Half-Life Extended melanocortin-4 receptor agonist compound

Abstract

The present invention relates to MC4R (Melanocortin-4 Receptor) agonist compounds with fatty acid binding technology optimized for improved in vivo stability, and pharmaceutical compositions, cosmetic compositions, and health functional foods containing the same.

Description

Technology Field

[0001] The present invention relates to MC4R (Melanocortin-4 Receptor) agonist compounds with fatty acid binding technology optimized for improved in vivo stability, and pharmaceutical compositions, cosmetic compositions, and health functional foods containing the same. Background Technology

[0002] Peptide-based drugs are being utilized in the treatment of various diseases due to their high target specificity and low toxicity. In particular, peptides have the advantage of possessing a relatively simple structure compared to protein-based drugs and can act rapidly in vivo due to their small molecular size. Thanks to these characteristics, peptide drugs have broad potential for application, including the treatment of cancer, metabolic diseases, neurological diseases, and infectious diseases.

[0003] In particular, among the peptides of the melanocortin hormone family, MSHs (SEQ No. 1), such as α-MSH and β-MSH, interact with MC4R (Melanocortin-4 Receptor) and play an important role in metabolic regulation and appetite suppression. Through their action on melanocortin receptors (MC1R to MC5R), they are involved in melanin production, inflammation regulation, and energy balance regulation. Among these, MC4R plays a central role in appetite suppression and energy expenditure regulation, and is attracting attention as a major target for the development of treatments for metabolic diseases and rare obesity.

[0004] However, naturally derived peptides such as α-MSH and β-MSH are easily degraded by endogenous enzymes and have short half-lives, which limits their clinical use. These drawbacks restrict the time the drug circulates in the bloodstream, necessitating frequent administration. Therefore, a technical approach is required to extend the half-life and enhance stability by modifying or chemically altering the natural sequences of α-MSH, β-MSH, and similar compounds.

[0005] Technologies such as fatty acid binding, polyethylene glycol (PEGylation), and albumin fusion protein have been utilized to extend the half-life and enhance the stability of peptide drugs. Among these, fatty acid binding technology is effective in extending the circulation time in the body by increasing the albumin binding affinity of peptide drugs. Fatty acid binding has established itself as a key technology, particularly in the development of treatments for metabolic diseases and hormone-based drugs. However, simply adopted fatty acid linkers (e.g., Glu) often fail to sufficiently increase the active exposure time (AUC) of the drug due to low in vivo stability. Due to this issue, structural optimization of the linker connecting the peptide and the fatty acid is essential to maximize the effectiveness of fatty acid binding technology. The problem to be solved

[0006] To overcome the shortcomings of existing MSH-based natural peptides, the present invention provides a new design method that optimizes fatty acid binding technology and enhances the in vivo stability of MC4R agonist compounds.

[0007] However, the problems that this invention seeks to solve are not limited to those mentioned above, and other unmentioned problems will be clearly understood by those skilled in the art from the description below. means of solving the problem

[0008] The first aspect of the present invention provides a compound comprising the following amino acid sequence, or a stereoisomer, solvate, or salt thereof:

[0009] KYRCEHFRWC(sequence number 2);

[0010] In the above sequence, the K at position 1 is an epsilon-amino group ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-GABA(γ-Aminobutyric Acid)-CO-(CH2) a - Connected to CO2H, where a is an integer from 14 to 18.

[0011] The second aspect of the present invention provides a pharmaceutical composition for the prevention or treatment of metabolic diseases, comprising one or more selected from the group consisting of the compound of the first aspect, stereoisomers thereof, solvates, and salts.

[0012] The third aspect of the present invention provides a composition for improving skin health comprising one or more selected from the group consisting of the compound of the first aspect, stereoisomers thereof, solvates, and salts.

[0013] The fourth aspect of the present invention provides a health functional food for preventing or improving metabolic diseases, comprising one or more selected from the group consisting of the compound of the first aspect, stereoisomers thereof, solvates, and salts. Effects of the invention

[0014] The present invention relates to an MC4R agonist compound containing GABA-fatty acid, which exhibits significantly improved in vivo stability and half-life compared to existing MC4R agonists. The compound of the present invention is designed to significantly increase resistance to degradation in vivo, particularly in liver metabolic enzymes and the intestinal environment, through the design of a peptide of SEQ ID NO. 2 and an optimal combination of fatty acid and GABA-based linker. Additionally, it is designed to allow the drug to circulate in the body for a longer period by strengthening albumin binding affinity. By extending the drug's half-life, the frequent administration required by existing technologies can be reduced, thereby increasing patient convenience in treatment.

[0015] The MC4R agonist compound of the present invention plays an essential role in metabolic regulation, such as appetite suppression and promotion of energy consumption, while maintaining high target specificity and stability, thereby maximizing weight loss and metabolic improvement effects. In particular, the GABA linker provides structural stability and enzymatic resistance to the peptide SEQ ID NO. 2 compared to existing linkers (such as NPV), resulting in significantly increased stability in the liver S9 and intestinal environment. This demonstrates that the compound of the present invention can significantly increase the in vivo exposure time compared to existing MC4R agonists, making a significant contribution to maximizing actual therapeutic effects. Brief explanation of the drawing

[0016] Figure 1 is a figure showing the results of a stability test in plasma of a compound (Examples 1-3) and a comparative example in one embodiment. Figure 2 is a figure showing the results of a stability test in S9 between a compound in one embodiment (Examples 1-3) and a comparative example. Figure 3 is a figure showing the results of a stability test in a compound (Examples 1-3) of one embodiment and an artificial serous fluid (SIF) of a comparative example. Figure 4 is a figure showing the results of a pharmacokinetic test of a compound according to one embodiment (Example 3) and a comparative example. Specific details for implementing the invention

[0017] Throughout this specification, when a part is described as being “connected” to another part, this includes not only cases where they are “directly connected” but also cases where they are “electrically connected” with other elements interposed between them.

[0018] Throughout this specification, when a component is described as being located “on” another component, this includes not only cases where a component is in contact with another component, but also cases where another component exists between the two components.

[0019] Throughout this specification, when a part is described as “comprising” a certain component, this means that, unless specifically stated otherwise, it does not exclude other components but may include additional components. Throughout this specification, terms such as “about,” “substantially,” etc., are used to mean at or near the stated value when inherent manufacturing and material tolerances are presented in the said sense, and are used to prevent unscrupulous infringers from unfairly exploiting disclosures in which precise or absolute values ​​are mentioned to aid in understanding this invention.

[0020] The terms “step” or “step of” used throughout this specification do not mean “step for”.

[0021] Throughout this specification, the term “combination(s) of these” included in the Markush-form expression means one or more mixtures or combinations selected from the group consisting of the components described in the Markush-form expression, and means including one or more selected from the group consisting of said components.

[0022] Throughout this specification, the description “A and / or B” means “A or B, or A and B”.

[0023] Hereinafter, embodiments and examples of the present invention will be described in detail with reference to the attached drawings. However, the present invention may not be limited to these embodiments and examples and the drawings.

[0025] The first aspect of the present invention provides a compound comprising the following amino acid sequence, or a stereoisomer, solvate, or salt thereof:

[0026] KYRCEHFRWC(sequence number 2);

[0027] In the above sequence, the K at position 1 is an epsilon-amino group ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-GABA(γ-Aminobutyric Acid)-CO-(CH2) a - Connected to CO2H, where a is an integer from 14 to 18.

[0028] In one embodiment of the present invention, K at position 1 may be L-type or D-type, C at position 4 may be L-type or D-type, F at position 7 may be L-type or D-type, and C at position 10 may be L-type or D-type.

[0029] In one embodiment of the present invention, K at position 1 may be of type D, C at position 4 may be of type D, F at position 7 may be of type D, and C at position 10 may be of type D.

[0030] In one embodiment of the present invention, a may be 14, 16, or 18. More specifically, a may be 18.

[0031] In one embodiment of the present invention, the C at position 4 and the C at position 10 may form a bond with each other. Specifically, the cysteine ​​at position 4 and the cysteine ​​at position 10 of the peptide of SEQ ID NO. 2 may form a disulfide bond. Such a disulfide bond causes the peptide to have a cyclic structure, thereby providing structural stability, which contributes to extending the in vivo stability and half-life of the peptide. The cyclic structure increases resistance to degradation by degrading enzymes, allowing the peptide to circulate stably in the body for a longer period of time. Furthermore, when the three-dimensional structure of the peptide is stabilized through the disulfide bond, the binding affinity with the target receptor (MC4R) increases, thereby maximizing biological activity.

[0032] In one embodiment of the present invention, the amino acid sequence may include an N-terminal amino acid that is acetylated, a C-terminal amino acid that is amidated, or an N-terminal amino acid that is acetylated and a C-terminal amino acid that is amidated. Specifically, in the peptide of SEQ ID NO. 2, the K at position 1 may have an N-terminal acetylation and the C at position 10 may have a C-terminal amidation to maximize stability and pharmacological efficacy. N-terminal acetylation modifies the epsilon-amino group of lysine (K) to adjust the charge of the peptide and increases resistance to enzymatic degradation, thereby extending the half-life in vivo. This allows the drug to circulate more stably in the body and provides the effect of improving binding affinity with the target receptor (MC4R). C-terminal amidation adjusts the polarity of the peptide and neutralizes the charge, helping to prevent unnecessary degradation of the drug in the body. In addition, C-terminal amidation optimizes the absorption and distribution of drugs in the body by enabling peptides to have higher physical stability. This combination of N-terminal acetylation and C-terminal amidation can simultaneously increase the stability and bioavailability of peptides.

[0033] The compound of the present invention comprises SEQ ID NO. 2 (KYRCEHFRWC), which can form a cyclic structure through a disulfide bond between cysteine ​​groups at positions 4 and 10. This cyclic structure significantly enhances the structural stability of the peptide and contributes to extending its half-life in vivo by increasing resistance to degradation enzymes. Additionally, the N-terminal lysine (K) is acetylated, which optimizes the physicochemical properties of the peptide and further strengthens its stability in the body. The compound increases binding affinity to the target receptor (MC4R) through a balance of polar and hydrophobic amino acids, and can provide excellent pharmacological efficacy in therapeutic areas such as metabolic diseases and rare obesity.

[0034] The above peptide of sequence number 2 may have conservative substitutions at 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids, but is not limited thereto.

[0035] In this specification, the term “conservative substitution” means substituting one amino acid with another amino acid having similar structural and / or chemical properties. The peptide may have, for example, one or more conservative substitutions while still retaining biological activity. Such amino acid substitutions may generally occur based on similarities in the polarity, charge, solubility, hydrophobicity, hydrophilicity, and / or amphipathic nature of the residues. For example, positively charged (basic) amino acids include arginine, lysine, and histidine; negatively charged (acidic) amino acids include glutamic acid and aspartic acid; aromatic amino acids include phenylalanine, tryptophan, and tyrosine; and hydrophobic amino acids include alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan. In addition, amino acids can be classified into those with electrically charged side chains and those with uncharged side chains. Amino acids with electrically charged side chains include aspartic acid, glutamic acid, lysine, arginine, and histidine. Amino acids with uncharged side chains can be further classified into nonpolar or polar amino acids. Nonpolar amino acids include glycine, alanine, valine, leucine, isoleucine, methionine, and proline, while polar amino acids include serine, threonine, cysteine, asparagine, and glutamine. Conservative substitution with amino acids having similar properties as described above can be expected to exhibit the same or similar activity.

[0036] Amino acid exchanges in proteins and peptides that do not alter the overall activity of the molecule are known in the art (H.Neurath, RLHill, The Proteins, Academic Press, New York, 1979). The most common exchanges are exchanges between amino acids Ala / Ser, Val / Ile, Asp / Glu, Thr / Ser, Ala / Gly, Ala / Thr, Ser / Asn, Ala / Val, Ser / Gly, Tyr / Phe, Ala / Pro, Lys / Arg, Asp / Asn, Leu / Ile, Leu / Val, and Gln / Glu.

[0037] In addition, the peptide may include non-standard amino acids that have undergone post-translational modification. Examples of post-translational modification may include phosphorylation, glycosylation, acylation (e.g., acetylation, myristoylation, and palmitoylation), amidation, alkylation, carboxylation, hydroxylation, glycation, biotinylation, ubiquitinylation, changes in chemical properties (e.g., beta-removal deimidation, deamidation), and structural changes (e.g., formation of disulfide bridges). In addition, it may include changes in amino acids resulting from chemical reactions occurring during the bonding process with crosslinkers to form peptide conjugates, such as changes in amino groups, carboxyl groups, or side chains.

[0038] In one embodiment, the peptide of SEQ ID NO. 2 may have an amino acid modified at the N-terminus, C-terminus, or both ends.

[0039] Here, the term "terminus" does not refer to a single amino acid at the very end of the C-terminus or N-terminus of the peptide, but may refer to up to five amino acids from the very end of the C-terminus or N-terminus. Additionally, the amino acid sequence may be modified by adding other amino acids, not limited to the types of amino acids described above. In this case, the addition or deletion of the amino acids may not affect the original functionality of the beta-melanocyte-stimulating hormone.

[0040] For example, the modification of the amino acids at the N-terminal, C-terminal, or both ends may be palmitoylation, myristoylation, acetylation, acylation, esterification, sulfonylation, alkylation, formylation, glycosylation, amidation, prenylation, or PEGylation of the amino acids. Specifically, the amino acid sequence may be such that the N-terminal amino acid is acetylated, the C-terminal amino acid is amidated, or the N-terminal amino acid is acetylated and the C-terminal amino acid is amidated. However, it is not limited thereto.

[0041] Even if the present specification describes a "peptide of a specific sequence number," if it has the same or corresponding activity as a peptide composed of the amino acid sequence of the said sequence number, it does not exclude meaningless sequence additions before or after the amino acid sequence of the said sequence number, naturally occurring mutations, or silent mutations thereof, and it is obvious that even in cases with such sequence additions or mutations, it falls within the scope of the present invention. That is, even if there are differences in some sequences, if it exhibits homology above a certain level and exhibits beta-melanocyte-stimulating hormone activity, it may fall within the scope of the present invention. Specifically, the above compound may include, but is not limited to, an amino acid sequence having 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% or more homology or identity with the amino acid sequence of SEQ ID NO. 2.

[0042] “Homology” or “identity” refers to the degree of relationship between two given amino acid sequences or base sequences and can be expressed as a percentage. The terms “homology” and “identity” are often used interchangeably.

[0043] Whether any two peptide sequences have homology, similarity, or identity can be determined using known computer algorithms, such as the “FASTA” program, using default parameters as in, for example, Pearson et al (1988) [Proc. Natl. Acad. Sci. USA 85]: 2444. Alternatively, it can be determined using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol. 48: 443-453), as performed in the Needleman program of the EMBOSS package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, Trends Genet. 16: 276-277) (version 5.0.0 or later). (Includes the GCG program package (Devereux, J., et al, Nucleic Acids Research 12: 387 (1984)), BLASTP, BLASTN, FASTA (Atschul, [S.] [F.,] [ET AL, J MOLEC BIOL 215]: 403 (1990); Guide to Huge Computers, Martin J. Bishop, [ED.,] Academic Press, San Diego, 1994, and [CARILLO ETA / .](1988) SIAM J Applied Math 48: 1073). For example, homology, similarity, or identity can be determined using BLAST from the National Center for Biotechnology Information Database, or ClustalW.

[0044] The homology, similarity, or identity of peptides can be determined by comparing sequence information using a GAP computer program, such as that described in, for example, Smith and Waterman, Adv. Appl. Math (1981) 2:482, or Needleman et al. (1970), J Mol Biol. 48: 443. In summary, the GAP program is defined as the total number of symbols in the shorter of the two sequences divided by the number of similarly arranged symbols (i.e., amino acids). The default parameters for the GAP program are (1) a univariate comparison matrix (containing values ​​of 1 for identity and 0 for non-identity) and, as disclosed by Schwartz and Dayhoff, eds., Atlas Of Protein Sequence And Structure, National Biomedical Research Foundation, pp. 353-358 (1979), or Gribskov et al. (1986) Nucl. Acids Res. 14: may include a weighted comparison matrix of 6745 (or an EDNAFULL (EMBOSS version of NCBI NUC4.4) substitution matrix); (2) a penalty of 3.0 for each gap and an additional penalty of 0.10 for each symbol in each gap (or a gap opening penalty of 10, a gap extension penalty of 0.5); and (3) no penalty for terminal gaps. Thus, as used herein, the terms “homology” or “identity” indicate relevance between sequences.

[0045] In one embodiment, the peptide may be in the form of its solvate. "Solvent" means that the peptide or its salt forms a complex with a solvent molecule.

[0046] The second aspect of the present invention provides a pharmaceutical composition for the prevention or treatment of metabolic diseases, comprising one or more selected from the group consisting of the compound of the first aspect, stereoisomers thereof, solvates, and salts.

[0047] The third aspect of the present invention provides a cosmetic composition for improving skin health comprising one or more selected from the group consisting of the compound of the first aspect, stereoisomers thereof, solvates, and salts.

[0048] The fourth aspect of the present invention provides a health functional food for preventing or improving metabolic diseases, comprising one or more selected from the group consisting of the compound of the first aspect, stereoisomers thereof, solvates, and salts.

[0049] In one embodiment of the present invention, the metabolic disease may be related to the regulation of the melanocortin-4 receptor (MC4R).

[0050] In one embodiment of the present invention, the metabolic disease may be selected from the group consisting of type 2 diabetes, obesity, insulin resistance, metabolic syndrome, hyperlipidemia, arteriosclerosis, inflammatory disease, Prader-Willi syndrome, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, cardiovascular disease and pulmonary disease, but is not limited thereto.

[0051] In this specification, the term “pharmaceutical composition” may refer to a molecule or compound that imparts several beneficial effects upon administration to a subject. Beneficial effects may include enabling diagnostic decisions; improvement of a disease, symptom, disorder, or pathological condition; reduction or prevention of the onset of a disease, symptom, disorder, or condition; and generally, a response to a disease, symptom, disorder, or pathological condition. In this invention, the pharmaceutical composition may impart beneficial effects on obesity or metabolic diseases.

[0052] In this specification, the term "prevention" refers to a method of partially or completely delaying or preventing the onset or recurrence of a disease, disorder, or its associated symptoms, preventing the acquisition or reacquisition of a disease or disorder, or reducing the risk of acquiring a disease or disorder. For example, the above prevention refers to any act of suppressing or delaying the occurrence of symptoms of obesity or metabolic disease through the administration of a composition according to the present invention.

[0053] In this specification, the term "treatment" includes the inhibition, alleviation, or elimination of the development of a disease, e.g., obesity or metabolic disease.

[0054] In this specification, the term "improvement" may mean any act that at least reduces the degree of parameters related to the alleviation or treatment of a condition, such as obesity or symptoms of metabolic disease.

[0055] The above "metabolic disease" may refer to a disease in which energy metabolism in the body occurs abnormally due to various causes such as excessive energy intake or hormonal imbalance, resulting in the excessive synthesis or accumulation of fat, or a disease related to the regulation of the Melanocortin-4 Receptor (MC4R). Accordingly, the above metabolic disease is not specifically limited in type, but may be one or more selected from the group consisting of type 2 diabetes, obesity, insulin resistance, metabolic syndrome, inflammatory disease, Prader-Willi syndrome, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, cardiovascular disease, and pulmonary disease.

[0056] In this specification, the term “Melanocortin-4 Receptor (MC4R)" is a heterotrimeric G-protein-coupled receptor that converts signals by activating adenylate cyclase. MC4R is expressed in the hypothalamic nucleus and other neuronal and non-neural tissues and regulates feeding behavior and energy homeostasis, integrating an agonist (appetite-suppressing) signal provided by α-melanocyte-stimulating hormone (α-MSH) and an antagonist (appetite-inducing) signal provided by agouti-associated peptide (AGPR).

[0057] The above melanocortins include agouti, agouti-related proteins, and their receptors, and integrate hormones, metabolism, and neural signals to regulate energy homeostasis and control appetite, energy expenditure, and body weight. MCs, including α-MSH, β-MSH, γ-MSH, and ACTH, are a family of peptide hormones derived from precursor proteins called pro-opiomelanocortins (POMCs).

[0058] In one embodiment, the compound may be in the form of a pharmaceutically acceptable salt thereof. The salt comprises common acid addition salts used in the pharmaceutical field, e.g., in the field of obesity or metabolic diseases, salts derived from inorganic acids such as hydrochloric acid, bromic acid, sulfuric acid, sulfamic acid, phosphoric acid, or nitric acid, and salts derived from organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, citric acid, maleic acid, malonic acid, methanesulfonic acid, tartaric acid, malic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, oxalic acid, or trifluoroacetic acid. Additionally, the salt may be a base addition salt such as ammonium, dimethylamine, monomethylamine, monoethylamine, or diethylamine. In addition, the salt comprises conventional metal salt forms, for example, salts derived from metals such as lithium, sodium, potassium, magnesium, or calcium. The acid addition salt, base addition salt, or metal salt may be prepared according to conventional methods. Pharmaceutically acceptable salts and general methodologies for preparing them are widely known in the relevant art. For example, reference may be made to the literature [P. Stahl, et al. Handbook of Pharmaceutical Salts: Properties, Selection and Use, 2nd Revised Edition (Wiley-VCH, 2011)]; [SM Berge, et al., "Pharmaceutical Salts," Journal of Pharmaceutical Sciences, Vol. 66, No. 1, January 1977].

[0059] The above pharmaceutical composition may be any one formulation selected from the group consisting of preparations for oral administration, preparations for oral application, preparations for injection, preparations for dialysis and perfusion, preparations for application to the bronchi and lungs, preparations for administration to the eyes, preparations for administration to the ears, preparations for application to the nose, preparations for application to the rectum, preparations for application to the vagina, and preparations for application to the skin.

[0060] The above composition may comprise a pharmaceutically effective amount of the peptide; and / or a pharmaceutically acceptable carrier.

[0061] In this specification, the term "pharmaceutical effective amount" may mean an amount sufficient to achieve the efficacy of preventing or treating obesity or metabolic diseases of the pharmaceutical composition.

[0062] The above pharmaceutically acceptable carriers are those commonly used in formulations and include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

[0063] When formulating the above pharmaceutical composition, it is prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Preparations for parenteral 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 may be used as non-aqueous solvents and suspension solvents. Witepsol, macrogol, Tween 61, cacao oil, lyulin oil, glycerogelatin, etc. may be used as bases for suppositories.

[0064] In order to increase stability or absorption, carbohydrates such as glucose, sucrose, or dextran, antioxidants such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins, or other stabilizers may be used as pharmaceutical agents in the above pharmaceutical composition.

[0065] In this specification, the term "cosmetic composition" refers to a composition that provides cosmetic or functional benefits when applied to the outside of the body, including the skin. The cosmetic composition may help improve the appearance of the skin, maintain and promote skin health, strengthen skin barrier function, or protect the skin.

[0066] In this specification, the term "improvement of skin health" may refer to any act that performs a beneficial action on the skin to maintain or improve the condition of the skin. Such improvement may include increasing the skin's moisture content, improving elasticity, reducing wrinkles, strengthening the skin barrier, evening out skin tone through the regulation of melanin synthesis, improving skin condition through anti-inflammatory effects, alleviating inflammatory skin diseases, promoting skin cell regeneration, preventing damage and skin aging through antioxidant action, etc. For example, improvement of skin health includes acts that maintain or increase the skin's moisture content, acts that alleviate wrinkles, or acts that reduce inflammatory diseases such as dermatitis.

[0067] The cosmetic composition according to the present invention may include an MC4R agonist compound specifically designed for improving skin health, or a stereoisomer, solvate, or salt thereof. The MC4R agonist can promote the metabolism of skin cells, improve skin condition through anti-inflammatory and antioxidant effects, and regulate melanin synthesis to provide skin tone uniformity and whitening effects. In addition, the composition can strengthen the skin barrier to protect the skin from the external environment, prevent skin damage, and enhance the skin's regenerative capacity.

[0068] The cosmetic composition according to the present invention can help prevent or improve various skin problems such as wrinkles, loss of elasticity, pigmentation, acne, and dryness. In particular, the composition of the present invention can be useful for maintaining and improving the overall health and appearance of the skin by promoting the recovery of the skin barrier and protecting the skin from external stimuli. Furthermore, its anti-inflammatory and antioxidant effects can contribute to alleviating the symptoms of inflammatory skin diseases and delaying skin aging.

[0069] In this specification, "prevention" may mean preventing damage caused by external or intrinsic factors that impair skin health, or delaying or suppressing the onset of skin diseases. Additionally, "improvement" may include all actions that optimize the condition of the skin and enhance its physical and physiological properties.

[0070] The above cosmetic composition may additionally include one or more selected from moisturizers, emulsifiers, softeners, stabilizers, viscosity modifiers, pH modifiers, antioxidants, soothing agents, preservatives, and fragrances. In addition, the above cosmetic composition may be used in a formulation selected from creams, lotions, fillers, gels, serums, oils, butters, balms, toners, essences, masks, and ampoules, but is not limited thereto.

[0071] One or more selected from the group consisting of compounds according to the present invention, stereoisomers thereof, solvates, and salts may be included in a food composition for the purpose of preventing and improving metabolic diseases, reducing body fat, or preventing and improving obesity. The food composition may be a health functional food.

[0072] In this specification, the term "health functional food" refers to a food manufactured or processed for the purpose of health supplementation using specific ingredients as raw materials or by methods such as extraction, concentration, purification, or mixing of specific ingredients contained in food raw materials, and refers to a food designed and processed to fully exert biological regulatory functions on the body, such as biological defense, regulation of biological rhythms, and prevention and recovery from disease, through said ingredients. The health functional food composition may perform functions related to the prevention and improvement of obesity, the prevention and improvement of metabolic diseases, etc.

[0073] There are no specific restrictions on the types of the above-mentioned foods. Examples of foods to which the above-mentioned substance may be added include formulations selected from the group consisting of powders, granules, tablets, capsules, pills, gels, jellies, suspensions, emulsions, syrups, tea bags, infused teas, and health drinks, and include all health foods in the conventional sense.

[0074] A health drink composition containing the compounds of the present invention may include various flavoring agents or natural carbohydrates as additional ingredients, similar to conventional beverages. The aforementioned natural carbohydrates may include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and natural sweeteners such as dextrin and cyclodextrin, or synthetic sweeteners such as saccharin and aspartame. The proportion of the natural carbohydrates is generally about 0.01 to 10 g, preferably about 0.01 to 0.1 g, per 100 ml of the composition of the present invention.

[0075] The above-mentioned health functional food may include food-grade acceptable food additives and may include a suitable carrier commonly used in the manufacture of health functional foods.

[0077] [Example]

[0078] Example: Synthesis of a compound

[0079] Solid Phase Peptide Synthesis (SPPS) was used to synthesize peptides according to one aspect.

[0080] Prepare the peptide of SEQ ID NO. 3, which is a form of the compound of SEQ ID NO. 2, wherein K at position 1 is an epsilon-amino group ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-GABA(γ-Aminobutyric Acid)-CO-(CH2) a - It was connected to CO2H, and the number of carbon atoms a was set to 14, 16, and 18, and these were named Examples 1, 2, and 3, respectively.

[0081] In addition, in the above examples, NPV (N-pentylvaleryl) was used instead of GABA, and compounds were prepared with fatty acid carbon atoms set to 14, 16, and 18, respectively, and named Comparative Examples 1, 2, and 3.

[0082] In addition, a peptide in which the K at position 20 of SEQ ID NO. 6 (chemical formula below) is not substituted (Comparative Example 4, Semaglutide backbone), Semaglutide (Comparative Example 5), and the K at position 20 of the peptide is ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-GABA(γ-Aminobutyric Acid)-CO-(CH2) 18 - A product that was made to be connected to CO2 (Comparative Example 6) was synthesized and selected as an additional comparative example.

[0083] [Chemical Formula]

[0084]

[0085] In addition, the peptides of SEQ ID NOs. 4 and 5 (without an increase in the half-life of the example) were prepared and named Comparative Examples 7 and 8, respectively.

[0087] Experimental Example 1: Evaluation of Plasma and Liver S9 Stability

[0088] 1-1. Experimental Method

[0089] Plasma stability tests and liver S9 stability tests were conducted using plasma and liver S9 fractions extracted from SD rats, respectively. The specific procedures for the test methods are as follows.

[0090] 1) Plasma stability test

[0091] Plasma collected from SD rats was prepared in an incubator controlled at 37°C. Test substances (Examples 1-3 and Comparative Examples 1-6) were added to the plasma at a concentration of 1,000 ng / mL each. Incubation was carried out with continuous stirring at 50 rpm, and 100 μL samples were collected at 0, 5, 15, 30, 60, and 120 minutes. Proteins were removed from the collected samples by adding 300 μL of acetonitrile, followed by centrifugation at 15,000 g for 10 minutes to separate the supernatant. The supernatant was concentrated and solvent removed using a Speed ​​Vacuum concentrator, and then reconstituted into a 50 μL mobile phase. The reconstituted samples were analyzed via HPLC-FLD to quantify the concentration of the test substances over time.

[0092] 2) Liver S9 stability test

[0093] The liver S9 stability test was performed under the same conditions as the plasma stability test. Liver S9 was thawed at room temperature and placed in an incubator controlled to 37°C, after which the test substance (Examples 1-3 and Comparative Examples 1-3, 5, 6) was added at a concentration of 1,000 ng / mL. Samples were collected at the same time intervals (0, 5, 15, 30, 60, 120 minutes), and protein removal and analysis were performed in the same manner as the plasma test.

[0095] 1-2. Check Results

[0096] The stability of the test substances in plasma and liver S9 was compared based on the residual amount over time (Figs. 1 and 2). The residual amount was expressed as a relative concentration over time, with the initial concentration set to 100%.

[0097] 1) Plasma stability

[0098] All test substances maintained a stable residual amount for 120 minutes, and no significant difference was observed between Examples 1-3 and Comparative Examples 1-6 regarding stability in plasma.

[0099] 2) Liver S9 stability

[0100] Examples 1-3 maintained high residual amounts even under liver S9 conditions, whereas Semaglutide-based Comparative Examples 5 and 6 were not detected in most samples within 120 minutes. Additionally, Comparative Example 1 showed levels similar to the Examples, while Comparative Examples 2 and 3 were confirmed to be highly unstable, as there were periods where no measurements were observed. Therefore, the NPV linker demonstrated that it is very easily degraded by liver metabolic enzymes or is unstable in this environment.

[0101] Overall, Examples 1-3 maintained a level of plasma stability similar to other substances, but in terms of hepatic S9 stability, they showed a level similar to Comparative Example 1 using an NPV linker, and higher stability compared to Comparative Examples 2 and 3 and Semaglutide-based substances. This suggests that Examples 1-3, particularly Example 3, have relatively high resistance to hepatic metabolic enzymes, indicating the possibility of having a longer half-life in vivo.

[0103] Experimental Example 2: Evaluation of the Stability of Simulated Intestinal Fluid (SIF)

[0104] 2-1. Experimental Method

[0105] The stability evaluation of SIF was performed using artificial serous fluid prepared according to the conditions specified in the U.S. Pharmacopoeia. The main experimental conditions and procedures are as follows.

[0106] 1) SIF manufacturing

[0107] SIF containing pancreatin (enzyme) was prepared according to United States Pharmacopoeia (USP) standards. The SIF was maintained at 37°C and adjusted to pH 6.8.

[0108] 2) Test substance and concentration

[0109] A total of nine substances were prepared for Examples 1-3 and Comparative Examples 1-6. Each substance was added to the test system at a concentration of 1,000 ng / mL.

[0110] 3) Sampling and Processing

[0111] Each substance was cultured at 37°C in a SIF environment, and 100 μL of samples were collected at 0, 5, 15, 30, 60, and 120 minutes as time progressed.

[0112] Proteins were removed from the collected samples by adding 300 μL of acetonitrile, and the supernatant was separated by centrifugation at 15,000 g for 10 minutes. The supernatant was concentrated using a Speed ​​Vacuum and reconstituted into 50 μL of mobile phase. Finally, the residual amount of each substance was analyzed using HPLC-FLD.

[0114] 2-2. Checking Results

[0115] The residual amount of each test substance over time was compared in a SIF environment, and relative stability was evaluated through Area Under Curve (AUC) values ​​(Fig. 3).

[0116] 1) Residual Amount Analysis

[0117] Based on 120 minutes, Example 3 recorded the highest residual amount among all test substances. Specifically, the examples combined with GABA-fatty acid showed differences in stability depending on the C14, C16, and C18 chain lengths, with the C18 chain exhibiting the highest stability.

[0118] On the other hand, Comparative Examples 1-3 using an NPV linker showed a lower residual amount compared to Examples 1-3. Also, Comparative Examples 4-6 based on Semaglutide showed a rapid decrease in residual amount after 60 minutes, and most were not detected at 120 minutes.

[0119] 2) AUC Comparison Results

[0120] Based on AUC values, stability in SIF is shown as follows (Table 1):

[0121] substance name Example 1 Example 2 Example 3 AUC 4545 4574 5079 substance name Comparative Example 1 Comparative Example 2 Comparative Example 3 AUC 3956 3337 2714 substance name Comparative Example 4 Comparative Example 5 Comparative Example 6 AUC 773.3 810.8 1048

[0123] In particular, the AUC value of Example 3 was recorded to be about 6 times higher than Comparative Example 5 and about 4.8 times higher than Comparative Example 6.

[0124] Meanwhile, although not shown in Fig. 3, Comparative Example 7, which is a form without fatty acid attachment of the Example, recorded an AUC value of 1575. That is, Example 3 showed an effect of increasing the half-life by approximately 3.2 times due to GABA-C18 attachment compared to Comparative Example 7. On the other hand, Comparative Example 6, which has GABA-C18 attachment, showed an effect of increasing the half-life by approximately 1.4 times compared to Comparative Example 4, which is a Semaglutidie backbone without fatty acid attachment. Therefore, it was confirmed that the peptide of SEQ ID NO. 2 shows a prominent effect of increasing the half-life due to GABA-fatty acid attachment compared to other linkers and other peptides.

[0125] Through this experiment, it was confirmed that Example 3 exhibited the highest stability under SIF conditions simulating a gastrointestinal environment. This implies that GABA-C18 maximizes the structural stability of the test substance and demonstrates an optimal effect, particularly in binding with the peptide of SEQ ID NO. 2. In particular, the substances of Comparative Examples 1-3, which were bound with an NPV linker, showed significantly reduced stability even when using a C18 chain, and exhibited results lower than those of Comparative Examples 3-6, which were based on Semaglutide. These results demonstrate that the binding of the peptide of SEQ ID NO. 2 with GABA-C18 specifically enhances stability and support the fact that GABA-C18 is an optimized linker for the peptide of SEQ ID NO. 2.

[0127] Experimental Example 3: Evaluation of pharmacokinetics in an animal model (intravenous administration)

[0128] Experiments were conducted on adult SD rats to confirm the pharmacokinetic characteristics upon intravenous administration of the examples and comparative examples. The experiment was designed to evaluate the distribution, metabolism, and excretion (ADME) characteristics of the test substances and to compare the half-life (t1 / 2) and the degree of exposure (AUC).

[0129] 3-1. Experimental Method

[0130] 1) Test substance

[0131] In this experiment, Example 3, which had the best effect in the SIF stability test, and Comparative Examples 7 and 8 were selected as subjects for evaluation for comparison.

[0132] 2) Method of administration

[0133] The substance was dissolved in physiological saline at a dose of 1 mg / kg and administered via intravenous bolus injection to adult SD rats (body weight 250–300 g).

[0134] 3) Sampling and Analysis

[0135] After administration, 200 μL of blood samples were collected by jugular vein at predetermined time intervals (0, 5, 15, 30, 60, 120, 240, 480, 720, 960, 1440 minutes). Proteins were removed from the collected samples using MeOH, and the supernatant was separated by centrifugation at 12,000 rpm. The blood concentration of the test substance was analyzed using LC-MS / MS on the supernatant.

[0136] The AUC, Cmax, Tmax, and half-life (t1 / 2) of each test group were calculated (Fig. 4).

[0138] 3-2. Checking Results

[0139] 1) Half-life (t 1 / 2 )

[0140] As shown in Table 2 below, Example 3 had an average half-life of about 647 minutes, which was the longest value among all comparative substances. This was longer than Comparative Example 7 (about 42 minutes) and Comparative Example 8 (about 88 minutes), suggesting that metabolic resistance was enhanced.

[0141] 2) Internal exposure (AUC)

[0142] The AUC value is an indicator of how long the test substance has existed in the body, and as shown in Table 2 below, Example 3 recorded the highest value at 7,693.96 μg·min / mL. In contrast, the AUC values ​​of Comparative Example 7 and Comparative Example 8 were 107.03 μg·min / mL and 131.52 μg·min / mL, respectively, confirming that exposure in the body was limited.

[0143] parameters unit Example 3 Comparative Example 7 Comparative Example 8 t 1 / 2 min 647.441 41.636 88.791 AUC 0-inf_obs μg·min / mL 7693.960 107.030 131.518 Cl _obs mL / min / kg 0.143 9.343 8.246

[0145] The foregoing description of the present invention is for illustrative purposes only, and those skilled in the art will understand that other specific forms can be easily modified without altering the technical concept or essential features of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive. For example, each component described as a single unit may be implemented in a distributed manner, and components described as distributed may likewise be implemented in a combined form.

[0146] The scope of the present invention is defined by the claims set forth below rather than by the detailed description above, and all modifications or variations derived from the meaning and scope of the claims and the concept of equivalents thereof should be interpreted as being included within the scope of the present invention.

Claims

Claim 1 A compound comprising the following amino acid sequence, or a stereoisomer, solvate, or salt thereof: KYRCEHFRWC (SEQ No. 2); wherein, in the above sequence, K at position 1 is an epsilon-amino group ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-GABA(γ-Aminobutyric Acid)-CO-(CH2) a - Connected to CO2H, where a is an integer from 14 to 18. Claim 2 A compound, or a stereoisomer, solvate, or salt thereof, wherein K at position 1 is L-type or D-type, C at position 4 is L-type or D-type, F at position 7 is L-type or D-type, and C at position 10 is L-type or D-type. Claim 3 A compound, or a stereoisomer, solvate, or salt thereof, wherein K at position 1 is of type D, C at position 4 is of type D, F at position 7 is of type D, and C at position 10 is of type D. Claim 4 In claim 1, the above a is a compound, or a stereoisomer, solvate, or salt thereof, wherein a is 14, 16, or 18. Claim 5 A compound, or a stereoisomer, solvate, or salt thereof, wherein the C at position 4 and the C at position 10 form a bond with each other. Claim 6 In claim 1, the amino acid sequence is a compound, or a stereoisomer, solvate, or salt thereof, wherein the N-terminal amino acid is acetylated, the C-terminal amino acid is amidated, or the N-terminal amino acid is acetylated and the C-terminal amino acid is amidated. Claim 7 A pharmaceutical composition for the prevention or treatment of metabolic diseases, comprising one or more selected from the group consisting of the compound of claim 1, stereoisomers thereof, solvates, and salts. Claim 8 A pharmaceutical composition according to claim 7, wherein the metabolic disease is selected from the group consisting of type 2 diabetes, obesity, insulin resistance, metabolic syndrome, hyperlipidemia, arteriosclerosis, inflammatory disease, Prader-Willi syndrome, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, cardiovascular disease, and pulmonary disease. Claim 9 A cosmetic composition for improving skin health comprising one or more selected from the group consisting of the compound of claim 1, stereoisomers thereof, solvates, and salts. Claim 10 A health functional food for the prevention or improvement of metabolic diseases, comprising one or more selected from the group consisting of the compound of claim 1, stereoisomers thereof, solvates, and salts.

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