as that of smegglutide. These results indicate that the peptides of the present invention exhibit good anti-obesity effects through amino acid alteration and fatty acid conjugation modification.

[0173] Example 7-3. Confirmation of fat reduction effect in each tissue To analyze the anti-obesity effect, fat mass in various tissues was measured after autopsy, further confirming the efficacy of peptides in obesity. Specifically, the epididymal, mesenteric adipose tissue, inguinal, and perirenal tissues of animals that had been bled to death were removed, and fat weight was measured and photographed at each site.

[0174] The results are confirmed as shown in Table 10 below. Figure 8 As shown. Specifically, regarding the amount of fat in various tissues, it was confirmed that smegglutinin, HU020K2FA, HU020M4, and HU020M5 all showed similar tendencies in weight changes among the various drugs. Based on these results, it was confirmed that the peptides of the present invention significantly reduce the fat weight of the epididymis, mesentery, groin, and perirenal region, which are significantly increased due to obesity, and are substances with excellent anti-obesity effects.

[0175] Table 10

[0176] Example 7-4. Confirmation of plasma obesity-related factor levels To analyze the anti-obesity effect, plasma was obtained from mice after necropsy, and obesity-related factors in the plasma were measured. In this case, the analytical method was the same as that described in Example 3.

[0177] The results are confirmed in Table 11 below. Figure 9 As shown. Specifically, smegglutinin, HU020K2FA, HU020M4, and HU020M5 were confirmed to exhibit similar tendencies in plasma levels of obesity-related factors. In particular, regarding plasma glucose (GLU), HU020K2FA, HU020M4, and HU020M5 all showed a more significant reduction effect than smegglutinin. Based on these results, it is confirmed that the peptides of the present invention reduce all plasma obesity-related factors, thereby demonstrating excellent therapeutic efficacy for blood fat-related diseases.

[0178] Table 11 Example 8. Confirmation of the good anti-obesity effect of the additional GLP analog in the liver The anti-obesity effect in the liver was analyzed using the same substance as in Example 7. Specifically, liver injury markers (AST / ALT), liver weight / lipidemia examination, liver neutral fat measurement, and oil red O staining analysis were performed.

[0179] Example 8-1. Analysis of liver injury markers AST / ALT To analyze the liver injury marker AST / ALT, blood biochemical tests were performed on blood from a treated obese mouse model. Specifically, collected blood was aliquoted into two tubes: approximately 200 μL of blood was injected into the EDTA tube, and the remainder was injected into a vacuum blood collection tube containing a coagulant. Glycated hemoglobin was measured on the same day in the EDTA tube. Blood from the vacuum blood collection tube was allowed to coagulate at room temperature for 15–20 minutes, then centrifuged for 10 minutes to obtain serum, which was then used for blood biochemical analysis.

[0180] The results are confirmed in Table 12 below. Figure 10 As shown. Specifically, studies have shown that when treated with HU020K2FA, HU020M4, and HU020M5, the elevated AST / ALT values ​​due to obesity were significantly reduced. Specifically, compared with smegglutinin, the peptides of the present invention showed a more statistically significant reduction, confirming them as substances exhibiting improved anti-obesity effects.

[0181] Table 12

[0182] Example 8-2. Examination of liver weight and steatohepatitis To examine liver weight / steatopathy, the increase in liver weight induced by obesity and the degree of steatopathy induction after liver H&E staining were graded from 0 to 3 to compare the drug's hepatotoxic effects. First, after the trial, to confirm the drug's hepatotoxic efficacy, the animals' livers were removed on the necropsy day. After measuring the weight of the removed liver, the median lobe was frozen in liquid nitrogen, and neutral fat was subsequently measured. The remaining liver was fixed in 10% neutral buffered formalin and subjected to histopathological examination.

[0183] For H&E analysis, liver tissue fixed in formalin was processed using an automated tissue processor and then embedded in paraffin to create paraffin blocks. These blocks were then prepared into tissue sections and stained with hematoxylin and eosin (H&E). Each stained tissue slide was observed and photographed using an optical microscope (BX61, Olympus, Japan) and (DP80, Olympus, Japan).

[0184] The quantitative histopathological evaluation of liver tissue involved scoring the degree of steatosis (0-3) on H&E-stained slides. The corresponding information for each scoring score is as follows.

[0185]

[0186] (* indicates the degree of fatty liver disease in the parenchymal cells. The parenchymal cells in the liver are mainly hepatocytes.) The results are confirmed in Table 13 below. Figures 11-12 As shown in the figure. Specifically, treatment with HU020K2FA, HU020M4, and HU020M5 significantly reduced liver weight and hepatic steatosis score, confirming their superior efficacy compared to smegglutide. In particular, HU020M4 and HU020M5 produced this effect at the same dosage as smegglutide, demonstrating their excellent efficacy in the liver.

[0187] Table 13

[0188] Example 8-3. Liver TG (Neutral Fat) Measurement To determine the level of neutral fat in the liver, neutral fat in liver tissue removed on the day of autopsy was analyzed using a assay kit (Biomax, Cat no. BM-TGR-100).

[0189] The results are confirmed in Table 14 below. Figure 13 As shown in the figure. Specifically, the results indicated that treatment with HU020K2FA, HU020M4, and HU020M5 significantly reduced liver triglycerides. In particular, it was confirmed that HU020M4 and HU020M5, at the same dosage as smegglutinin, could reduce the increased triglycerides in the liver to the level of normal mice (control group), thus confirming their superior anti-obesity effect compared to smegglutinin.

[0190] Table 14

[0191] Example 8-4. ORO Analysis Finally, to confirm the anti-obesity effect in the liver, Oil Red O staining analysis was performed. ORO staining is a staining method used to detect neutral fat in fat-related diseases, and is used to differentiate between fat embolism, liposarcoma, hepatic steatosis, and accumulation diseases. It can also detect lipid complexes such as phospholipids. For ORO analysis, frozen tissue sections need to be prepared. Liver tissue is refrigerated in 30% sucrose for 3 days to create embedded frozen blocks (OCT blocks), which are then used to prepare cryosections and stained with Oil Red O (ORO).

[0192] The results are confirmed in Table 15 below. Figures 14-15 As shown. Specifically, when treated with HU020K2FA, HU020M4, and HU020M5, the percentage of ORO-positive areas in the liver was significantly reduced, and almost no orange to dark red neutral fat was observed by the naked eye. In particular, this effect was confirmed to be superior to smegglutinin regardless of the type or dosage of HU020K2FA, HU020M4, and HU020M5.

[0193] Table 15

[0194] Example 9. Confirmation of the sustained anti-obesity effect of the additional GLP analogue Using HU020M4, which has the best efficacy among existing QD (once a day) administration methods, we analyzed whether a sustained anti-obesity effect would occur with prolonged administration. In this case, the experiment used the same experimental method as before, and conducted a probability test of administration of smegglutinin and Q2D (once every 2 days) as control groups on the same DIO obese mice.

[0195] Example 9-1. Comparison of weight loss effect and cumulative feed intake The analysis results are shown in Table 16 below. Figure 16 As shown.

[0196] Table 16

[0197] Specifically, all treatment groups showed statistically significant weight loss compared to G2 (obese mice) (P<0.05). When semaglutide and HU020M4 were administered at the same dose of 25 nmol / kg once every two days (Q2D), the weight loss effect was particularly significant with HU020M4 (P<0.05). Furthermore, the semaglutide 25 nmol / kg / Q2D treatment group showed comparable levels of weight loss to the HU020M4 10 nmol / kg / Q2D treatment group (P>0.05).

[0198] These results confirm that, due to the 1.5-fold longer half-life of HU020M4 compared to smegglutinin, the same level of weight loss can be achieved even when the drug is administered at half the concentration. Furthermore, when administered at the same concentration of 25 nmol / kg / Q2D, an even greater weight loss effect was confirmed.

[0199] In conclusion, the HU020M4 of the present invention exhibits superior anti-obesity activity compared to smegglutinin, not only when administered once a day but also when administered once every two days, as confirmed by animal experimental data.

[0200] Example 9-2. Confirmation of plasma obesity-related factor levels The analysis results are shown in Table 17 below. Figure 17 As shown in the figure. Specifically, the levels of obesity-related factors in plasma decreased after treatment with both smegglutide and HU020M4, but the decrease in most factors induced by HU020M4 was comparable to or even better than that smegglutide.

[0201] Table 17

[0202] Example 9-3. Analysis of liver injury markers AST / ALT The analysis results are confirmed in Table 18 below. Figure 18 As shown in the figure, plasma levels of obesity-related factors decreased after treatment with both smegglutide and HU020M4. In particular, HU020M4 treatment showed superior reduction effects on all indicators, including GLU, TCHO, TG, LDL, and HDL, compared to smegglutide.

[0203] Table 18

[0204] The above description of the present invention is merely illustrative. Those skilled in the art should understand that modifications can be readily implemented in other specific forms without altering the technical concept or essential features of the invention. Therefore, the embodiments listed above should be understood in all respects as exemplary, not restrictive.

[0205] Industrial availability This invention relates to a pharmaceutical composition for the prevention or treatment of obesity, comprising a GLP (glucagon-like peptide) analog as an active ingredient. The GLP analog of this invention, as analogs of GLP-1 and GLP-2, has been shown to have a longer duration of activity in vivo due to its increased half-life, thereby exhibiting excellent efficacy in the prevention or treatment of metabolic diseases. Specifically, the peptide of this invention reduces individual body weight and fat, decreases blood glucose, total cholesterol, high-density lipoprotein, low-density lipoprotein, and triglyceride levels in the blood, while also reducing liver ASL and AST levels, liver weight, degree of hepatic steatosis, and liver triglyceride levels. Therefore, the peptide of this invention can be utilized as an excellent agent for the prevention or treatment of metabolic diseases and possesses industrial applicability.

Claims

1. A peptide, characterized in that, It is represented by the following formula I: HX2EGX5FTX8DX 41 , 39 , 36 , 34 , 43 , 32 , 42 , 31 , 40 , 38 , 35 , 33 X 11 X 12 X 13 X 14 X 15 X 16 X 17 X 18 X 19 X 20 X 21 FIX 24 X 25 LX 27 X 28 X 29 X 30 X 31 X 32 X 33 X 34 X 35 X 36 X37X 38 X 39 X 40 X 41 X 42 X 43 [Formula I] In equation I, X2 is either Gly or Aib; X5 is either Thr or Ser; X8 is either Ser or Asp; X 10 For Leu or Val; X 11 For Ser or Asn; X 12 For Thr or Ser; X 13 For Tyr or Ile; X 14 For Leu or Met; X 15 For Asp or Glu; X 16 It is any one of the choices from the group consisting of Ala, Gly, and Asn; X 17 For Leu or Gln; X 18 and X 19 For Ala or His; X 20 For Arg or Cys; X 21 For Asp or Glu; X 24 For Asn or Lys; X 25 For Trp or Thr; X 27 For Ile or Val; X 28 For Gln or Lys; X 29 For Thr or Gly; X 30 For Lys or Arg; X 31 For Ile or missing; X 32 Thr or missing; X 33 For ASP or missing; X 34 ~X 43 For missing or Lys.

2. The peptide according to claim 1, characterized in that, X5 is Thr, X8 and X 11 For Ser, X 12 For Thr, X 13 For Tyr, X 14 For Leu, X 15 For Asp, X 16 For Ala, X 17 For Leu, X 19 For Ala, X 21 For Asp, X 25 For Trp, X 27 For Ile, X 28 For Gln, X 29 For Thr, X 30 For Lys, X 31 For Ile, X 32 For Thr, X 33 For Asp, X 34 ~X 43 or X 36 ~X 43 Missing.

3. The peptide according to claim 1 or 2, characterized in that, X 10 For Leu, X 18 For Ala, X 34 and X 35 For Lys.

4. The peptide according to any one of claims 1 to 3, characterized in that, When the X 20 When it is Cys, PEG (Polyethylene glycol) is bound to Cys.

5. The peptide according to any one of claims 1 to 4, characterized in that, From the X 24 and X 35 ~X 43 The group consists of one or more amino acids selected from the group that are linked to fatty acids.

6. The peptide according to claim 5, characterized in that, The fatty acid is C. 16 ~C 20 .

7. The peptide according to claim 6, characterized in that, The fatty acid is any one selected from the group consisting of palmitic acid, octadecanedioic acid, eicosanedioic acid, margaric acid, heptadecanedioic acid, hexadecanedioic acid, stearic acid, nonadecylic acid, nonadecanedioic acid, and eicosanedioic acid.

8. The peptide according to any one of claims 1 to 7, characterized in that, The peptide comprises any one selected from the group consisting of amino acid sequences represented by sequence numbers 1 to 9.

9. A pharmaceutical composition for the prevention or treatment of metabolic diseases, characterized in that, The peptide described in any one of claims 1 to 8 may be included as an active ingredient.

10. The pharmaceutical composition according to claim 9, characterized in that, The metabolic disease is any one or more selected from the group consisting of obesity, diabetes, non-alcoholic fatty liver disease, hepatosteosis, dyslipidemia, arteriosclerosis, insulin resistance syndrome, and complications thereof.

11. The pharmaceutical composition according to claim 9 or 10, characterized in that, The peptide is persistent.

12. The pharmaceutical composition according to any one of claims 9 to 11, characterized in that, The composition is administered by any method selected from the group consisting of subcutaneous administration, intravenous administration and oral administration.

13. The pharmaceutical composition according to any one of claims 9 to 12, characterized in that, The composition activates GLP-1 and GLP-2.

14. The pharmaceutical composition according to any one of claims 9 to 13, characterized in that, The composition has any one or more characteristics selected from the group consisting of the following features: a) Reduce individual weight and fat; b) Lower blood sugar, total cholesterol, high-density lipoprotein, low-density lipoprotein, and triglycerides; as well as c) Reduce liver ASL and AST levels, liver weight, degree of liver steatosis, and liver neutral fat.

15. A kit for the prevention or treatment of metabolic diseases, characterized in that, The invention includes compositions comprising any one of the peptides described in claims 1 to 8 as active ingredients, and the description thereof.

16. A food composition for preventing or improving metabolic diseases, characterized in that, The peptide described in any one of claims 1 to 8 is included as an active ingredient.

17. The food composition according to claim 16, characterized in that, The food in question is a health-functional food.

18. A cosmetic composition for preventing or improving obesity, characterized in that, The peptide described in any one of claims 1 to 8 is included as an active ingredient.

19. A method for the prevention, improvement, or treatment of metabolic diseases, characterized in that, The method includes the step of administering a pharmaceutically effective amount of a composition comprising a peptide as an active ingredient, according to any one of claims 1 to 8, to an individual in need of the composition.

20. The use of a composition for the prevention, improvement, or treatment of metabolic diseases, characterized in that, The peptide described in any one of claims 1 to 8 is included as an active ingredient.

21. Use of a composition in the preparation of an agent for the prevention, improvement or treatment of metabolic diseases, characterized in that, The peptide described in any one of claims 1 to 8 is included as an active ingredient.

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