Use of CRF2 receptor agonist and acylated conjugate thereof

Abstract

Provided are CRF2 receptor agonists as defined by Chemical Formulas 1 and 2 and fatty acid conjugates thereof.

Description

Uses of CRF2 receptor agonists and their acylated conjugates

[0001] The present invention relates to a novel CRF2 receptor agonist. More specifically, the present invention relates to a CRF2 receptor agonist and a fatty acid conjugate thereof.

[0002] Urocortin 2 (UCN2) is an endogenous peptide of 38 amino acids. UCN2 is one of three known endogenous urocortins found in mammals and is a member of the CRF family of hormones, which also includes corticotropin-releasing factor (CRF). CRF family hormones play a crucial role in regulating physiological and behavioral responses to stress.

[0003] The agonist role of CRF family hormones is mediated by their receptor, the CRF receptor. CRF receptors are G-protein-coupled receptors and come in two types: type 1 (CRF1 receptor) and type 2 (CRF2), which have different pharmacological properties and target agonists. The CRF1 receptor binds to CRF and UCN1 with high affinity, but not UCN2 or UCN3, whereas the CRF2 receptor binds to both UCN1 and UCN3 with greater affinity than CRF. The CRF1 receptor activates the hypothalamic-pituitary-adrenal axis, stimulating cortisol secretion from the adrenal gland and mediating stress-related behaviors. In contrast, the CRF2 receptor plays a key role in regulating energy homeostasis in the stress response, which includes appetite suppression, mobilization of energy stores, and increased energy expenditure [Bale et al., Annu. Rev. Pharmacol. Toxicol. 44: 525~557 (2004)].

[0004] In animal studies, activating the CRF2 receptor with urocortin has been shown to improve cardiovascular function through decreased vascular resistance and inotropic and lusitropic effects on myocardial contractility. In clinical studies, UCN2 and UCN3 have been shown to exert direct vasodilatory effects in healthy volunteers and patients with heart failure (reviewed in [Stirrat et al., Br. J. Clin. Pharmacol., 2016, 82, 974-982]), while UCN2 has also been shown to increase cardiac output and decrease vascular resistance in patients with heart failure (reviewed in Davis et al., Eur. Heart J., 2007, 28, 2589-2597 and Chan et al., JACC: Heart Failure, 2013, 1, 433-441).

[0005] Recent studies have shown that UCN2 and / or UCN3 gene transfer in mice improves not only cardiac function but also glucose uptake (see [Giamouridis et al., JACC: Basic to Translational Science, 2018, 3, 2]). Furthermore, subcutaneous delivery of PEGylated UCN2 has been shown to improve glucose tolerance and increase glucose uptake in skeletal muscle, while reducing body weight through food restriction (see Borg et al., Diabetes, 2019, 1403–1414). These results suggest that urocortin may be useful not only for the treatment of cardiovascular diseases but also for diseases such as diabetes and obesity.

[0006] Nevertheless, the extremely short half-life of urocortin remains a major limitation to its therapeutic use (see Davies et al., JACC, 2007, 49, 461–471). To date, urocortin treatment cannot be sustained without chronic intravenous infusion.

[0007] Therefore, novel CRF2 receptor agonists that exhibit stability and long lifespan, and preferably selectively act on only the CRF2 receptor among the CRF1 and CRF2 receptors, are expected to be very useful in the treatment and prevention of diseases, including metabolic diseases such as cardiovascular disease, obesity, and diabetes. However, the demand for such novel CRF2 receptor agonists is currently not sufficiently met.

[0008] The technical challenge of the present invention is to provide a novel agent that can selectively activate human CRF2 receptor and has improved stability and lifespan in a liquid environment compared to urocortin 2.

[0009] In one aspect of the present invention, a peptide defined by chemical formula 1 and a pharmaceutically acceptable salt thereof are provided as CRF2 receptor agonists.

[0010] [Chemical Formula 1]

[0011]

[0012] X0 is absent or lysyl; X1 is isoleucyl or phenylalanyl; X2 is valyl or threonyl; X3 is leucyl, valyl or seryl; X5 is leucyl or valyl; X7 is valyl or isoleucyl; X8 is prolyl or seryl; X9 is isoleucyl or phenylalanyl; X 10 Silver glycyl, histidyl or tyrosyl; X 13 is arginyl, glutaminyl, asparagine, glutamyl, aspartyl, alanyl or lysyl; X 15 is leucine or tryptophan; X 17 is glutamyl, prolyl or lysyl; X 18 is glutaminyl, arginyl, isoleucyl, prolyl or lysyl; X 19 is alanyl, glutamyl, prolyl or tryptophanyl; X 20 is glutaminyl, asparagine, glutamyl, aspartyl, alanyl, arginyl or lysyl; X 21is alanyl or glutaminyl; X 22 is arginyl, lysyl, glutaminyl, asparagine, aspartyl, alanyl or glutamyl; X 23 Silver alanyl or lysyl; X 24 is alanyl, leucyl or glutamyl; X 25 is arginyl or lysyl; X 26 is glutamyl or glutaminyl; X 28 Silver alanyl or valyl; X 29 is threonyl, glutamyl or alanyl; X 30 is threonyl, glutamyl or lysyl; X 32 is alanyl or arginyl; X 33 is arginyl, glutamyl, lysyl, glutaminyl, asparagine, aspartyl, or alanyl; X 34 is isoleucine or methionyl; X 35 is lysyl or phenylalanyl; X 36 is alanyl, glutamyl, glutaminyl, asparagine, arginyl, lysyl or aspartyl; X 37 represents arginyl, glutamyl, aspartyl or glutaminyl. Z 39 is OH or NH2.

[0013] The peptide of chemical formula 1 can be used as an agonist of the CRF2 receptor.

[0014] In another aspect of the present invention, a fatty acid conjugate of the aforementioned peptide is disclosed. This peptide-fatty acid conjugate has a structure represented by chemical formula 2.

[0015] [Chemical Formula 2]

[0016]

[0017] In chemical formula 2, R A is a fatty acid group having 14 to 24 carbon atoms and is covalently bonded to linker L through its acyl group;

[0018] L is a linker selected from the group consisting of one to four amino acid residues, one to three iminobis(ethyleneoxy)acetyl groups, one to three 4-iminobutanoyl groups, one to three 5-iminopentanoyl groups, one to three 6-iminohexanoyl groups, a triazole group, one to three oxyethylene groups, and any combination thereof, and Δ is a peptide of the aforementioned chemical formula 1, wherein among the peptide residues, X0, X 13 , X 17 , X 18 , X 20 , X 22 , X 23 , X 25 , X 30 , X 33 and X 36 At least one of which is a lysyl residue, the peptide-fatty acid conjugate is Δ of the above X0, X 13 , X 17 , X 18 , X 20 , X 22 , X 23 , X 25 , X 30 , X 33 or X 36 L is covalently linked to the ε-amino group of the side chain of one lysyl residue.

[0019] The CRF2 receptor agonist of the present invention can stimulate the human CRF2 receptor, and furthermore, has excellent selectivity for the CRF2 receptor due to very low human CRF1 receptor agonistic activity. The CRF2 receptor agonist of the present invention in the form of a fatty acid conjugate has a significantly improved in vivo lifespan compared to urocortin 2.

[0020] Figure 1 is a graph showing the weight loss effect in diet-induced obesity (DIO) mice when the peptide-fatty acid conjugate of the present invention was administered at a dose of 100 nmol / kg every other day (Q2D) compared to the negative control group DIO mice administered with a vehicle (buffer solution) as a percentage. In Figure 1 *** The mark indicates that the change in body weight of the mice was statistically significant (p<0.001) compared to the negative control DIO mice when statistically tested by one-way ANOVA.

[0021] Hereinafter, embodiments of the present invention will be described in detail. Prior to this, it should be noted that the terms and words used in this specification and claims should not be construed as limited to their conventional or dictionary meanings. Based on the principle that the inventor can appropriately define the concept of a term to best explain his or her invention, they should be interpreted in a way that aligns with the technical spirit of the present invention.

[0022] Therefore, the embodiments described in this specification are merely examples presented for the purpose of helping to understand the present invention and do not represent all of the technical ideas of the present invention. Therefore, it should be understood that there may be various equivalents and modified examples that can replace them at the time of filing this application.

[0023] Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by those skilled in the art. While preferred methods and samples are described herein, similar or equivalent methods are also included within the scope of the present invention. Numerical values ​​described herein are considered to include the meaning of "about" even if not explicitly stated. The numerical range indicated using the term "to" herein includes the ranges that include the values ​​described before and after the term "to" as the lower and upper limits, respectively.

[0024] Definition of terms

[0025] As used herein, "agonist" means any compound that activates a receptor, including but not limited to antibodies. For example, CRF receptor agonists include, but are not limited to, CRF and CRF analogs.

[0026] “CRF receptor” refers to either the CRF1 receptor or the CRF2 receptor.

[0027] "CRF receptor agonist" means a compound or molecule capable of activating either CRF1 or CRF2 or both. Activation of the CRF receptor can be measured as described below.

[0028] As used herein, the term "CRF1 receptor" encompasses all isoforms of the CRF1 receptor, which may be derived from any animal. The CRF1 receptor has been previously referred to as the receptor for CRF-RA, PC-CRF, CRF, or CRH.

[0029] The definition of CRF1 receptor includes, but is not limited to, receptors for which the encoding cDNA or genomic sequence has been deposited in a sequence database. The accession numbers for these receptors are as follows: X72304, E11431, L23332, I92584, T37068, T28968, Q81952, L23333, NM_004382, AF180301, T28970, L25438, L24096, I92586, Q81954, AH006791, NM_007762, X72305, AF054582, Y14036, AF229359, AF229361, AB055434, and L41563. Giga: Nucleotide and protein sequences of these receptors can be obtained from GenBank or Derwent.

[0030] As used herein, the term "CRF2 receptor" encompasses all isoforms of the CRF2 receptor that can be derived from any animal. The CRF2 receptor has also been previously referred to as HM-CRF and CRF-RB.

[0031] The definition of CRF2 receptor includes, but is not limited to, receptors for which the encoding cDNA or genomic sequence has been deposited in a sequence database. The accession numbers for these receptors are as follows: U34587, E12752, NM_001883, T12247, T66508, AF011406, AF019381, U16253, T12244, T28972, U17858, NM_009953, Y14037, and AF229360. The nucleotide and protein sequences of these receptors can be obtained from GenBank or Derwent.

[0032] The term "selective agonist" or "selective" agonist means that the agonist has significantly greater activity at a particular receptor(s) compared to other receptors, but this does not mean that the agonist is completely inactive at said other receptors.

[0033] In this specification, human urocortin 2 (UCN2) refers to a C-terminally amidated peptide having the following sequence.

[0034] IVLSLDVPIGLLQILLEQARARAAREQATTNARILARV

[0035] In this specification, "fatty acid having 14 to 24 carbon atoms" or "C" as the same meaning 14 ~C 24 "Fatty acid" means a carboxylic acid having 14 to 24 carbon atoms, saturated carbon atoms except for the carboxyl carbon atom. "Saturated" means that the fatty acid does not contain carbon-carbon double or triple bonds. In the present invention, a fatty acid having 14 to 24 carbon atoms or C 14 ~C24 Fatty acids include saturated monoacids or saturated diacids. C that can be used in the present invention 14 ~C 24 Examples of fatty acids include myristic acid (tetradecanoic acid) (C), including branched and substituted derivatives. 14 Ilsan), tetradecanedioic acid (C 14 (C) Palmitic acid (hexadecanoic acid) (C 16 Ilsan), hexadecanedioic acid (C 16 (C) (Heptadecanoic acid) (C ... 17 Ilsan), heptadecanedioic acid (C 17 (C) ... 18 Ilsan), octadecanedioic acid (C 18 (C) Nonadecylic acid (nonadecanoic acid) (C 19 Ilsan), nonadecandioic acid (C 19 (C) ... 20 Ilsan), eicosandioic acid (C 20 (C) ... 21 Ilsan), Heneikosandiosan (C 21 (C) ... 22 Ilsan), Dokosan Diosan (C 22 (C) (Lignoceric acid (tetracosanoic acid) (C) 24 Ilsan), tetracosanedioic acid (C 24 There is a (discrete)

[0036] The term "EC" as used herein 50 " refers to the concentration of an agonist that produces 50% of the assay endpoint activity (e.g., cAMP activation) when performing an agonist activity test with respect to a CRF2 receptor agonist.

[0037] As used herein, the term "pharmaceutically acceptable salt" for a compound refers to a salt that does not impair the desired biological activity of the compound and does not cause inappropriate toxicity, irritation, or allergic reactions when used in contact with human or animal tissues. Pharmaceutically acceptable salts are well known in the art. For example, reference may be made to prior literature, such as the paper by Berge et al., J. Pharmaceutical Sciences (1977), Vol. 66, pp. 1-19. Some examples of pharmaceutically acceptable salts include acid addition salts such as hydrochloride, hydrobromide, phosphate, sulfate, perchlorate, acetate, oxalate, maleate, tartrate, citrate, succinate, malonate, adipate, alginate, ascorbate, aspartate, benzoate, benzenesulfonate, and hydrosulfide, and base addition salts such as alkali metal salts, alkaline earth metal salts, ammonium salts, and quaternary ammonium (N + (C 1~4 There are alkyl)4), and examples of alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, and magnesium.

[0038] Throughout this specification, when describing the sequences of peptides and proteins, the conventional one-letter and three-letter codes for naturally occurring regular amino acids are used, and in addition, three-letter codes for unnatural amino acids such as Aib (2-aminoisobutyric acid), Sar (N-methylglycine), and AMG (α-methyl-glutamic acid) may also be used.

[0039] Amino acid sequences of peptides and proteins are presented in this specification in left-to-right order from the N-terminus to the C-terminus, unless otherwise specified. In addition, amino acids referred to by abbreviations in this specification are written as follows according to the IUPAC-IUB nomenclature. In addition, in order to distinguish amino acid residues existing in the internal position of a peptide or protein from free amino acids, the corresponding radical name of the amino acid (i.e., the chemical formula of the free amino acid minus one water molecule) is used (also written in parentheses). For example, alanine refers to a free molecule with the chemical formula NH2CH(CH3)COOH, but its radical alanyl refers to the -NHCH(CH3)C(=O)- functional group when referred to as a member of an amino acid sequence in this specification. When a single-letter code is written as A, it refers to alanyl in the amino acid sequence.

[0040] Alanine A (alanyl) Arginine R (arginyl)

[0041] Asparagine N (asparagineyl) aspartate D (aspartyl)

[0042] Cysteine ​​C (cysteineyl) glutamic acid E (glutamyl)

[0043] Glutamine Q (glutamic acid) Glycine G (glycyl acid)

[0044] Histidine H (histidyl) Isoleucine I (isoleucyl)

[0045] Leucine L (leucyl), Lysine K (lysyl), Methionine M (methionyl)

[0046] Phenylalanine F (phenylalanyl), proline P (prolyl), serine S (seryl)

[0047] Threonine T (threonyl) Tryptophan W (tryptophanyl)

[0048] Tyrosine Y (tyrosyl) Valine V (valyl)

[0049] CRF2 receptor agonist peptide

[0050] In one aspect of the present invention, a CRF2 receptor agonist peptide defined by chemical formula 1 and a pharmaceutically acceptable salt thereof are provided.

[0051]

[0052] In the chemical formula above, Z0 is hydrogen;

[0053] X0 is absent or is lysyl;

[0054] X1 is isoleucyl or phenylalanyl;

[0055] X2 is valyl or threonyl;

[0056] X3 is Ryusil, Valil or Seril;

[0057] X5 is Ryusil or Valil;

[0058] X7 is valyl or isoleucyl;

[0059] X8 is prolyl or seryl;

[0060] X9 is isoleucyl or phenylalanyl;

[0061] X 10 Silver glycyl, histidyl or tyrosyl;

[0062] X 13 is arginyl, glutaminyl, asparagine, glutamyl, aspartyl, alanyl or lysyl;

[0063] X 15 is leucine or tryptophan;

[0064] X 17 is glutamyl, prolyl or lysyl;

[0065] X 18 is glutaminyl, arginyl, isoleucyl, prolyl or lysyl;

[0066] X 19 is alanyl, glutamyl, prolyl or tryptophanyl;

[0067] X 20 is glutaminyl, asparagine, glutamyl, aspartyl, alanyl, arginyl or lysyl;

[0068] X 21 is alanyl or glutaminyl;

[0069] X 22 is arginyl, lysyl, glutaminyl, asparagine, aspartyl, alanyl or glutamyl;

[0070] X 23 alanyl or lysyl;

[0071] X 24 is alanyl, leucyl or glutamyl;

[0072] X 25 is arginyl or lysyl;

[0073] X 26 is glutamyl or glutaminyl;

[0074] X 28 is alanyl or valyl;

[0075] X 29 is threonyl, glutamyl or alanyl;

[0076] X 30 is threonyl, glutamyl or lysyl;

[0077] X 32 is alanyl or arginyl;

[0078] X 33 is arginyl, glutamyl, glutaminyl, asparagine, aspartyl, alanyl or lysyl;

[0079] X 34 is isoleucine or methionyl;

[0080] X 35 is lysyl or phenylalanyl;

[0081] X 36 is alanyl, glutamyl, glutaminyl, asparagine, arginyl, lysyl or aspartyl;

[0082] X 37 is arginyl, glutamyl, aspartyl or glutaminyl;

[0083] Z 39 is OH or NH2.

[0084] This peptide can optionally be amidated at the C-terminus.

[0085] In one specific embodiment, the peptide of formula 1 is a peptide wherein X1 is isoleucyl; X2 is valyl; X3 is leucyl; X8 is prolyl; X 10 This glycyrrhiza; X 18 This is glutaminyl.

[0086] In another specific embodiment, the peptide of formula 1 is X 23 and X 24 Ga Alanyl; X 25 Ga arginyl, X 26 This is glutamyl.

[0087] In one embodiment, the CRF2 receptor agonist peptide of the present invention forms an intramolecular lactam ring. Specifically, the lactam ring is formed between the 17th and 20th residues, and the 23rd and 26th residues. In a more specific embodiment, X 20 is lysyl, and the side chain of the lysyl residue and the side chain of the 17th residue glutamyl form an intramolecular lactam ring. In another specific embodiment, X 23 is lysyl, and X 26 is glutamyl, and the side chain of the lysyl residue and the side chain of the glutamyl residue form an intramolecular lactam ring.

[0088] The peptide chain of the CRF2 receptor agonist of the present invention can be synthesized using standard manual or automated solid-phase synthesis procedures. Automated peptide synthesizers are commercially available from, for example, Applied Biosystems (Foster City, California) and Protein Technologies Inc. (Tucson, Arizona). Reagents for solid-phase synthesis are readily available from commercial sources. Solid-phase synthesis equipment can be used according to the manufacturer's instructions for blocking interfering groups, protecting amino acids during the reaction, coupling, deprotection, and capping unreacted amino acids.

[0089] CRF2 receptor agonist peptide-fatty acid conjugate

[0090] In another aspect of the present invention, a CRF2 receptor agonist peptide-fatty acid conjugate is disclosed. In the form of the CRF2 receptor agonist of the present invention, the peptide-fatty acid conjugate is covalently linked to the epsilon (ε)-amino group of the side chain of any lysine residue of the CRF2 receptor agonist peptide moiety, either directly or via a linker. 14 ~C 24 Contains fatty acids.

[0091] The CRF2 receptor agonist peptide peptide-fatty acid conjugate of the present invention has a structure represented by chemical formula 2.

[0092]

[0093] In chemical formula 2, Δ is the CRF2 receptor agonist peptide described above, and its residues X0, X 13 , X 17 , X 18 , X 20 , X 22 , X 23 , X 25 , X 30 , X 33 and X 36 At least one of them is a lysyl residue, wherein X0, X 13 , X 17 , X 18 , X 20 , X 22 , X 23 , X 25 , X 30 , X 33 or X 36 A linker L is covalently linked to the ε-amino group of the side chain of one lysyl residue.

[0094] Specifically, in the peptide-fatty acid conjugate of chemical formula 2, R A is a fatty acid group having 14 to 24 carbon atoms and is covalently bonded to linker L through its acyl group;

[0095] L is a linker selected from the group consisting of one to four amino acid residues, one to three iminobis(ethyleneoxy)acetyl groups, one to three 4-iminobutanoyl groups, one to three 5-iminopentanoyl groups, one to three 6-iminohexanoyl groups, a triazole group, one to three oxyethylene groups, and any combination thereof;

[0096] Δ is a peptide having a structure of chemical formula 3, which is the same as the CRF2 receptor agonist peptide described above.

[0097]

[0098] (Sequence number 50)

[0099] In the chemical formula above, Z0 is hydrogen;

[0100] X0 is absent or is lysyl;

[0101] X1 is isoleucyl or phenylalanyl;

[0102] X2 is valyl or threonyl;

[0103] X3 is Ryusil, Valil or Seril;

[0104] X5 is Ryusil or Valil;

[0105] X7 is valyl or isoleucyl;

[0106] X8 is prolyl or seryl;

[0107] X9 is isoleucyl or phenylalanyl;

[0108] X 10 Silver glycyl, histidyl or tyrosyl;

[0109] X 13 is arginyl, glutaminyl, asparagine, glutamyl, aspartyl, alanyl or lysyl;

[0110] X 15 is leucine or tryptophan;

[0111] X 17 is glutamyl, prolyl or lysyl;

[0112] X 18 is glutaminyl, arginyl, isoleucyl, prolyl or lysyl;

[0113] X 19 is alanyl, glutamyl, prolyl or tryptophanyl;

[0114] X 20 is glutaminyl, asparagine, glutamyl, aspartyl, alanyl, arginyl or lysyl;

[0115] X 21 is alanyl or glutaminyl;

[0116] X 22 is arginyl, lysyl, glutaminyl, asparagine, aspartyl, alanyl or glutamyl;

[0117] X 23 alanyl or lysyl;

[0118] X 24 is alanyl, leucyl or glutamyl;

[0119] X 25 is arginyl or lysyl;

[0120] X 26 is glutamyl or glutaminyl;

[0121] X 28 is alanyl or valyl;

[0122] X 29 is threonyl, glutamyl or alanyl;

[0123] X 30 is threonyl, glutamyl or lysyl;

[0124] X 32 is alanyl or arginyl;

[0125] X 33 is arginyl, glutamyl, glutaminyl, asparagine, aspartyl, alanyl or lysyl;

[0126] X 34 is isoleucine or methionyl;

[0127] X 35 is lysyl or phenylalanyl;

[0128] X 36 is alanyl, glutamyl, glutaminyl, asparagine, arginyl, lysyl or aspartyl;

[0129] X 37 is arginyl, glutamyl, aspartyl or glutaminyl;

[0130] Z 39 is OH or NH2.

[0131] In one embodiment of the CRF2 receptor agonist peptide-fatty acid conjugate of the present invention, the CRF2 receptor agonist peptide of formula 3 is further defined as follows:

[0132] X1 is isolates;

[0133] X3 is Ryusil or Seril;

[0134] X5 is Ryu Sil;

[0135] X7 is Valil;

[0136] X8 is prolyl;

[0137] X9 is isolates;

[0138] X 10 Silver glycyl or tyrosyl;

[0139] X 13 Silver Lysyl;

[0140] X 15 is Ryu Sil;

[0141] X 17 is glutamyl;

[0142] X 18 is glutaminyl or arginyl;

[0143] X 19 is alanine;

[0144] X 20 Silver arginyl or lysyl;

[0145] X 21 Silver alanyl;

[0146] X 22 is arginyl or lysyl;

[0147] X 23 Silver alanyl;

[0148] X 24 is alanine;

[0149] X 25 is arginyl;

[0150] X 26 is glutamyl;

[0151] X 29 is threonyl;

[0152] X 30 is threonyl;

[0153] X 32 is alanine;

[0154] X 33 is glutamyl or lysyl;

[0155] X 35 is Ryu Sil;

[0156] X 36 is glutamyl;

[0157] X 37 is arginyl or glutaminyl,

[0158] At this time L is X0, X 13 , X 20 , X 22 or X 33 It is covalently linked to the ε-amino group of the side chain of one lysyl residue.

[0159] In one specific embodiment, L of the CRF2 receptor agonist peptide-fatty acid conjugate is X0, X 13 , X 20 , X 22 or X 33 is covalently linked to the ε-amino group of the CRF2 receptor agonist peptide-fatty acid conjugate of the present invention. In another embodiment of the present invention, Δ is a peptide having a structure of chemical formula 4.

[0160]

[0161] In the above chemical formula 4, Z0 is hydrogen;

[0162] X0 is absent or is lysyl;

[0163] X1 is isoleucyl or phenylalanyl;

[0164] X2 is valyl or threonyl;

[0165] X3 is Ryusil, Valil or Seril;

[0166] X8 is prolyl or seryl;

[0167] X 10 Silver glycyl, histidyl or tyrosyl;

[0168] X 13 is glutaminyl, asparagineyl or lysyl;

[0169] X 18 is glutaminyl, arginyl or lysyl;

[0170] X 19 is alanyl, glutamyl or tryptophanyl;

[0171] X 20 Silver arginyl or lysyl;

[0172] X 21 is alanyl or glutaminyl;

[0173] X 22 is arginyl, lysyl or glutamyl;

[0174] X 23 alanyl or lysyl;

[0175] X 24 is alanyl, leucyl or glutamyl;

[0176] X 25 is arginyl or lysyl;

[0177] X 26 is glutamyl or glutaminyl;

[0178] X 28 is alanyl or valyl;

[0179] X 29 is threonyl, glutamyl or alanyl;

[0180] X 30 is threonyl, glutamyl or lysyl;

[0181] X 32 is alanyl or arginyl;

[0182] X 33 is arginyl, glutamyl or lysyl;

[0183] X 34 is isoleucine or methionyl;

[0184] X 35 is lysyl or phenylalanyl;

[0185] X 36 alanyl, glutamyl or aspartyl;

[0186] X 37 is arginyl, glutamyl, aspartyl or glutaminyl;

[0187] Z 39 is OH or NH2.

[0188] X0, X in the CRF2 receptor agonist peptide-fatty acid conjugate of chemical formula 4 13 , X 20 , X 22 or X 33 At least one of them is a lysyl residue, and the linker L is X0, X 13 , X 20 , X 22 or X 33 It is covalently linked to the ε-amino group of the side chain of one lysyl residue.

[0189] In one specific embodiment, the CRF2 receptor agonist peptide-fatty acid conjugate R of the present invention A C of 14 ~C 24 Saturated fatty acids C 14 Ilsan, Saturated C 14 Discrete, saturated C 16 Ilsan, Saturated C 16 Discrete, saturated C 18 Ilsan, Saturated C 18 Discrete, saturated C 19 Discrete, saturated C 20 Ilsan, Saturated C 20 Discrete, saturated C 22is selected from the group consisting of isocyanates, and branched and substituted derivatives thereof. In a more specific embodiment, C 14 ~C 24 The fatty acid is selected from the group consisting of myristic acid, tetradecanedioic acid, palmitic acid, hexadecanedioic acid, stearic acid, octadecanedioic acid, nonadecanedioic acid, arachidic acid, eicosanedioic acid and docosanedioic acid. More specifically, C 14 ~C 24 The fatty acid is octadecanedioic acid or eicosanedioic acid.

[0190] In one embodiment, L may comprise one to three units derived from [2-(2-aminoethoxy)ethoxy]acetyl, 4-aminobutyric acid, 5-aminovaleric acid, or 6-aminohexanoic acid.

[0191] In one embodiment, the peptide-fatty acid conjugate described above has a structure represented by chemical formula 5.

[0192] X3 is Ryusil, Valil or Seril;

[0193] X8 is prolyl or seryl;

[0194] X 10 Silver glycyl, histidyl or tyrosyl;

[0195] X 13 is glutaminyl, asparagineyl or lysyl;

[0196] X 18 is glutaminyl, arginyl or lysyl;

[0197] X 19 is alanyl, glutamyl or tryptophanyl;

[0198] X 20 Silver arginyl or lysyl;

[0199] X 21 is alanyl or glutaminyl;

[0200] X 22 is arginyl, lysyl or glutamyl;

[0201] X 23 alanyl or lysyl;

[0202] X 24 is alanyl, leucyl or glutamyl;

[0203] X 25 is arginyl or lysyl;

[0204] X 26 is glutamyl or glutaminyl;

[0205] X 28 is alanyl or valyl;

[0206] X 29 is threonyl, glutamyl or alanyl;

[0207] X 30 is threonyl, glutamyl or lysyl;

[0208] X 32 is alanyl or arginyl;

[0209] X 33 is arginyl, glutamyl or lysyl;

[0210] X 34 is isoleucine or methionyl;

[0211] X 35 is lysyl or phenylalanyl;

[0212] X 36 alanyl, glutamyl or aspartyl;

[0213] X 37 is arginyl, glutamyl, aspartyl or glutaminyl;

[0214] Z 39 is OH or NH2.

[0215] X0, X in the CRF2 receptor agonist peptide-fatty acid conjugate of chemical formula 4 13 , X 20 , X 22 or X 33 At least one of them is a lysyl residue, and the linker L is X0, X 13 , X 20 , X 22 or X 33It is covalently linked to the ε-amino group of the side chain of one lysyl residue.

[0216] In one specific embodiment, the CRF2 receptor agonist peptide-fatty acid conjugate R of the present invention A C of 14 ~C 24 Saturated fatty acids C 14 Ilsan, Saturated C 14 Discrete, saturated C 16 Ilsan, Saturated C 16 Discrete, saturated C 18 Ilsan, Saturated C 18 Discrete, saturated C 19 Discrete, saturated C 20 Ilsan, Saturated C 20 Discrete, saturated C 22 is selected from the group consisting of isocyanates, and branched and substituted derivatives thereof. In a more specific embodiment, C 14 ~C 24 The fatty acid is selected from the group consisting of myristic acid, tetradecanedioic acid, palmitic acid, hexadecanedioic acid, stearic acid, octadecanedioic acid, nonadecanedioic acid, arachidic acid, eicosanedioic acid and docosanedioic acid. More specifically, C 14 ~C 24 The fatty acid is octadecanedioic acid or eicosanedioic acid.

[0217] In one embodiment, L may comprise one to three units derived from [2-(2-aminoethoxy)ethoxy]acetyl, 4-aminobutyric acid, 5-aminovaleric acid, or 6-aminohexanoic acid.

[0218] In one embodiment, the peptide-fatty acid conjugate described above has a structure represented by chemical formula 5.

[0219]

[0220] In chemical formula 5, i is an integer from 12 to 22, and j is an integer from 0 to 3.

[0221] Sp is absent or selected from the functional groups of the chemical formula 6 below.

[0222]

[0223] Here, * indicates the direction in which the lysyl residue ε-amino group of Δ is connected, i.e., the indicated carbonyl carbon within the Sp functional group is amide-bonded to the lysyl residue ε-amino group.

[0224] In one specific embodiment, i is 18 and j is 1.

[0225] In another specific embodiment, Sp am.

[0226]

[0227] pharmaceutical composition

[0228] In another aspect of the present invention, a pharmaceutical composition comprising the aforementioned CRF2 receptor agonist and a pharmaceutically acceptable excipient is disclosed. The pharmaceutical composition of the present invention can be used for activating the CRF2 receptor in the body, for example, for the prevention and treatment of obesity. In one embodiment, a pharmaceutical composition for the prevention or treatment of obesity is disclosed, comprising a therapeutically effective amount of the aforementioned peptide-fatty acid conjugate and a pharmaceutically acceptable excipient.

[0229] In the pharmaceutical composition of the present invention, the CRF2 receptor agonist described above may be present in the form of a pharmaceutically acceptable salt. Pharmaceutically acceptable salts include, for example, base addition salts and acid addition salts. Pharmaceutically acceptable base addition salts may be in the form of metal or amine bases, such as alkali metal and alkaline earth metal bases, or organic amines. Pharmaceutically acceptable salts of the compound may also be prepared using pharmaceutically acceptable cations. Pharmaceutically acceptable acid addition salts include salts of inorganic or organic acids.

[0230] In the present invention, a pharmaceutically acceptable excipient refers to an inactive ingredient approved by a relevant administrative agency (e.g., the Ministry of Food and Drug Safety of the Republic of Korea, the Federal Food and Drug Administration (FDA) of the United States) as being suitable for use together with an active pharmaceutical ingredient for the purpose of manufacturing a medicine for treating a disease in humans or livestock. Such pharmaceutically acceptable excipients include, but are not limited to, carriers, lubricants, fluidizing agents, disintegrating agents, sweeteners, diluents, preservatives, coloring agents, flavoring agents, surfactants, wetting agents, dispersing agents, suspending agents, stabilizers, isotonic agents, solvent emulsifiers, and adjuvants.

[0231] The pharmaceutical composition of the present invention may be in a form suitable for oral administration, such as a tablet, capsule, pill, powder, sustained-release preparation, solution, or suspension; in a form suitable for parenteral injection, such as a sterile solution, suspension, or emulsion; in a form suitable for topical administration, such as an ointment or cream; or in a form suitable for rectal administration, such as a suppository.

[0232] The pharmaceutical composition according to the present invention can be prepared in a conventional manner, for example, by conventional mixing, dissolving, granulating, sugar-coated tablet preparation, powdering, emulsifying, encapsulating, encapsulating, or lyophilizing processes. The appropriate formulation will vary depending on the chosen route of administration.

[0233] Pharmaceutical compositions suitable for oral administration can be readily formulated by combining the CRF2 receptor agonists disclosed herein with pharmaceutically acceptable excipients, such as carriers well known in the art. Using such excipients and carriers, the compositions can be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, etc. for oral ingestion by a patient to be treated with the CRF2 receptor agonists disclosed herein. Oral pharmaceutical preparations can be obtained by adding the CRF2 receptor agonists of the present invention together with solid excipients, grinding the resulting mixture if necessary, adding suitable auxiliaries if necessary, and then processing the granulated mixture to form tablets or dragee cores. Suitable excipients include, for example, fillers and cellulose preparations. If desired, a disintegrant may be added.

[0234] For pharmaceutical compositions for oral administration of a therapeutically effective amount of a CRF2 receptor agonist of the present disclosure, the compositions are generally in the form of solid (e.g., tablets, capsules, pills, powders, or troches) or liquid formulations (e.g., aqueous suspensions, solutions, elixirs, or syrups).

[0235] When administered in tablet form, the composition may additionally contain a functional solid and / or solid carrier such as gelatin or an adjuvant. Compositions in the form of tablets, capsules, and powders may contain from about 1 to about 95 wt% of the CRF2 receptor agonist of the present invention, preferably from about 15 to about 90 wt% of the CRF2 receptor agonist of the present invention, based on the total weight of the composition. An example of a tablet composition may contain, for example, up to about 80 wt% of the active pharmaceutical ingredient, from about 10 wt% to about 90 wt% of the binder, from about 0 wt% to about 85 wt% of the diluent, from about 2 wt% to about 10 wt% of the disintegrant, and from about 0.25 wt% to about 10 wt% of the lubricant.

[0236] When administered in liquid or suspension form, a functional liquid and / or liquid carrier such as water, petroleum, or animal or vegetable oil may be added. The liquid composition may further comprise saline, a sugar alcohol solution, dextrose or other sugar solution, or glycol. When administered in liquid or suspension form, the composition may contain from about 0.5% to about 90% by weight of the CRF2 receptor agonist of the present invention, preferably from about 1% to about 50% by weight of the CRF2 receptor agonist of the present invention. In one contemplated embodiment, the liquid carrier is non-aqueous or substantially non-aqueous. For administration in liquid form, the composition may be supplied as a rapidly dissolving solid formulation for dissolution or suspension immediately prior to administration.

[0237] When the pharmaceutical composition of the present invention is administered intravenously, transdermally, or subcutaneously, it is in the form of a parenteral aqueous solution that does not contain a pyrogen. The preparation of such parenteral solutions, taking into account pH, isotonicity, stability, etc., is within the skill of the art. Preferred compositions for intravenous, transdermal, or subcutaneous injection generally contain an isotonic vehicle. Such compositions can be prepared for administration as a solution of the free base or a pharmacologically acceptable salt in water suitably mixed with a surfactant such as hydroxypropyl cellulose. Dispersions in glycerol, liquid polyethylene glycol, and mixtures thereof, as well as dispersions in oils, can also be prepared. Under normal storage and use conditions, such preparations may optionally contain a preservative to prevent the growth of microorganisms.

[0238] Injectable compositions may include sterile aqueous solutions, suspensions, or dispersions, and sterile powders for the extemporaneous preparation of sterile injectable solutions, suspensions, or dispersions. Sterile injectable solutions are prepared by incorporating the active compound in the required amount in an appropriate solvent with various other ingredients, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterile active ingredients into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In embodiments of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying techniques, which produce a powder of the active ingredient and any additional required ingredients from a previously sterile-filtered solution thereof.

[0239] To achieve controlled release of the active compound upon contact with body fluids in the gastrointestinal tract and to provide substantially constant and effective levels of the active compound in the plasma, sustained-release or sustained-release formulations may be prepared. For example, release may be controlled by one or more of dissolution, diffusion, and ion exchange. Furthermore, sustained-release approaches may enhance absorption through saturable or restricted pathways within the gastrointestinal tract. For example, for this purpose, the CRF2 receptor agonist may be embedded in a polymer matrix comprising a biodegradable polymer, a water-soluble polymer, or a mixture thereof, and optionally a suitable surfactant. Embedding in this context may mean incorporating microparticles into the polymer matrix. Controlled-release formulations may also be obtained by encapsulating dispersed microparticles or emulsified microdroplets using known dispersion or emulsion coating techniques.

[0240] The pharmaceutical composition of the present invention may be formulated for parenteral administration by injection (e.g., bolus injection or continuous infusion). The injectable formulation may be presented in unit dosage form (e.g., ampoules or multi-dose containers) with an added preservative. The composition may take the form of a suspension, solution, or emulsion in an oily or aqueous vehicle, and may contain formulatory agents such as suspending, stabilizing, and / or dispersing agents.

[0241] The pharmaceutical composition may be presented in unit dosage form suitable for single administration of precise dosages.

[0242] A pharmaceutical composition containing the compound of the present invention can be used according to the method described below.

[0243] Treatment methods

[0244] In another aspect, the present invention provides a method for treating a disease treatable by activation of the CRF2 receptor. The method comprises administering to a patient a therapeutically effective amount of a CRF2 receptor agonist of the present invention, such as a CRF2 receptor agonist as defined by Formula 1 or 2, or a peptide-fatty acid conjugate as defined by Formula 2, or a pharmaceutical composition comprising a CRF2 receptor agonist peptide or a CRF2 receptor agonist peptide-fatty acid conjugate of the present invention. In one embodiment, the disease is obesity.

[0245] As used herein, unless specifically stated otherwise, a “therapeutically effective amount” of a compound refers to an amount of the compound sufficient to delay or minimize one or more symptoms associated with a disease, disorder, or condition, or to provide a therapeutic effect for a disease, disorder, or condition. The term “therapeutically effective amount” encompasses an amount that improves overall healing, an amount that alleviates or avoids a symptom or cause of a disease or condition, and an amount that enhances the therapeutic effectiveness of another therapeutic agent.

[0246] The amount of a compound, such as a CRF2 receptor agonist, administered may vary depending on the subject being treated, the subject's age, health, sex, and body weight, the type (if any) of concurrent treatment, the severity of the pain, the nature of the desired effect, the treatment regimen and frequency, and the judgment of the prescribing physician. The frequency of administration may also vary depending on the pharmacodynamic effects on arterial oxygen tension. However, the most desirable dosage can be tailored to the individual subject, as understood and determined by those skilled in the art without undue experimentation. This typically involves adjusting the standard dose (e.g., reducing the dose if the patient is underweight).

[0247] Although individual case needs may vary, it is common sense in the art to determine the optimal range of therapeutically effective doses of compounds, such as CRF2 receptor agonists. When administered to humans in the therapeutic or prophylactic treatment of conditions and disorders using the compounds of the present invention, for example, a typical dosage of the compounds of the present invention may be from about 0.05 mg / kg / day to about 50 mg / kg / day, for example, at least 0.05 mg / kg, at least 0.08 mg / kg, at least 0.1 mg / kg, at least 0.2 mg / kg, at least 0.3 mg / kg, at least 0.4 mg / kg, or at least 0.5 mg / kg, preferably not more than 50 mg / kg, not more than 40 mg / kg, not more than 30 mg / kg, not more than 20 mg / kg, or not more than 10 mg / kg, which may be, for example, from about 2.5 mg / day (0.5 mg / kg × 5 kg) to about 5000 mg / day (50 mg / kg × 100 kg). For example, the dosage of the compound may be from about 0.1 mg / kg / day to about 50 mg / kg / day, from about 0.05 mg / kg / day to about 10 mg / kg / day, from about 0.05 mg / kg / day to about 5 mg / kg / day, from about 0.05 mg / kg / day to about 3 mg / kg / day, from about 0.07 mg / kg / day to about 3 mg / kg / day, from about 0.09 mg / kg / day to about 3 mg / kg / day, from about 0.05 mg / kg / day to about 0.1 mg / kg / day, from about 0.1 mg / kg / day to about 1 mg / kg / day, from about 1 mg / kg / day to about 10 mg / kg / day, from about 1 mg / kg / day to about 5 mg / kg / day, from about 1 mg / kg / day to about 3 mg / kg / day, from about 3 mg / day to about The dose may be 500 mg / day, about 5 mg / day to about 250 mg / day, about 10 mg / day to about 100 mg / day, about 3 mg / day to about 10 mg / day, or about 100 mg / day to about 250 mg / day. These doses may be administered as a single dose or divided into multiple doses.

[0248] In another aspect of the present invention, a method for activating an intracellular CRF2 receptor is provided. The method comprises contacting a cell in which the CRF2 receptor is to be activated with an effective amount of a CRF2 receptor agonist of the present invention, e.g., a CRF2 receptor agonist defined by Formula 1 or Formula 2.

[0249] In another aspect of the present invention, the use of a peptide-fatty acid conjugate of the present invention as a medicament is provided. In one embodiment, the use of a peptide-fatty acid conjugate of the present invention is provided for the treatment or prevention of obesity.

[0250] In another aspect of the present invention, the peptide-fatty acid conjugate of the present invention is used in the manufacture of a pharmaceutical. In one embodiment, the peptide-fatty acid conjugate of the present invention is used in the manufacture of a pharmaceutical for the treatment or prevention of cancer.

[0251] [Example]

[0252] Hereinafter, the present invention will be described in more detail through the following examples and experimental examples. However, these examples and experimental examples are intended only to aid understanding of the present invention and are not intended to limit the scope of the present invention in any way. Various changes and modifications may be made to the present examples, and such changes and modifications also fall within the scope of the appended claims.

[0253] [Synthesis Example 1] Preparation of CRF2 Receptor Agonist Peptide

[0254] CRF2 receptor agonist peptides were synthesized using an automated peptide synthesizer (Symphony X, Gyros Protein Tech.) utilizing solid-phase synthesis. Rink amide resin was used for C-terminal amidation. Each peptide was synthesized from the C-terminus to the N-terminus by a conventional method using an excess of Fmoc (9H-fluoren-9-ylmethoxycarbonyl)-protected amino acid and an excess of HOBt (1-hydroxybenzotriazole) DIC (diisopropylcarbodiimide) relative to the peptide resin.

[0255] The CRF2 receptor agonist peptides obtained in this way and their sequences are shown in Table 1. The notations E* and K* in SEQ ID NO. 2 and SEQ ID NO. 4 indicate that the corresponding residues form an intramolecular lactam ring.

[0256]

[0257] [Synthesis Example 2] Preparation of CRF2 Receptor Agonist Peptide-Fatty Acid Conjugate

[0258] Similar to the peptide synthesis described above, the synthesis was performed by applying an acylated lysine intermediate instead of lysine in the synthesis of the acylated lysine residue. The C-terminus was amidated, and the acylated lysyl intermediate was the compound below, which contains an eicosanedioic acid and gamma glutamic acid linker and two units of [2-(2-aminoethoxy)ethoxy]acetyl spacer. was used.

[0259] The CRF2 receptor agonist peptide-fatty acid conjugates obtained in this way and their sequences are shown in Table 2. In Table 2, (K) for each conjugate indicates that eicosanedioic acid acylation is performed at the corresponding lysine residue position, and the notations E* and K* in SEQ ID NO. 52 and SEQ ID NO. 54 indicate that the corresponding residues form an intramolecular lactam ring.

[0260]

[0261] [Experimental Example 1] Effect of peptide-fatty acid conjugate on CRF2 receptor

[0262] In this experimental example, the selective agonistic activity of the peptide-fatty acid conjugates of the above-mentioned SEQ ID NOs: 51 to 100 and 116 to 130 toward the CRF2 receptor was evaluated. A method for measuring in vitro CRF receptor stimulating activity was used using cell lines transformed with the CRF2 receptor or the CRF1 receptor. Specifically, each of the transformed cell lines is a cell line transformed to express the human CRF2 receptor in a CHO (Chinese hamster ovary) cell line (CHO-K1 / CRF2R cells) and a cell transformed to express the human CRF1 receptor gene (CHO-K1 / CRF1R cells).

[0263] In order to measure the activity of the peptide-fatty acid conjugate prepared in the above synthetic example, human UCN2 was serially diluted from 10 nM to 0.000169 nM in 3-fold increments, and the conjugates of SEQ ID NOs: 51 to 100 and 116 to 130 were serially diluted from 50 nM to 0.000847 nM in 3-fold increments. The culture medium was removed from the cultured CHO cells expressing the human CRF2 receptor, and the serially diluted substances were added to the cells, followed by incubation at room temperature for 15 minutes. Then, the Eu-cAMP tracer mix and the anti-cAMP detection mix containing the cell lysis buffer were sequentially added to lyse the cells, and the cells were reacted at room temperature for 60 minutes under light-shielding conditions. The cAMP accumulated in the cell lysate upon completion of the reaction was measured by fluorescence, and the EC 50 After calculating the values, they were compared with each other. The relative titers compared to human UCN2 are shown in Table 2 below.

[0264] To measure the activity of the peptide-fatty acid conjugates prepared in the above examples against the CRF1 receptor, native human UCN1 was serially diluted from 1 nM to 0.000977 nM in 2-fold increments, and the conjugates of SEQ ID NOS: 51 to 100 and 116 to 130 were serially diluted from 1,000 nM to 0.97656 nM in 2-fold increments. Alternatively, they were serially diluted from 10,000 nM to 9.7656 nM in 2-fold increments. The culture medium was removed from the cultured CHO cells expressing the human CRF1 receptor, and the serially diluted substances were added to the cells, followed by incubation at room temperature for 15 minutes. Then, the Eu-cAMP tracer mix and the anti-cAMP detection mix containing the cell lysis buffer were sequentially added to lyse the cells, and the cells were reacted at room temperature for 60 minutes under light-shielding conditions. The cAMP accumulated in the cell lysate after the above reaction was completed was measured by fluorescence, and this was used to determine the EC 50 After calculating the values, they were compared with each other. The relative titers compared to human UCN1 are shown in Table 3 below.

[0265] In vitro activity compared to native peptideCHO-K1 / CRF2R cellsCHO-K1 / CRF1R cellsCRF2 receptor activity against CRF1 receptorNon-seq valueRelative to human UCN2 activity (%)Relative to human UCN1 activity (%)519.20.19148525.60.11450539.90.25239545.90.0401505538.20.03311615611.20.172655710.30.0861205810.00.112895917.20.0228006012.30.01210596130.80.0853636225.40. 1611586312.70.577226422.00.1491476517.00.00627876637.01.77521679.50.00422566839.00.03012866939.10.03013217011.10.00620137129.10.00557387222.90.01022247341.10.07 35607427.70.01028207543.30.0775647678.30.1515177767.40.01069787858.70.02325257969.10.1953548053.80.04312538168.20.1394938248.60.0509818335.40.0625768458.90.024 24938561.40.02722778660.80.01833658722.40.1201868873.80.2053608932.00.00934949034.50.0408729131.50.1372299236.00.01721139320.50.001193289430.40.002122409511.7Detected Not detected ∞9627.10.00134185971.4 Not detected ∞983.8 Not detected ∞9910.30.00910981002.50.000∞11642.2 Not detected ∞1170.6 Not detected ∞11834.00.006539211928.80.00644931203.90.0057471210.2 Not detected ∞1228.00.000222771235.80.002332812443.90.0001052811251.40.001189612611.4 Not detected ∞1270.2Not detected∞1281.20.00038271290.60.00156813038.9Not detected∞.

[0266] Thus, it was confirmed that the peptide-fatty acid conjugates of sequence numbers 51 to 100 and 116 to 130 of the present invention manufactured in the examples have a certain or higher activity on both CRF2 and CRF1 receptors, but have a strong selective action on CRF2 or have stimulatory activity only on CRF2 receptors.

[0267] From the above experimental results, it was confirmed that the CRF2 receptor agonists of the present invention can activate the CRF2 receptor.

[0268] [Experimental Example 2] Weight loss effect of peptide-fatty acid conjugates in high-fat diet-induced obese mice.

[0269] In this experimental example, the weight loss effect of the peptide-fatty acid conjugate of the present invention was verified in mice.

[0270] Diet-induced obese (DIO) C57BL / 6J mice were divided into negative control and experimental groups, six per group, and the experimental group was repeatedly subcutaneously administered with a buffer solution (pH 7.3, Dulbecco's phosphate-buffered saline (PBS)) of the peptide-fatty acid conjugate of SEQ ID NO: 55, 71, 76, 77, 86, 95, 96, 116, 118, or 130 at a concentration of 100 nmol / kg at alternate-day intervals for two weeks. The negative control group was repeatedly subcutaneously administered with the same buffer at the same volume. After two weeks of repeated administration, the body weights of the mice were measured, and the change in body weight compared to the negative control group was measured, and statistical analysis was performed using one-way ANOVA.

[0271] As shown in Figure 1, in all mice administered the peptide-fatty acid conjugate of the present invention, statistically significantly (p) higher than the negative control group ***<0.001) body weight was reduced, confirming the weight loss efficacy of the peptide-fatty acid conjugate of the present invention.

[0272] Although the present invention has been described through limited embodiments as described above, the technical idea and scope of the present invention are not limited to these embodiments, and the invention of the patent claims described below, as well as various modifications or variations of the invention described in the patent claims that are obvious to a person having ordinary skill in the technical field to which the present invention pertains, are also included in the scope of the present invention within the equivalent scope of the invention described in the patent claims.

Claims

1. A peptide of chemical formula 1 or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] In the chemical formula above, Z0 is hydrogen; X0 is absent or is lysyl; X1 is isoleucyl or phenylalanyl; X2 is valyl or threonyl; X3 is Ryusil, Valil or Seril; X5 is Ryusil or Valil; X7 is valyl or isoleucyl; X8 is prolyl or seryl; X9 is isoleucyl or phenylalanyl; X 10 Silver glycyl, histidyl or tyrosyl; X 13 is arginyl, glutaminyl, asparagine, glutamyl, aspartyl, alanyl or lysyl; X 15 is leucine or tryptophan; X 17 is glutamyl, prolyl or lysyl; X 18 is glutaminyl, arginyl, isoleucyl, prolyl or lysyl; X 19 is alanyl, glutamyl, prolyl or tryptophanyl; X 20 is glutaminyl, asparagine, glutamyl, aspartyl, alanyl, arginyl or lysyl; X 21 is alanyl or glutaminyl; X 22 is arginyl, lysyl, glutaminyl, asparagine, aspartyl, alanyl or glutamyl; X 23 alanyl or lysyl; X 24 is alanyl, leucyl or glutamyl; X 25 is arginyl or lysyl; X 26 is glutamyl or glutaminyl; X 28 Silver alanyl or valyl; X 29 is threonyl, glutamyl or alanyl; X 30 is threonyl, glutamyl or lysyl; X 32 is alanyl or arginyl; X 33 is arginyl, glutamyl, glutaminyl, asparagine, aspartyl, alanyl or lysyl; X 34 is isoleucine or methionyl; X 35 is lysyl or phenylalanyl; X 36 is alanyl, glutamyl, glutaminyl, asparagine, arginyl, lysyl or aspartyl; X 37 is arginyl, glutamyl, aspartyl or glutaminyl; Z 39 is OH or NH2.

2. In paragraph 1, X1 is isoleucyl; X2 is valyl; X3 is leucyl; X8 is prolyl; X 10 Silver glycyrrhizin; X 18 A peptide characterized in that it is glutaminyl.

3. In paragraph 2, X 23 and X 24 is alanyl; X 25 is arginyl, X 26 A peptide characterized by being glutamyl; 4. In paragraph 1, X 20 A peptide characterized in that the side chain of the lysyl residue and the side chain of the 17th residue, glutamyl, form an intramolecular lactam ring.

5. In paragraph 1, X 23 is lysyl, and X 26 A peptide characterized in that the side chain of the lysyl residue and the side chain of the glutamyl residue form an intramolecular lactam ring.

6. Peptide-fatty acid conjugate of chemical formula 2 or a pharmaceutically acceptable salt thereof: [Chemical Formula 2] In the above formula, R A is a fatty acid group having 14 to 24 carbon atoms and is covalently bonded to linker L through its acyl group; L is a linker selected from the group consisting of one to four amino acid residues, one to three iminobis(ethyleneoxy)acetyl groups, one to three 4-iminobutanoyl groups, one to three 5-iminopentanoyl groups, one to three 6-iminohexanoyl groups, a triazole group, one to three oxyethylene groups, and any combination thereof; Δ is a peptide of chemical formula 3, [Chemical Formula 3] In the chemical formula above, Z0 is hydrogen; X0 is absent or is lysyl; X1 is isoleucyl or phenylalanyl; X2 is valyl or threonyl; X3 is Ryusil, Valil or Seril; X5 is Ryusil or Valil; X7 is valyl or isoleucyl; X8 is prolyl or seryl; X9 is isoleucyl or phenylalanyl; X 10 Silver glycyl, histidyl or tyrosyl; X 13 is arginyl, glutaminyl, asparagine, glutamyl, aspartyl, alanyl or lysyl; X 15 is leucine or tryptophan; X 17 is glutamyl, prolyl or lysyl; X 18 is glutaminyl, arginyl, isoleucyl, prolyl or lysyl; X 19 is alanyl, glutamyl, prolyl or tryptophanyl; X 20 is glutaminyl, asparagine, glutamyl, aspartyl, alanyl, arginyl or lysyl; X 21 is alanyl or glutaminyl; X 22 is arginyl, lysyl, glutaminyl, asparagine, aspartyl, alanyl or glutamyl; X 23 alanyl or lysyl; X 24 is alanyl, leucyl or glutamyl; X 25 is arginyl or lysyl; X 26 is glutamyl or glutaminyl; X 28 Silver alanyl or valyl; X 29 is threonyl, glutamyl or alanyl; X 30 is threonyl, glutamyl or lysyl; X 32 is alanyl or arginyl; X 33 is arginyl, glutamyl, glutaminyl, asparagine, aspartyl, alanyl or lysyl; X 34 is isoleucine or methionyl; X 35 is lysyl or phenylalanyl; X 36 is alanyl, glutamyl, glutaminyl, asparagine, arginyl, lysyl or aspartyl; X 37 is arginyl, glutamyl, aspartyl or glutaminyl; Z 39 is OH or NH2, But at this time X0, X 13 , X 17 , X 18 , X 20 , X 22 , X 23 , X 25 , X 30 , X 33 and X 36 At least one of them is a lysyl residue, wherein X0, X 13 , X 17 , X 18 , X 20 , X 22 , X 23 , X 25 , X 30 , X 33 or X 36 L is covalently linked to the ε-amino group of the side chain of one lysyl residue.

7. In the 6th paragraph, the peptide-fatty acid conjugate is characterized by having a structure of chemical formula 5: [Chemical Formula 5] In chemical formula 5, i is an integer from 12 to 22; j is an integer from 0 to 3; Sp is absent or selected from the functional groups of the chemical formula 6 below, [Chemical Formula 6] Here, * indicates the direction in which the lysyl residue of Δ is connected to the ε-amino group.

8. A peptide-fatty acid conjugate in which i is 18 and j is 1 in paragraph 7.

9. In paragraph 7, Sp A peptide-fatty acid conjugate.

10. In the 6th paragraph, L is X0, X 13 , X 20 , X 22 or X 33 A peptide-fatty acid conjugate characterized by being covalently linked to the ε-amino group of .

11. In paragraph 6, X1 is isolates; X3 is Ryusil or Seril; X5 is Ryu Sil; X7 is Valil; X8 is prolyl; X9 is isolates; X 10 Silver glycyl or tyrosyl; X 13 Silver Lysyl; X 15 is Ryu Sil; X 17 is glutamyl; X 18 is glutaminyl or arginyl; X 19 is alanine; X 20 Silver arginyl or lysyl; X 21 Silver alanyl; X 22 is arginyl or lysyl; X 23 Silver alanyl; X 24 is alanine; X 25 is arginyl; X 26 is glutamyl; X 29 is threonyl; X 30 is threonyl; X 32 is alanine; X 33 is glutamyl or lysyl; X 35 is Ryu Sil; X 36 is glutamyl; X 37 is arginyl or glutaminyl, L is X0, X 20 , X 22 or X 33 A peptide-fatty acid conjugate characterized in that it is covalently linked to the ε-amino group of the side chain of one lysyl residue.

12. A pharmaceutical composition for preventing or treating obesity, comprising a therapeutically effective amount of a peptide-fatty acid conjugate according to claim 6 and a pharmaceutically acceptable excipient.

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