VIPR2 antagonist peptide

Abstract

To provide novel VIPR2 antagonist peptides useful for use as pharmaceuticals, diagnostic agents, and / or research reagents.SOLUTION: The present invention provides a cyclic peptide comprising an amino acid sequence represented by formula (1): c[XN-X4-X5-X6-c(X7-X8-X9-X10]-X11)-X12-X13-X14-X15-X16 (1) or formula (2): c[XN-X4-X5-X6-c(X7-X8-X9-X10)]-X11-X12-X13-X14-X15-X16 (2) or a pharmaceutically acceptable salt thereof.SELECTED DRAWING: Figure 1

Description

Cross Reference

[0001] This application claims priority in Japan based on Japanese Patent Application No. 2020-059721, filed on March 30, 2020, the entire contents of which are incorporated herein by reference in their entirety. In addition, the entire contents of all patents, patent applications, and literature cited in this application are incorporated herein by reference in their entirety. [Technical Field]

[0002] The present invention relates to peptides that inhibit the activity of VIPR2, and more particularly to cyclic peptides that have a specific amino acid sequence and a cyclic structure. [Background technology]

[0003] Vasoactive intestinal peptide receptor 2 (VIPR2), also known as VPAC2, is a class B G protein-coupled receptor (GPCR). It is expressed throughout the body, including in the central nervous system, peripheral nervous system, sensory organs, digestive system, and reproductive organs. It exerts diverse physiological functions through interaction with its ligands, vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP). For example, VIP and PACAP signaling via VIPR2 and its receptor subtypes, vasoactive intestinal peptide receptor 1 (VIPR1 or VPAC1) and pituitary adenylate cyclase-activating polypeptide type-1 receptor (PAC1), is known to have neuroprotective effects (Non-Patent Documents 1 and 2). Interestingly, recent studies have suggested that high expression or overactivation of VIPR2 may be involved in the development of schizophrenia and autism spectrum disorder (Non-Patent Documents 3, 4, and 5). It has also been reported that inhibition of VIP signaling suppresses cancer growth and activates the immune system (Non-patent Documents 6 and 7).

[0004] In 2011, Vacic et al. reported that some schizophrenia and autism spectrum disorder patients have duplications in the gene region encoding VIPR2, and that their lymphocytes exhibit increased VIPR2 mRNA expression levels and are hypersensitive to VIP (Non-Patent Document 3). In 2015, Ago et al. reported that when newborn mice were administered the VIPR2-selective agonist Ro25-1553 to artificially induce VIPR2 hyperactivity, adult mice exhibited cognitive decline (Non-Patent Document 4). Furthermore, in 2019, Tian et al. generated transgenic mice with duplications in the gene region encoding VIPR2 and reported that the mice exhibited symptoms such as dopamine dysfunction, cognitive dysfunction, and impaired social behavior (Non-Patent Document 5). These findings strongly suggest that excessive signaling via VIPR2 is involved in the development of psychiatric disorders such as schizophrenia and autism spectrum disorder.

[0005] Schizophrenia is said to affect 24 million people worldwide. It is a mental illness that not only reduces an individual's social function by causing positive symptoms (such as hallucinations and delusions), negative symptoms (such as loss of motivation), and cognitive impairment, but also has a high suicide rate. While the heritability is high at 80%, no specific disease-causing genes common to all patients have been identified, and the development of new drugs and treatment methods is desired. Autism spectrum disorder is said to affect approximately 1-2 people per 1,000 worldwide, and causes impairments in social and interpersonal communication skills. Because congenital factors play a large role, no clear prevention or treatment methods have been established.

[0006] As mentioned above, it has been reported that some patients with schizophrenia and some patients with autism spectrum disorder overexpress VIPR2, suggesting that overactivation of VIPR2 may be involved in the onset of the pathology. In other words, if genome analysis confirms that there is duplication in the gene region encoding VIPR2, administration of a VIPR2 inhibitor can be expected to be effective as a means of preventing and / or treating schizophrenia and autism spectrum disorder.

[0007] In 2018, the present inventors reported the discovery of a cyclic antagonist peptide, VIpep-3:Ac-c(Cys-Pro-Pro-Tyr-Leu-Pro-Arg-Arg-Leu-Cys)-Thr-Leu-Leu-Leu-Arg-Ser-OH (SEQ ID NO: 11), which binds to a recombinant protein of the extracellular domain of VIPR2 and inhibits the VIPR2-mediated signaling pathway of VIP and PACAP in a cellular assay (Non-Patent Document 8). [Prior art documents] [Non-patent literature]

[0008] [Non-Patent Document 1] Vaudry D, Falluel-Morel A, Bourgault S, Basille M, Burel D, Wurtz O, Fournier A, Chow BK, Hashimoto H, Galas L, and Vaudry H. Pharmacol Rev. 2009,61(3):283-357. [Non-patent document 2] Passemard S, Sokolowska P, Schwendimann L, and Gressens P. Curr Pharm Des. 2011,17(10):1036-1039. [Non-patent document 3] Vacic V,McCarthy S,Malhotra D,Murray F,Chou HH,Peoples A,Makarov V,Yoon S,Bhandari A,Corominas R,Iakoucheva LM,Krastoshevsky O,Krause V,Larach-Walters V,Welsh DK,Craig D,Kelsoe JR,Aquila, Gallery, SMD M,Morris DW,Gill M,Corvin A,Insel PA,McClellan J,King MC,Karayiorgou M,Levy DL,DeLisi LE,and Sebat J. Nature. 2011,471(7339):499-503.

Fashion 4

Wood 5

Wood 6

Waterfall 7

[0009] The above-mentioned VIpep-3 discovered by the inventors is a cyclic antagonist peptide that tightly binds to the extracellular domain of human VIPR2 and inhibits the VIP-VIPR2 signaling pathway. However, because it is composed only of natural amino acids, there are concerns that it has low resistance to protease degradation.

[0010] The present invention has been made in view of the above problems, and aims to provide a novel VIPR2 antagonist peptide suitable for use as a pharmaceutical, diagnostic agent, and / or research reagent. [Means for solving the problem]

[0011] The present inventors discovered a group of novel VIPR2 antagonist peptides during optimization studies of VIpep-3. To solve the above problems, they investigated (1) amino acid residue substitution, (2) molecular weight reduction by deletion of N-terminal and / or C-terminal amino acid residues, and (3) introduction of a bicyclic structure. As a result, they discovered a group of novel VIPR2 antagonist peptide sequences that are resistant to protease degradation and have VIPR2 antagonist activity, leading to the completion of the present invention.

[0012] That is, the present invention includes the following embodiments. [1] Formula (1): c[X N -X 4 -X 5 -X 6 -c(X 7 -X 8 -X 9 -X 10 ]-X 11 )-X 12 -X 13 -X 14 -X 15 -X 16 (1) or formula (2): c[X N -X 4 -X 5 -X 6 -c(X 7 -X 8 -X 9 -X 10 )]-X 11 -X 12 -X 13 -X 14 -X 15 -X 16 (2) or a pharmacologically acceptable salt thereof.

[0013] Here, in formula (1), X N and X 10 and X 7 and X11 and in formula (2), X N and X 7 and X 10 are independently covalently bonded to each other to form two cyclic structures in the molecule. These covalent bonds may be direct covalent bonds between the main chain and the side chain or between the side chains, or may be indirect covalent bonds via a linker. In the above formulas (1) and (2), X N is X 1 -X 2 -X 3 and X 1 , X 2 , X 3 , X 7 , X 10 and X 11 When involved in the cyclization within the peptide molecule, each independently represents an amino acid residue having an amino group, a carboxy group, a thiol group, an allyl group, an alkynyl group, an azide group, or a halogen atom, or a derivative thereof; when not involved in the cyclization within the peptide molecule, X 1 and X 2 each independently represents any amino acid residue or deletion, and X 3 and X 11 are each independently any amino acid residue. 5 , X 9 , X 12 and X 13 Each of X independently represents an amino acid residue having a hydrocarbon group which may have a substituent, or a derivative thereof. 4 represents an amino acid residue having an optionally substituted aromatic carbocyclic group or a derivative thereof, and X 6 and X 8 represents any amino acid residue, and X 14 and X 15 and X 16 each independently represents an arbitrary amino acid residue or deletion, and the N-terminal amino group and the C-terminal carboxy group may be modified or deleted.

[0014] Preferred or alternative embodiments of the present invention and the X in the above cyclic peptides 1 From X16 Preferred aspects of each amino acid residue are described in detail below, but each preferred aspect is independent of each other, and each embodiment can be combined arbitrarily. [Effects of the Invention]

[0015] The present invention provides novel cyclic peptides that are useful for using VIPR2 antagonists as pharmaceuticals, diagnostic agents, and / or research reagents. [Brief explanation of the drawings]

[0016] [Figure 1] Figure 1 shows the amino acid sequence of VIpep-3, which is involved in its VIPR2 binding activity (pharmacophore) and its cyclic structure regulation (SS bond). It also shows an outline of the structure of the peptide of the present invention, which is a modified version of VIpep-3. [Figure 2] FIG. 2 shows examples of amino acids used in substitution studies at each amino acid position of VIpep-3, and the positions of the amino acid residues examined for bicyclization. [Figure 3] Figure 3 shows an outline of the competitive binding assay using cell-based ELISA to confirm the binding activity of peptides with substituted amino acid residues to VIPR2. In the figure, SA represents streptavidin, B represents biotin, and HRP represents horseradish peroxidase. [Figure 4] FIG. 4 shows the results of comparing the binding activity to VIPR2 of a group of peptide sequences subjected to amino acid residue substitution studies and representative examples of the peptides of the present invention with that of the parent peptide VIpep-3. [Figure 5] FIG. 5 shows the results of adding a representative example of the peptide of the present invention and VIP to VIPR2-expressing cells and evaluating changes in intracellular calcium concentration, which is one of the signals downstream of VIPR2. [Figure 6] FIG. 6 shows the results of comparing the resistance of VIpep-3 and representative examples of the peptides of the present invention to protease degradation when mixed with rat plasma. [Figure 7]Figure 7 shows the results of adding a representative example of the peptide of the present invention and VIP to VIPR1-expressing cells and evaluating changes in intracellular calcium concentration, which is one of the signals downstream of VIPR1; adding a representative example of the peptide of the present invention and VIP to VIPR2-expressing cells and evaluating changes in intracellular calcium concentration, which is one of the signals downstream of VIPR2; and adding a representative example of the peptide of the present invention and PACAP to PAC1-expressing cells and evaluating changes in intracellular calcium concentration, which is one of the signals downstream of PAC1. [Figure 8] Figure 8 shows the results of assessing the phosphorylation of cAMP response element binding protein (CREB) (one of the VIPR2 downstream signals) in cells of the prefrontal cortex of ICR mice (12 days old) after subcutaneous administration of Ro25-1553 (indicated as "Vehicle" in the figure), a VIP analog and a selective VIPR2 agonist, or a mixture of a representative example of the peptide of the present invention and Ro25-15532 (indicated as "peptide concentration 1 nmol / g" or "10 nmol / g" in the figure). [Figure 9] Figure 9(A) shows the results of evaluating the novel object recognition function of mice administered Ro25-1553, a VIP analog and VIPR2-selective agonist, and Figure 9(B) shows the results of evaluating the novel object recognition function of mice administered Ro25-1553 alone or a mixture of a representative example of the peptide of the present invention and Ro25-15532. [Figure 10] FIG. 10 shows the structural formula of a representative example of the peptide of the present invention used in the Examples. DETAILED DESCRIPTION OF THE INVENTION

[0017] Next, preferred embodiments of the present invention will be described with reference to the drawings. Note that the embodiments described below do not limit the invention according to the claims, and not all of the elements and combinations thereof described in the embodiments are necessarily essential to the solution of the present invention. In addition, the disclosures of all patent and non-patent documents cited in this specification are incorporated herein by reference in their entirety.

[0018] (definition) As used herein, a peptide refers to two or more amino acids bonded together via amide bonds (peptide bonds), and can be, for example, 2 to 20 amino acids bonded together via amide bonds. In addition, according to the convention for peptide notation, the left end is the N-terminus (amino terminus) and the right end is the C-terminus (carboxy terminus). The first carbon atom adjacent to the carbonyl group that forms the peptide bond is referred to as the Cα carbon.

[0019] As used herein, the term "any amino acid or derivative thereof" is used in its broadest sense and includes not only naturally occurring amino acids, but also artificial amino acids with unnatural structures, chemically synthesized compounds having properties known in the art that are characteristic of amino acids, and carboxylic acids having functional groups. Examples of unnatural amino acids include D-amino acids, α / α-disubstituted amino acids with a backbone structure that differs from that of natural amino acids (such as α-methylated amino acids such as 2-aminoisobutyric acid), N-alkyl-amino acids (such as N-methylated amino acids), N-substituted glycines (peptoids), amino acids with extended backbones (such as homo-β and homo-γ amino acids), amino acids with a side chain structure that differs from that of natural amino acids (such as cyclohexylalanine, allylglycine, 2-(2-pyridyl)-glycine, and 3-(1H-benzimidazol-2-yl)-alanine), amino acids with partially substituted side chains (such as norleucine, diaminopropanoic acid, and 3-(2-pyridyl)-alanine), amino acids with extra functional groups in the side chain; amino acids with extra C, alkyl groups, or methyl groups in the side chain (such as homonorleucine and γ-methylleucine), amino acids with halogen atoms (F, Cl, Br, I) in the side chain (such as 3-chloro-alanine), and amino acids with halogen atoms (F, Examples of amino acids include, but are not limited to, carboxylic acids having an extra N or amino group in the side chain (such as 3-azidoalanine and ornithine), amino acids having an extra O or methoxy group in the side chain (such as O-methyl-serine and O-methyl-threonine), amino acids having an extra hydroxy group in the side chain (such as 3-hydroxy-phenylalanine), amino acids having an extra carboxy group (-COOH) in the side chain (such as 3-carboxy-phenylalanine), amino acids having an extra S in the side chain (such as ethionine), amino acids in which the carboxylic acid functional group in the side chain is protected with an ester (such as aspartic acid-4-methyl ester), and amino acids in which the thio group (-S-) in the side chain has been oxidized to a sulfinyl group (-S(=O)-) or a sulfonyl group (-S(=O)2-) (methionine sulfoxide).

[0020] In this specification, the hydrocarbon group which may have a substituent includes, for example, C 1, C 2, C 3, C 4, C 5, C 6, C 7, C 8, C 9, C 10, C 11, C 12, C 13, C 14, C 15, C 16, C 17, C 18, C 19, C 20, C 21, C 22, C 23, C 24, C 25, C 26, C 27, C 28, C 29, C 30, C 31, C 32, C 33, C 34, C 35, C 36, C 37, C 38, C 39, C 40, C 41, C 42, C 43, C 44, C 45, C 46, C 47, C 48, C 49 ...9, C 49, C 49, C 49, C 49, C 49, C 49, C 49, C 49, C 49, C 49, C 49, C 49, C 49, C 49, C 49, C 49, C 49, C 49, C 49, C 49, C 49, C 49, C 49, C 4 1-10 C alkyl groups such as ethenyl, 1-propenyl, 2-propenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 3-methyl-2-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 4-methyl-3-pentenyl, 1-hexenyl, 3-hexenyl, and 5-hexenyl 2-10 Alkenyl groups, such as ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, 5-hexynyl, and 4-methyl-2-pentynyl. 2-10 Alkynyl groups, C3-10 cycloalkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl, bicyclo[3.2.1]octyl, and adamantyl, C4 groups such as cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl 3-10The term "cycloalkenyl group" refers to a cycloalkenyl group, and examples of the aromatic carbocyclic group which may have a substituent include a phenyl group and a naphthyl group. Examples of the aromatic heterocyclic group which may have a substituent include 5- or 6-membered monocyclic aromatic heterocyclic groups such as pyridyl, thienyl, furyl, pyrrolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyrazinyl, pyrimidinyl, pyridazinyl, 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, triazolyl, tetrazolyl, and triazinyl, as well as benzofuranyl, benzothienyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, benzotriazolyl, imidazopyridinyl, and thienopyryl. inyl, furopyridinyl, pyrrolopyridinyl, pyrazolopyridinyl, oxazolopyridinyl, thiazolopyridinyl, imidazopyrazinyl, imidazopyrimidinyl, thienopyrimidinyl, furopyrimidinyl, pyrrolopyrimidinyl, pyrazolopyrimidinyl, oxazolopyrimidinyl, thiazolopyrimidinyl, pyrazolotriazinyl, naphtho[2,3-b]thienyl, phenoxathiinyl, indolyl, iridinyl It refers to 8- to 14-membered fused polycyclic (preferably bi- or tricyclic) aromatic heterocyclic groups such as isoindolyl, 1H-indazolyl, purinyl, isoquinolyl, quinolyl, phthalazinyl, naphthyridinyl, quinoxalinyl, quinazolinyl, cinnolinyl, carbazolyl, β-carbolinyl, phenanthridinyl, acridinyl, phenazinyl, phenothiazinyl, and phenoxazinyl, but is not limited to these.

[0021] As used herein, "VIPR2" refers to a protein found in mammals such as mice, rats, dogs, monkeys, and humans.

[0022] As used herein, the term "having VIPR2 antagonist activity" refers to the following: (1) inhibiting the binding of a biotin-labeled version of the previously reported VIPR2 antagonist peptide VIpep-3 to a VIPR2 recombinant protein in an in vitro test; (2) inhibiting the binding of a biotin-labeled version of the previously reported VIPR2 antagonist peptide VIpep-3 to VIPR2-expressing cells; (3) binding to a VIPR2 recombinant protein in a concentration-dependent manner; (4) inhibiting the binding of a biotin-labeled version of the previously reported VIPR2 antagonist peptide VIpep-3 to VIPR2-expressing cells; (5) binding to VIPR2 in a concentration-dependent manner, (6) suppressing an increase in cAMP concentration in VIPR2-expressing cells when added prior to or co-added with VIP, and (7) suppressing β-arrestin recruitment in VIPR2-expressing cells when added prior to or co-added with VIP, etc. If a compound exhibits any one of these effects, it is said to have VIPR2 antagonist activity. The presence or absence of VIPR2 antagonist activity can be confirmed by those skilled in the art according to known methods, such as by referring to Non-Patent Document 7, but is not limited thereto.

[0023] (cyclic peptide) The structure of a cyclic peptide according to one embodiment of the present invention is described with reference to Figure 1. Figure 1 schematically illustrates the characteristics (pharmacophore) of VIpep-3 that are the basis for cyclic peptide design and the structure related to the function of cyclic structure regulation (SS bond), as well as the relationship with the cyclic peptide of this embodiment, which is a modified version of VIpep-3. The first to thirteenth amino acid residues of VIpep-3 correspond to the first to thirteenth amino acid residues of the cyclic peptide of this embodiment, but the first amino acid or the first and second amino acid residues may be deleted in the cyclic peptide of this embodiment. In the cyclic peptide of this embodiment, the cyclization between the first and tenth cysteine residues in VIpep-3 can be represented as: (1) two cyclic structures formed by bond A between the first and tenth amino acid residues and bond B between the seventh and eleventh amino acid residues; (2) two cyclic structures formed by bond C between the second and tenth amino acid residues and bond D between the seventh and eleventh amino acid residues; or two cyclic structures formed by bond G between the second and tenth amino acid residues and bond H between the seventh and eleventh amino acid residues; or (3) two cyclic structures formed by bond G between the third and tenth amino acid residues. (3) two cyclic structures formed by bond E between the 7th and 11th amino acid residues and bond F between the 7th and 11th amino acid residues, or two cyclic structures formed by bond I between the 3rd and 10th amino acid residues and bond J between the 7th and 11th amino acid residues, (4) two cyclic structures formed by bond K between the 1st, 7th, and 10th amino acid residues, (5) two cyclic structures formed by bond L or bond N between the 2nd, 7th, and 10th amino acid residues, and (6) two cyclic structures formed by bond M or bond O between the 3rd, 7th, and 10th amino acid residues. Examples of amino acids used for substitution at each amino acid position of VIpep-3 and the positions of the amino acid residues examined for bicyclization are shown in Figure 2.

[0024] [1] The cyclic peptide shown in FIG. 1 is more specifically represented by the following formula (1): c[X N -X 4 -X 5 -X 6 -c(X7 -X 8 -X 9 -X 10 ]-X 11 )-X 12 -X 13 -X 14 -X 15 -X 16 (1), or The following formula (2): c[X N -X 4 -X 5 -X 6 -c(X 7 -X 8 -X 9 -X 10 )]-X 11 -X 12 -X 13 -X 14 -X 15 -X 16 It can be expressed as (2).

[0025] Here, in formula (1), X N and X 10 and X 7 and X 11 In addition, in formula (2), X N and X 7 and X 10 and independently form two cyclic structures in the molecule by covalent bonding. These covalent bonds may be direct covalent bonds between the main chain and the side chain, or between the side chains, or may be indirect bonds via a linker.

[0026] In the above formulas (1) and (2), X N is X 1 -X 2 -X 3 and X 1 , X 2 , X 3 , X 7 , X 10 and X 11When involved in the cyclization within the peptide molecule, each independently represents an amino acid residue having an amino group, a carboxy group, a thiol group, an allyl group, an alkynyl group, an azide group, or a halogen atom, or a derivative thereof; when not involved in the cyclization within the peptide molecule, X 1 and X 2 each independently represents any amino acid residue or deletion, and X 3 and X 11 are each independently any amino acid residue. 5 , X 9 , X 12 and X 13 Each of X independently represents an amino acid residue having a hydrocarbon group which may have a substituent, or a derivative thereof. 4 represents an amino acid residue having an optionally substituted aromatic carbocyclic group or a derivative thereof, and X 6 and X 8 represents any amino acid residue, and X 14 and X 15 and X 16 each independently represents an arbitrary amino acid residue or deletion, and the N-terminal amino group and the C-terminal carboxyl group may be modified or deleted.

[0027] [2] A preferred embodiment of formula (1) is a compound represented by the following formula (3): c[X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -c(X 7 -X 8 -X 9 -X 10 ]-X 11 )-X 12 -X 13 (3) In formula (3), X is a cyclic peptide having an amino acid sequence represented by the formula (3) or a pharmacologically acceptable salt thereof. 1 and X 10 and X 7 and X 11 are independently covalently bonded to form two cyclic structures within the molecule, and the covalent bonds are1 and the main chain or side chain of X 10 and the side chain of X 7 and the side chain of X 11 The side chain of X 1 ~X 13 is as described in [1].

[0028] In one embodiment, X 1 represents cysteine, D-cysteine, 3-mercaptopropanoic acid, 2,3-diaminopropanoic acid, D-2,3-diaminopropanoic acid, β-alanine, aspartic acid, or glutamic acid; X 10 represents cysteine, D-cysteine, 2,3-diaminopropanoic acid, D-2,3-diaminopropanoic acid, aspartic acid, or glutamic acid; X 2 and X 3 each independently represents any amino acid residue, preferably proline; X 4 and X 8 each independently represents an amino acid residue having an aromatic carbocyclic group, preferably tyrosine, phenylalanine, or a derivative thereof; X 5 , X 9 , X 12 and X 13 each independently represents leucine, isoleucine, or a derivative thereof; X 7 and X 11 each independently represent lysine, ornithine, arginine, 2,3-diaminopropanoic acid, 2,4-diaminobutanoic acid, aspartic acid, glutamic acid, cysteine, homocysteine, or a derivative thereof.

[0029] [3] where X 1 and X 10 The bond between the Cα carbon atoms is represented by the following formula (4): Cα 1 -(CH2) A -L 1 -(CH2) B -Cα 10 (4) In the formula, A and B each independently represent an integer of 0 to 2. The sum of A and B is an integer of 1 to 4. The linker L 1 are S, SS, CH2-CH2, S-CH2, CH2-S, O-CH2, CH2-O, CH=CH, NH-C(=O), N(CH3)-C(=O), NH-C(=S), N(CH3)-C(=S), OC(=O), C(=O)-NH, C(=O)-N(CH3), C(=S)-NH, C(=S )-N(CH3), C(=O)-O, CH2-CH2-CH2, S-CH2-CH2, CH2-CH2-S, CH2-S-CH2, O-CH2-CH2, CH2-CH2-O, CH2-O-CH2, S-CH2-S, SC(=CH2)-S, S-(CH2)2-S, or triazole.

[0030] Among these, L 1 preferably represents SS, NH—C(═O), CH—S—CH, CH═CH, or CH—CH, and more preferably represents L 1 represents SS or NH-C(=O). 1 represents SS, for example, X 1 and X 10 are both amino acid residues or derivatives thereof having a thiol group, and form a disulfide bond under oxidative conditions. 1 represents NH-C(=O), for example, X 1 is an amino acid residue having an amino group in the side chain, and X 10 are amino acid residues having a carboxy group in the side chain, and have a structure cyclized by dehydration condensation of these amino and carboxy groups. Examples of amino acid residues having a thiol group in the side chain include cysteine, D-cysteine, 3-mercaptopropanoic acid, homocysteine, and D-homocysteine. Examples of amino acid residues having an amino group in the side chain include 2,3-diaminopropanoic acid, D-2,3-diaminopropanoic acid, and β-alanine. Examples of amino acid residues having a carboxy group in the side chain include aspartic acid and glutamic acid.

[0031] [4] Another preferred embodiment of the formula (1) is a compound represented by the formula (5): c[X 2 -X 3 -X 4 -X 5 -X 6 -c(X 7 -X 8 -X 9 -X 10 ]-X 11 )-X 12 -X 13 (5) In formula (5), X is a cyclic peptide having an amino acid sequence represented by the formula (5) or a pharmacologically acceptable salt thereof. 2 and X 10 and X 7 and X 11 are independently covalently bonded to form two cyclic structures within the molecule, and the covalent bonds are 2 and the main chain or side chain of X 10 and the side chain of X 7 and the side chain of X 11 The side chain of X 2 ~X 13 is as described in [1].

[0032] [5] where X 2 and X 10 The bond between the Cα carbon atoms is represented by the following formula (6): Cα 2 -(CH2) A -L 2 -(CH2) B -Cα 10 (6) In the formula, A and B each independently represent an integer of 0 to 6, and the sum of A and B is an integer of 4 to 7. 2are S, SS, CH2-CH2, S-CH2, CH2-S, O-CH2, CH2-O, CH=CH, NH-C(=O), N(CH3)-C(=O), NH-C(=S), N(CH3)-C(=S), OC(=O), C(= O)-NH, C(=O)-N(CH3), C(=S)-NH, C(=S)-N(CH3), C(=O)-O, CH2-CH2-CH2, S-CH2-CH2, CH2-CH2-S, CH2-S-CH2, O-CH2-CH 2, CH2-CH2-O, CH2-O-CH2, S-CH2-S, SC(=CH2)-S, S-(CH2)2-S, S-(CH2)3-S, S-(CH2)4-S, S-(CH2)5-S, S-CH2-CH=CH-CH2-S, S-CH2-C(=O)-NH, NH-C(=O)-CH2-S, S-CH2-C6H4-CH2-S (the bonding position of the methylene group to the phenylene ring may be ortho, meta or para), S-CH2-C 10 H6-CH2-S, S-CH2-C(=O)-CH2-S, or S-CH2-C(=CH2)-CH2-S, triazole, or dithiotetrafluorobenzene.

[0033] Among these, L 2 Preferably, represents S—(CH2)2—S, S—(CH2)3—S, S—(CH2)4—S, or S—(CH2)5—S. For example, X 2 and X 10 are both amino acid residues or derivatives thereof having a thiol group, and a thioether bond can be formed between the thiol group of these residues and a halogenated alkyl linker such as 1,2-diiodoethane, 1,3-diiodopropane, or 1,4-diiodobutane.

[0034] [6] Yet another preferred embodiment of the formula (1) is a compound represented by the formula (7): c[X 3 -X 4 -X 5 -X 6 -c(X 7 -X 8 -X 9 -X 10 ]-X 11 )-X12 -X 13 (7) In formula (7), X is a cyclic peptide having an amino acid sequence represented by the formula (7) or a pharmacologically acceptable salt thereof. 3 and X 10 and X 7 and X 11 are independently covalently bonded to form two cyclic structures within the molecule, and the covalent bonds are 3 and the main chain or side chain of X 10 and the side chain of X 7 and the side chain of X 11 The side chain of X 3 ~X 13 is as described in [1].

[0035] In one embodiment, X 3 represents cysteine, D-cysteine, homocysteine, D-homocysteine, 3-mercaptopropanoic acid, 6-mercaptohexanoic acid, or 8-mercaptohexanoic acid, and X 10 represents cysteine, D-cysteine, homocysteine, or D-homocysteine, and X 4 and X 8 each independently represents an amino acid residue having an aromatic carbocyclic group, preferably tyrosine, phenylalanine, or a derivative thereof; X 5 , X 9 , X 12 and X 13 each independently represents leucine, isoleucine, or a derivative thereof; X 7 and X 11 each independently represent lysine, ornithine, arginine, 2,3-diaminopropanoic acid, 2,4-diaminobutanoic acid, aspartic acid, glutamic acid, or a derivative thereof.

[0036] [7] where X 3 and X 10 The bond between the Cα carbon atoms is represented by the following formula (8): Cα 3 -(CH2) A -L 3 -(CH2) B -Cα10 (8) In the formula, A and B each independently represent an integer of 0 to 10, the sum of A and B is an integer of 7 to 10, and L 3 are S, SS, CH2-CH2, S-CH2, CH2-S, O-CH2, CH2-O, CH=CH, NH-C(=O), N(CH3)-C(=O), NH-C(=S), N(CH3)-C(=S), OC(=O), C(=O)-NH, C(=O)-N(CH3), C (=S)-NH, C(=S)-N(CH3), C(=O)-O, CH2-CH2-CH2, S-CH2-CH2, CH2-CH2-S, CH2-S-CH2, O-CH2-CH2, CH2-CH2-O, CH2-O-CH2, S-CH2-S, SC(=CH2)-S , S-(CH2)2-S, S-(CH2)3-S, S-(CH2)4-S, S-(CH2)5-S, S-(CH2)6-S, S-(CH2)7-S, S-(CH2)8-S, S-CH2-CH=CH-CH2-S, S-CH2-C(=O)-NH, NH-C(=O)-CH2-S, S-CH2-C6H4-CH2-S (the bonding position of the methylene group to the phenylene ring may be ortho, meta or para), S-CH2-C6H4-C6H4-CH2-S, S-CH2-C5NH3-C5NH3-CH2-S, S-CH2-C 10 H6-CH2-S, S-CH2-C(=O)-CH2-S, or S-CH2-C(=CH2)-CH2-S, triazole, or dithiotetrafluorobenzene.

[0037] Among these, L 3 is preferably S—(CH)—S, S—(CH)—S, S—(CH)—S, S—(CH)—S, or S—CH—C—H—CH—S (the bonding position of the methylene group to the phenylene ring may be ortho, meta, or para). For example, X 3 and X 10are both amino acid residues or derivatives thereof having a thiol group, and a thioether bond can be formed between the thiol group of these residues and a halogenated alkyl linker such as 1,3-diiodopropane, 1,4-diiodobutane, 1,5-diiodopentane, 1,6-diiodohexane, 1,2-bisbromomethylbenzene, 1,3-bisbromomethylbenzene, or 1,4-bisbromomethylbenzene.

[0038] [8] Other preferred embodiments of the above formulas (1), (3), (5), and (7) include X 7 and X 11 The bond between the Cα carbon atoms is represented by the following formula (9): Cα 7 -(CH2) A -L 7 -(CH2) B -Cα 11 (9) In the formula, A and B each independently represent an integer of 0 to 7, the sum of A and B is an integer of 2 to 7, and L 7 are S, SS, CH2-CH2, S-CH2, CH2-S, O-CH2, CH2-O, CH=CH, NH-C(=O), N(CH3)-C(=O), NH-C(=S), N(CH3)-C(=S), OC(=O), C(=O)-NH, C(=O)- N(CH3), C(=S)-NH, C(=S)-N(CH3), C(=O)-O, CH2-CH2-CH2, S-CH2-CH2, CH2-CH2-S, CH2-S-CH2, O-CH2-CH2, CH2-CH2-O, CH2-O-CH2, S-C H2-S, SC(=CH2)-S, S-(CH2)2-S, S-(CH2)3-S, S-(CH2)4-S, S-CH2-CH=CH-CH2-S, S-CH2-C(=O)-NH, NH-C(=O)-CH2-S, S-CH2-CH4-CH2-S (the bonding position of the methylene group to the phenylene ring may be ortho, meta, or para), S-CH2-C(=O)-CH2-S, or S-CH2-C(=CH2)-CH2-S, triazole, or dithiotetrafluorobenzene.

[0039] Among these, L 7 Preferably, L represents NH—C(═O), SS, or S—(CH)—S. 7 represents NH-C(=O), for example, X 7 is an amino acid residue having an amino group in the side chain, and X 11 is an amino acid residue having a carboxyl group in the side chain, and the structure is cyclized by dehydration condensation of these amino and carboxyl groups. 7 represents SS, for example, X 7 and X 11 are both amino acid residues or derivatives thereof having a thiol group, and form a disulfide bond under oxidative conditions. 7 represents S-(CH2)3-S, for example, X 7 and X 11 are both amino acid residues or derivatives thereof having a thiol group, and a thioether bond can be formed between the thiol group of these residues and a halogenated alkyl linker such as 1,3-diiodopropane. Examples of amino acid residues having an amino group in the side chain include lysine and ornithine. Examples of amino acid residues having a carboxy group in the side chain include aspartic acid and glutamic acid. Examples of amino acid residues having a thiol group in the side chain include cysteine, D-cysteine, homocysteine, D-homocysteine, etc.

[0040] [9] A preferred embodiment of the formula (2) is the formula (10): c[X 3 -X 4 -X 5 -X 6 -c(X 7 -X 8 -X 9 -X 10 )]-X 11 -X 12 -X 13 (10) In formula (10), X is a cyclic peptide having an amino acid sequence represented by the formula (10) or a pharmacologically acceptable salt thereof. 3 and X7 and X 10 is a compound that forms two ring structures within a molecule by covalent bonding, and X 3 ~X 13 is as described in [1].

[0041] In one embodiment, X 3 represents cysteine, D-cysteine, homocysteine, D-homocysteine, or 3-mercaptopropanoic acid, and X 7 and X 10 each independently represents cysteine, D-cysteine, homocysteine, or D-homocysteine, and X 4 and X 8 each independently represents an amino acid residue having an aromatic carbocyclic group, preferably tyrosine, phenylalanine, or a derivative thereof; X 5 , X 9 , X 12 and X 13 each independently represents leucine, isoleucine, or a derivative thereof; X 11 represents serine, threonine, diaminopropanoic acid, diaminobutanoic acid, or a derivative thereof.

[0042]

[10] In the cyclic peptide represented by the above formula (2) or formula (10), X 1 and X 7 and X 10 , X 2 and X 7 and X 10 , or X 3 and X 7 and X 10 The bond between the Cα carbon atoms is represented by the following formula (11): [ka] In the formula, N represents 1, 2, or 3, A, B, and D each independently represent an integer of 1 or 2, and Y represents [(S-CH3)3-C6H3], [(S-CH2-CH2-C(=O))3-C3N3H6], [(S-CH2-C(=O)-NH)3-C6H3], [(NH-C(=O))3-C6H3], [(S-CH3)3-C3N3H6], or [(S-CH2-C(=O))3-C3N3H6].

[0043] Among these, it is preferable that Y represents [(S-CH3)3-C6H3]. For example, X 3 3-mercaptopropanoic acid (Mpa), X 7 and X 10 When both of the amino acid residues have a thiol group, such as cysteine (Cys), the peptide can be cyclized by a thioether bond between the thiol group of these residues and a halogenated alkyl linker, such as 1,3,5-tris(bromomethyl)benzene.

[0044]

[11] In addition, in the cyclic peptide represented by the above formula (2) or formula (10), X 1 and X 7 and X 10 , X 2 and X 7 and X 10 , or X 3 and X 7 and X 10 The bond between the Cα carbon atoms is the following formula (12): [ka] In the formula, N represents 1, 2, or 3, A, B, D, E, and F each independently represent an integer of 1 or 2, and Z and Z′ each independently represent S, SS, S-CH2-S, SC(=CH2)-S, S-(CH2)2-S, S-(CH2)3-S, S-(CH2)4-S, S-CH2-CH=CH-CH2-S, S-CH2-C(=O)-NH, NH-C(=O)-CH2-S, S-CH2-CH4-CH2-S (the bonding position of the methylene group to the phenylene ring may be ortho, meta, or para), S-CH2-C6H4-C6H4-CH2-S, S-CH2-C 10 H6-CH2-S, S-CH2-C(=O)-CH2-S, or S-CH2-C(=CH2)-CH2-S, or dithiotetrafluorobenzene.

[0045]

[12] In the cyclic peptides shown in the above formulas (1) to (3), (5), (7) and (10), X 5 , X 9 , X 12 and X 13 are each independently represented by the following formula (13): [ka] In the formula, the wavy line represents the point of attachment to the carbonyl group or nitrogen atom that forms an amide bond in the main chain, and R 1 , R 2 , R 6 and R 7 each independently represents a hydrogen atom or a methyl group, R 3 , R 4 , R 5 each independently represents a hydrogen atom, a methyl group, or a halogen atom (F, Cl, Br, I), and n represents an integer of 0 to 10. Among these, particularly preferred are leucine, N-methylated leucine, α-methylated leucine, isoleucine, β-homoleucine, norvaline, norleucine, 2-aminoheptanoic acid, 2-aminooctanoic acid, and 2-aminononanoic acid.

[0046] Or X5 , X 9 , X 12 and X 13 is expressed by the following formula (14): [ka] In the formula, the wavy line represents the point of attachment to the carbonyl group or nitrogen atom of the main chain that forms an amide bond, Z represents C or N, and R 8 and R 9 represents a hydrogen atom or a methyl group, and n represents an integer of 1 to 6. Among these, particularly preferred are cyclopropylalanine and cyclobutylalanine.

[0047]

[13] In the cyclic peptides shown in the above formulas (1) to (3), (5), (7) and (10), X 4 is expressed by the following formula (15): [ka] In the formula, the wavy line represents the point of attachment to the carbonyl group or nitrogen atom that forms an amide bond in the main chain, and R 10 , R 11 , R 12 , R 13 and R 14 each independently represents a hydrogen atom, a hydroxy group, a hydroxymethyl group, a methoxy group, a methoxymethyl group, an amino group, an aminomethyl group, a monomethylated amino group, a dimethylated amino group, a trimethylated amino group, a monomethylated aminomethyl group, a dimethylated aminomethyl group, a trimethylated aminomethyl group, an acetyl group, an amido group, a methyl group, a tert-butyl group, a halogenated methyl group, or a halogen atom (F, Cl, Br, I); R 15 and R 16 represents a hydrogen atom or a methyl group.

[0048] Among these, X 4preferably represents tyrosine, 3-hydroxyphenylalanine, 4-fluorophenylalanine, 4-aminophenylalanine, 4-aminomethylphenylalanine, 4-amidophenylalanine, N-methylated tyrosine, α-methylated tyrosine, O-methyl-tyrosine, 3-methoxy-phenylalanine, 4-acetyl-phenylalanine.

[0049]

[14] In addition, in the cyclic peptides shown in the above formulas (1) to (3), (5), (7) and (10), X 8 is preferably an amino acid residue having an aromatic carbocyclic group which may have a substituent, or a derivative thereof.

[0050]

[15] Aromatic carbocyclic group X 8 As a specific embodiment, the following formula (16): [ka] In the formula, the wavy line represents the point of attachment to the carbonyl group or nitrogen atom that forms an amide bond in the main chain, and R 17 , R 18 , R 19 , R 20 and R 21 each independently represents a hydrogen atom, a hydroxy group, a hydroxymethyl group, a methoxy group, a methoxymethyl group, an amino group, an aminomethyl group, a monomethylated amino group, a dimethylated amino group, a trimethylated amino group, a monomethylated aminomethyl group, a dimethylated aminomethyl group, a trimethylated aminomethyl group, an acetyl group, an amido group, a methyl group, a tert-butyl group, a halogenated methyl group, or a halogen atom (F, Cl, Br, I); R 22 and R 23 represents a hydrogen atom or a methyl group.

[0051] Among these, X 8More preferably, represents tyrosine, 3-hydroxyphenylalanine, 4-fluorophenylalanine, 4-aminophenylalanine, 4-aminomethylphenylalanine, 4-amidophenylalanine, N-methylated tyrosine, α-methylated tyrosine, O-methyl-tyrosine, 3-methoxy-phenylalanine, 4-acetyl-phenylalanine.

[0052]

[16] Furthermore, in the cyclic peptides shown in the above formulas (1) to (3), (5), (7) and (10), X 6 preferably represents glycine, N-methylated glycine, alanine, N-methylated alanine, D-alanine, N-methylated D-alanine, 2-aminoisobutyric acid, N-methylated 2-aminoisobutyric acid, 2-azetidine-2-carboxylic acid, D-2-azetidine-2-carboxylic acid, proline, D-proline, thioproline, D-thioproline, 3,4-dehydroproline, D-3,4-dehydroproline, pipecolic acid, D-pipecolic acid, or a derivative thereof.

[0053] Among these, X 6 More preferably, represents proline, N-methylated glycine, N-methylated alanine, 2-azetidine-2-carboxylic acid, pipecolic acid.

[0054]

[17] In one embodiment of the present invention, the cyclic peptide has the following formula (17): c[X 1 -Pro 2 -X 3 -Tyr 4 -Leu 5 -Pro 6 -c(X 7 -X 8 -Leu 9 -Cys 10 ]-X 11 )-X 12 -X 13 It consists of the amino acid sequence represented by (17). In equation (17), X 1 represents cysteine, Mpa (3-mercaptopropionic acid) or D-cysteine, and X 3represents N-methylated glycine, N-methylated alanine, 2-azetidine-2-carboxylic acid, proline, hydroxyproline, 3,4-dehydroproline, pipecolic acid, serine, or lysine; X 8 represents tyrosine, proline or arginine, X 7 and X 11 represents any combination of lysine and aspartic acid, ornithine and glutamic acid, aspartic acid and lysine, glutamic acid and ornithine, lysine and glutamic acid, or glutamic acid and lysine; X 12 and X 13 each independently represents leucine, isoleucine, or norleucine; X 1 and Cys 10 forms a disulfide bond between each side chain, and X 7 and X 11 form an amide bond between the respective side chains, so that the peptide of formula (17) has two cyclic structures in the molecule, and the N-terminal amino group and the C-terminal carboxy group may be modified or deleted.

[0055]

[18] A more preferred embodiment of the cyclic peptide is represented by the formula (17), wherein X 1 represents cysteine, and X 3 represents proline or serine, and X 7 represents lysine, and X 8 represents tyrosine, and X 11 represents aspartic acid, and X 12 and X 13 each independently represents leucine, isoleucine, or norleucine. The N-terminal amino group is acetylated, and the C-terminal carboxy group is amidated.

[0056] Examples of individual cyclic peptides included in the above formula (17) or formula (18) include the following. Ac-c[Cys 1 -Pro 2 -Pro 3 -Tyr 4 -Leu 5 -Pro 6-c(Lys 7 -Tyr 8 -Leo 9 -Cys 10 ]-Asp 11 )-Leu 12 -Ile 13 -NH2(10) Ac-c[Cys 1 -Pro 2 -Ser 3 -Tyr 4 -Leo 5 -Pro 6 -c(Lys 7 -Tyr 8 -Leo 9 -Cys 10 ]-Asp 11 )-Leu 12 -Ile 13 -NH2 Ac-c[Cys 1 -Pro2-Pro 3 -Tyr 4 -Leo 5 -Pro 6 -vase 7 -Tyr 8 -Leo 9 -Cys 10 ]-Glu 11 )-Leu 12 -Ile 13 -NH2 Ac-c[Cys 1 -Pro 2 -Ser 3 -Tyr 4 -Leo 5 -Pro 6 -vase 7 -Tyr 8 -Leo 9 -Cys 10 ]-Glu 11 )-Leu 12 -Ile 13 -NH2 Ac-c[Cys 1 -Pro 2 -Pro 3 -Tyr 4 -Leo 5 -Pro 6 -c(Asp 7- Tower 8 -Leu 9 -Cys 10 ]-Court 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Asp 7 - Tower 8 -Leu 9 -Cys 10 ]-Court 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Tower 8 -Leu 9 -Cys 10 ]-Orn 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Tower 8 -Leu 9 -Cys 10 ]-Orn 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Tower 8-Leu 9 -Cys 10 ]-Asp 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Tower 8 -Leu 9 -Cys 10 ]-Asp 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Orn 7 - Tower 8 -Leu 9 -Cys 10 ]-Glu 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Orn 7 - Tower 8 -Leu 9 -Cys 10 ]-Glu 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Asp 7 - Tower 8 -Leu 9-Cys 10 ]-Court 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Asp 7 - Tower 8 -Leu 9 -Cys 10 ]-Court 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Tower 8 -Leu 9 -Cys 10 ]-Orn 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Tower 8 -Leu 9 -Cys 10 ]-Orn 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Tower 8 -Leu 9 -Cys 10]-Asp 11 )-No 12 -Ile 13 -NH2 Ac-c[Cys 1 -Pro 2 -Ser 3 -Tyr 4 -Leo 5 -Pro 6 -c(Lys 7 -Tyr 8 -Leo 9 -Cys 10 ]-Asp 11 )-No 12 -Ile 13 -NH2 Ac-c[Cys 1 -Pro 2 -Pro 3 -Tyr 4 -Leo 5 -Pro 6 -vase 7 -Tyr 8 -Leo 9 -Cys 10 ]-Glu 11 )-No 12 -Ile 13 -NH2 Ac-c[Cys 1 -Pro 2 -Ser 3 -Tyr 4 -Leo 5 -Pro 6 -vase 7 -Tyr 8 -Leo 9 -Cys 10 ]-Glu 11 )-No 12 -Ile 13 -NH2 Ac-c[Cys 1 -Pro 2 -Pro 3 -Tyr 4 -Leo 5 -Pro 6 -c(Asp 7 -Tyr 8 -Leo 9 -Cys 10 ]-Lys 11)-No 12 -Ile 13 -NH2 Ac-c[Cys 1 -Pro 2 -Ser 3 -Tyr 4 -Leo 5 -Pro 6 -c(Asp 7 -Tyr 8 -Leo 9 -Cys 10 ]-Lys 11 )-No 12 -Ile 13 -NH2 Ac-c[Cys 1 -Pro 2 -Pro 3 -Tyr 4 -Leo 5 -Pro 6 -c(Glu 7 -Tyr 8 -Leo 9 -Cys 10 ]-Orn 11 )-No 12 -Ile 13 -NH2 Ac-c[Cys 1 -Pro 2 -Ser 3 -Tyr 4 -Leo 5 -Pro 6 -c(Glu 7 -Tyr 8 -Leo 9 -Cys 10 ]-Orn 11 )-No 12 -Ile 13 -NH2 Ac-c[Cys 1 -Pro 2 -Pro 3 -Tyr 4 -Leo 5 -Pro 6 -c(Lys 7 -Tyr 8 -Leo 9 -Cys 10 ]-Asp 11 )-No 12-Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Tower 8 -Leu 9 -Cys 10 ]-Asp 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Orn 7 - Tower 8 -Leu 9 -Cys 10 ]-Glu 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Orn 7 - Tower 8 -Leu 9 -Cys 10 ]-Glu 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Asp 7 - Tower 8 -Leu 9 -Cys 10 ]-Court 11 ) - Place 12 -Leu 13-NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Asp 7 - Tower 8 -Leu 9 -Cys 10 ]-Court 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Tower 8 -Leu 9 -Cys 10 ]-Orn 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Tower 8 -Leu 9 -Cys 10 ]-Orn 11 ) - Place 12 -Leu 13 -NH2

[0057] Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Court 7 - Rec 8 -Leu 9 -Cys 10 ]-Asp 11 )-Leu 12 -Where13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec 4 -Leu 5 - Pro 6 -c (Court 7 - Rec 8 -Leu 9 -Cys 10 ]-Asp 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Orn 7 - Rec 8 -Leu 9 -Cys 10 ]-Glu 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec 4 -Leu 5 - Pro 6 -c (Orn 7 - Rec 8 -Leu 9 -Cys 10 ]-Glu 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Asp 7 - Rec 8 -Leu 9 -Cys 10 ]-Court 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec 4 -Leu 5 - Pro 6 -c (Asp 7 - Rec 8 -Leu 9 -Cys 10 ]-Court 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Glu 7 - Rec 8 -Leu 9 -Cys 10 ]-Orn 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec 4 -Leu 5 - Pro 6 -c (Glu 7 - Rec 8 -Leu 9 -Cys 10 ]-Orn 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Court 7 - Rec 8 -Leu 9 -Cys 10 ]-Asp 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec 4 -Leu 5 - Pro 6 -c (Court 7 - Rec 8 -Leu 9 -Cys 10 ]-Asp 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Orn 7 - Rec 8 -Leu 9 -Cys 10 ]-Glu 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec 4 -Leu 5 - Pro 6 -c (Orn 7 - Rec 8 -Leu 9 -Cys 10 ]-Glu 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Asp 7 - Rec 8 -Leu 9 -Cys 10 ]-Court 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons1 - Pro 2 - Star 3 - Rec 4 -Leu 5 - Pro 6 -c (Asp 7 - Rec 8 -Leu 9 -Cys 10 ]-Court 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Glu 7 - Rec 8 -Leu 9 -Cys 10 ]-Orn 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec 4 -Leu 5 - Pro 6 -c (Glu 7 - Rec 8 -Leu 9 -Cys 10 ]-Orn 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Court 7 - Rec 8 -Leu 9 -Cys 10 ]-Asp 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro2 - Star 3 - Rec 4 -Leu 5 - Pro 6 -c (Court 7 - Rec 8 -Leu 9 -Cys 10 ]-Asp 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Orn 7 - Rec 8 -Leu 9 -Cys 10 ]-Glu 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec 4 -Leu 5 - Pro 6 -c (Orn 7 - Rec 8 -Leu 9 -Cys 10 ]-Glu 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Asp 7 - Rec 8 -Leu 9 -Cys 10 ]-Court 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star3 - Rec 4 -Leu 5 - Pro 6 -c (Asp 7 - Rec 8 -Leu 9 -Cys 10 ]-Court 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Glu 7 - Rec 8 -Leu 9 -Cys 10 ]-Orn 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec 4 -Leu 5 - Pro 6 -c (Glu 7 - Rec 8 -Leu 9 -Cys 10 ]-Orn 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Court 7 - Rec 8 -Leu 9 -Cys 10 ]-Asp 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec4 -Leu 5 - Pro 6 -c (Court 7 - Rec 8 -Leu 9 -Cys 10 ]-Asp 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Orn 7 - Rec 8 -Leu 9 -Cys 10 ]-Glu 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec 4 -Leu 5 - Pro 6 -c (Orn 7 - Rec 8 -Leu 9 -Cys 10 ]-Glu 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Asp 7 - Rec 8 -Leu 9 -Cys 10 ]-Court 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec 4 -Leu5 - Pro 6 -c (Asp 7 - Rec 8 -Leu 9 -Cys 10 ]-Court 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Glu 7 - Rec 8 -Leu 9 -Cys 10 ]-Orn 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec 4 -Leu 5 - Pro 6 -c (Glu 7 - Rec 8 -Leu 9 -Cys 10 ]-Orn 11 ) - Place 12 -Leu 13 -NH2

[0058] Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Court 7 - Pro 8 -Leu 9 -Cys 10 ]-Asp 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec4 -Lion 5 -Pro 6 -c(Lys 7 -Pro 8 -Lion 9 -Cys 10 ]-Asp 11 )-Lion 12 -Ile 13 NH2 Ac-c[Cys 1 -Pro 2 -Pro 3 -Arg 4 -Lion 5 -Pro 6 -horn 7 -Pro 8 -Lion 9 -Cys 10 ]-Glu 11 )-Lion 12 -Ile 13 NH2 Ac-c[Cys 1 -Pro 2 -Serum 3 -Arg 4 -Lion 5 -Pro 6 -horn 7 -Pro 8 -Lion 9 -Cys 10 ]-Glu 11 )-Lion 12 -Ile 13 NH2 Ac-c[Cys 1 -Pro 2 -Pro 3 -Arg 4 -Lion 5 -Pro 6 -c(Asp 7 -Pro 8 -Lion 9 -Cys 10 ]-Lys 11 )-Lion 12 -Ile 13 NH2 Ac-c[Cys 1 -Pro 2 -Serum 3 -Arg 4 -Lion5 -Pro 6 -c(Asp 7 -Pro 8 -Lion 9 -Cys 10 ]-Lys 11 )-Lion 12 -Ile 13 NH2 Ac-c[Cys 1 -Pro 2 -Pro 3 -Arg 4 -Lion 5 -Pro 6 -c(Glu 7 -Pro 8 -Lion 9 -Cys 10 ]-Orn 11 )-Lion 12 -Ile 13 NH2 Ac-c[Cys 1 -Pro 2 -Serum 3 -Arg 4 -Lion 5 -Pro 6 -c(Glu 7 -Pro 8 -Lion 9 -Cys 10 ]-Orn 11 )-Lion 12 -Ile 13 NH2 Ac-c[Cys 1 -Pro 2 -Pro 3 -Arg 4 -Lion 5 -Pro 6 -c(Lys 7 -Pro 8 -Lion 9 -Cys 10 ]-Asp 11 )-Lion 12 -Lion 13 NH2 Ac-c[Cys 1 -Pro 2 -Serum 3 -Arg 4 -Lion 5 -Pro6 -c (Court 7 - Pro 8 -Leu 9 -Cys 10 ]-Asp 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Orn 7 - Pro 8 -Leu 9 -Cys 10 ]-Glu 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec 4 -Leu 5 - Pro 6 -c (Orn 7 - Pro 8 -Leu 9 -Cys 10 ]-Glu 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Asp 7 - Pro 8 -Leu 9 -Cys 10 ]-Court 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec 4 -Leu 5 - Pro 6 -c (Asp7 - Pro 8 -Leu 9 -Cys 10 ]-Court 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Glu 7 - Pro 8 -Leu 9 -Cys 10 ]-Orn 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec 4 -Leu 5 - Pro 6 -c (Glu 7 - Pro 8 -Leu 9 -Cys 10 ]-Orn 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Court 7 - Pro 8 -Leu 9 -Cys 10 ]-Asp 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec 4 -Leu 5 - Pro 6 -c (Court 7 - Pro8 -Lion 9 -Cys 10 ]-Asp 11 )-No 12 -Ile 13 NH2 Ac-c[Cys 1 -Pro 2 -Pro 3 -Arg 4 -Lion 5 -Pro 6 -horn 7 -Pro 8 -Lion 9 -Cys 10 ]-Glu 11 )-No 12 -Ile 13 NH2 Ac-c[Cys 1 -Pro 2 -Serum 3 -Arg 4 -Lion 5 -Pro 6 -horn 7 -Pro 8 -Lion 9 -Cys 10 ]-Glu 11 )-No 12 -Ile 13 NH2 Ac-c[Cys 1 -Pro 2 -Pro 3 -Arg 4 -Lion 5 -Pro 6 -c(Asp 7 -Pro 8 -Lion 9 -Cys 10 ]-Lys 11 )-No 12 -Ile 13 NH2 Ac-c[Cys 1 -Pro 2 -Serum 3 -Arg 4 -Lion 5 -Pro 6 -c(Asp 7 -Pro 8 -Lion9 -Cys 10 ]-Court 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Glu 7 - Pro 8 -Leu 9 -Cys 10 ]-Orn 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec 4 -Leu 5 - Pro 6 -c (Glu 7 - Pro 8 -Leu 9 -Cys 10 ]-Orn 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Court 7 - Pro 8 -Leu 9 -Cys 10 ]-Asp 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec 4 -Leu 5 - Pro 6 -c (Court 7 - Pro 8 -Leu 9 -Cys10 ]-Asp 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Orn 7 - Pro 8 -Leu 9 -Cys 10 ]-Glu 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec 4 -Leu 5 - Pro 6 -c (Orn 7 - Pro 8 -Leu 9 -Cys 10 ]-Glu 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Asp 7 - Pro 8 -Leu 9 -Cys 10 ]-Court 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec 4 -Leu 5 - Pro 6 -c (Asp 7 - Pro 8 -Leu 9 -Cys 10 ]-Court11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Rec 4 -Leu 5 - Pro 6 -c (Glu 7 - Pro 8 -Leu 9 -Cys 10 ]-Orn 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Rec 4 -Leu 5 - Pro 6 -c (Glu 7 - Pro 8 -Leu 9 -Cys 10 ]-Orn 11 ) - Place 12 -Leu 13 -NH2

[0059] Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Tower 8 -Leu 9 -Cys 10 ]-Glu 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Tower 8 -Leu 9 -Cys 10]-Glu 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Tower 8 -Leu 9 -Cys 10 ]-Glu 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Tower 8 -Leu 9 -Cys 10 ]-Glu 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Tower 8 -Leu 9 -Cys 10 ]-Glu 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Tower 8 -Leu 9 -Cys 10 ]-Glu 11)-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Tower 8 -Leu 9 -Cys 10 ]-Glu 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Tower 8 -Leu 9 -Cys 10 ]-Glu 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Rec 8 -Leu 9 -Cys 10 ]-Glu 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Rec 8 -Leu 9 -Cys 10 ]-Glu 11 )-Leu 12-Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Rec 8 -Leu 9 -Cys 10 ]-Glu 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Rec 8 -Leu 9 -Cys 10 ]-Glu 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Rec 8 -Leu 9 -Cys 10 ]-Glu 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Rec 8 -Leu 9 -Cys 10 ]-Glu 11 )-Leu 12 -Leu 13-NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Rec 8 -Leu 9 -Cys 10 ]-Glu 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Rec 8 -Leu 9 -Cys 10 ]-Glu 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Pro 8 -Leu 9 -Cys 10 ]-Glu 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Pro 8 -Leu 9 -Cys 10 ]-Glu 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Pro 8 -Leu 9 -Cys 10 ]-Glu 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Pro 8 -Leu 9 -Cys 10 ]-Glu 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Pro 8 -Leu 9 -Cys 10 ]-Glu 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Pro 8 -Leu 9 -Cys 10 ]-Glu 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Pro 8 -Leu 9 -Cys 10 ]-Glu 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Court 7 - Pro 8 -Leu 9 -Cys 10 ]-Glu 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Tower 8 -Leu 9 -Cys 10 ]-Court 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Tower 8 -Leu 9 -Cys 10 ]-Court 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Tower 8 -Leu 9 -Cys 10 ]-Court 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Tower 8 -Leu 9 -Cys 10 ]-Court 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Tower 8 -Leu 9 -Cys 10 ]-Court 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Tower 8 -Leu 9 -Cys 10 ]-Court 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Tower 8 -Leu 9 -Cys 10 ]-Court 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Tower 8 -Leu 9 -Cys 10 ]-Court 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Rec 8 -Leu 9 -Cys 10 ]-Court 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Rec 8 -Leu 9 -Cys 10 ]-Court 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower4 -Leu 5 - Pro 6 -c (Glu 7 - Rec 8 -Leu 9 -Cys 10 ]-Court 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Rec 8 -Leu 9 -Cys 10 ]-Court 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Rec 8 -Leu 9 -Cys 10 ]-Court 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Rec 8 -Leu 9 -Cys 10 ]-Court 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu5 - Pro 6 -c (Glu 7 - Rec 8 -Leu 9 -Cys 10 ]-Court 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Rec 8 -Leu 9 -Cys 10 ]-Court 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Pro 8 -Leu 9 -Cys 10 ]-Court 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Pro 8 -Leu 9 -Cys 10 ]-Court 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro6 -c (Glu 7 - Pro 8 -Leu 9 -Cys 10 ]-Court 11 ) - Place 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Pro 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Pro 8 -Leu 9 -Cys 10 ]-Court 11 ) - Place 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Pro 8 -Leu 9 -Cys 10 ]-Court 11 )-Leu 12 -Where 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu 7 - Pro 8 -Leu 9 -Cys 10 ]-Court 11 )-Leu 12 -Leu 13 -NH2 Ac-c[Cons 1 - Pro 2 - Star 3 - Tower 4 -Leu 5 - Pro 6 -c (Glu7 -Pro 8 -Leu 9 -Cys 10 ]-Lys 11 )-Nle 12 -Ile 13 -NH2 Ac-c[Cys 1 -Pro 2 -Ser 3 -Tyr 4 -Leu 5 -Pro 6 -c(Glu 7 -Pro 8 -Leu 9 -Cys 10 ]-Lys 11 )-Nle 12 -Leu 13 -NH2

[0060] The peptides of the present invention include peptides that have homology to the amino acid sequences represented by [1] to

[17] above, in which one to several amino acids have been deleted, added, and / or substituted, as long as they have binding activity to VIPR2. As used herein, when referring to a "peptide in which one to several amino acids have been deleted, added, and / or substituted," the number of amino acids is not particularly limited as long as the peptide has VIPR2 binding activity, but is preferably one to five, and more preferably one or two. The deletions, additions, and / or substitutions may be at the terminals or in the middle of the peptide, and may occur at one or more positions.

[0061] Such amino acid sequences in which one to several amino acids have been deleted, added, and / or substituted in the above amino acid sequences include those that have a homology of at least 50% or more, preferably 70% or more, more preferably 80% or more, and particularly preferably 90% or more with the above amino acid sequence when calculated using BLAST (Basic Local Alignment Search Tool at the National Center for Biological Information) or the like (for example, using default, i.e., initial setting parameters).

[0062] It is also known that peptides and protein domains exist that have a high degree of structural similarity even when the homology in the primary structure is low. Therefore, peptides that have at least 50% or more, preferably 70% or more, more preferably 80% or more, and particularly preferably 90% or more structural homology with the above-mentioned amino acid sequences and have VIPR2 binding activity are also included in this embodiment. The homology of the three-dimensional structure between such peptides can be determined by predicting the three-dimensional structure of the peptide from the amino acid sequence of an unknown peptide using a homology modeling method or the like, as follows: For example, when an arbitrary amino acid sequence (target sequence) similar to the cyclic peptide (reference peptide) of this embodiment is given, an alignment (a juxtaposition of sequences) between the target sequence and the reference sequence is generated. Using an alignment calculated by FASTA, PSI-BLAST, LIBRA, or the like, the correspondence between each amino acid in the target sequence and the reference sequence is determined. Based on this relationship, the three-dimensional coordinates of each amino acid in the target sequence are generated from the three-dimensional coordinates of the reference peptide. When constructing 3D coordinates, structurally inappropriate gaps, collisions, and distortions between amino acid residues can occur. These structural distortions are eliminated through energy minimization calculations. To facilitate this process, some modeling software performs this stepwise process rather than simultaneously for all atoms in the peptide. That is, first, the α-carbon atoms that form the peptide backbone, then the main-chain atoms including the α-carbon atoms, and finally the entire peptide including the side-chain atoms. Once an alignment of the target sequence is obtained in this way, its 3D structure can be predicted and constructed. The 3D structural homology can be compared using indices such as RMSD (Root Mean Square Deviation), which is the difference between the XYZ coordinates when optimally superimposed.

[0063] The peptide of the present invention also includes various derivatives and / or modifications thereof, so long as they solve the problems of the present invention.Such derivatives include those in which the saturated fatty chain of a peptide is replaced with an unsaturated fatty chain, those in which some of the atoms of a peptide are replaced with other atoms including radioactive or non-radioactive isotope atoms, those in which the amide bond of a peptide is replaced with a thioamide bond (-NH-C(=S)-), those in which the amide bond of a peptide is replaced with an alkene (-C=C-), those in which the amide bond of a peptide is replaced with an alkyl (-CC-), those in which the amide bond of a peptide is replaced with a hydroxyethylene (-C(-OH)-C-), and those in which the amide bond of a peptide is replaced with an amide Examples of such modifications include those in which the α-carbon of the peptide is disubstituted, those in which the amide bond of the peptide is N-alkylated, those in which some of the functional groups of the peptide are halogenated, cyanated, nitrated, oxalated, hydroxylated, aminated, deaminated, etc. Peptides that have been modified by dehydrogenation, amidation, acetylation, methoxylation, prenylation, alkylation, etc. (for example, peptides in which some of the amino groups have been acetylated, formylated, myristoylated, palmitoylated, pyroglutamated, alkylated, or deaminated; peptides in which some of the carboxyl groups have been N-pyrrolidinylated or N-piperidinylated; peptides in which some of the carboxyl groups have been converted into amides (amide, methylamide, ethylamide, p-nitroanilide, β-naphthylamide, etc.) or esters (methyl ester, ethyl ester, thioester, etc.) Examples of peptides include, but are not limited to, peptides in which S is a sulfoxide S(=O) or sulfone S(=O)2, peptides multimerized via a chemical linker, peptides labeled with biotin, peptides labeled with fluorescence, peptides labeled with luminescence, and peptides fused with alkyl chains, polyethylene glycol, antibodies, lectins, sugar chains, enzymes, membrane-permeable peptides, low-molecular-weight compounds, or molecules that induce protein ubiquitination.

[0064] The peptide of the present invention also encompasses peptide salts. Examples of peptide salts include salts with physiologically acceptable bases or acids, such as addition salts with inorganic acids (hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, etc.), addition salts with organic acids (p-toluenesulfonic acid, methanesulfonic acid, oxalic acid, p-bromophenylsulfonic acid, carboxylic acids, succinic acid, citric acid, benzoic acid, acetic acid, etc.), addition salts with inorganic bases (ammonium hydroxide, alkali or alkaline earth metal hydroxides, carbonates, bicarbonates, etc.), and addition salts with amino acids.

[0065] The peptide of the present invention may be a prodrug. A prodrug refers to a compound that is converted into the peptide of the present invention by a reaction with an enzyme, gastric acid, or the like under physiological conditions in a living body, i.e., a compound that is converted into the peptide of the present invention by enzymatic oxidation, reduction, hydrolysis, or the like, or a compound that is converted into the peptide of the present invention by hydrolysis, etc., with gastric acid, or the like.

[0066] Prodrugs of the peptide of the present invention include compounds in which the amino group of the peptide of the present invention is acylated, alkylated, or phosphorylated (for example, compounds in which the amino group of the peptide of the present invention is eicosanoylated, alanylated, pentylaminocarbonylated, (5-methyl-2-oxo-1,3-dioxolen-4-yl)methoxycarbonylated, tetrahydrofuranylated, pyrrolidylmethylated, pivaloyloxymethylated, or tert-butylated), compounds in which the hydroxy group of the peptide of the present invention is acylated, alkylated, phosphorylated, or borated (for example, compounds in which the hydroxy group of the peptide of the present invention is acetylated, palmitoylated, propanoylated, pivaloylated, succinylated, fumarylated, alanylated, or dimethylaminomethylcarbonylated), and compounds in which the hydroxy group or carboxy group of the peptide of the present invention is esterified or amidated (for example, compounds in which the hydroxy group or carboxy group of the peptide of the present invention is C 1-6Examples of such compounds include, but are not limited to, alkyl esterification, phenyl esterification, carboxymethyl esterification, dimethylaminomethyl esterification, pivaloyloxymethyl esterification, ethoxycarbonyloxyethyl esterification, phthalidyl esterification, (5-methyl-2-oxo-1,3-dioxolen-4-yl)methyl esterification, cyclohexyloxycarbonylethyl esterification, or methylamidated compounds. These compounds can be produced from the peptides of the present invention by methods known per se.

[0067] The prodrug of the peptide of the present invention may be one that is converted into the peptide of the present invention under physiological conditions as described in "Drug Development," Hirokawa Shoten, 1990, Vol. 7, Molecular Design, pp. 163-198.

[0068] In the present specification, the prodrug may be in the form of a salt, and such salts include those exemplified as salts of the peptide of the present invention.

[0069] The peptide of the present invention may be in the form of a crystal, and both a single crystalline form and a mixture of crystalline forms are encompassed within the scope of the peptide of the present invention. Crystals can be produced by crystallization using a crystallization method known per se.

[0070] The peptide of the present invention may be in the form of a pharmaceutically acceptable cocrystal or cocrystal salt. Here, a cocrystal or a cocrystal salt refers to a crystalline substance composed of two or more distinct solids at room temperature, each of which has different physical properties (e.g., structure, melting point, heat of fusion, hygroscopicity, solubility, stability, etc.). A cocrystal or a cocrystal salt can be prepared by a known cocrystallization method.

[0071] (Action and effect of cyclic peptides) As shown in the Examples below, Seq-1 to 10, which are representative examples of the amino acid sequence group represented by this embodiment, exhibit binding activity to VIPR2-expressing cells, and Seq-1, 6, 9, and 10 have antagonist activity to VIPR2-expressing cells and resistance to protease degradation. Because the amino acid sequence group represented by this embodiment has similar amino acid sequences and three-dimensional structural characteristics to Seq-1 to 10, it is highly likely that they also have resistance to protease degradation, VIPR2-binding activity, and VIPR2 antagonist activity.

[0072] The VIPR2-binding activity of the peptides shown in the Examples below demonstrates the following: (1) Deletion of the three C-terminal residues of VIpep-3 (amino acid residues 14-16: Leu-Arg-Ser) does not abolish VIPR2-binding activity. (2) Cross-linking between positions 7 and 11 improves VIPR2-binding activity. (3) VIpep-3 is cyclized by an S-type disulfide bond formed between the cysteine residues at positions 1 and 10. However, cyclization for VIPR2-binding activity is not limited to positions 1 and 10. Alternatively, cross-linking between positions 2 and 10 and between positions 3 and 10 can be achieved by introducing an amino acid residue with a long side chain at positions 2 or 3 to directly cross-link the cysteine residue at position 10, or by indirectly cross-linking the cysteine residues using linkers of different lengths and structures. This can be achieved by bringing the interatomic distances between positions 2 and 10 and between positions 3 and 10 closer to the interatomic distance between positions 1 and 10. This can be achieved by replacing the interatomic distances between positions 2 and 10 and between positions 3 and 10. (4) Peptides bicyclized between three amino acid residues also exhibit VIPR2 binding activity, and the cross-linking pattern is considered to be acceptable for any combination of positions 1-7-10, positions 2-7-10, and positions 3-7-10. The above (3) and (4) will be explained in more detail. As shown in the examples below, the cyclic structure formed between the first to third amino acid residues and the tenth amino acid residue of VIpep-3: Cα 3 -NH-C(=O)-Cα 2 -NH-C(=O)-Cα 1 -CH2-SS-CH2-Cα 10 For example, the ring structure formed between the third and tenth amino acid residues having a side chain of an unnatural structure: Cα3 -CH2-CH2-CH2-CH2-CH2-CH2-CH2-SS-CH2-CH2-Cα 10 or the like. That is, the amino acid residues 1 to 3 of VIpep-3 are not significantly involved in VIPR2 binding activity, suggesting that restriction of the cyclic size of the peptide is important for VIPR2 binding activity. The peptides of the present invention can be cyclized not only between positions 1 and 10, but also between positions 2 and 10 and between positions 3 and 10. As long as the cyclic size of the peptide falls within a certain range, it is believed that they can accept a wide range of chemical structures, such as amide bonds, disulfide bonds, thioether bonds, C=C bonds, CC bonds, bonds via triazole, and bonds via dithiotetrafluorobenzene. This property is believed to be maintained even when cyclization is via position 7 (between positions 1, 7, and 10, between positions 2, 7, and 10, and between positions 3, 7, and 10).

[0073] The amino acid sequence group represented by [1] to

[16] above has similar amino acid sequence and three-dimensional structural characteristics to VIpep-3, which has antagonist activity against VIPR2 in humans, mice, and rats. Therefore, it is highly likely that they will also bind to VIPR2 in mammals other than humans, such as mice and rats, and inhibit its function.

[0074] (Method for producing cyclic peptides) The peptide of this embodiment can be produced by known peptide production methods, such as chemical synthesis methods including a liquid phase method, a solid phase method, or a hybrid method that combines a liquid phase method and a solid phase method.

[0075] The solid-phase method can be performed using a commercially available automated synthesizer. For example, the hydroxyl group of a hydroxyl-containing resin is esterified with the carboxyl group of a first amino acid (usually the C-terminal amino acid of the target peptide) whose α-amino group is protected with a protecting group such as an Fmoc group. Known dehydration condensation agents such as 1-mesitylenesulfonyl-3-nitro-1,2,4-triazole (MSNT), dicyclohexylcarbodiimide (DCC), and diisopropylcarbodiimide (DIPCDI) can be used as esterification catalysts. Next, the protecting group of the α-amino group of the first amino acid is removed, and a second amino acid in which all functional groups except the carboxyl group of the main chain are protected is added, activating the carboxyl group, and the first and second amino acids are bonded. Furthermore, the α-amino group of the second amino acid is deprotected, and a third amino acid in which all functional groups except the carboxyl group of the main chain are protected is added, activating the carboxyl group, and the second and third amino acids are bonded. This process is repeated to synthesize a peptide of the desired length. The linear peptide is cleaved from the resin and purified, after which the functional groups for cyclizing the peptide are deprotected and the peptide is cyclized according to a standard method.

[0076] Examples of resins for solid-phase synthesis include Merrifield resin, MBHA resin, Cl-Trt resin, SASRIN resin, Wang resin, Rink amide resin, HMFS resin, Amino-PEGA resin (Merck), and HMPA-PEGA resin (Merck). These resins may be washed with a solvent (dimethylformamide (DMF), 2-propanol, methylene chloride, etc.) before use. Examples of protecting groups for the α-amino group include benzyloxycarbonyl (Cbz) group, tert-butoxycarbonyl (Boc) group, fluorenylmethoxycarbonyl (Fmoc) group, benzyl group, allyl group, and allyloxycarbonyl (Alloc) group. The Cbz group can be deprotected using hydrofluoric acid or hydrogenation, the Boc group can be deprotected using trifluoroacetic acid (TFA), and the Fmoc group can be deprotected by treatment with piperidine. The α-carboxyl group can be protected using methyl esters, ethyl esters, benzyl esters, tert-butyl esters, cyclohexyl esters, etc. As for other functional groups of amino acids, the hydroxyl group of serine, threonine, etc. can be protected with a benzyl group or a tert-butyl group, and the hydroxyl group of tyrosine, etc. can be protected with a 2-bromobenzyloxycarbonyl group or a tert-butyl group. The amino group of the side chain of lysine, etc., and the carboxyl group of glutamic acid, aspartic acid, etc. can be protected in the same way as the α-amino group and the α-carboxyl group.

[0077] Carboxy group activation can be achieved using a condensation agent, such as dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIPCDI), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC or WSC), (1H-benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate (BOP), or 1-[bis(dimethylamino)methyl]-1H-benzotriazolium-3-oxide hexafluorophosphate (HBTU). Cleavage of the peptide chain from the resin can be achieved by treatment with an acid such as TFA or hydrogen fluoride (HF).

[0078] The peptide of this embodiment is cyclized in one aspect. As used herein, cyclization refers to the covalent bonding of two or more amino acids separated by one or more amino acids within a peptide molecule, either directly or indirectly via a linker, to form one or more ring structures within the molecule. Cyclization can be performed according to the techniques described in Non-Patent Documents 9, 10, and 11. Examples of cyclization include an amide bond between an amino group and a carboxyl group, a disulfide bond between thiol groups, a thioether bond between a thiol group and a halogen group, a thioether bond formed by a thiol-ene reaction between a thiol group and an allyl group, a C=C bond formed by an olefin metathesis reaction between allyl groups (the C=C bond may be converted to a C=C bond by a reduction reaction), a triazole-mediated bond formed by a click reaction between an alkynyl group and an azide group, and a thioether bond between a linker having a halogen group and two thiol groups, but are not limited to these. The covalent bond for cyclization, either direct or indirect via a linker, may be between main chains, between main chains and side chains, between side chains and main chains, or between side chains and side chains.

[0079] For example, the peptide of the present invention can be cyclized by (1) using cysteine, D-cysteine, homocysteine, or D-homocysteine having a thiol group as amino acid 1 and amino acid 2, respectively, independently, and using a disulfide bond formed between the thiol groups of these amino acids; (2) using an amino acid having a halogen atom (chloro, bromo, or iodo) as a nucleophilic group (e.g., 3-chloroalanine) or a carboxylic acid having a halogen atom (chloro, bromo, or iodo) (e.g., 3-chloropropanoic acid) as amino acid 1 and using a disulfide bond formed between the thiol groups of these amino acids; (3) a thioether bond formed between amino acid 1 and amino acid 2 having a thiol group and a linker having a halogen atom (chloro, bromo, or iodo) as a nucleophilic group (e.g., 1,3-diiodopropane, 1,4-diiodobutane, 1,5-diiodopentane, 1,6-diiodohexane, 1,2-bis(bromomethyl)benzene, 1,3-bis(bromomethyl)benzene, 1,4-bis(bromomethyl)benzene, etc.); (4) A covalent bond via a triazole formed by a click reaction between β-azidoalanine having an azido group as amino acid 1 and 2-amino-5-hexynoic acid having an alkynyl group as amino acid 2 can be used. (5) A thioether bond formed by a thiol-ene reaction between an amino acid having an allyl group (e.g., allylglycine, D-allylglycine, homoallylglycine, D-homoallylglycine, etc.) or a carboxylic acid having an allyl group (e.g., 3-butenoic acid) as amino acid 1 and an amino acid having a thiol group can be used. (6) A C=C bond formed by an olefin metathesis reaction between the allyl groups of an amino acid having an allyl group or a carboxylic acid having an allyl group as amino acid 1 and amino acid 2 can be used. (7) A C-C bond formed by reducing a C=C bond formed by an olefin metathesis reaction can be used. (8) An amino acid having an amino group (e.g., 2,3-diaminopropanoic acid, 2,An amide bond can be used between an amino acid 1 having a carboxyl group (e.g., 4-diaminobutanoic acid, ornithine, lysine, or the D-form amino acids thereof) and an amino acid 2 having a carboxyl group (e.g., aspartic acid, glutamic acid, or the D-form amino acids thereof). (9) An amide bond can be used between an N-terminal amino group (e.g., β-alanine or γ-aminobutyric acid) and an amino acid having a carboxyl group. Either of these amino acids 1 and 2 can be located at the N-terminus. Furthermore, (10) a thioether bond can be used between an amino acid 1, amino acid 2, or amino acid 3 having a thiol group and a linker (e.g., 1,3,5-tris(bromomethyl)benzene) having a nucleophilic halogen atom (chloro, bromo, or iodo). Either of amino acid 1, amino acid 2, or amino acid 3 can be located at the N-terminus.

[0080] (Medicines, diagnostic agents, and research reagents containing cyclic peptides) The pharmaceutical composition of the present invention contains the above-described amino acid sequence as an active ingredient, and by binding to VIPR2, the peptide inhibits the binding of a natural ligand to VIPR2, thereby suppressing VIPR2-mediated signal transduction. The administration form of the pharmaceutical composition is not particularly limited, and may be oral or parenteral. Examples of parenteral administration include transmucosal administration (intranasal, oral, ocular, pulmonary, vaginal, or rectal administration), injection (intravenous injection, subcutaneous injection, intramuscular injection, etc.), and transdermal administration. The peptide in the pharmaceutical composition can be modified in various ways in consideration of its susceptibility to metabolism and excretion. For example, adding an alkyl chain, polyethylene glycol, or sugar chain to the peptide can increase its blood residence time and reduce its antigenicity. Alternatively, biodegradable polymers such as polylactic acid glycol (PLGA), porous hydroxyapatite, liposomes, surface-modified liposomes, emulsions prepared with unsaturated fatty acids, nanoparticles, nanospheres, etc. may be used as sustained-release bases, and peptides may be encapsulated in these. For transdermal administration, a weak electric current can be passed through the skin surface to allow penetration through the stratum corneum (iontophoresis method).

[0081] The above-mentioned pharmaceutical compositions may contain the active ingredient as is, or may be formulated by adding pharmaceutically acceptable carriers, excipients, additives, etc. Dosage forms include, for example, liquids (e.g., injections), dispersions, suspensions, tablets, pills, powders, suppositories, powders, fine granules, granules, capsules, syrups, lozenges, inhalants, ointments, eye drops, nasal drops, ear drops, and poultices. These formulations may also be controlled-release formulations such as immediate-release formulations or sustained-release formulations (e.g., sustained-release microcapsules). Formulation can be carried out by conventional methods using, for example, excipients, binders, disintegrants, lubricants, solubilizers, solubilizers, colorants, flavorings, stabilizers, emulsifiers, absorption enhancers, surfactants, pH adjusters, preservatives, antioxidants, etc. as appropriate. Examples of ingredients used in formulations include, but are not limited to, purified water, saline, phosphate buffer, dextrose, glycerol, ethanol and other pharmaceutically acceptable organic solvents, animal and vegetable oils, lactose, mannitol, glucose, sorbitol, crystalline cellulose, hydroxypropyl cellulose, starch, corn starch, anhydrous silicic acid, magnesium aluminum silicate, collagen, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymers, sodium carboxymethylcellulose, sodium polyacrylate, sodium alginate, water-soluble dextran, sodium carboxymethyl starch, pectin, methylcellulose, ethylcellulose, xanthan gum, gum arabic, tragacanth, casein, agar, polyethylene glycol, diglycerin, glycerin, propylene glycol, petrolatum, paraffin, octyldodecyl myristate, isopropyl myristate, higher alcohols, stearyl alcohol, stearic acid, and human serum albumin.When a peptide is poorly absorbed transmucosally, absorption enhancers that improve the absorption of poorly absorbed drugs may be used, including surfactants such as polyoxyethylene lauryl ethers, sodium lauryl sulfate, and saponin; bile salts such as glycocholic acid, deoxycholic acid, and taurocholic acid; chelating agents such as EDTA and salicylic acids; fatty acids such as caproic acid, capric acid, lauric acid, oleic acid, linoleic acid, and mixed micelles; enamine derivatives, N-acyl collagen peptides, N-acyl amino acids, cyclodextrins, chitosans, and nitric oxide donors.

[0082] Pills or tablets can also be coated with sugar, gastric, or enteric coating materials. Injections can contain distilled water for injection, physiological saline, propylene glycol, polyethylene glycol, vegetable oils, alcohols, etc. Furthermore, wetting agents, emulsifiers, dispersants, stabilizers, solubilizers, solubilizers, preservatives, etc. can also be added. If necessary, conventional additives such as preservatives, antioxidants, colorants, sweeteners, adsorbents, wetting agents, etc. can also be used appropriately and in appropriate amounts.

[0083] The pharmaceutical composition of the present invention is useful for treating central nervous system diseases in which activation of VIPR2 is involved, such as psychiatric disorders (schizophrenia, schizoaffective disorder, schizophreniform disorder, delusional disorder, etc.), childhood psychiatric disorders (attention deficit disorder, attention deficit / hyperactivity disorder, conduct disorder, autism, etc.), neurodegenerative disorders, neural stem cell disorders, neural precursor disorders, ischemic disorders, neurotraumatic disorders, affective disorders, psychomotor disorders, sleep disorders (hypersomnia, circadian rhythm sleep disorders, insomnia, parasomnia, sleep deprivation, etc.), and psychiatric disorders such as anxiety (acute sleep disorders). stress disorder, generalized anxiety disorder, social anxiety disorder, panic disorder, post-traumatic stress disorder, agoraphobia, obsessive-compulsive disorder, etc.), factitious psychosis (acute hallucinatory mania, etc.), impulse control disorder (compulsive gambling, intermittent explosive disorder, etc.), mood disorders (bipolar I disorder, bipolar II disorder, mania, mixed affective state, etc.), major depression, chronic depression, seasonal depression, psychotic depression, seasonal depression, cognitive disorders (amnesia, senile dementia, HIV-associated dementia, Alzheimer's disease, Huntington's disease, dementia with Lewy bodies) , vascular dementia, drug-related dementia, tardive dyskinesia, myoclonic spasms, dystonia, delirium, Pick's disease, Creutzfeldt-Jakob disease, HIV disease, Gilles de la Tourette syndrome, epilepsy, muscle spasms, mild cognitive impairment, etc.), mental retardation (spasticity, Down syndrome, fragile X syndrome, etc.); premenstrual syndrome (PMS), premenstrual dysphoric disorder (PDD), postpartum depression, neuronal damage disorders (eye damage, ocular retinopathy or macular degeneration, tinnitus, hearing impairment, cerebral edema, etc.), Parkinson's disease, The compounds are useful for the prevention and / or treatment of, but are not limited to, Parkinson's disease-like disorders, migraines, epilepsy, Alzheimer's disease, brain injury, stroke, cerebrovascular diseases (cerebral arteriosclerosis, cerebral amyloid angiopathy, hereditary cerebral hemorrhage, cerebral hypoxia-ischemia, etc.), drug addiction (narcotic addiction, alcoholism, amphetamine addiction, cocaine addiction, nicotine addiction, drug withdrawal syndrome, etc.), eating disorders (anorexia, bulimia, binge eating disorder, bulimia, obesity, compulsive eating disorder, pagophagia, etc.), etc. Additional diseases include, but are not limited to, use in suppressing the growth of VIPR2-expressing cancers and stimulating immunity by inhibiting VIPR2 function.

[0084] The pharmaceutical composition of the present invention may be used in combination with other medicines or treatments such as various chemotherapy, surgical treatment, and radiotherapy that are useful for the above-mentioned diseases.

[0085] When the pharmaceutical composition of the present invention is administered to mammals (e.g., humans, mice, rats, guinea pigs, rabbits, dogs, horses, monkeys, pigs, etc.), particularly humans, the dosage varies depending on the symptoms, the patient's age, sex, weight, and sensitivity, the administration method, the administration interval, the type of active ingredient, and the type of formulation, and is not particularly limited. For example, 30 μg to 1000 mg, 100 μg to 500 mg, or 100 μg to 100 mg can be administered once or in divided doses.

[0086] The following examples are merely illustrative and are intended to explain the present invention in detail together with the above-described embodiments, but are not intended to limit the present invention. Those skilled in the art can modify the present invention in various aspects without departing from the spirit of the present invention, and such modifications are also included in the scope of the present invention. [Example]

[0087] The abbreviations used in this specification have the following meanings. VIPR2:Vasoactive intestinal peptide receptor 2 VIP:Vasoactive intestinal peptide BSA: Bovine serum albumin RP-HPLC: Reverse-phase high performance liquid chromatography HRP: Horseradish peroxidase SA:Streptavidin HBSS:Hank's balanced salt solution ELISA:Enzyme-linked immunosorbent assay Ac: Acetyl Cys:L-Cysteine hmC:L-homocystein Mpa:3-mercaptopropanoic acid Pro:L-Proline Tyr:L-Tyrosine Leu:L-Leucine Ile:L-Isoleucine Arg:L-Arginine Asp:L-Aspartic acid Glu:L-Glutamic acid Dap:(S)-2,3-Diaminopropanonic acid Dab:(S)-2,4-Diaminobutanoic acid Tyr(Ome):O-methyl-L-Tyrosine 4aF:4-amino-L-Phenylalanine 4AcF:4-acetyl-L-Phenylalanine 4amdF:4-amide-L-Phenylalanine 3hF:3-hydroxy-L-Phenylalanine 3(Ome)F:3-methoxy-L-Phenylalanine 4amF:4-aminomethyl-L-Phenylalanine 4fF:4-fluoro-L-Phenylalanine Gly:Glycine Ala:L-Alanine Aze:L-Azetidine carboxylic acid thioP:L-Thioproline Dhp:3,4-dehydro-L-Proline Pip:L-Pipecolic acid Val:L-Valine Nle:L-Norleucine Ahep:(S)-2-Aminoheptanonic acid Aoc: (S)-2-Aminooctanonic acid Anon: (S)-2-Aminononanonic acid CprA: L-Cyclopropylalanine CbuA: L-Cyclobutylalanine βhmLeu: β-homo―L-Leucine M6a: 6-Mercaptohexanoic acid M8a: 8-Mercaptooctanoic acid DIBut: 1,4-Diiodobutane DIPen: 1,5-Diiodopentane DIHex: 1,6-Diiodohexane DBoXy: 1,2-Bis(bromomethyl)benzene DBmXy: 1,3-Bis(bromomethyl)benzene DBpXy: 1,4-Bis(bromomethyl)benzene TBMB: 1,3,5-Tris(bromomethyl)benzene Nme-X: N-methylated amino acid X aMe-X: α-methylated amino acid X D X: D-amino acid X c[XY], c(XY): Cyclization between amino acid X to amino acid Y

[0088] (Peptide synthesis) Chemical synthesis of all peptides used in this example was outsourced to Scrum Corporation (Tokyo, Japan) and performed using standard solid-phase synthesis techniques with a 9-fluorenylmethoxycarbonyl (Fmoc) group as the α-amino group protecting group on a Syro II (Biotage) automated synthesizer. The C-terminal side-chain-protected amino acid-resin was placed in a synthesis column and the system was set up. The next amino acid protected with an Fmoc group was then activated with 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU) / diisopropylethylamine (DIEA), and then loaded onto the column for reaction. After the reaction was complete, the column was washed and the Fmoc group was deprotected with 20% piperidine. This process was repeated to elongate the peptide chain. After the Fmoc group of the final amino acid was deprotected, the peptide resin was removed from the system.

[0089] Peptide cyclization was performed based on the techniques described in Non-Patent Documents 8, 9, 10, and 11. The cyclization of Seq-1, 6, and 9 is shown below as an example. For Seq-1 and Seq-6, linear side-chain-protected peptide resin was swollen in dimethylformamide (DMF) and reacted in a 2% hydrazine solution for 5–10 minutes to deprotect the lysine side-chain protecting group (Dde) and the aspartic acid side-chain protecting group (ODmab). The coupling reagents Oxima Pure and diisopropylcarbodiimide (DIC) were then added and reacted at 50°C for 3 hours. After washing the resin, TFA was added to deprotect the thiol protecting group and simultaneously cleave the monocyclic peptide from the resin. The monocyclic peptide was purified by RT-HPLC using a SunFire C18 column (10 × 150 mm) (Waters) and then lyophilized.

[0090] For Seq-1, the monocyclic peptide was dissolved in a mixture of Tris-HCl buffer (pH 8.5) and acetonitrile, followed by the addition of DMSO and stirring at room temperature for 36 hours to cyclize the peptide via a disulfide bond. For Seq-6, the monocyclic peptide was dissolved in dimethyl sulfoxide (DMSO), and then cyclized overnight at room temperature with 3 equivalents of 1,3-bisbromomethylbenzene dissolved in DMF, 0.1 M NaHCO3 containing 10 mM tris(2-carboxyethyl)phosphine (TCEP), and acetonitrile. This mixture was then cyclized via a thioether bond between the thiol group and the alkyl halide linker. Bicyclic peptides Seq-1 and 6 were purified by RT-HPLC using a SunFire C18 column (10 × 150 mm) (Waters) and then lyophilized. The molecular weights of the final peptides were determined using a microflex (Bruker).

[0091] For Seq-9, after washing the resin, TFA was added to deprotect the thiol protecting groups and simultaneously cleave the linear peptide from the resin. It was purified by RT-HPLC using a SunFire C18 column (10 × 150 mm) (Waters) and then lyophilized. The linear peptide was dissolved in dimethyl sulfoxide (DMSO) and incubated with 3 equivalents of 1,3,5-tris(bromomethyl)benzene (TBMB) in DMF, 0.1 M NaHCO3 containing 10 mM tris(2-carboxyethyl)phosphine (TCEP), and acetonitrile at 80°C for 3 hours to cyclize the peptide via a thioether bond between the thiol group and the alkyl halide linker. The bicyclic peptide was purified by RT-HPLC using a SunFire C18 column (10 × 150 mm) (Waters) and then lyophilized. The molecular weight of the final peptide was determined using a microflex (Bruker) to identify the target product. The theoretical molecular weight, measured molecular weight, purity, cyclization type, and sequence of the peptides synthesized in this example are shown in Table 1. The amino acid sequences of Seq-1 to Seq-9 are shown below, and their structural formulas are shown in Figure 7. In Table 1 and the amino acid sequences below, amino acids without a D-form designation are L-forms.

[0092] [Table 1-1]

[0093] [Table 1-2]

[0094] Seq-1:c(Mpa-Pro-Pro-Tyr-Leu-Pro-c[Lys-Tyr-Leu-Cys)-Asp]-Leu-Ile-NH2 (SEQ ID NO: 1) Seq-2: c(Mpa-NmeTyr-Leu-Pro-c[Lys-Tyr-Leu-Cys)-Asp]-Leu-Ile-NH2 (SEQ ID NO: 2) Seq-3: c(Mpa-NmeTyr-Leu-Pro-c[Lys-Tyr-Leu-Cys)-Asp]-Leu-Ile-NH2 (SEQ ID NO: 3) Seq-4: c(Mpa-NmeTyr-Leu-Pro-c[Lys-Tyr-Leu-Cys)-Asp]-Leu-Ile-NH2 (SEQ ID NO: 4) Seq-5: c(Mpa-NmeTyr-Leu-Pro-c[Lys-Tyr-Leu-Cys)-Asp]-Leu-Ile-NH2 (SEQ ID NO: 5) Seq-6: c(Mpa-NmeTyr-Leu-Pro-c[Lys-Tyr-Leu-Cys)-Asp]-Leu-Ile-NH2 (SEQ ID NO: 6) Seq-7: c(Mpa-NmeTyr-Leu-Pro-c[Lys-Tyr-Leu-Cys)-Asp]-Leu-Ile-NH2 (SEQ ID NO: 7) Seq-8: c(Mpa-NmeTyr-Leu-NmeGly-c(Cys)-Tyr-Leu-Cys)-Ser-Leu-Ile-NH2 (SEQ ID NO: 8) Seq-9: c(Mpa-NmeTyr-Leu-NmeGly-c(Cys)-Tyr-Leu-Cys)-Dap-Leu-Ile-NH2 (SEQ ID NO: 9) Seq-10:Ac-c[Cys 1 -Pro 2 -Pro 3 -Tyr 4 -Leu 5 -Pro 6 -c(Lys 7 -Tyr 8 -Leu 9 -Cys 10 ]-Asp 11 )-Leu 12 -Ile 13 -NH2 (SEQ ID NO: 10)

[0095] Seq-1 and Seq-10 are peptides obtained by removing the three C-terminal residues (Leu-Arg-Ser) of VIpep-3 and bicyclizing them via an SS bond between positions 1 and 10 and an amide bond between positions 7 and 11. Seq-2 to 7 are peptides obtained by removing the two N-terminal residues (Cys-Pro) and the three C-terminal residues (Leu-Arg-Ser) of VIpep-3 and bicyclizing them via a thioether bond between positions 3 and 10 via a chemical linker and an amide bond between positions 7 and 11. Seq-8 to 9 are peptides obtained by removing the two N-terminal residues (Cys-Pro) and the three C-terminal residues (Leu-Arg-Ser) of VIpep-3 and bicyclizing them via a thioether bond between positions 3, 7, and 10 via a TBMB linker.

[0096] (Establishment of a competitive binding assay using cell ELISA) To confirm the binding activity of the amino acid-substituted peptides to VIPR2, we performed a competitive binding assay using a cell-based ELISA, as shown in Figure 3. The cell-based ELISA is briefly described below. Human VIPR2-expressing cell lines (catalog no. HTS079RTA, Eurofins) were seeded into 96-well plates using the medium provided by the manufacturer and cultured overnight at 37°C in a CO2 incubator. After washing the plate with HBSS containing 0.1% BSA, a mixture of Biotin-VIpep-3 (1000 nM) diluted in HBSS containing 0.1% BSA and the amino acid-substituted peptide or VIpep-3 at the desired concentration was added at 50 μL per well. After incubation on ice for 60 minutes, the plate was washed with HBSS containing 0.1% BSA. Biotin-VIpep-3 bound to VIPR2-expressing cells adhered to the plate was detected using SA-HRP (catalog no. ab7403, Abcam). HRP was quantified by measuring chemiluminescence using SuperSignal ELISA Pico Chemiluminescent Substrate (Catalog No. 37070, Thermo Fisher Scientific). Biotin-VIpep-3 binding to cell surface VIPR2 competes with the binding of the amino acid-substituted peptides and VIpep-3 present in the solution, resulting in concentration-dependent inhibition. Thus, the binding activity of the amino acid-substituted peptides and VIpep-3 to VIPR2 was detected as competitive inhibition of Biotin-VIpep-3 binding. The luminescence values of wells without Biotin-VIpep-3 were set as 100% inhibitory activity, and the luminescence values of wells without amino acid-substituted peptides or VIpep-3 were set as 0% inhibitory activity. The 50% inhibitory activity of the amino acid-substituted peptides and VIpep-3 was then calculated relative to the 50% inhibitory activity of VIpep-3.

[0097] (Evaluation of the binding activity of amino acid-substituted peptides to VIPR2) Figure 4 shows the results of a competitive binding test performed with VIPR2-expressing cells (N=4). Representative examples of the peptides of the present invention (Seq-1 to 9) all competed with Biotin-VIpep-3. These results demonstrate that the peptides of the present invention, which have amino acid substitutions and bicyclization, have VIPR2 binding activity. In particular, the binding activity of Seq-1 to 4 and Seq-6 was equal to or greater than that of VIpep-3.

[0098] (Evaluation of antagonistic activity of the peptide of the present invention against VIPR2) Changes in intracellular calcium concentration are known to be a downstream signal of VIPR2. Fluo-8, an indicator reagent for measuring changes in intracellular calcium concentration, was introduced into a human VIPR2-expressing cell line (Catalog No. HTS079RTA, Eurofins) using the ScreeQuest™ Fluo-8 No Wash Calcium Assay Kit (Catalog No. 36315, AAT Bioquest). The natural ligand VIP (final concentration 150 nM) was co-added with VIpep-3 (final concentration 750 nM) or representative peptides of the present invention (Seq-1, 5-9) (final concentration 750 nM), and the changes in fluorescence intensity were observed over time using a fluorescence microscope. The difference between the fluorescence intensity before and after VIP addition was taken as 100%, and was compared with the difference in fluorescence intensity after co-addition of VIpep-3 or the peptides of the present invention.

[0099] As shown in Figure 5, VIpep-3 and representative examples of the peptides of the present invention (Seq-1, 5 to 9) inhibited the receptor agonist activity of VIP against VIPR2. In particular, the inhibitory activity of Seq-1 and Seq-6 was equal to or greater than that of VIpep-3.

[0100] (Evaluation of Resistance of the Peptide of the Invention to Protease Degradation) To confirm the protease degradation resistance of the peptides of the present invention compared with VIpep-3, VIpep-3 or representative peptides of the present invention (Seq-1, 6, 9, and 10) were mixed with rat plasma and incubated for a set period of time. The remaining peptides were evaluated by the presence or absence of peaks on RP-HPLC. 20 μL of rat plasma was added to 1 μL of each 10 mM peptide and incubated at 37°C for a set period of time (0 or 24 hours). Then, 200 μL of 80% acetonitrile was added to VIpep-3, and 200 μL of acetonitrile was added to the other peptides. After thorough mixing, the mixture was left on ice for 10 minutes and centrifuged at 15,000 rpm at 4°C for 10 minutes to remove plasma proteins. The supernatant was collected, and the remaining peptides were analyzed by RP-HPLC using a SunFire C18 column (5 μm, 4.6 × 150 mm) (Solution A: 0.1% TFA / water, Solution B: 0.1% TFA / acetonitrile, 20-min gradient from 80% Solution A to 10% Solution A at 1 mL / min). As shown in Figure 6, the peak derived from VIpep-3 significantly decreased after 24 hours of incubation, suggesting that some portion of the sequence had been degraded by proteases. On the other hand, the peaks derived from representative peptides of the present invention (Seq-1, 6, 9, and 10) remained nearly the same after 24 hours compared to the time of incubation at 0 hours. These results demonstrate that the peptides of the present invention, which contain amino acid substitutions and bicyclization, are resistant to protease degradation.

[0101] (Evaluation of the selective antagonist activity of the peptide of the present invention against VIPR2) A representative example of the peptide of the present invention (Seq-10) was added at any concentration to VIPR1-expressing cells transfected with an intracellular calcium indicator, VIPR2-expressing cells transfected with an intracellular calcium indicator, or PAC1-expressing cells transfected with an intracellular calcium indicator, and the cells were incubated for 30 minutes. Subsequently, VIP (25 nM and 150 nM) was added to VIPR1 and VIPR2 cells, and PACAP (35 nM) was added to PAC1 cells. The change in intracellular calcium concentration over the next 2 minutes was measured using a FLIPR Tetra. The inhibitory effect of the antagonist peptide was calculated by defining the change in intracellular calcium concentration resulting from the addition of only the VIP or PACAP ligand as 0% inhibition and the change in intracellular calcium concentration resulting from the addition of neither the antagonist peptide nor the ligand as 100% inhibition.

[0102] As shown in FIG. 7, the antagonist activity of a representative example of the peptide of the present invention (Seq-10) was significantly stronger against the VIP-VIPR2 interaction than against the VIP-VIPR1 interaction or the PACAP-PAC1 interaction.

[0103] (Evaluation of in vivo VIPR2 antagonist activity of the peptide of the present invention) Ro25-1553 (Peptide Institute, Inc.), a VIP analog and selective VIPR2 agonist (0.4 nmol / g body weight), was dissolved in PBS and mixed with a representative peptide of the present invention (Seq-10) dissolved in DMSO (1 nmol / g body weight or 10 nmol / g body weight) at a 9:1 ratio (final DMSO concentration of 10%) or DMSO (vehicle group). These solutions were subcutaneously administered to ICR mice (12-day-old, male). One hour later, the brains were removed, and the prefrontal cortex was isolated. Intracellular phosphorylation of cyclic AMP-responsive element-binding protein (CREB), a downstream signaling pathway of VIPR2, was assessed by Western blotting. Antibodies used for Western blotting were anti-phospho-CREB antibody (#9198) and anti-CREB antibody (#9197) purchased from Cell Signaling Technology.

[0104] As shown in Figure 8, CREB phosphorylation was clearly suppressed in the presence of Seq-10, demonstrating that representative examples of the peptides of the present invention have antagonist activity against VIPR2 in vivo.

[0105] (Evaluation of Improvement in Recognition Function for Novel Objects in Mice by Administration of the Peptide of the Present Invention) As reported in Non-Patent Document 12, Ro25-1553 was administered subcutaneously once daily for 14 days to day-old mice, and then continued to be raised (8 weeks of age) to demonstrate a clear decline in novel object recognition function (Figure 9(A)). We evaluated whether the peptide of the present invention could improve novel object recognition function, which is impaired by VIPR2 activation. Ro25-1553 (Peptide Institute, Inc.), a VIP analog and selective VIPR2 agonist (0.07 nmol / g body weight), was dissolved in PBS and mixed with a representative peptide of the present invention (Seq-10) dissolved in DMSO (1 nmol / g body weight) or DMSO (vehicle group) at a 99:1 ratio (final DMSO concentration: 1%). These solutions were administered subcutaneously once daily for 14 days to day-old ICR mice (male), and then continued to be raised until they were 8 weeks of age when they underwent a novel object recognition test. The novel object recognition test was conducted during the light phase (8:00-20:00) according to Non-Patent Document 12. First, in a soundproof laboratory set at 30 lux, test mice were habituated to a test cage (30 cm × 30 cm × 35 cm) made of acrylic-modified polyvinyl chloride, lined only with sterilized wooden soft-tip pads (Sankyo Laboservice Co., Ltd.), for 10 min per day for 3 consecutive days. On the fourth day, two different objects (objects a and b were randomly selected from a golf ball, a Lego block, a plastic cylinder, and an electrical outlet) were placed 8 cm from the wall, and the mice were allowed to freely explore for 10 min (training trial). Twenty-four hours later, object b was replaced with a novel object c, and the mice were allowed to freely explore for 5 min (test trial). The behavior of the animals in the training and test trials was videotaped, and the exploration time for each of the two objects was measured. The discrimination index was calculated as the percentage (%) of the exploration time difference between object c and object a relative to the total exploration time in the test trial.

[0106] As shown in Figure 9(B), mice administered Ro25-1553 showed a clear decline in novel object recognition function. On the other hand, mice administered a mixture of Ro25-1553 and a representative example of the peptide of the present invention (Seq-10) showed a significant improvement in the decline in novel object recognition function (p value of Student's t-test: 0.001 or less). Furthermore, their recognition function was nearly equivalent to that of normal mice. These results demonstrate that administration of the peptide of the present invention can significantly prevent or ameliorate the decline in novel object recognition function in mice caused by VIPR2 activation. [Industrial Applicability]

[0107] The peptides of the present invention are resistant to protease degradation and inhibit VIPR2 signaling mediated by a natural ligand by binding to VIPR2, and are therefore considered to be useful for the prevention and treatment of diseases associated with VIPR2 activation, such as schizophrenia and autism spectrum disorder.

[0108] The peptide of the present invention can be used not only as a drug itself, but also as a delivery molecule to tissues expressing VIPR2, or as a molecule for detecting the expression of VIPR2.

[0109] Furthermore, the peptide of the present invention is expected to be more effective in preventing and treating schizophrenia, autism spectrum disorder, and the like when used in combination with pharmaceuticals or therapies having other mechanisms of action.

Claims

1. Formula (1): c[X Ｎ -X ４ -X ５ -X ６ -c(X ７ -X ８ -X ９ -X １０ ]-X １１ )-X １２ -X １３ -X １４ -X １５ -X １６ (1)or Formula (2): c[X Ｎ -X ４ -X ５ -X ６ -c(X ７ -X ８ -X ９ -X １０ )]-X １１ -X １２ -X １３ -X １４ -X １５ -X １６ (2) (In formula (1), X N and X １０ and X ７ and X １１ and in formula (2), X N and X ７ and X １０ are independently covalently bonded to each other to form two cyclic structures in the molecule, and these covalent bonds may be between the main chain and the side chain or between the side chains, X N is X １ -X ２ -X ３ represents X １ , X ２ , X ３ , X ７ , X １０ and X １１ When involved in the cyclization within the peptide molecule, each independently represents an amino acid residue having an amino group, a carboxy group, a thiol group, an allyl group, an alkynyl group, an azide group, or a halogen atom, or a derivative thereof; when not involved in the cyclization within the peptide molecule, X １ and X ２ each independently represents any amino acid residue or deletion, and X ３ and X １１ each independently represents any amino acid residue, X ５ , X ９ , X １２ and X １３ each independently represents an amino acid residue having a hydrocarbon group which may have a substituent, or a derivative thereof; X ４ represents an amino acid residue having an optionally substituted aromatic carbocyclic group, or a derivative thereof; X ６ and X ８ represents any amino acid residue, X １４ and X １５ and X １６ each independently represents any amino acid residue or deletion, The N-terminal amino group and the C-terminal carboxy group may be modified or deleted. or a pharmacologically acceptable salt thereof.

2. Formula (3): c[X １ -X ２ -X ３ -X ４ -X ５ -X ６ -c(X ７ -X ８ -X ９ -X １０ ]-X １１ )-X １２ -X １３ (3) (In the formula, X １ and X １０ and X ７ and X １１ means that two cyclic structures are formed in the molecule by independently forming covalent bonds, and the covalent bonds are １ The main chain or side chain of X １０ and the side chain of X ７ The side chain of X １１ is formed between the side chain of X １ ~X １３ is as defined in claim 1. or a pharmacologically acceptable salt thereof.

3. X １ and X １０ The bond between the Cα carbon atoms is represented by the following formula (4): Cα １ -(CH ２ ) Ａ -L １ -(CH ２ ) Ｂ -Cα １０ (4) (In the formula, A and B each independently represent an integer of 0 to 2, the sum of A and B is an integer of 1 to 4, and L １ is S, S-S, CH ２ -CH ２ , S-CH ２ , C.H. ２ -S, O-CH ２ , C.H. ２ -O, CH=CH, NH-C(=O), N(CH ３ )-C(=O), NH-C(=S), N(CH ３ )-C(=S), O-C(=O), C(=O)-NH, C(=O)-N(CH ３ ), C(=S)-NH, C(=S)-N(CH ３ ), C(=O)-O, CH ２ -CH ２ -CH ２ , S-CH ２ -CH ２ , C.H. ２ -CH ２ -S, CH ２ -S-CH ２ , O-CH ２ -CH ２ , C.H. ２ -CH ２ -O, CH ２ -O-CH ２ , S-CH ２ -S, S-C (=CH ２ )-S, S-(CH ２ ) ２ -S, or triazole.

3. The cyclic peptide according to claim 1 or 2, comprising a structure represented by the following formula: or a pharmacologically acceptable salt thereof.

4. Formula (5): c[X ２ -X ３ -X ４ -X ５ -X ６ -c(X ７ -X ８ -X ９ -X １０ ]-X １１ )-X １２ -X １３ (5) (In the formula, X ２ and X １０ and X ７ and X １１ means that two cyclic structures are formed in the molecule by independently forming covalent bonds, and the covalent bonds are ２ The main chain or side chain of X １０ and the side chain of X ７ The side chain of X １１ is formed between the side chain of X ２ ~X １３ is as defined in claim 1. or a pharmacologically acceptable salt thereof.

5. X ２ and X １０ The bond between the Cα carbon atoms is represented by the following formula (6): Cα ２ -(CH ２ ) Ａ -L ２ -(CH ２ ) Ｂ -Cα １０ (6) (In the formula, A and B each independently represent an integer of 0 to 6, the sum of A and B is an integer of 4 to 7, and L ２ is S, S-S, CH ２ -CH ２ , S-CH ２ , C.H. ２ -S, O-CH ２ , C.H. ２ -O, CH=CH, NH-C(=O), N(CH ３ )-C(=O), NH-C(=S), N(CH ３ )-C(=S), O-C(=O), C(=O)-NH, C(=O)-N(CH ３ ), C(=S)-NH, C(=S)-N(CH ３ ), C(=O)-O, CH ２ -CH ２ -CH ２ , S-CH ２ -CH ２ , C.H. ２ -CH ２ -S, CH ２ -S-CH ２ , O-CH ２ -CH ２ , C.H. ２ -CH ２ -O, CH ２ -O-CH ２ , S-CH ２ -S, S-C (=CH ２ )-S, S-(CH ２ ) ２ -S, S-(CH ２ ) ３ -S, S-(CH ２ ) ４ -S, S-(CH ２ ) ５ -S, S-CH ２ -CH=CH-CH ２ -S, S-CH ２ -C(=O)-NH, NH-C(=O)-CH ２ -S, S-CH ２ -C ６ H ４ -CH ２ -S (the bonding position of the methylene group to the phenylene ring may be ortho, meta or para), S-CH ２ -C 10 H ６ -CH ２ -S, S-CH ２ —C(═O)—CH ２ -S, or S-CH ２ -C(=CH ２ )-CH ２ represents —S, triazole, or dithiotetrafluorobenzene. The cyclic peptide according to claim 1 or 4, which comprises a structure represented by the following formula: or a pharmacologically acceptable salt thereof.

6. Formula (7): c[X ３ -X ４ -X ５ -X ６ -c(X ７ -X ８ -X ９ -X １０ ]-X １１ )-X １２ -X １３ (7) (In the formula, X ３ and X １０ and X ７ and X １１ means that two cyclic structures are formed in the molecule by independently forming covalent bonds, and the covalent bonds are ３ The main chain or side chain of X １０ and the side chain of X ７ The side chain of X １１ is formed between the side chain of X ３ ~X １３ is as defined in claim 1. or a pharmacologically acceptable salt thereof.

7. X ３ and X １０ The bond between the Cα carbon atoms is represented by the following formula (8): Cα ３ -(CH ２ ) Ａ -L ３ -(CH ２ ) Ｂ -Cα １０ (8) (In the formula, A and B each independently represent an integer of 0 to 10, the sum of A and B is an integer of 7 to 10, and L ３ is S, S-S, CH ２ -CH ２ , S-CH ２ , C.H. ２ -S, O-CH ２ , C.H. ２ -O, CH=CH, NH-C(=O), N(CH ３ )-C(=O), NH-C(=S), N(CH ３ )-C(=S), O-C(=O), C(=O)-NH, C(=O)-N(CH ３ ), C(=S)-NH, C(=S)-N(CH ３ ), C(=O)-O, CH ２ -CH ２ -CH ２ , S-CH ２ -CH ２ , C.H. ２ -CH ２ -S, CH ２ -S-CH ２ , O-CH ２ -CH ２ , C.H. ２ -CH ２ -O, CH ２ -O-CH ２ , S-CH ２ -S, S-C (=CH ２ )-S, S-(CH ２ ) ２ -S, S-(CH ２ ) ３ -S, S-(CH ２ ) ４ -S, S-(CH ２ ) ５ -S, S-(CH ２ ) ６ -S, S-(CH ２ ) ７ -S, S-(CH ２ ) ８ -S, S-CH ２ -CH=CH-CH ２ -S, S-CH ２ -C(=O)-NH, NH-C(=O)-CH ２ -S, S-CH ２ -C ６ H ４ -CH ２ -S (the bonding position of the methylene group to the phenylene ring may be ortho, meta or para), S-CH ２ -C ６ H ４ -C ６ H ４ -CH ２ -S, S-CH ２ -C ５ NH ３ -C ５ NH ３ -CH ２ -S, S-CH ２ -C 10 H ６ -CH ２ -S, S-CH ２ —C(═O)—CH ２ -S, or S-CH ２ -C(=CH ２ )-CH ２ represents —S, triazole, or dithiotetrafluorobenzene. The cyclic peptide according to claim 1 or 6, which comprises a structure represented by the following formula: or a pharmacologically acceptable salt thereof.

8. X ７ and X １１ The bond between the Cα carbon atoms is represented by the following formula (9): Cα 7 -(CH ２ ) Ａ -L ７ -(CH ２ ) Ｂ -Cα １１ (9) (In the formula, A and B each independently represent an integer of 0 to 7, the sum of A and B is an integer of 2 to 7, and L ７ is S, S-S, CH ２ -CH ２ , S-CH ２ , C.H. ２ -S, O-CH ２ , C.H. ２ -O, CH=CH, NH-C(=O), N(CH ３ )-C(=O), NH-C(=S), N(CH ３ )-C(=S), O-C(=O), C(=O)-NH, C(=O)-N(CH ３ ), C(=S)-NH, C(=S)-N(CH ３ ), C(=O)-O, CH ２ -CH ２ -CH ２ , S-CH ２ -CH ２ , C.H. ２ -CH ２ -S, CH ２ -S-CH ２ , O-CH ２ -CH ２ , C.H. ２ -CH ２ -O, CH ２ -O-CH ２ , S-CH ２ -S, S-C (=CH ２ )-S, S-(CH ２ ) ２ -S, S-(CH ２ ) ３ -S, S-(CH ２ ) ４ -S, S-CH ２ -CH=CH-CH ２ -S, S-CH ２ -C(=O)-NH, NH-C(=O)-CH ２ -S, S-CH ２ -C ６ H ４ -CH ２ -S (the bonding position of the methylene group to the phenylene ring may be ortho, meta or para), S-CH ２ —C(═O)—CH ２ -S, or S-CH ２ -C(=CH ２ )-CH ２ represents —S, triazole, or dithiotetrafluorobenzene. The cyclic peptide according to any one of claims 1 to 7, comprising a structure represented by the following formula: or a pharmacologically acceptable salt thereof.

9. Formula (10): c[X ３ -X ４ -X ５ -X ６ -c(X ７ -X ８ -X ９ -X １０ )]-X １１ -X １２ -X １３ (10) (In the formula, X ３ and X ７ and X １０ means that the molecule has two ring structures formed by forming a covalent bond, and X ３ ~X １３ is as defined in claim 1. or a pharmacologically acceptable salt thereof.

10. X １ and X ７ and X １０ , X ２ and X ７ and X １０ , or X ３ and X ７ and X １０ The bond between the Cα carbon atoms is represented by the following formula (11): 【Chemistry 7】 (In the formula, N represents 1, 2, or 3; A, B, and D each independently represent an integer of 1 or 2; Y represents [(S-CH ３ ) ３ -C ６ H ３ ], [(S-CH ２ -CH ２ -C(=O) ３ -C ３ N ３ H ６ ], [(S-CH ２ -C(=O)-NH) ３ -C ６ H ３ ], [(NH-C(=O)) ３ -C ６ H ３ ], [(S-CH ３ ) ３ -C ３ N ３ H ６ ], or [(S-CH ２ -C(=O) ３ -C ３ N ３ H ６ ].) The cyclic peptide according to claim 1 or 9, which comprises a structure represented by the following formula: or a pharmacologically acceptable salt thereof.

11. X １ and X ７ and X １０ , X ２ and X ７ and X １０ , or X ３ and X ７ and X １０ The bond between the Cα carbon atom and the Cα carbon atom of the formula (12): 【Chemistry 8】 (In the formula, N represents 1, 2, or 3; A, B, D, E, and F each independently represent an integer of 1 or 2; Z and Z′ each independently represent S, S—S, or S—CH ２ -S, S-C (=CH ２ )-S, S-(CH ２ ) ２ -S, S-(CH ２ ) ３ -S, S-(CH ２ ) ４ -S, S-CH ２ -CH=CH-CH ２ -S, S-CH ２ -C(=O)-NH, NH-C(=O)-CH ２ -S, S-CH ２ -C ６ H ４ -CH ２ -S (the bonding position of the methylene group to the phenylene ring may be ortho, meta or para), S-CH ２ -C ６ H ４ -C ６ H ４ -CH ２ -S, S-CH ２ -C 10 H ６ -CH ２ -S, S-CH ２ —C(═O)—CH ２ -S, or S-CH ２ -C(=CH ２ )-CH ２ -S, or dithiotetrafluorobenzene. The cyclic peptide according to claim 1 or 9, which comprises a structure represented by the following formula: or a pharmacologically acceptable salt thereof.

12. X ５ , X ９ , X １２ and X １３ are each independently represented by the following formula (13): 【Chemistry 9】 (In the formula, the wavy line represents the point of attachment to the carbonyl group or nitrogen atom that forms the amide bond of the main chain, and R １ , R ２ , R ６ and R ７ each independently represents a hydrogen atom or a methyl group, R ３ , R ４ , R ５ each independently represents a hydrogen atom, a methyl group, or a halogen atom (F, Cl, Br, I), and n represents an integer of 0 to 10; or The following formula (14): 【Chemistry 10】 (wherein the wavy line represents the point of attachment to the carbonyl group or nitrogen atom of the main chain that forms an amide bond, Z represents C or N, and R ８ and R ９ represents a hydrogen atom or a methyl group, and n represents an integer of 1 to 6; The cyclic peptide according to any one of claims 1 to 11, or a pharmacologically acceptable salt thereof, which is an amino acid residue represented by the following formula:

13. X ４ is expressed by the following formula (15): 【Chemistry 11】 In the formula, the wavy line represents the point of attachment to the carbonyl group or nitrogen atom that forms an amide bond in the main chain, and R １０ , R １１ , R １２ , R １３ and R １４ each independently represents a hydrogen atom, a hydroxy group, a hydroxymethyl group, a methoxy group, a methoxymethyl group, an amino group, an aminomethyl group, a monomethylated amino group, a dimethylated amino group, a trimethylated amino group, a monomethylated aminomethyl group, a dimethylated aminomethyl group, a trimethylated aminomethyl group, an acetyl group, an amido group, a methyl group, a tert-butyl group, a halogenated methyl group, or a halogen atom (F, Cl, Br, I); R １５ and R １６ represents a hydrogen atom or a methyl group; The cyclic peptide according to any one of claims 1 to 12, or a pharmacologically acceptable salt thereof, which is an amino acid residue represented by the following formula:

14. X ８ represents an amino acid residue having an aromatic carbocyclic group which may have a substituent, or a derivative thereof, or a cyclic peptide or a pharmacologically acceptable salt thereof according to any one of claims 1 to 13.

15. X ８ is expressed by the following formula (16): 【Chemistry 12】 In the formula, the wavy line represents the point of attachment to the carbonyl group or nitrogen atom that forms an amide bond in the main chain, and R １７ , R １８ , R １９ , R ２０ and R ２１ each independently represents a hydrogen atom, a hydroxy group, a hydroxymethyl group, a methoxy group, a methoxymethyl group, an amino group, an aminomethyl group, a monomethylated amino group, a dimethylated amino group, a trimethylated amino group, a monomethylated aminomethyl group, a dimethylated aminomethyl group, a trimethylated aminomethyl group, an acetyl group, an amido group, a methyl group, a tert-butyl group, a halogenated methyl group, or a halogen atom (F, Cl, Br, I); R ２２ and R ２３ represents a hydrogen atom or a methyl group; or an amino acid residue having a basic side chain, or a derivative thereof. The cyclic peptide according to any one of claims 1 to 14, or a pharmacologically acceptable salt thereof.

16. X ６ represents glycine, N-methylated glycine, alanine, N-methylated alanine, D-alanine, N-methylated D-alanine, 2-aminoisobutyric acid, N-methylated 2-aminoisobutyric acid, 2-azetidine-2-carboxylic acid, D-2-azetidine-2-carboxylic acid, proline, D-proline, thioproline, D-thioproline, 3,4-dehydroproline, D-3,4-dehydroproline, pipecolic acid, D-pipecolic acid, or a derivative thereof. The cyclic peptide or pharmacologically acceptable salt thereof according to any one of claims 1 to 15.

17. Formula (17): c[X １ -Pro ２ -X ３ -Tyr ４ -Leu ５ -Pro ６ -c(X ７ -X ８ -Leu ９ -Cys １０ ]-X １１ )-X １２ -X １３ (17) (In the formula, X １ represents cysteine, Mpa (3-mercaptopropionic acid) or D-cysteine, X ３ represents N-methylated glycine, N-methylated alanine, 2-azetidine-2-carboxylic acid, proline, hydroxyproline, 3,4-dehydroproline, pipecolic acid, serine, or lysine; X ８ represents tyrosine, proline or arginine, X ７ and X １１ represents any combination of lysine and aspartic acid, ornithine and glutamic acid, aspartic acid and lysine, glutamic acid and ornithine, lysine and glutamic acid, or glutamic acid and lysine, X １２ and X １３ each independently represents leucine, isoleucine, or norleucine, X １ and Cys １０ forms a disulfide bond between each side chain, and X ７ and X １１ form an amide bond between the respective side chains, so that the peptide of formula (17) has two cyclic structures in the molecule, and the N-terminal amino group and the C-terminal carboxy group may be modified or deleted.

18. X １ represents cysteine, X ３ represents proline or serine, X ７ represents lysine, X ８ represents tyrosine, X １１ represents aspartic acid, X １２ and X １３ are each independently leucine, isoleucine or norleucine, the N-terminal amino group is acetylated and the C-terminal carboxy group is amidated, or a cyclic peptide or a pharmacologically acceptable salt thereof according to claim 17.

19. A cyclic peptide or a pharmacologically acceptable salt thereof having VIPR2 antagonist activity, wherein one to several amino acids are deleted, added, and / or substituted in the amino acid sequence of the peptide according to any one of claims 1 to 18, or the peptide has at least 50% or more sequence homology with the amino acid sequence, and / or at least 50% or more conformational homology with the peptide according to any one of claims 1 to 18.

20. A cyclic peptide having an amino acid sequence set forth in any one of SEQ ID NOs: 1 to 10, or an amino acid sequence in which one or several amino acids have been deleted, added, and / or substituted in the amino acid sequence set forth in any one of SEQ ID NOs: 1 to 10, and having VIPR2 antagonist activity, or a pharmacologically acceptable salt thereof.

21. A derivative and / or modified form of the peptide according to any one of claims 1 to 20.

22. A pharmaceutical, diagnostic agent, and / or research reagent comprising the peptide according to any one of claims 1 to 21.

Images