Peptide having inhibitory activity on formation of leukocyte extracellular traps

Abstract

Provided is a novel peptide molecule having an inhibitory activity on the formation of leukocyte extracellular traps. In the description, a peptide molecule having a structure containing an amino acid sequence represented by formula (1) or an amino acid sequence represented by formula (2) and having an activity to inhibit the formation of leukocyte extracellular traps, or a pharmacologically acceptable salt of the peptide molecule is disclosed.　X1-KCRR-X2-Q-X3-R-X4-KK (1) R1-CH2CH2-CO-KCRR-X2-Q-X3-R-X4-KK (2) (In each of the formulae, X1, X2, X3, X4 and R1 are as defined in the description.)

Description

Peptides that inhibit extracellular trap formation in leukocytes

[0001] The present invention relates to peptides having activity of inhibiting the formation of extracellular traps in leukocytes.

[0002] Neutrophil extracellular traps (NETs) are extracellular structures that are formed when neutrophils are activated by bacterial infection, resulting in loss of segmented nuclear morphology and chromatin distribution, followed by loss of the nuclear membrane, resulting in the chromatin structure being mixed with cytoplasm and granular components. When the cell membrane is ruptured, these net-like structures are released, capturing bacteria, fungi, parasites, and viruses and exerting antibacterial effects. Dysregulation of NETs has been shown to be involved in various inflammatory diseases, cardiovascular diseases, autoimmune diseases, as well as malignant tumor metastasis. Diseases thought to involve dysregulation of NETs include a wide range of representative diseases, including autoimmune diseases such as systemic lupus erythematosus (SLE), rheumatoid arthritis, ANCA-associated vasculitis, and psoriasis; perioperative acute kidney injury due to ischemia-reperfusion; thrombotic diseases such as disseminated intravascular coagulation (DIC) and deep vein thrombosis associated with sepsis and sickle cell disease; cardiovascular diseases such as arteriosclerosis, acute myocardial infarction, and cerebral infarction; acute inflammatory diseases such as acute pancreatitis and gout attacks; delayed wound healing due to diabetes; and the progression and metastasis of malignant tumors. Furthermore, because the number of patients affected is enormous, the socioeconomic impact is significant, and the development of therapies targeting NETs is of great medical and therapeutic value.

[0003] Among these diseases, for example, acute kidney injury (AKI) after rhabdomyolysis (PHR) is limited to specialized treatment options, such as massive fluid infusion and renal replacement therapy, which must be performed at a medical institution. Therefore, the development of new treatments and preventative measures is awaited. ARK is a form of crush syndrome (crush syndrome) with an extremely poor prognosis and is one of the causes of many crush syndrome casualties worldwide. In Japan, the 1995 Great Hanshin-Awaji Earthquake resulted in over 370 crush syndrome patients, with approximately 50 deaths. Muscle tissue destruction is generally known to occur due to factors other than trauma, such as drug-induced and ischemic causes. In all cases, skeletal muscle tissue ruptures, and muscle-derived components leak into the bloodstream, causing various organ damage, a condition known as rhabdomyolysis. Approximately 13% to 50% of patients with rhabdomyolysis develop acute kidney injury (AKI), and it is known that patients with AKI have a dramatically worse prognosis than those without. However, there are no portable AKI prevention methods that can be used quickly and easily at disaster sites, and there are no effective treatments other than symptomatic treatments such as fluid infusion and hemodialysis. AKI is therefore considered to be a disease with high unmet medical needs.

[0004] Psoriasis, one of the diseases associated with dysregulated NETs, ​​is an intractable chronic autoimmune disease characterized by the formation of raised erythema and scales due to the excessive proliferation of skin epithelial cells. Psoriasis treatments include topical application of medications to the affected area, phototherapy, which uses artificial ultraviolet light to improve symptoms, and oral therapies such as immunosuppressants and retinoids. New biologic therapies include various monoclonal antibodies, including TNFα inhibitors, which were approved for the treatment of psoriasis in 2010. While these treatments can alleviate symptoms, there is still no cure. Furthermore, new biologics are expensive, not always effective in all patients, and the effects of long-term administration remain unclear. While increased sun exposure is a traditional treatment for psoriasis, excessive sun exposure can increase the risk of skin cancer. Therefore, psoriasis is considered a disease with high unmet medical needs. Thus, the development of drugs with novel mechanisms of action for psoriasis is urgently needed.

[0005] Lactoferrin is known to inhibit the formation of extracellular traps by leukocytes (Patent Document 1). However, human lactoferrin is a polypeptide with 691 amino acid residues and a molecular weight of approximately 80,000. Therefore, using human lactoferrin as a drug poses various difficulties, including manufacturing, formulation, and stability issues. Furthermore, it has been reported that lactoferrin fragments with a specific amino acid sequence inhibit the formation of extracellular traps by leukocytes (Patent Document 2). However, Patent Document 2 does not confirm the effects of these lactoferrin fragments in vivo, and it is unclear whether these lactoferrin fragments will exert the desired effects when administered to the body. Furthermore, the lactoferrin fragments described in Patent Document 2 are based on the amino acid sequences of lactoferricins derived from bovine, human, monkey, goat, sheep, horse, camel, dog, mouse, pig, and rat. Patent Document 2 does not disclose or suggest the use of amino acid residues not present at specific positions in the amino acid sequences of existing mammalian lactoferrins.

[0006] Patent Document 1: International Publication No. 2014 / 168253 Patent Document 2: International Publication No. 2016 / 056665

[0007] In view of the above background, an object of the present invention is to provide a novel peptide molecule that has activity of inhibiting the formation of extracellular traps in leukocytes.

[0008] The present invention relates to, for example, the following [1] to

[20] . [1] A peptide molecule or a pharmaceutically acceptable salt thereof, which contains an amino acid sequence represented by the following formula (1) or has a structure containing an amino acid sequence represented by the following formula (2), and has activity of inhibiting the formation of extracellular traps in leukocytes: X1-KCRR-X2-Q-X3-R-X4-KK (1) R 1 -CH2CH2-CO-KCRR-X2-Q-X3-R-X4-KK (2) [In each formula, X1 is R 1 an α-amino acid residue having a side chain represented by —CH—, R 1is a hydrogen atom, an alkyl group, an alicyclic hydrocarbon group, an aromatic hydrocarbon group, a heteroalicyclic group, or a heteroaromatic group, and X2 represents a structure represented by the following formula (3): (In the formula, R 2 is a hydrogen atom, an alkyl group, an aromatic hydrocarbon group, or a heteroaromatic group, and n is 1 or 2. X3 represents a structure represented by the following formula (4): (In the formula, R 3 is a hydrogen atom, an alkyl group, an alicyclic hydrocarbon group, an aromatic hydrocarbon group, a heteroalicyclic group, or a heteroaromatic group; X4 is a structure represented by the following formula (5): (In the formula, R 4 is a hydrogen atom, or -R 5 -Z-R 6 Or -Z-R 7 R 5 is an alkylene group, and R 6 is an alkyl group, or R 5 and R 6 form a heterocycle together with Z to which they are attached, and R 7 is an alkyl group, Z represents an oxygen atom or a sulfur atom, and R 1 , R 2 , and R 3 The heteroalicyclic group or heteroaromatic group in the formula (1) is one in which the atoms constituting the ring contain 1 to 4 heteroatoms selected from oxygen atoms, sulfur atoms, and nitrogen atoms, with the proviso that the amino acid sequence represented by formula (1) is not any of FKCRRWQWRMKK, AKCRRWQWRMKK, FKCRRFQWRMKK, AKCRRFQWRMKK, FKCRRWQVRMKK, AKCRRWQVRMKK, FKCRRFQVRMKK, and AKCRRFQVRMKK.] [2] R 1 is a hydrogen atom, an alkyl group, a monocyclic or bicyclic cycloalkyl group, a monocyclic or bicyclic cycloalkenyl group, a monocyclic or bicyclic aromatic hydrocarbon group, or a monocyclic or bicyclic heteroaromatic group; R 2 is a hydrogen atom, an alkyl group, a monocyclic or bicyclic aromatic hydrocarbon group, or a monocyclic or bicyclic heteroaromatic group, and R3 is a hydrogen atom, an alkyl group, a monocyclic or bicyclic cycloalkyl group, a monocyclic or bicyclic cycloalkenyl group, a monocyclic or bicyclic aromatic hydrocarbon group, or a monocyclic or bicyclic heteroaromatic group, and R 1 , R 2 , and R 3 The monocyclic or bicyclic heteroaromatic group in the formula (I) contains 1 to 4 heteroatoms selected from oxygen, sulfur, and nitrogen atoms as ring constituent atoms. 1 is a hydrogen atom, an alkyl group having 1 to 9 carbon atoms, a monocyclic cycloalkyl group having 3 to 10 carbon atoms, a monocyclic cycloalkenyl group having 3 to 10 carbon atoms, a monocyclic aromatic hydrocarbon group, a monocyclic heteroaromatic group, or a bicyclic group having a cyclic structure represented by the following formula (6): (wherein ring A is a ring that makes the structure represented by formula (6) a 9- to 11-membered bicyclic ring structure, the atoms constituting ring A include at least one carbon atom and may include one or two heteroatoms selected from oxygen atoms, sulfur atoms, and nitrogen atoms, Z1, Z2, Z3, and Z4 are each independently a carbon atom or a nitrogen atom, and the number of nitrogen atoms in Z1, Z2, Z3, and Z4 is a maximum of 2, represents a single bond or a double bond, and the bonding point to the CH2 moiety in the side chain of X1 described in [1] above or in the structure represented by formula (2) described in [1] above may be ring A or a ring having Z1.) R 2 is a hydrogen atom, an alkyl group having 1 to 9 carbon atoms, a monocyclic aromatic hydrocarbon group having 3 to 10 carbon atoms, a monocyclic heteroaromatic group having 3 to 10 carbon atoms, or a bicyclic group having a cyclic structure represented by the following formula (7): (wherein ring B is a ring that makes the structure represented by formula (7) a 9- to 11-membered bicyclic aromatic ring, and the atoms constituting ring B include at least one carbon atom and may include one or two heteroatoms selected from an oxygen atom, a sulfur atom, and a nitrogen atom; Z1, Z2, Z3, and Z4 are each independently a carbon atom or a nitrogen atom, and the number of nitrogen atoms in Z1, Z2, Z3, and Z4 is a maximum of two; and the bonding point to the CH2 moiety in the structure represented by formula (3) described in [1] above may be ring B or a ring having Z1.) and R 3 is a hydrogen atom, an alkyl group having 1 to 9 carbon atoms, a monocyclic cycloalkyl group having 3 to 10 carbon atoms, a monocyclic cycloalkenyl group having 3 to 10 carbon atoms, a monocyclic aromatic hydrocarbon group, a monocyclic heteroaromatic group, or a bicyclic group having a cyclic structure represented by the following formula (8): (wherein ring C is a ring that makes the structure represented by formula (8) a 9- to 11-membered bicyclic ring structure, and the atoms constituting ring C include at least one carbon atom and may include one or two heteroatoms selected from an oxygen atom, a sulfur atom, and a nitrogen atom; Z1, Z2, Z3, and Z4 are each independently a carbon atom or a nitrogen atom, and the number of nitrogen atoms in Z1, Z2, Z3, and Z4 is a maximum of 2; represents a single bond or a double bond, and the point of attachment to the CH2 moiety in the structure represented by formula (4) described in [1] above may be a C ring or a ring having Z1.) The peptide molecule described in [1] above or a pharmaceutically acceptable salt thereof. [4] R 1 is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, a monocyclic cycloalkyl group having 3 to 7 carbon atoms, or an aromatic hydrocarbon group or bicyclic heteroaromatic group having a cyclic structure represented by the following formula (6-1): (In the formula, the ring A1 is a ring that makes the structure represented by formula (6-1) a 9- to 10-membered bicyclic aromatic ring, or is absent; the atoms constituting the ring A1 include at least one carbon atom and may include one or two heteroatoms selected from an oxygen atom, a sulfur atom, and a nitrogen atom; the bonding point to the CH2 moiety in the side chain of X1 described in [1] above or in the structure represented by formula (2) described in [1] above may be the ring A1 or a benzene ring.) R 2 is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, or an aromatic hydrocarbon group or bicyclic heteroaromatic group having a cyclic structure represented by the following formula (7-1): (wherein ring B1 is a ring that makes the structure represented by formula (7-1) a 9- to 10-membered bicyclic aromatic ring, or is absent; the atoms constituting ring B1 include at least one carbon atom and may include one or two heteroatoms selected from oxygen atoms, sulfur atoms, and nitrogen atoms; the point of attachment to the CH2 moiety in the structure represented by formula (3) described in [1] above may be ring B1 or a benzene ring.) and R 3 is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, a monocyclic cycloalkyl group having 3 to 7 carbon atoms, or an aromatic hydrocarbon group or bicyclic heteroaromatic group having a cyclic structure moiety represented by the following formula (8-1): (wherein the C1 ring is a ring that results in the structure represented by formula (8-1) being a 9- to 10-membered bicyclic aromatic ring, or is absent; the atoms constituting the C1 ring include at least one carbon atom and optionally include one or two heteroatoms selected from an oxygen atom, a sulfur atom, and a nitrogen atom; and the attachment point to the CH2 moiety in the structure represented by formula (4) described in [1] above may be the C1 ring or a benzene ring.) The peptide molecule described in [1] above or a pharmaceutically acceptable salt thereof. [5] The peptide molecule described in any one of [1] to [4] above or a pharmaceutically acceptable salt thereof, comprising the amino acid sequence represented by formula (1), wherein X1 is an alanine residue, a phenylalanine residue, a tryptophan residue, a cyclohexylalanine residue, or a 2-naphthylalanine residue. [6] The peptide molecule described in any one of [1] to [5] above or a pharmaceutically acceptable salt thereof, wherein X2 is an alanine residue, a tryptophan residue, or a homophenylalanine residue. [7] The peptide molecule or pharmaceutically acceptable salt thereof according to any one of [1] to [6] above, wherein X3 is an alanine residue, a tryptophan residue, a cyclohexylalanine residue, or a 2-naphthylalanine residue. [8] The peptide molecule or pharmaceutically acceptable salt thereof according to any one of [1] to [7] above, wherein X4 is not a methionine residue. [9] R 4 is a hydrogen atom, or -R 5 -Z-R 6 Or -Z-R 7 R 5 is an alkylene group having 1 to 6 carbon atoms, and R 6 is an alkyl group having 1 to 6 carbon atoms, or R 5 and R 6 form a 4- to 7-membered heterocyclic ring together with Z to which they are attached, and R 7

[10] The peptide molecule or pharmaceutically acceptable salt thereof according to [8] above, wherein R is an alkyl group having 1 to 6 carbon atoms. 4 But, -R 5 -Z-R 6 R 5 is an alkylene group having 1 to 3 carbon atoms, and R 6is an alkyl group having 1 to 3 carbon atoms, or R 5 and R 6 and R are taken together with Z to which they are attached to form a 5- to 6-membered heterocycle, or a pharmaceutically acceptable salt thereof. 5 is an alkylene group having 1 to 3 carbon atoms, and R 6

[10] The peptide molecule or pharmaceutically acceptable salt thereof according to

[10] above, wherein X is an alkyl group having 1 to 3 carbon atoms, and Z is an oxygen atom.

[12] The peptide molecule or pharmaceutically acceptable salt thereof according to

[11] above, wherein X is an O-methylhomoserine residue.

[13] R 1 is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, a monocyclic cycloalkyl group having 3 to 7 carbon atoms, or an aromatic hydrocarbon group or bicyclic heteroaromatic group having a cyclic structure represented by the following formula (6-1): (In the formula, the ring A1 is a ring that makes the structure represented by formula (6-1) a 9- to 10-membered bicyclic aromatic ring, or is absent; the atoms constituting the ring A1 include at least one carbon atom and may include one or two heteroatoms selected from an oxygen atom, a sulfur atom, and a nitrogen atom; the bonding point to the CH2 moiety in the side chain of X1 described in [1] above or in the structure represented by formula (2) described in [1] above may be the ring A1 or a benzene ring.) R 2 is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, or an aromatic hydrocarbon group or bicyclic heteroaromatic group having a cyclic structure represented by the following formula (7-1): (In the formula, ring B1 is a ring that makes the structure represented by formula (7-1) a 9- to 10-membered bicyclic aromatic ring, or is absent; the atoms constituting ring B1 include at least one carbon atom and may include one or two heteroatoms selected from oxygen atoms, sulfur atoms, and nitrogen atoms; and the bonding point to the CH2 moiety in the structure represented by formula (3) described in [1] above may be ring B1 or a benzene ring.) R 3 is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, a monocyclic cycloalkyl group having 3 to 7 carbon atoms, or an aromatic hydrocarbon group or bicyclic heteroaromatic group having a cyclic structure moiety represented by the following formula (8-1): (In the formula, the C1 ring is a ring that makes the structure represented by formula (8-1) a 9- to 10-membered bicyclic aromatic ring, or is absent; the atoms constituting the C1 ring include at least one carbon atom and may include one or two heteroatoms selected from oxygen atoms, sulfur atoms, and nitrogen atoms; and the bonding point to the CH2 moiety in the structure represented by formula (4) described in [1] above may be the C1 ring or a benzene ring.) R 4 But, -R 5 -Z-R 6 R 5 is an alkylene group having 1 to 3 carbon atoms, and R 6

[14] The peptide molecule or pharmaceutically acceptable salt thereof according to [1] above, wherein R is an alkyl group having 1 to 3 carbon atoms, and Z is an oxygen atom. 5 and R 6 and X1 are taken together with the Z to which they are attached to form a 5- to 6-membered heteroaromatic ring, or a pharmaceutically acceptable salt thereof.

[15] The peptide molecule according to any one of [1] to [7] above, comprising the amino acid sequence represented by formula (1), wherein X1 is not a phenylalanine residue, or a pharmaceutically acceptable salt thereof.

[16] The peptide molecule according to any one of [1] to

[15] above, comprising 11 or 12 amino acid residues, or a pharmaceutically acceptable salt thereof.

[17] An inhibitor of extracellular trap formation by leukocytes, comprising the peptide molecule according to any one of [1] to

[16] above or a pharmaceutically acceptable salt thereof.

[18] A pharmaceutical composition for treating or preventing a disease associated with extracellular trap formation by leukocytes, comprising the peptide molecule according to any one of [1] to

[16] above or a pharmaceutically acceptable salt thereof.

[19] A pharmaceutical composition for treating or preventing acute kidney injury, comprising the peptide molecule or a pharmaceutically acceptable salt thereof according to any one of [1] to

[16] above.

[20] The pharmaceutical composition according to

[19] above, wherein the acute kidney injury is post-rhabdomyolytic acute kidney injury.

[0009] The present invention provides novel peptide molecules having inhibitory activity against leukocyte extracellular trap formation. According to one aspect of the present invention, the peptide molecules may have good in vitro activity with respect to inhibitory activity against leukocyte extracellular trap formation. According to one aspect of the present invention, the peptide molecules may have good in vivo activity and / or in vivo stability with respect to inhibitory activity against leukocyte extracellular trap formation. According to one aspect of the present invention, the peptide molecules may exhibit good safety or tolerability. According to one aspect of the present invention, it may be possible to provide a pharmaceutical containing, as an active ingredient, a peptide molecule having inhibitory activity against leukocyte extracellular trap formation.

[0010] This graph shows the inhibitory activity of test substances (F1A, K2A, C3A, R4A, R5A, W6A, Q7A, W8A, R9A, M10A, K11A, K12A, and FK-12 amide) on neutrophil extracellular trap (NET) formation. NET formation was induced by stimulating human neutrophils with 25 nM phorbol 12-myristate 13-acetate (PMA). Each test substance was added to the culture medium at a concentration of 40 μg / mL (22.9 μM). The vertical axis represents the relative amount of DNA released, with the value in the absence of test substance taken as 1. *: p < 0.05, **: p < 0.01. This graph shows the inhibitory activity of test substances (F1W, F1Cha, F1Nal, W6Hph, W6Cha, and FK-12 amide) on NET formation. NET formation was induced by stimulating human neutrophils with 25 nM PMA. Each test substance was added to the culture medium at a concentration of 120 μg / mL (68.7 μM). The vertical axis represents the relative amount of DNA released, with the value in the absence of test substance taken as 1. ***: p < 0.001. This graph shows the NET formation inhibitory activity of test substances (Q7E, Q7N, Q7D, Q7Orn, and FK-12 amide). NET formation was induced by stimulating human neutrophils with 25 nM PMA. Each test substance was added to the culture medium at a concentration of 120 μg / mL (68.7 μM). The vertical axis represents the relative amount of DNA released, with the value in the absence of test substance taken as 1. ***: p < 0.001. This graph shows the inhibitory activity of test substances (W8Nal, W8Cha, M10Nle, M10Hse(Me), M10Thi, and FK-12 amide) on NET formation. NET formation was induced by stimulating human neutrophils with 25 nM PMA. Each test substance was added to the culture medium at a concentration of 120 μg / mL (68.7 μM). The vertical axis represents the relative amount of DNA released, with the value in the absence of test substance taken as 1. ***: p < 0.001. This graph shows the effect of test substances (Q7Orn, M10Hse(Me), and FK-12 amide) on blood urea nitrogen (BUN) in a mouse model of glycerol-induced rhabdomyolysis. Each test substance was intraperitoneally administered at 720 μg / 100 μL. The control group received 100 μL of PBS intraperitoneally (Control). n represents the number of individuals.*: p < 0.05, **: p < 0.01. Graph showing the effect of test substances (Q7Orn, M10Hse(Me), and FK-12 amide) on blood creatinine (Cr) levels in a mouse model of glycerol-induced rhabdomyolysis. 720 μg / 100 μL of each test substance was administered intraperitoneally. 100 μL of PBS was administered intraperitoneally to a control group (Control). n represents the number of animals. **: p < 0.01, ***: p < 0.001. Graph showing the results of evaluation of urinary occult blood (UOB) for the test substance-administered group (M10Hse(Me)) and the control group (Control) in a mouse model of glycerol-induced rhabdomyolysis. n represents the number of animals. *: p < 0.05. 1 is a graph showing the time course of BUN for the test substance administration group (M10Hse(Me)) and the control group (Control) in a glycerol-induced rhabdomyolysis model mouse. In the test substance administration group, 720 μg / 100 μL of the test substance was intraperitoneally administered 6 hours after glycerol administration. In the control group, 100 μL of PBS was intraperitoneally administered 6 hours after glycerol administration. n represents the number of individuals (upper row: test substance administration group, lower row: control group). *: p < 0.05, **: p < 0.01. FIG. 1 is a graph showing the time course of Cr values ​​for the test substance administration group (M10Hse(Me)) and the control group (Control) in a glycerol-induced rhabdomyolysis model mouse. In the test substance administration group, 720 μg / 100 μL of the test substance was intraperitoneally administered 6 hours after glycerol administration. To the control group, 100 μL of PBS was administered intraperitoneally 6 hours after glycerol administration. n represents the number of individuals (upper row: test substance-administered group, lower row: control group). *: p < 0.05. This graph shows the number of survivors for the test substance-administered and control groups in a glycerol-induced rhabdomyolysis model mouse. To the test substance-administered group, 720 μg / 100 μL of the test substance (M10Hse(Me)) was administered intraperitoneally 6 hours after glycerol administration. To the control group, 100 μL of PBS was administered intraperitoneally 6 hours after glycerol administration. This is a microscopic photograph of skin tissue from the control group in an imiquimide (IMQ)-induced psoriasis model mouse.The control group received intraperitoneal administration of 100 μL of PBS once daily for 7 days. These are microscopic photographs of skin tissue from the test substance administration group in imiquimide (IMQ)-induced psoriasis model mice. The test substance administration group received intraperitoneal administration of 720 μg / 100 μL of M10Hse(Me) once daily for 7 days. These are graphs showing the epidermal thickness for the control group (Control) and the test substance administration group (M10Hse(Me)) in imiquimide (IMQ)-induced psoriasis model mice. These are graphs showing the thickness of the entire skin tissue (epidermis + dermis + subcutaneous tissue) for the control group (Control) and the test substance administration group (M10Hse(Me)) in imiquimide (IMQ)-induced psoriasis model mice.

[0011] As used herein, an amino acid refers to an organic compound containing an amino group and a carboxy group, and may be a natural amino acid or an unnatural amino acid. A natural amino acid refers to a naturally occurring, unmodified amino acid. Furthermore, as used herein, an amino acid may be in the L-form, the D-form, or the DL-form (racemic form), unless otherwise specified. Preferably, the L-form is used unless otherwise specified. As used herein, amino acids may be represented by the one-letter or three-letter symbols shown in Table 1.

[0012]

[0013] Unless otherwise specified, the amino acid sequences described herein are written in the direction from the N-terminus (amino terminus) to the C-terminus (carboxyl terminus). In the peptide molecules described herein, the term "amino acid" refers to an amino acid residue. As used herein, the term "amino acid residue" refers to a portion of a peptide or protein molecule that corresponds to one unit of an amino acid constituting the peptide or protein. More specifically, it refers to a divalent group derived from an α-amino acid, as represented by the following formula (A):

[0014] In formula (A), R 0 is the side chain of an amino acid, for example, a hydrogen atom for Gly, or a methyl group for Ala.

[0015] As used herein, the term "alkyl group" refers to a linear or branched saturated hydrocarbon group. Examples of alkyl groups include, but are not limited to, alkyl groups having 1 to 12 carbon atoms. In the present invention, the alkyl group may be an alkyl group having 1 to 10 carbon atoms, an alkyl group having 1 to 8 carbon atoms, an alkyl group having 1 to 6 carbon atoms, an alkyl group having 1 to 4 carbon atoms, or an alkyl group having 1 to 3 carbon atoms. Specific examples of alkyl groups include, for example, a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, an isobutyl group, a sec-butyl group, a tert-butyl group, a pentyl group, an isopentyl group, a neopentyl group, and a hexyl group. As used herein, the term "alicyclic hydrocarbon group" refers to a non-aromatic cyclic saturated or unsaturated hydrocarbon group. An alicyclic hydrocarbon group may be monocyclic or have multiple rings (e.g., bicyclic), and may be substituted or unsubstituted. Examples of alicyclic hydrocarbon groups include, but are not limited to, cycloalkyl groups and cycloalkenyl groups. The number of carbon atoms in the alicyclic hydrocarbon group is not particularly limited, but is, for example, 3 or more, 4 or more, or 5 or more, and 12 or less, 10 or less, 8 or less, 7 or less, or 6 or less. In this specification, "cycloalkyl group" means a cyclic saturated hydrocarbon group. The cycloalkyl group may be monocyclic or may have multiple rings (e.g., bicyclic). Examples of cycloalkyl groups include, but are not limited to, cycloalkyl groups having 3 to 12 carbon atoms. The number of carbon atoms in the cycloalkyl group is not particularly limited, but is, for example, 3 or more, 4 or more, or 5 or more, and 12 or less, 10 or less, 8 or less, 7 or less, or 6 or less. In the present invention, the cycloalkyl group may be a cycloalkyl group having 3 to 10 carbon atoms, a cycloalkyl group having 3 to 8 carbon atoms, a cycloalkyl group having 3 to 7 carbon atoms, or a cycloalkyl group having 3 to 6 carbon atoms. Specific examples of the cycloalkyl group include a cyclodecyl group, a cyclononyl group, a cyclooctyl group, a cycloheptyl group, a cyclohexyl group, a cyclopentyl group, a cyclobutyl group, a cyclopropyl group, a decahydronaphthalenyl group, a bicyclo[4.3.0]nonyl group, a norbornyl group, an isobornyl group, and an adamantyl group.As used herein, the term "cycloalkenyl group" refers to a non-aromatic, cyclic, unsaturated hydrocarbon group having at least one carbon-carbon double bond. The cycloalkenyl group may be monocyclic or may have multiple rings (e.g., bicyclic). Examples of cycloalkenyl groups include, but are not limited to, cycloalkenyl groups having 3 to 12 carbon atoms. The number of carbon atoms in the cycloalkenyl group is not particularly limited, but may be, for example, 3 or more, 4 or more, or 5 or more, and 12 or less, 10 or less, 8 or less, 7 or less, or 6 or less. In the present invention, the cycloalkenyl group may be a cycloalkenyl group having 3 to 10 carbon atoms, a cycloalkenyl group having 3 to 8 carbon atoms, a cycloalkenyl group having 3 to 7 carbon atoms, or a cycloalkenyl group having 3 to 6 carbon atoms. Specific examples of cycloalkenyl groups include cyclodecenyl groups, cyclononenyl groups, cyclooctenyl groups, cycloheptenyl groups, cyclohexenyl groups, cyclopentenyl groups, cyclobutenyl groups, cyclopropenyl groups, hexahydronaphthalenyl groups, octahydronaphthalenyl groups, tetrahydroindenyl groups, and hexahydroindenyl groups.

[0016] As used herein, the term "aromatic hydrocarbon group" refers to an aromatic hydrocarbon group of a monocyclic or fused ring structure (e.g., bicyclic), which may be substituted or unsubstituted. Examples of aromatic hydrocarbon groups include, but are not limited to, aromatic hydrocarbon groups having 6 to 20 carbon atoms. In the present invention, the aromatic hydrocarbon group may be an aromatic hydrocarbon group having 6 to 15 carbon atoms or an aromatic hydrocarbon group having 6 to 12 carbon atoms. Specific examples of aromatic hydrocarbon groups include a phenyl group, a naphthyl group, a tolyl group, a phenanthryl group, a tetrahydronaphthyl group, an indenyl group, and an indanyl group. As used herein, the term "heteroaromatic group" refers to an aromatic group of a monocyclic or fused ring structure (e.g., bicyclic), in which the atoms constituting the ring contain heteroatoms, which may be substituted or unsubstituted. For example, the number of atoms constituting the ring of the heteroaromatic group is 5 or more or 6 or more, and 20 or less, 15 or less, 12 or less, 10 or less, 8 or less, 7 or less, or 6 or less. In the present invention, the heteroaromatic group may contain 1 to 4, 1 to 3, 1 to 2, or 1 heteroatom selected from oxygen, sulfur, and nitrogen atoms constituting its ring. Specific examples of heteroaromatic groups include pyridyl, pyrrolyl, pyrazyl, pyrimidyl, pyridazyl, pyrazolyl, imidazolyl, indolyl, puryl, quinolyl, isoquinolyl, benzimidazolyl, carbazolyl, triazolyl, tetrazolyl, furanyl, thienyl, thiazolyl, isothiazolyl, benzothiazolyl, thiadiazolyl, oxazolyl, isoxazolyl, and benzoxazolyl. In this specification, the term "heteroalicyclic group" refers to a non-aromatic group having a monocyclic or fused ring structure (e.g., bicyclic) and containing heteroatoms constituting the ring, and may be substituted or unsubstituted. For example, the number of atoms constituting the ring of the heteroalicyclic group is 3 or more, 4 or more, or 5 or more, and 20 or less, 15 or less, 12 or less, 10 or less, 8 or less, 7 or less, or 6 or less. In the present invention, the atoms constituting the ring of the heteroalicyclic group may include 1 to 4, 1 to 3, 1 to 2, or 1 heteroatom selected from oxygen atoms, sulfur atoms, and nitrogen atoms.Specific examples of the heteroalicyclic group include a piperazyl group, a piperidyl group, a pyrrolidyl group, a pyrrolinyl group, a pyrazolidinyl group, a pyrazolinyl group, a morpholyl group, an oxetanyl group, a thietanyl group, a tetrahydrofuranyl group, a tetrahydrothienyl group, a tetrahydropyranyl group, a tetrahydrothiopyranyl group, a pyranyl group, a thiopyranyl group, a dioxanyl group, and a morpholinyl group. When the aromatic hydrocarbon group, heteroaromatic group, or heteroalicyclic group has a substituent, examples of the substituent include a linear or branched alkyl group having 1 to 6 carbon atoms (e.g., methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, or hexyl group), a linear or branched alkoxy group having 1 to 6 carbon atoms, an amino group, a carboxyl group, an ester group, a carbamoyl group, an amido group, a nitro group, a sulfo group, a sulfonamido group, an oxo group, and / or a halogen atom (e.g., a fluorine atom, a chlorine atom, a bromine atom, or an iodine atom).

[0017] One aspect of the present invention relates to a peptide molecule. This peptide molecule has a structure containing an amino acid sequence represented by the following formula (1) or an amino acid sequence represented by the following formula (2), and has activity of inhibiting the formation of extracellular traps in leukocytes: X1-KCRR-X2-Q-X3-R-X4-KK (1) R 1 -CH2CH2-CO-KCRR-X2-Q-X3-R-X4-KK (2) [In each formula, X1 is R 1 an α-amino acid residue having a side chain represented by —CH—, R 1 is a hydrogen atom, an alkyl group, an alicyclic hydrocarbon group, an aromatic hydrocarbon group, a heteroalicyclic group, or a heteroaromatic group, and X2 represents a structure represented by the following formula (3): (In the formula, R 2 is a hydrogen atom, an alkyl group, an aromatic hydrocarbon group, or a heteroaromatic group, and n is 1 or 2. X3 represents a structure represented by the following formula (4): (In the formula, R 3is a hydrogen atom, an alkyl group, an alicyclic hydrocarbon group, an aromatic hydrocarbon group, a heteroalicyclic group, or a heteroaromatic group; X4 is a structure represented by the following formula (5): (In the formula, R 4 is a hydrogen atom, or -R 5 -Z-R 6 Or -Z-R 7 R 5 is an alkylene group, and R 6 is an alkyl group, or R 5 and R 6 form a heterocycle together with Z to which they are attached, and R 7 represents an alkyl group, and Z represents an oxygen atom or a sulfur atom. 1 , R 2 , and R 3 The heteroalicyclic group or heteroaromatic group in the formula (I) is one in which the atoms constituting the ring contain 1 to 4 heteroatoms selected from oxygen atoms, sulfur atoms, and nitrogen atoms. In the present invention, the amino acid sequence represented by formula (1) does not correspond to any of FKCRRWQWRMKK (SEQ ID NO: 1), AKCRRWQWRMKK (SEQ ID NO: 2), FKCRRFQWRMKK (SEQ ID NO: 3), AKCRRFQWRMKK (SEQ ID NO: 4), FKCRRWQVRMKK (SEQ ID NO: 5), AKCRRWQVRMKK (SEQ ID NO: 6), FKCRRFQVRMKK (SEQ ID NO: 7), and AKCRRFQVRMKK (SEQ ID NO: 8).

[0018] In one embodiment of the present invention, in X in formula (1) or in formula (2), R 1 may be a hydrogen atom, an alkyl group, a monocyclic or bicyclic cycloalkyl group, a monocyclic or bicyclic cycloalkenyl group, a monocyclic or bicyclic aromatic hydrocarbon group, or a monocyclic or bicyclic heteroaromatic group. Here, the monocyclic or bicyclic heteroaromatic group has ring atoms containing 1 to 4 heteroatoms selected from oxygen atoms, sulfur atoms, and nitrogen atoms. In one embodiment of the present invention, X in formula (1) or R in formula (2) 1may be a hydrogen atom, an alkyl group having 1 to 9 carbon atoms, a monocyclic cycloalkyl group having 3 to 10 carbon atoms, a monocyclic cycloalkenyl group having 3 to 10 carbon atoms, a monocyclic aromatic hydrocarbon group, a monocyclic heteroaromatic group, or a bicyclic group having a cyclic structure represented by the following formula (6): (wherein ring A is a ring that makes the structure represented by formula (6) a 9- to 11-membered bicyclic ring structure, the atoms constituting ring A include at least one carbon atom and may include one or two heteroatoms selected from oxygen atoms, sulfur atoms, and nitrogen atoms, Z1, Z2, Z3, and Z4 are each independently a carbon atom or a nitrogen atom, and the number of nitrogen atoms in Z1, Z2, Z3, and Z4 is a maximum of 2, represents a single bond or a double bond, and the point of attachment to the CH2 moiety in the side chain of X1 in formula (1) or in the structure represented by formula (2) may be ring A or the ring having Z1.) Examples of the bicyclic group having a cyclic structure represented by formula (6) include bicyclic aromatic hydrocarbon groups, bicyclic heteroaromatic groups, bicyclic cycloalkyl groups, and bicyclic cycloalkenyl groups such as naphthyl group, tetrahydronaphthyl group, indenyl group, indanyl group, indolyl group, pryl group, quinolyl group, isoquinolyl group, benzimidazolyl group, benzothiazolyl group, benzoxazolyl group, decahydronaphthalenyl group, bicyclo[4.3.0]nonyl group, hexahydronaphthalenyl group, octahydronaphthalenyl group, tetrahydroindenyl group, and hexahydroindenyl group.

[0019] In one embodiment of the present invention, in X in formula (1) or in formula (2), R 1 may be a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, a monocyclic cycloalkyl group having 3 to 7 carbon atoms (preferably a cyclohexyl group), or an aromatic hydrocarbon group or bicyclic heteroaromatic group having a cyclic structure represented by the following formula (6-1): (In the formula, ring A1 is a ring that makes the structure represented by formula (6-1) a 9- to 10-membered bicyclic aromatic ring, or is absent; the atoms constituting ring A1 include at least one carbon atom and may include one or two heteroatoms selected from oxygen atoms, sulfur atoms, and nitrogen atoms; and the point of attachment to the CH2 moiety in the side chain of X1 in formula (1) or in the structure represented by formula (2) may be ring A1 or a benzene ring.) Examples of the aromatic hydrocarbon group or bicyclic heteroaromatic group having a cyclic structure represented by formula (6-1) include a phenyl group, a naphthyl group, a tetrahydronaphthyl group, an indenyl group, an indanyl group, an indolyl group, a quinolyl group, an isoquinolyl group, a benzimidazolyl group, a benzothiazolyl group, and a benzoxazolyl group. Of these, a phenyl group, a naphthyl group, an indenyl group, an indolyl group, a quinolyl group, an isoquinolyl group, or a benzimidazolyl group is preferred, and a phenyl group, a naphthyl group, or an indolyl group is more preferred. In one embodiment of the present invention, X1 in formula (1) may be an alanine residue, a phenylalanine residue, a tryptophan residue, a cyclohexylalanine residue, or a 2-naphthylalanine residue. In one embodiment of the present invention, X1 in formula (1) is not a phenylalanine residue. In one embodiment of the present invention, R1 in formula (2) is not a phenyl group.

[0020] In one embodiment of the present invention, in formula (3), R 2 may be a hydrogen atom, an alkyl group, a monocyclic or bicyclic aromatic hydrocarbon group, or a monocyclic or bicyclic heteroaromatic group. Here, the monocyclic or bicyclic heteroaromatic group has ring atoms containing 1 to 4 heteroatoms selected from oxygen, sulfur, and nitrogen atoms. In one embodiment of the present invention, in formula (3), R 2 may be a hydrogen atom, an alkyl group having 1 to 9 carbon atoms, a monocyclic aromatic hydrocarbon group having 3 to 10 carbon atoms, a monocyclic heteroaromatic group having 3 to 10 carbon atoms, or a bicyclic group having a cyclic structure represented by the following formula (7): (wherein ring B is a ring that forms a 9- to 11-membered bicyclic aromatic ring in the structure represented by formula (7), and the atoms constituting ring B include at least one carbon atom and optionally one or two heteroatoms selected from an oxygen atom, a sulfur atom, and a nitrogen atom; Z1, Z2, Z3, and Z4 are each independently a carbon atom or a nitrogen atom, and the number of nitrogen atoms in Z1, Z2, Z3, and Z4 is a maximum of two; and the bonding point to the CH2 moiety in the structure represented by formula (3) may be ring B or the ring having Z1.) Examples of bicyclic groups having a cyclic structure represented by formula (7) include bicyclic aromatic hydrocarbon groups or bicyclic heteroaromatic groups such as naphthyl, tetrahydronaphthyl, indenyl, indanyl, indolyl, pryl, quinolyl, isoquinolyl, benzimidazolyl, benzothiazolyl, and benzoxazolyl groups.

[0021] In one embodiment of the present invention, in formula (3), R 2 can be a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, or an aromatic hydrocarbon group or bicyclic heteroaromatic group having a cyclic structure represented by the following formula (7-1): (In the formula, ring B1 is a ring that forms a 9- to 10-membered bicyclic aromatic ring in the structure represented by formula (7-1), or is absent; the atoms constituting ring B1 include at least one carbon atom and optionally one or two heteroatoms selected from oxygen, sulfur, and nitrogen atoms; and the bonding point to the CH2 moiety in the structure represented by formula (3) may be ring B1 or a benzene ring.) Examples of the aromatic hydrocarbon group or bicyclic heteroaromatic group having a cyclic structure represented by formula (7-1) include a phenyl group, a naphthyl group, a tetrahydronaphthyl group, an indenyl group, an indanyl group, an indolyl group, a quinolyl group, an isoquinolyl group, a benzimidazolyl group, a benzothiazolyl group, and a benzoxazolyl group, and the like are preferred, and a phenyl group, a naphthyl group, an indenyl group, an indolyl group, a quinolyl group, an isoquinolyl group, or a benzimidazolyl group is more preferred. In one embodiment of the present invention, in formula (1) or (2), X2 may be an alanine residue, a tryptophan residue, or a homophenylalanine residue.

[0022] In one embodiment of the present invention, in formula (4), R 3 may be a hydrogen atom, an alkyl group, a monocyclic or bicyclic cycloalkyl group, a monocyclic or bicyclic cycloalkenyl group, a monocyclic or bicyclic aromatic hydrocarbon group, or a monocyclic or bicyclic heteroaromatic group. Here, the monocyclic or bicyclic heteroaromatic group has ring atoms containing 1 to 4 heteroatoms selected from oxygen, sulfur, and nitrogen atoms. In one aspect of the present invention, in formula (4), R 3 may be a hydrogen atom, an alkyl group having 1 to 9 carbon atoms, a monocyclic cycloalkyl group having 3 to 10 carbon atoms, a monocyclic cycloalkenyl group having 3 to 10 carbon atoms, a monocyclic aromatic hydrocarbon group, a monocyclic heteroaromatic group, or a bicyclic group having a cyclic structure represented by the following formula (8): (wherein ring C is a ring that makes the structure represented by formula (8) a 9- to 11-membered bicyclic ring structure, and the atoms constituting ring C include at least one carbon atom and may include one or two heteroatoms selected from an oxygen atom, a sulfur atom, and a nitrogen atom; Z1, Z2, Z3, and Z4 are each independently a carbon atom or a nitrogen atom, and the number of nitrogen atoms in Z1, Z2, Z3, and Z4 is a maximum of 2; represents a single bond or a double bond, and the point of attachment to the CH2 moiety in the structure represented by formula (4) may be either the C-ring or the ring having Z1.) Examples of the bicyclic group having a cyclic structure represented by formula (8) include bicyclic aromatic hydrocarbon groups, bicyclic heteroaromatic groups, bicyclic cycloalkyl groups, and bicyclic cycloalkenyl groups such as naphthyl group, tetrahydronaphthyl group, indenyl group, indanyl group, indolyl group, pryl group, quinolyl group, isoquinolyl group, benzimidazolyl group, benzothiazolyl group, benzoxazolyl group, decahydronaphthalenyl group, bicyclo[4.3.0]nonyl group, hexahydronaphthalenyl group, octahydronaphthalenyl group, tetrahydroindenyl group, and hexahydroindenyl group.

[0023] In one embodiment of the present invention, in formula (4), R 3 may be a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, a monocyclic cycloalkyl group having 3 to 7 carbon atoms (preferably a cyclohexyl group), or an aromatic hydrocarbon group or bicyclic heteroaromatic group having a cyclic structure represented by the following formula (8-1): (In the formula, the C1 ring is a ring that forms a 9- to 10-membered bicyclic aromatic ring in the structure represented by formula (8-1), or is absent; the atoms constituting the C1 ring include at least one carbon atom and optionally one or two heteroatoms selected from an oxygen atom, a sulfur atom, and a nitrogen atom; and the bonding point to the CH2 moiety in the structure represented by formula (4) may be the C1 ring or a benzene ring.) Examples of the aromatic hydrocarbon group or bicyclic heteroaromatic group having a cyclic structure represented by formula (8-1) include a phenyl group, a naphthyl group, a tetrahydronaphthyl group, an indenyl group, an indanyl group, an indolyl group, a quinolyl group, an isoquinolyl group, a benzimidazolyl group, a benzothiazolyl group, and a benzoxazolyl group, and the like are preferred, and a naphthyl group, an indenyl group, an indolyl group, a quinolyl group, an isoquinolyl group, or a benzimidazolyl group is more preferred. In one embodiment of the present invention, in formula (1) or (2), X3 can be an alanine residue, a tryptophan residue, a cyclohexylalanine residue, or a 2-naphthylalanine residue.

[0024] In one embodiment of the present invention, in formula (5), R 4 is a hydrogen atom or -R 5 -Z-R 6 Or -Z-R 7 and R 5 is an alkylene group having 1 to 6 carbon atoms, and R 6 is an alkyl group having 1 to 6 carbon atoms, and R 7 In one embodiment of the present invention, in formula (5), R 4 is a hydrogen atom or -R 5 -Z-R 6 Or -Z-R 7 and R 5 and R 6 form a 4- to 7-membered heterocyclic ring together with Z to which they are attached, and R 7 In one embodiment of the present invention, in formula (5), R 4 is -R 5 -Z-R6 and R 5 is an alkylene group having 1 to 3 carbon atoms, and R 6 In one embodiment of the present invention, in formula (5), R 4 is -R 5 -Z-R 6 and R 5 is an alkylene group having 1 to 3 carbon atoms, and R 6 may be an alkyl group having 1 to 3 carbon atoms, and Z may be an oxygen atom. 4 is -R 5 -Z-R 6 and R 5 and R 6 can form a 5- or 6-membered heterocycle together with Z to which they are attached. Examples of such 5- or 6-membered heterocycles include furan, thiophene, pyran, thiopyran, tetrahydrofuran, tetrahydrothiophene, tetrahydropyran, and tetrahydrothiopyran, and are preferably furan, thiophene, pyran, or thiopyran, and more preferably thiophene. In one embodiment of the present invention, in formula (1) or (2), X4 is not a methionine residue. In one embodiment of the present invention, in formula (1) or (2), X4 can be an O-methylhomoserine residue or a 2-thienylalanine residue, and is preferably an O-methylhomoserine residue.

[0025] Although examples of each group are given individually above, the examples of each group may be in any combination. For example, R 1 Example of R 2 Combination with examples of R 1 Example of R 3 Combination with examples of R 1 Example of R 4 Combination with examples of R 2 Example of R 3 Combination with examples of R 2 Example of R 4 Combination with examples of R 3 Example of R 4Any combination of groups is disclosed herein, such as the combination with the examples of: Such combinations are not limited to combinations of two groups, but can also be combinations of three or more groups.

[0026] In one embodiment of the present invention, in the amino acid sequence represented by formula (1), X1 is an alanine residue, a phenylalanine residue, a tryptophan residue, a cyclohexylalanine residue, or a 2-naphthylalanine residue, X2 is an alanine residue, a tryptophan residue, or a homophenylalanine residue, X3 is an alanine residue, a tryptophan residue, a cyclohexylalanine residue, or a 2-naphthylalanine residue, and X4 is an alanine residue, a methionine residue, an O-methylhomoserine residue, or a 2-thienylalanine residue. However, as described above, the amino acid sequence represented by formula (1) does not correspond to any of FKCRRWQWRMKK, AKCRRWQWRMKK, FKCRRFQWRMKK, AKCRRFQWRMKK, FKCRRWQVRMKK, AKCRRWQVRMKK, FKCRRFQVRMKK, and AKCRRFQVRMKK. In one aspect of the present invention, in the amino acid sequence in the structure represented by formula (2), X2 is an alanine residue, tryptophan residue, or homophenylalanine residue, X3 is an alanine residue, tryptophan residue, cyclohexylalanine residue, or 2-naphthylalanine residue, X4 is an alanine residue, methionine residue, O-methylhomoserine residue, or 2-thienylalanine residue, and in the structure represented by formula (2), R 1 is a hydrogen atom, a phenyl group, a 2-indolyl group, a cyclohexyl group, or a 2-naphthyl group.

[0027] In one embodiment of the present invention, in the amino acid sequence represented by formula (1), X1 is an alanine residue, phenylalanine residue, tryptophan residue, cyclohexylalanine residue, or 2-naphthylalanine residue, X2 is an alanine residue, tryptophan residue, or homophenylalanine residue, X3 is an alanine residue, tryptophan residue, cyclohexylalanine residue, or 2-naphthylalanine residue, and X4 is an O-methylhomoserine residue. In one embodiment of the present invention, in the amino acid sequence in the structure represented by formula (2), X2 is an alanine residue, tryptophan residue, or homophenylalanine residue, X3 is an alanine residue, tryptophan residue, cyclohexylalanine residue, or 2-naphthylalanine residue, X4 is an O-methylhomoserine residue, and in the structure represented by formula (2), R 1 is a hydrogen atom, a phenyl group, a 2-indolyl group, a cyclohexyl group, or a 2-naphthyl group.

[0028] In one embodiment of the present invention, the amino acid sequence represented by formula (1) may be selected from the following: WKCRRWQWRMKK (SEQ ID NO: 9) XKCRRWQWRMKK (SEQ ID NO: 10) (X represents cyclohexylalanine) XKCRRWQWRMKK (SEQ ID NO: 11) (X represents 2-naphthylalanine) FKCRRXQWRMKK (SEQ ID NO: 12) (X represents homophenylalanine) FKCRRWQXRMKK (SEQ ID NO: 13) (X represents 2-naphthylalanine) FKCRRWQXRMKK (SEQ ID NO: 14) (X represents cyclohexylalanine) FKCRRWQWRXKK (SEQ ID NO: 15) (X represents O-methylhomoserine) FKCRRWQWRXKK (SEQ ID NO: 16) (X represents 2-thienylalanine)

[0029] In one embodiment of the present invention, the structure represented by formula (2) can be selected from the following: The amino acid sequence in the structure represented by formula (2) is KCRRWQWRMKK (SEQ ID NO: 17), and preferably R 1is a 2-indolyl group, a cyclohexyl group, or a 2-naphthyl group. The amino acid sequence in the structure represented by formula (2) is KCRRXQWRMKK (SEQ ID NO: 18) (X represents homophenylalanine), and preferably, R 1 is a phenyl group. The amino acid sequence in the structure represented by formula (2) is KCRRWQXRMKK (SEQ ID NO: 19) (X represents 2-naphthylalanine), and preferably, R 1 is a phenyl group. The amino acid sequence in the structure represented by formula (2) is KCRRWQXRMKK (SEQ ID NO: 20) (X represents cyclohexylalanine), and preferably, R 1 is a phenyl group. The amino acid sequence in the structure represented by formula (2) is KCRRWQWRXKK (SEQ ID NO: 21) (X represents O-methylhomoserine), and preferably, R 1 is a phenyl group. The amino acid sequence in the structure represented by formula (2) is KCRRWQWRXKK (SEQ ID NO: 22) (X represents 2-thienylalanine), and preferably, R 1 is a phenyl group.

[0030] In the present invention, a peptide molecule has a structure containing the amino acid sequence represented by formula (1) or the amino acid sequence represented by formula (2), and may contain an additional amino acid sequence on the N-terminal side of the amino acid sequence represented by formula (1) and / or on the C-terminal side of the amino acid sequence represented by formula (1) or formula (2), as long as the peptide molecule has the activity of inhibiting the formation of extracellular traps in leukocytes. Such additional amino acid sequences can be appropriately designed with reference to WO 2016 / 056665, etc. In the present invention, the number of amino acid residues in the peptide molecule is preferably 11 or 12. That is, the peptide molecule preferably does not contain any additional amino acid residues other than those specified in the structure containing the amino acid sequence represented by formula (1) or the amino acid sequence represented by formula (2).

[0031] In the present invention, the structure of the N-terminal side of a peptide molecule containing the amino acid sequence represented by formula (1) is not particularly limited, and may be, for example, a hydrogen atom (i.e., unmodified) or a structure to which a modifying group has been introduced by a conventionally known method. Examples of modifying groups at the N-terminus include alkyl groups having 1 to 20 carbon atoms, cycloalkyl groups having 1 to 20 carbon atoms, alkenyl groups having 1 to 20 carbon atoms, alkynyl groups having 1 to 20 carbon atoms, aromatic hydrocarbon groups having 6 to 20 carbon atoms, heterocyclic groups, groups represented by the following formula (B), sulfonyl groups, carboxyl groups, glyoxyl groups, formyl groups; polyethylene glycol groups (PEGylated), polyoxyethylene glycol groups, polypropylene glycol groups; tert-butoxycarbonyl groups (Boc groups), benzyloxycarbonyl groups, and the like. Examples of protecting groups include a methyl group (Z group), a fluorenylmethoxycarbonyl group (Fmoc group), and the like; cycloalkyloxycarbonyl groups such as a cyclopentyloxycarbonyl group, a cyclohexyloxycarbonyl group, an adamantyloxycarbonyl group, a norbornyloxycarbonyl group, and an isobornyloxycarbonyl group; protecting groups derived from amino acids such as pyroglutamic acid and morotanoic acid; carbamate protecting groups; and protecting groups derived from sulfonic acids such as benzenesulfonic acid and phosphoric acid. The number of carbon atoms in the alkyl group, cycloalkyl group, alkenyl group, or alkynyl group that may be present on the N-terminal side of the peptide molecule is, for example, 1 to 20, and preferably 1 to 10. The alkyl group, alkenyl group, or alkynyl group that may be present on the N-terminal side may have a linear or branched chain structure. More specifically, examples of the alkyl group, cycloalkyl group, alkenyl group, or alkynyl group that may be present on the N-terminal side include a methyl group, an ethyl group, a propyl group, an isopropyl group, a cyclopropyl group, a butyl group, an isobutyl group, a sec-butyl group, a tert-butyl group, an amyl group, an isoamyl group, a tert-amyl group, a hexyl group, a cyclohexyl group, a heptyl group, an octyl group, a 2-ethylhexyl group, a nonyl group, and a decyl group.

[0032] The number of carbon atoms in the aromatic hydrocarbon group that may be present at the N-terminus of the peptide molecule is, for example, 6 to 20, and more specific examples include a phenyl group, a naphthyl group, a tolyl group, and a phenanthryl group. Examples of heterocyclic groups that may be present at the N-terminus of the peptide molecule include substituents having a monocyclic, fused bicyclic, or fused tricyclic structure containing 1 to 3 heteroatoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom in the ring, and more specific examples include a pyrrolidyl group, a pyrrole group, a piperidyl group, a pyridyl group, an imidazolyl group, a pyrazolyl group, an oxazolyl group, a thiazolyl group, a morpholyl group, an indolyl group, a benzimidazolyl group, a quinolyl group, a carbazolyl group, a tetrahydrofuranyl group, a tetrahydrothiophenyl group, a furanyl group, a thiophenyl group, a tetrahydropyranyl group, and a tetrahydrothiopyranyl group. These aromatic hydrocarbon groups and heterocyclic groups may be substituted with further substituents such as a linear or branched alkyl group having 1 to 6 carbon atoms, a linear or branched alkoxy group having 1 to 6 carbon atoms, an amino group, a carboxyl group, an ester group, a carbamoyl group, an amide group, a nitro group, a sulfo group, a sulfonamide group, an oxo group, and / or a halogen atom (e.g., a fluorine atom, a chlorine atom, a bromine atom, an iodine atom), etc. The modifying group at the N-terminus may be, for example, a functional group represented by the following formula (B):

[0033] However, in formula (B), X 0 is a single bond, an oxygen atom, or a sulfur atom, or a divalent linking group selected from the group consisting of alkylene groups having 1 to 3 carbon atoms (e.g., methylene, ethylene, trimethylene, and propylene groups), oxyalkylene groups having 1 to 3 carbon atoms (e.g., oxymethylene, oxyethylene, oxytrimethylene, and oxypropylene groups), and alkyleneoxy groups having 1 to 3 carbon atoms (e.g., methyleneoxy, ethyleneoxy, trimethyleneoxy, and propyleneoxy groups), which optionally have a substituent selected from the group consisting of amino groups, acetylamino groups, and propionylamino groups; R 10 represents an alkyl group, a cycloalkyl group, an alkenyl group, or an alkynyl group having 1 to 20 carbon atoms, which may have a substituent;

[0034] wherein R is selected from the group consisting of 11 ~R 30 are each independently selected from the group consisting of a hydrogen atom, a halogen atom (e.g., a fluorine atom, a chlorine atom, a bromine atom, an iodine atom), an alkyl group having 1 to 3 carbon atoms (i.e., a methyl group, an ethyl group, a propyl group), an alkoxy group having 1 to 3 carbon atoms (i.e., a methoxy group, an ethoxy group, a propoxy group), a hydroxyl group, and an amino group.

[0035] In the above formula (B), R 10 is an alkyl group, a cycloalkyl group, an alkenyl group, or an alkynyl group having 1 to 20 carbon atoms, which may have a substituent; 10 The number of carbon atoms in R is preferably 2 to 12. 10 The alkyl group, alkenyl group, or alkynyl group represented by R may have a linear or branched chain structure. 10 Examples of the substituent in the formula (B) include a hydroxy group, an alkoxy group having 1 to 5 carbon atoms (e.g., a methoxy group, an ethoxy group), an amino group, a carboxyl group, an ester group, a carbamoyl group, an amide group, a nitro group, a sulfo group, and a halogen atom (e.g., a fluorine atom, a chlorine atom, a bromine atom, an iodine atom). 0 is a single bond or a divalent linking group selected from the group consisting of alkylene groups having 1 to 3 carbon atoms and oxyalkylene groups having 1 to 3 carbon atoms, which may have a substituent selected from the group consisting of an amino group and an acetylamino group.

[0036] In one embodiment, the group represented by the formula (B) above is an acyl group. Examples of the acyl group include acyl groups derived from various carboxylic acids. More specifically, it may be an acyl group having an aliphatic chain, an aromatic ring, or a heterocyclic ring, or may be an acyl group derived from a compound selected from the group consisting of amino acids, vitamins having an acyl group, and nucleic acid bases having an acyl group. R in the formula (B) above 10

[0039] More specific examples of the acyl group, which is an alkyl group, cycloalkyl group, alkenyl group, or alkynyl group having 1 to 20 carbon atoms and which may have a substituent, include, but are not limited to, an acetyl group, a propionyl group, a butyryl group, an isobutyryl group, a valeryl group, an isovaleryl group, a pivaloyl group, a caproyl group, a caprinoyl group, a methylhexanoyl group, a cyclopropanecarbonyl group, an aminocyclopropanecarbonyl group, a cyclohexanecarbonyl group, a cyclohexylacetyl group, a cyclopentylpropionyl group, a cyclohexylpropionyl group, a cyclopentylbutanoyl group, a cyclohexylbutanoyl group, an adamantylacetyl group, a lauroyl group, a myristoyl group, a palmitoyl group, a stearoyl group, an oxalyl group, a malonyl group, a succinyl group, a glutaryl group, an adipoyl group, a glycol group, a lactoyl group, a glyceroyl group, a pyruvoyl group, and an acetoacetyl group. Examples of vitamins having an acyl group include nicotinic acid, pantothenic acid, biotin, pteroylglutamic acid (folic acid), orotic acid, fluoroorotic acid, α-lipoic acid, pyridoxic acid, biocytin, pteroic acid, 10-formylpteroic acid, 7,8-dihydrofolic acid, homopteroic acid, pterin-6-carboxylic acid, dihydrolipoic acid, hydroorotic acid, etc. The nucleic acid base having an acyl group refers to a base component constituting a nucleotide and a derivative thereof, and preferred examples include pyrimidine derivatives, such as 5-carboxymethyluracil and 5-carboxythiouracil.

[0037] Examples of sulfonyl groups that can be present at the N-terminus of a peptide molecule include those having a structure in which the carbonyl structure in the above-mentioned acyl group is converted to a sulfone structure. Polyethylene glycol groups that can be present at the N-terminus of a peptide molecule have a structure in which polyethylene glycol or its analogs are linked via an ester bond, an amine (-NH-), an acyl group (e.g., an acyl group having 1 to 12 carbon atoms), or a combination thereof. The number of carbon atoms in the polyethylene glycol group can be, for example, 2 to 20 (i.e., -(C2H4O) n -, where n=1 to 10), preferably 4 to 16 (i.e., -(CHO)n -, where n is 2 to 8. The end of the polyethylene glycol group opposite to the end linked to the N-terminus of the peptide molecule may be modified with an amino group or a protecting group generally used for protecting a hydroxyl group, such as an alkyl group having 1 to 6 carbon atoms (e.g., methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, sec-butyl, tert-butyl, amyl, isoamyl, tert-amyl, or hexyl).

[0038] In the present invention, the structure of the C-terminal side of a peptide molecule having a structure containing the amino acid sequence represented by formula (1) or the amino acid sequence represented by formula (2) is also not particularly limited, and may be a structure modified with a protecting group commonly used for protecting a carboxylic acid. More specifically, the C-terminal structure of the peptide molecule may be, for example, a carboxyl group (—COOH), a carboxylate (—COO - ), amide (-CONH2), alkylamide (-CONHR 31 , -CONR 3 1 R 32 ), ester (-COOR 31 ), acyloxyalkyl (—R 33 -OCOR 31 ), a phthalidyl group optionally substituted with an alkyl or alkoxy group having 1 to 4 carbon atoms (for example, a phthalidyl group, a dimethylphthalidyl group, a dimethoxyphthalidyl group), or a (5-methyl-2-oxo-1,3-dioxolen-4-yl)methyl group. Of these, the C-terminal structure of the peptide molecule is preferably an amide. R in the above alkylamide, ester, and acyloxyalkyl 31 and R 32are each independently an alkyl group or cycloalkyl group having 1 to 6 carbon atoms such as a methyl group, an ethyl group, a propyl group, an isopropyl group, a cyclopropyl group, a butyl group, an isobutyl group, a sec-butyl group, a tert-butyl group, an amyl group, an isoamyl group, a tert-amyl group, a hexyl group, or a cyclohexyl group; an aromatic hydrocarbon group having 6 to 10 carbon atoms such as a phenyl group or a naphthyl group; an aralkyl group having 7 to 18 carbon atoms such as a benzyl group, a phenethyl group, or a benzhydryl group; a sugar such as glucose; or a polyethylene glycol group which may be modified with an alkyl group having 1 to 6 carbon atoms (for example, a methyl group, an ethyl group, a propyl group, an isopropyl group, a cyclopropyl group, a butyl group, an isobutyl group, a sec-butyl group, a tert-butyl group, an amyl group, an isoamyl group, a tert-amyl group, or a hexyl group). 33 is an alkylene group having 1 to 4 carbon atoms, such as a methylene group, an ethylene group, an n-propylene group, an isopropylene group, an n-butylene group, an isobutylene group, an s-butylene group, or a t-butylene group.

[0039] In the present invention, the peptide molecule may be in the form of a pharmaceutically acceptable salt. As used herein, a "pharmaceutically acceptable salt" refers to a metal salt, an ammonium salt, an organic acid salt, an inorganic acid salt, or a salt with an organic or inorganic base that does not produce undesirable physiological effects after administration to a patient or subject. More specific examples include sodium salts, potassium salts, calcium salts, magnesium salts, barium salts, aluminum salts, zinc salts, ammonium salts, methylamine salts, ethylamine salts, aniline salts, dimethylamine salts, diethylamine salts, pyrrolidine salts, piperidine salts, morpholine salts, piperazine salts, trimethylamine salts, triethylamine salts, ethanolamine salts, diethanolamine salts, triethanolamine salts, hydrochloride, hydrobromide, nitrate, sulfate, phosphate, formate, acetate, trifluoroacetate, phthalate, fumarate, oxalate, tartrate, maleate, citrate, succinate, malate, methanesulfonate, benzenesulfonate, and p-toluenesulfonate.

[0040] In the present invention, the peptide molecule has the activity of inhibiting extracellular trap formation in leukocytes. Extracellular trap formation has been reported in various leukocytes, including neutrophils (Brinkmann, V., et al., Science 2004;303:1532-1535), basophils (Yousefi, S., et al., Nat Med 2008;14:949-953), mast cells (von Koeckritz-Blickwede M, et al., Blood 2008;111:3070-3080), and monocytes (Webster SJ, et al., J Immunol 2010;185:2968-2979), such as macrophages (Chow, OA, et al., Cell Host & Microbe, Volume 8, Issue 5, 445-454, November 18, 2010). It has been reported that the extracellular traps formed by these leukocytes share a common characteristic of releasing fibrillar components composed primarily of DNA and granule proteins (Simon, D., et al., Allergy 68 (2013) 409-416). In the present invention, peptide molecules aggregate and / or condense the fibrillar components, thereby inhibiting their release. Therefore, it has been found that the peptide molecules aggregate and / or condense fibrillar components released upon extracellular trap formation not only from neutrophils used in the Examples but also from other leukocytes (e.g., basophils, mast cells, and monocytes (e.g., macrophages)), thereby inhibiting the formation of extracellular traps by these leukocytes. In one aspect of the present invention, the peptide molecules may have an inhibitory activity against extracellular trap formation by leukocytes equivalent to or greater than that of FKCRRWQWRMKK (SEQ ID NO: 1), a partial amino acid sequence of bovine lactoferrin, or its C-terminal amidated form.

[0041] In the present invention, with regard to the activity of inhibiting leukocyte extracellular trap formation, the leukocytes are derived from organisms that form leukocyte extracellular traps, preferably from vertebrates, and more preferably from mammals. Examples of mammals include monkeys, cows, goats, sheep, humans, camels, horses, dogs, cats, mice, and rats, with humans being preferred. Furthermore, the leukocytes are derived from the above-mentioned sources and are preferably any one selected from the group consisting of neutrophils, eosinophils, basophils, monocytes, macrophages, and mast cells, more preferably any one selected from the group consisting of neutrophils, basophils, monocytes, macrophages, and mast cells, and even more preferably neutrophils. In the present invention, the activity of inhibiting leukocyte extracellular trap formation can be confirmed by culturing leukocytes in the presence and absence of a test substance (peptide molecule) and examining the culture system for a decrease in extracellular trap formation in the presence of the test substance by microscopic observation. Leukocyte culture can be performed appropriately using known techniques depending on the type of leukocyte. In such a test, leukocytes are preferably treated with an extracellular trap formation promoter, such as phorbol 12-myristate 13-acetate (PMA) or lipopolysaccharide (LPS). Alternatively, the culture supernatant can be collected after leukocyte culture and the DNA concentration in the supernatant measured to confirm whether the DNA concentration is reduced in the presence of the test substance compared to the absence of the test substance. DNA concentration can be easily measured using a commercially available kit, such as Quant-iT Picogreen dsDNA assay reagent (Life Technologies).Regarding the culture conditions for each leukocyte and the method for confirming extracellular traps, the following can be referred to: neutrophils (Brinkmann, V., et al., Science 2004;303:1532-1535 and A.K. Gupta. FEBS letters 2010;584:3193-3197, D.J. Novo. Antimicrob Agents Chemother. 2000;44(4):827-34), basophils (Yousefi, S., et al., Nat Med 2008;14:949-953), mast cells (von Koeckritz-Blickwede M, et al., Blood 2008;111:3070-3080), monocytes (Webster SJ, et al., J Immunol 2010;185:2968-2979), for example, macrophages (Chow, OA, et al., Cell Host & Microbe, Volume 8, Issue 5, pp. 445-454, November 18, 2010. In the present invention, the peptide molecule may be modified with a compound, as long as it has the activity of inhibiting the formation of extracellular traps by leukocytes. For example, the peptide molecule may be a modified protein bound with polyethylene glycol (Japanese Patent Nos. 4195486, 4261531, and WO 2009 / 113743), or a fusion protein fused with another protein or a fragment thereof (e.g., blood-stable proteins such as IgG or albumin or a fragment thereof) (Japanese Patent Application No. 2012-98085).

[0042] In the present invention, peptide molecules can be produced by known methods. Specifically, peptide molecules can be produced by chemical synthesis using known peptide synthesis methods. In such production methods, the raw materials, reagents, solid-phase resins, solvents, and other materials used may be commercially available products, or can be synthesized by those skilled in the art using organic chemistry techniques. Commercially available amino acids containing protecting groups can be used as they are.

[0043] One aspect of the present invention relates to an agent for inhibiting leukocyte extracellular trap formation, comprising a peptide molecule or a pharmaceutically acceptable salt thereof. The peptide molecule is as described above, has a structure comprising the amino acid sequence represented by formula (1) or the amino acid sequence represented by formula (2), and has the activity of inhibiting leukocyte extracellular trap formation. Another aspect of the present invention relates to a pharmaceutical composition for treating or preventing a disease associated with leukocyte extracellular trap formation, comprising a peptide molecule or a pharmaceutically acceptable salt thereof. The peptide molecule is as described above, has a structure comprising the amino acid sequence represented by formula (1) or the amino acid sequence represented by formula (2), and has the activity of inhibiting leukocyte extracellular trap formation. As used herein, "treatment" means amelioration of symptoms in a subject. Accordingly, a pharmaceutical composition for treating a disease associated with leukocyte extracellular trap formation may be one that improves the symptoms of the disease. As used herein, "prevention" means inhibition of disease progression in a subject. Accordingly, a pharmaceutical composition for preventing a disease associated with leukocyte extracellular trap formation may be one that inhibits the progression of the disease. In the present invention, the subject (patient) is an organism suffering from or at risk of suffering from a disease associated with leukocyte extracellular trap formation, preferably a vertebrate, more preferably a mammal. Examples of mammals include mammals selected from the group consisting of humans, cows, horses, goats, sheep, dogs, and cats, preferably humans.

[0044] In the present invention, the term "diseases associated with leukocyte extracellular trap formation" is not particularly limited as long as an increase in leukocyte extracellular trap formation is observed in the patient's body. Examples of such diseases include psoriasis, vasculitis syndrome, ANCA-associated vasculitis (e.g., Wegener's granulomatosis, microscopic polyangiitis, allergic granulomatous vasculitis), acute kidney injury (AKI) (e.g., acute kidney injury with ischemia-reperfusion injury, post-rhabdomyolysis acute kidney injury), systemic lupus erythematosus (SLE), rheumatoid arthritis, localized Shwartzman reaction, appendicitis, Aspergillus infection, pneumonia, and lung cancer. These include acute inflammatory diseases such as Streptococcus pneumoniae infection, necrotizing fasciitis, streptococcal infection, sepsis, preeclampsia, Crohn's disease, schistosomiasis, periodontitis, tuberculosis, mastitis, malaria, cystic fibrosis, acute pancreatitis, and gout attacks; delayed wound healing due to diabetes; progression or metastasis of malignant tumors; and thrombotic diseases such as deep vein thrombosis, myocardial infarction, cerebral infarction, tumor-associated thrombosis, sickle cell disease, and disseminated intravascular coagulation (DIC). Vasculitis syndromes are classified into large-, medium-, and small-vasculitis depending on the size of the affected vessels. While the aforementioned ANCA-associated vasculitis and NETs-related diseases, such as SLE, are classified as small-vasculitis, DIC is frequently associated with the aggravation of polyarteritis nodosa, a medium-sized vasculitis (Guidelines for the Diagnosis and Treatment of Cardiovascular Disease (2006-2007 Joint Research Group Report)), sepsis, and solid tumors. In these diseases, cytotoxicity due to the formation of extracellular traps (e.g., NETs) by leukocytes leads to vascular endothelial damage, resulting in organ dysfunction. Furthermore, in the above-mentioned thrombotic diseases, the formation of extracellular traps (e.g., NETs) by leukocytes triggers the promotion of the thrombus formation cascade. In the present invention, the peptide molecules prevent vascular endothelial damage by inhibiting the formation, release, and diffusion of leukocyte extracellular traps (e.g., NETs), and further exhibit organ-protecting effects by inhibiting the thrombus formation cascade, which is believed to be effective in treating the above-mentioned diseases.Preferably, the disease to be treated is psoriasis, vasculitis syndrome, ANCA-associated vasculitis (e.g., Wegener's granulomatosis, microscopic polyangiitis, allergic granulomatous vasculitis), acute kidney injury (AKI) (e.g., acute kidney injury associated with ischemia-reperfusion injury, post-rhabdomyolytic acute kidney injury), systemic lupus erythematosus (SLE), or localized Shwartzman reaction, more preferably psoriasis or acute kidney injury (AKI), and even more preferably post-rhabdomyolytic acute kidney injury.

[0045] In the present invention, the leukocyte extracellular trap formation inhibitor and the pharmaceutical composition for treating or preventing diseases associated with leukocyte extracellular trap formation may contain pharmaceutical additives such as pharmaceutically acceptable carriers, solvents, diluents, excipients, stabilizers, buffers, binders, coating agents, disintegrants, surfactants, lubricants, flow enhancers, colorants, and flavorings. Pharmaceutical additives such as pharmaceutically acceptable carriers, diluents, excipients, stabilizers, disintegrants, and binders are well known in the art. The route of administration of the inhibitor and pharmaceutical composition is not particularly limited as long as it is a commonly used route, and specific examples include oral, sublingual, nasal, pulmonary, gastrointestinal, transdermal, ophthalmic, intravenous, subcutaneous, intramuscular, intraperitoneal, local injection, and surgical implantation. The inhibitor and pharmaceutical composition may be in the form of a solid formulation such as a capsule, tablet, or powder; a liquid formulation such as a solution, suspension, or emulsion; a spray; or a semi-liquid formulation such as an ointment, cream, or paste. The peptide molecule may also be an injectable preparation, and in the case of an injectable preparation, it may be a solid preparation, including a lyophilized preparation, or a liquid preparation. For oral administration, the inhibitor and the pharmaceutical composition are preferably solid preparations. For transdermal administration, the peptide molecule may be prepared as a liquid preparation (including a spray), such as a solution, suspension, or emulsion, or as a semi-liquid preparation, such as an ointment, cream, or paste, or as a poultice preparation. In the present invention, the peptide molecule may be added to food or feed and ingested by humans or non-human target animals as food or feed. Methods for producing such food or feed are also known to those skilled in the art. Furthermore, solution preparations can also be formulated as an injectable preparation. Furthermore, the peptide molecule may be added directly or after formulation to nutrients, foods, beverages, etc.

[0046] In the present invention, a therapeutically or prophylactically effective amount of a peptide molecule or a pharmaceutically acceptable salt thereof can be administered to a subject by administering to the subject a leukocyte extracellular trap formation inhibitor and a pharmaceutical composition for treating or preventing a disease associated with leukocyte extracellular trap formation. The term "therapeutically effective amount" refers to an amount that produces a therapeutic effect depending on the disease, route of administration, or form of administration, and is determined appropriately based on the symptoms, gender, and age of the subject, as well as other factors. The term "prophylactically effective amount" refers to an amount that produces a prophylactic effect depending on the disease, route of administration, or form of administration, and is determined appropriately based on the symptoms, gender, and age of the subject, as well as other factors. For example, in the case of oral administration, a therapeutically or prophylactically effective amount can be 0.001 g / kg / day or more, 0.005 g / kg / day or more, 0.008 g / kg / day or more, or 0.01 g / kg / day or more, and can be 10 g / kg / day or less, 5 g / kg / day or less, or 3 g / kg / day or less. When administered orally to humans, the therapeutically or prophylactically effective amount can be, for example, 5 mg or more, 10 mg or more, 15 mg or more, 20 mg or more, 25 mg or more, or 30 mg or more per day, and 15,000 mg or less, 12,000 mg or less, 10,000 mg or less, 8,000 mg or less, or 6,000 mg or less. When administered transdermally, the therapeutically or prophylactically effective amount can be 0.001 g / kg / day or more, 0.005 g / kg / day or more, 0.008 g / kg / day or more, or 0.01 g / kg / day or more, and 10 g / kg / day or less, 5 g / kg / day or less, or 3 g / kg / day or less. When administered to humans by transdermal administration, the therapeutically or prophylactically effective amount can be, for example, 5 mg or more, 10 mg or more, 15 mg or more, 20 mg or more, 25 mg or more, or 30 mg or more per day, and 15,000 mg or less, 12,000 mg or less, 10,000 mg or less, 8,000 mg or less, or 6,000 mg or less.Furthermore, for example, in the case of administration by injection such as intravenous injection, the therapeutically or prophylactically effective amount can be 0.0005 g / kg / day or more, 0.001 g / kg / day or more, 0.005 g / kg / day or more, 0.008 g / kg / day or more, or 0.01 g / kg / day or more, and can be 10 g / kg / day or less, 5 g / kg / day or less, or 3 g / kg / day or less. When administered to humans by injection such as intravenous injection, the therapeutically or prophylactically effective amount can be, for example, 1 mg or more, 5 mg or more, 10 mg or more, 15 mg or more, 20 mg or more, 25 mg or more, or 30 mg or more per day, and can be 15,000 mg or less, 12,000 mg or less, 10,000 mg or less, 8,000 mg or less, or 6,000 mg or less. The inhibitor or pharmaceutical composition can be administered to a subject in need of treatment or prevention of a disease associated with leukocyte extracellular trap formation at such a daily dose, either all at once or in divided doses. The dosage and frequency of administration of the inhibitor and pharmaceutical composition vary depending on various factors, such as the subject's species, body weight, sex, age, disease progression, and administration route. A skilled artisan, such as a physician, veterinarian, dentist, or pharmacist, can determine the dosage taking these factors into consideration. The above-mentioned therapeutically effective amount, prophylactically effective amount, dosage, and administration frequency are merely exemplary values, and values ​​greater than or less than these values ​​may still achieve therapeutic or prophylactic effects. Therefore, values ​​greater than or less than the above-mentioned therapeutically effective amount, prophylactically effective amount, dosage, and administration frequency may also be included within the therapeutically effective amount, prophylactically effective amount, dosage, and administration frequency of the inhibitor and pharmaceutical composition.

[0047] Another aspect of the present invention relates to a method for treating or preventing a disease associated with leukocyte extracellular trap formation, comprising administering a peptide molecule or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the peptide molecule or a pharmaceutically acceptable salt thereof, to a subject in need of such treatment or prevention. The peptide molecule is as described above, has a structure containing the amino acid sequence represented by formula (1) or the amino acid sequence represented by formula (2), and has activity of inhibiting leukocyte extracellular trap formation. Specific diseases are as described above. Any of the above-described features may be applied to the therapeutic or prophylactic method. For example, the therapeutic or prophylactic method comprises administering a therapeutically or prophylactically effective amount of the peptide molecule or a pharmaceutically acceptable salt thereof to a subject in need of such treatment or prevention. Another aspect of the present invention relates to the peptide molecule or a pharmaceutically acceptable salt thereof for use in the treatment or prevention of a disease associated with leukocyte extracellular trap formation. Another aspect of the present invention relates to the use of said peptide molecule or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment or prevention of a disease associated with the formation of extracellular traps in leukocytes.

[0048] The present invention will be described in more detail below with reference to specific examples, but the scope of the present invention is not limited to these examples in any way.

[0049] [Animals (mice)] Wild-type mice (C57BL6j) were purchased from Sankyo Labo Service Corporation, Inc. and used. The mice were kept in a specific pathogen-free facility (SPF) at the Keio University Animal Care and Use Center. All mice used in the experiments were 8 weeks old and male. Experiments were conducted in accordance with procedures approved by the Keio University Animal Care and Use Committee.

[0050] [Human blood samples] Blood samples were collected from healthy volunteers who provided their consent. Sample donors were adults who were not taking regular oral medications. The research protocol was approved by the Keio University Ethics Committee in accordance with the ethical guidelines set out in the Declaration of Helsinki.

[0051] [Isolation of human peripheral blood neutrophils] Human polymorphonuclear neutrophils (hPMNs) were isolated from peripheral blood samples from healthy volunteers by gravity centrifugation using Mono-Poly Resolving Medium (DS Pharma Biomedical) according to the protocol provided with the product. Red blood cells were lysed by hypotonic hemolysis using chilled water.

[0052] [Peptide Synthesis] Test peptides were synthesized using the Fmoc method using a peptide synthesizer, Prelude (Gyros Protein Technologies). The target peptides were automatically synthesized using Fmoc-NH-SAL resin (44 mg, 0.02 mmol, Watanabe Chemical Industry Co., Ltd.), starting with the removal of the Fmoc group. Commercially available amino acids containing protecting groups were used as is. The condensation reaction of each amino acid was carried out as follows: In N,N-dimethylformamide (DMF, 2.0 mL), Fmoc-amino acids (0.2 mmol) were sequentially reacted with the resin or the elongating peptide in the presence of N,N-diisopropylethylamine (DIEA, 0.4 mmol) at room temperature for 30 minutes using O-(7-aza-1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU, 0.2 mmol) and 1-hydroxy-7-azabenzotriazole (HOAt, 0.2 mmol) as coupling reagents. After each amino acid coupling step, the Fmoc group was deprotected by reaction with 20% piperidine / DMF solution (2.5 mL) at room temperature for 20 minutes. The resin-bound synthetic peptide was reacted with a mixture of trifluoroacetic acid (TFA), m-cresol, thioanisole, and 1,2-ethanedithiol (4.3 mL, 40:1:1:1, v:v:v:v) at room temperature for 150 minutes to deprotect the side chains of the synthetic peptide and cleave it from the resin, yielding a crudely purified peptide. The crudely purified peptide was purified by preparative reversed-phase high-performance liquid chromatography (RP-HPLC) (mobile phase: water / acetonitrile with 0.1% TFA) to obtain the synthetic peptide (TFA salt) as a white solid. The purity of the synthetic peptide was analyzed by RP-HPLC. A C18 reversed-phase column (4.6 x 150 mm) was used: COSMOSIL 5C4-AR-300 (Nacalai Tesque) for W8A and M10Hse(Me), and COSMOSIL 5C18-AR-II (Nacalai Tesque) for the other peptides. The analytical conditions were as follows:

[0053] Acetonitrile mobile phase gradient: 10-30%, 20 min (W8A), 5-65%, 40 min (other peptides). Flow rate: 1.0 mL / min. Detection: UV 220 nm.

[0054] The yield of the synthetic peptide obtained as a white solid was calculated as the TFA salt, and mass spectrometry was performed (HRMS (TOF MS ES+) or LRMS (TOF MS MALDI+)). The results are shown below together with the abbreviations and amino acid sequences of the synthetic peptides.

[0055] FK-12 amide: FKCRRWQWRMKK-NH2 (SEQ ID NO: 23) Yield: 35%; HRMS m / z [M+H] + found 1751.9578 (calculated for C 80 H 127 N 28 O 13 S2 1751.9579); HPLC purity: 97.7% (t R = 14.63 min)

[0056] F1A: AKCRRWQWRMKK-NH2 (SEQ ID NO: 24) Yield: 48%; HRMS m / z [M+H] + found 1675.9332 (calculated for C 74 H 123 N 28 O 13 S2 1675.9266); HPLC purity: 98.8% (t R = 13.11 min)

[0057] K2A: FACRRWQWRMKK-NH2 (SEQ ID NO: 25) Yield: 60%; HRMS m / z [M+H] + found 1694.9061 (calculated for C 77 H 120 N 27 O 13 S2 1694.9000); HPLC purity: 99.1% (t R = 14.19 min)

[0058] C3A: FKARRWQWRMKK-NH2 (Allocation number: 26) Yield: 41%; HRMS m / z [M+H] + found 1720.9630 (calcd. for C 80 H 127 N 28 O 13 S 1720.9698); HPLC purity: 99.9% (t R = 15.43 min)

[0059] R4A: FKCARWQWRMKK-NH2 (Allocation number: 27) Yield: 64%; HRMS m / z [M+H] + found 1666.8990 (calcd. for C 77 H 120 N 25 O 13 S2 1666.8939); HPLC purity: 99.8% (t R = 14.69 min)

[0060] R5A: FKCRAWQWRMKK-NH2 (Allocation number: 28) Yield: 36%; HRMS m / z [M+H] + found 1666.8899 (calcd. for C 77 H 120 N 25 O 13 S2 1666.8939); HPLC purity: 99.6% (t R = 14.42 min)

[0061] W6A: FKCRRAQWRMKK-NH2 (Allocation number: 29) Yield: 24%; HRMS m / z [M+H] + found 1636.9172 (calcd. for C 72 H 122 N 27 O 13 S2 1636.9157); HPLC purity: 99.4% (t R = 12.02 min)

[0062] Q7A: FKCRRWAWRMKK-NH2 (Allocation number: 30) Yield: 36%; HRMS m / z [M+H] +found 1694.9371 (calcd. for C 78 H 124 N 27 O 12 S2 1694.9364); HPLC purity: 98.4% (t R = 14.67 min)

[0063] W8A: FKCRRWQARMKK-NH2 (Allocation number: 31) Yield: 44%; HRMS m / z [M+H] + found 1636.9219 (calcd. for C 72 H 122 N 27 O 13 S2 1636.9157); HPLC purity: 99.7% (t R = 8.91 min)

[0064] R9A: FKCRRWQWAMKK-NH2 (Allocation number: 32) Yield: 80%; HRMS m / z [M+H] + found 1666.8912 (calcd. for C 77 H 120 N 25 O 13 S2 1666.8939); HPLC purity: 99.1% (t R = 14.98 min)

[0065] M10A: FKCRRWQWRAKK-NH2 (Allocation number: 33) Yield: 63%; HRMS m / z [M+H] + found 1691.9570 (calcd. for C 78 H 123 N 28 O 13 S 1691.9545); HPLC purity: 99.8% (t R = 13.60 min)

[0066] K11A: FKCRRWQWRMAK-NH2 (Allocation number: 34) Yield: 50%; HRMS m / z [M+H] + found 1694.9033 (calcd. for C 77 H 120 N27 O 13 S2 1694.9000); HPLC purity: 99.3% (t R = 15.17 min)

[0067] K12A: FKCRRWQWRMKA-NH2 (SEQ ID NO: 35) Yield: 45%; HRMS m / z [M+H] + found 1694.8993 (calculated for C 77 H 120 N 27 O 13 S2 1694.9000); HPLC purity: 99.8% (t R = 15.08 min)

[0068] F1W: WKCRRWQWRMKK-NH2 (SEQ ID NO: 36) Yield: 19%; LRMS m / z [M+H] + found 1690.84 (calculated for C 82 H 128 N 29 O 13 S2 1790.97); HPLC purity: 98.1% (t R = 15.09 min)

[0069] F1Cha: XKCRRWQWRMKK-NH2 (SEQ ID NO: 37) (X represents cyclohexylalanine) Yield: 33%; LRMS m / z [M+H] + found 1758.68 (calculated for C 80 H 133 N 28 O 13 S2 1758.01); HPLC purity: 99.2% (t R = 15.41 min)

[0070] F1Nal: XKCRRWQWRMKK-NH2 (SEQ ID NO: 38) (X represents 2-naphthylalanine) Yield: 25%; LRMS m / z [M+H] + found 1802.69 (calculated for C 84 H 129 N 28 O 13S2 1801.97); HPLC purity: 98.6% (t R = 16.13 min)

[0071] W6Hph: FKCRRXQWRMKK-NH2 (SEQ ID NO: 39) (X represents homophenylalanine) Yield: 26%; LRMS m / z [M+H] + found 1727.44 (calculated for C 79 H 128 N 27 O 13 S2 1726.96); HPLC purity: 97.7% (t R = 15.09 min)

[0072] W6Cha: FKCRRXQWRMKK-NH2 (SEQ ID NO: 40) (X represents cyclohexylalanine). Yield: 30%; LRMS m / z [M+H] + found 1718.91 (calculated for C 78 H 132 N 27 O 13 S2 1718.99); HPLC purity: 99.8% (t R = 15.60 min)

[0073] Q7E: FKCRRWEWRMKK-NH2 (SEQ ID NO: 41) Yield: 21%; LRMS m / z [M+H] + found 1752.71 (calculated for C 80 H 126 N 27 O 14 S2 1752.94); HPLC purity: 100% (t R = 14.68 min)

[0074] Q7N: FKCRRWNWRMKK-NH2 (SEQ ID NO: 42) Yield: 25%; LRMS m / z [M+H] + found 1737.92 (calculated for C 79 H 125 N 28 O 13 S2 1737.94); HPLC purity: 76.4% (t R= 14.36 min)

[0075] Q7D: FKCRRWDWRMKK-NH2 (SEQ ID NO: 43) Yield: 17%; LRMS m / z [M+H] + found 1738.42 (calculated for C 79 H 124 N 27 O 14 S2 1738.93); HPLC purity: 98.8% (t R = 14.43 min)

[0076] Q7Orn: FKCRRWXWRMKK-NH2 (SEQ ID NO: 44) (X represents ornithine) Yield: 24%; LRMS m / z [M+H] + found 1738.00 (calcd. for C 80 H 129 N 28 O 12 S2 1737.98); HPLC purity: 98.7% (t R = 14.26 min)

[0077] W8Nal: FKCRRWQXRMKK-NH2 (SEQ ID NO: 45) (X represents 2-naphthylalanine) Yield: 31%; LRMS m / z [M+H] + found 1763.40 (calcd. for C 82 H 128 N 27 O 13 S2 1762.96); HPLC purity: 98.5% (t R = 15.89 min)

[0078] W8Cha: FKCRRWQXRMKK-NH2 (SEQ ID NO: 46) (X represents cyclohexylalanine). Yield: 51%; LRMS m / z [M+H] + found 1718.67 (calculated for C 78 H 132 N 27 O 13 S2 1718.99); HPLC purity: 98.4% (t R = 15.51 min)

[0079] M10Nle: FKCRRWQWRXKK-NH2 (SEQ ID NO: 47) (X represents norleucine) Yield: 30%; LRMS m / z [M+H] + found 1734.32 (calculated for C 81 H 129 N 28 O 13 S 1734.00); HPLC purity: 100% (t R = 15.19 min)

[0080] M10Hse(Me): FKCRRWQWRXKK-NH2 (SEQ ID NO: 48) (X represents O-methylhomoserine). Yield: 46%; LRMS m / z [M+H] + found 1736.57 (calculated for C 80 H 127 N 28 O 14 S 1735.98); HPLC purity: 99.1% (t R = 13.61 min)

[0081] M10Thi: FKCRRWQWRXKK-NH2 (SEQ ID NO: 49) (X represents 2-thienylalanine) Yield: 17%; LRMS m / z [M+H] + found 1775.06 (calcd. for C 82 H 125 N 28 O 13 S2 1775.21 [avg.]); HPLC purity: 99.6% (t R = 15.15 min)

[0082] [Statistical Processing] Data obtained in the following examples are presented as mean ± standard error. Statistical significance was verified using an unpaired Student's t-test or one-way ANOVA followed by Dunnett's multiple comparison test. A P value of less than 0.05 was considered significant. JMP 10 software (SAS Institute Inc.) was used for statistical analysis.

[0083] Example 1: Induction of neutrophil extracellular traps (NETs) and inhibition of NET formation by pretreatment. Human segmented neutrophils, 1 x 10 5 Neutrophils were cultured in 400 μL of DMEM-based cell culture medium containing 2% human serum. For pretreatment, test substances were added to the culture medium and incubated for 30 minutes at 37°C under 5% CO2. Next, 25 nM phorbol 12-myristate 13-acetate (PMA; Sigma-Aldrich) was added to the culture medium and the neutrophils were stimulated for 3 hours. After 3 hours, the NET-DNA concentration in the culture medium was measured using Quant-iT Picogreen dsDNA assay reagent (Life Technologies). Fluorescence intensity was measured using a Gen5 microplate reader (BioTek) with excitation at 480 nm and detection at 520 nm. The test substances F1A, K2A, C3A, R4A, R5A, W6A, Q7A, W8A, R9A, M10A, K11A, K12A, and FK-12 amide were added to the culture medium at a concentration of 40 μg / mL (22.9 μM). The relative DNA release levels are shown in Figure 1 (n=3), with the level in the absence of test substance taken as 1. FK-12 amide showed no significant difference compared to the absence of test substance. On the other hand, F1A, W6A, and M10A significantly inhibited DNA release (NETs formation) from neutrophils compared to the absence of test substance. Q7A and W8A also showed a greater inhibitory effect on NETs formation than FK-12 amide.

[0084] The detection results for the test substances F1W, F1Cha, F1Nal, W6Hph, W6Cha, and FK-12 amide added to the culture medium at a concentration of 120 μg / mL (68.7 μM) are shown in Figure 2 (n = 3) as relative DNA release, with the value for the culture medium without test substance set to 1. F1W, F1Cha, and F1Nal exhibited greater inhibitory effects on NETs formation than FK-12 amide. W6Hph exhibited a similar inhibitory effect to FK-12 amide. The detection results for the test substances Q7E, Q7N, Q7D, Q7Orn, and FK-12 amide added to the culture medium at a concentration of 120 μg / mL (68.7 μM) are shown in Figure 3 (n = 3) as relative DNA release, with the value for the culture medium without test substance set to 1. None of the test substances exhibited a greater inhibitory effect on NETs formation than FK-12 amide. The test substances W8Nal, W8Cha, M10Nle, M10Hse(Me), M10Thi, and FK-12 amide were added to the culture medium at a concentration of 120 μg / mL (68.7 μM). The relative DNA release levels are shown in Figure 4 (n = 3), with the level in the absence of test substance taken as 1. W8Nal and M10Thi exhibited greater inhibitory effects on NETs formation than FK-12 amide. W8Cha, M10Nle, and M10Hse(Me) exhibited inhibitory effects on NETs formation comparable to those of FK-12 amide.

[0085] Example 2: Evaluation of renal function in a glycerol-induced rhabdomyolysis model To confirm the effects of the test substances in vivo, a test was performed in a glycerol-induced rhabdomyolysis model as follows. 75 μL of 50% glycerol was intramuscularly injected into each thigh muscle of wild-type mice (total of 150 μL per mouse). Immediately thereafter, 720 μg / 100 μL of the test substances (Q7Orn, M10Hse(Me), and FK-12 amide) were intraperitoneally administered. 100 μL of PBS was intraperitoneally administered to the control group instead of the test substances. 24 hours later, the blood urea nitrogen (BUN) and blood creatinine (Cr) levels of the mice were measured. BUN was measured by the urease-GIDH method, and Cr was measured by the enzymatic method. The results are shown in Figures 5 and 6. In addition, a qualitative urine test using TESTAP was performed as a marker for myoglobinuria to evaluate the presence and level of urinary occult blood (UOB). The results are shown in Figure 7. M10Hse(Me) demonstrated a renal function protective effect superior to FK-12 amide, showing significant differences in blood BUN, blood Cr, and UOB compared to the control group. On the other hand, Q7Orn, which showed a similar in vitro activity to FK-12 amide in inhibiting NETs formation, failed to reduce either blood BUN or blood Cr. This contrasts with M10Hse(Me), which demonstrated a similar in vitro activity to FK-12 amide in inhibiting NETs formation and a renal function protective effect superior to FK-12 amide in vivo. Furthermore, when a similar test was performed using M10Nle at 720 μg / 100 μL PBS, all mice died, a result that contrasts with the favorable safety and tolerability of M10Hse(Me).

[0086] Further studies were performed as follows. Wild-type mice were intramuscularly injected with 75 μL of 50% glycerol into both thigh muscles (total 150 μL per mouse). Six hours later, the test substance (M10Hse(Me)) was intraperitoneally administered at 720 μg / 100 μL PBS. A control group received 100 μL of PBS intraperitoneally instead of the test substance. Blood BUN and Cr levels were measured 6, 24, 48, and 96 hours after glycerol administration. The results are shown in Figures 8 and 9. Considering the effects of blood sampling on the mice, different groups of mice were used for each measurement. Measurements at 24, 48, and 96 hours after glycerol administration showed significant differences in both blood BUN and Cr levels in M10Hse(Me) compared to the control group.

[0087] Example 3: Evaluation of Survival Rate in a Glycerol-Induced Rhabdomyolysis Model. Survival rate was evaluated using five mice. Wild-type mice were intramuscularly injected with 75 μL of 50% glycerol into each thigh muscle (total 150 μL per mouse). Six hours later, the test substance (M10Hse(Me)) was intraperitoneally administered at 720 μg / 100 μL PBS. The control group received intraperitoneal administration of 100 μL PBS instead of the test substance. In the control group, one death was confirmed 24 hours and one death was confirmed 36 hours after glycerol administration. In contrast, the test substance-administered group showed a 100% survival rate. The results are shown in Figure 10.

[0088] Example 4: Evaluation of the Therapeutic Effect of Imiquimide (IMQ) on Psoriasis in a Model of Psoriasis Induced by Imiquimide (IMQ) Wild-type mice were shaved the day before the experiment, and 50 mg of commercially available IMQ cream (5% Veselna Cream; Mochida Pharmaceutical) was applied once daily for 7 consecutive days to create an IMQ-induced psoriasis model. The test substance-treated group received intraperitoneal administration of 720 μg / 100 μL M10Hse(Me) in PBS once daily for 7 days (n=5). The control group received intraperitoneal administration of 100 μL PBS once daily for 7 days (n=4). On day 8, the dorsal skin of each mouse was excised after triple anesthesia, fixed with paraformaldehyde, and stained with HE. Measurements of epidermal thickness and total skin tissue thickness (epidermis + dermis + subcutaneous tissue) revealed significant suppression of epidermal thickening in the test substance-treated group. There was no significant difference in the overall thickness of the skin tissue. The results are shown in Figures 11 to 14.

Claims

1. It has a structure containing an amino acid sequence represented by the following formula (1) or an amino acid sequence represented by the following formula (2), and A peptide molecule or a pharmaceutically acceptable salt thereof, which has activity of inhibiting the formation of extracellular traps in leukocytes. X 1 -KCRR-X 2 -Q-X 3 -R-X 4 -KK (1) R 1 -CH 2 CH 2 -CO-KCRR-X 2 -Q-X 3 -R-X 4 -KK (2) [In each formula, X 1 But, R 1 -CH 2 an α-amino acid residue having a side chain represented by -, R 1 is a hydrogen atom, an alkyl group, an alicyclic hydrocarbon group, an aromatic hydrocarbon group, a heteroalicyclic group, or a heteroaromatic group, X 2 represents a structure represented by the following formula (3): (In the formula, R 2 is a hydrogen atom, an alkyl group, an aromatic hydrocarbon group, or a heteroaromatic group, and n is 1 or 2. X 3 represents a structure represented by the following formula (4): (In the formula, R 3 is a hydrogen atom, an alkyl group, an alicyclic hydrocarbon group, an aromatic hydrocarbon group, a heteroalicyclic group, or a heteroaromatic group. X 4 represents a structure represented by the following formula (5): (In the formula, R 4 is a hydrogen atom, or -R 5 -Z-R 6 Or -Z-R 7 represents the structure shown in R 5 is an alkylene group, and R 6 is an alkyl group, or R 5 and R 6 form a heterocyclic ring together with Z to which they are attached, R 7 is an alkyl group, Z represents an oxygen atom or a sulfur atom. and R 1 , R 2 , and R 3 The heteroalicyclic group or heteroaromatic group in the formula (I) contains 1 to 4 heteroatoms selected from oxygen, sulfur, and nitrogen atoms as ring constituent atoms, However, the amino acid sequence represented by formula (1) is not any of FKCRRWQWRMKK, AKCRRWQWRMKK, FKCRRFQWRMKK, AKCRRFQWRMKK, FKCRRWQVRMKK, AKCRRWQVRMKK, FKCRRFQVRMKK, and AKCRRFQVRMKK.]

2. R 1 is a hydrogen atom, an alkyl group, a monocyclic or bicyclic cycloalkyl group, a monocyclic or bicyclic cycloalkenyl group, a monocyclic or bicyclic aromatic hydrocarbon group, or a monocyclic or bicyclic heteroaromatic group, R 2 is a hydrogen atom, an alkyl group, a monocyclic or bicyclic aromatic hydrocarbon group, or a monocyclic or bicyclic heteroaromatic group, R 3 is a hydrogen atom, an alkyl group, a monocyclic or bicyclic cycloalkyl group, a monocyclic or bicyclic cycloalkenyl group, a monocyclic or bicyclic aromatic hydrocarbon group, or a monocyclic or bicyclic heteroaromatic group, and R 1 , R 2 , and R 3 The monocyclic or bicyclic heteroaromatic group in the formula (I) contains 1 to 4 heteroatoms selected from oxygen, sulfur, and nitrogen atoms as ring constituent atoms.

2. The peptide molecule of claim 1 or a pharmaceutically acceptable salt thereof.

3. R 1 is a hydrogen atom, an alkyl group having 1 to 9 carbon atoms, a monocyclic cycloalkyl group having 3 to 10 carbon atoms, a monocyclic cycloalkenyl group having 3 to 10 carbon atoms, a monocyclic aromatic hydrocarbon group, a monocyclic heteroaromatic group, or a bicyclic group having a cyclic structure represented by the following formula (6): (wherein ring A is a ring that makes the structure represented by formula (6) a 9- to 11-membered bicyclic ring structure, and the atoms constituting ring A include at least one carbon atom and may include one or two heteroatoms selected from an oxygen atom, a sulfur atom, and a nitrogen atom; Z 1 , Z 2 , Z 3 and Z 4 are each independently a carbon atom or a nitrogen atom, Z 1 , Z 2 , Z 3 and Z 4 The number of nitrogen atoms is at most two, represents a single bond or a double bond, X according to claim 1 1 or in the side chain of the structure represented by formula (2) of claim 1 2 The point of attachment to the moiety can be either the A ring or the Z 1 It may also be a ring having the following structure: R 2 is a hydrogen atom, an alkyl group having 1 to 9 carbon atoms, a monocyclic aromatic hydrocarbon group having 3 to 10 carbon atoms, a monocyclic heteroaromatic group having 3 to 10 carbon atoms, or a bicyclic group having a cyclic structure represented by the following formula (7): (wherein ring B is a ring that makes the structure represented by formula (7) a 9- to 11-membered bicyclic aromatic ring, and the atoms constituting ring B include at least one carbon atom and may include one or two heteroatoms selected from an oxygen atom, a sulfur atom, and a nitrogen atom; Z 1 , Z 2 , Z 3 and Z 4 are each independently a carbon atom or a nitrogen atom, Z 1 , Z 2 , Z 3 and Z 4 The number of nitrogen atoms is at most two, CH in the structure represented by formula (3) according to claim 1 2 The point of attachment to the moiety is either the B ring or the Z 1 It may also be a ring having the following structure: and R 3 is a hydrogen atom, an alkyl group having 1 to 9 carbon atoms, a monocyclic cycloalkyl group having 3 to 10 carbon atoms, a monocyclic cycloalkenyl group having 3 to 10 carbon atoms, a monocyclic aromatic hydrocarbon group, a monocyclic heteroaromatic group, or a bicyclic group having a cyclic structure represented by the following formula (8): (In the formula, Ring C is a ring that makes the structure represented by formula (8) a 9- to 11-membered bicyclic ring structure, and the atoms constituting Ring C include at least one carbon atom and may include one or two heteroatoms selected from an oxygen atom, a sulfur atom, and a nitrogen atom, Z 1 , Z 2 , Z 3 and Z 4 are each independently a carbon atom or a nitrogen atom, Z 1 , Z 2 , Z 3 and Z 4 The number of nitrogen atoms is at most two, represents a single bond or a double bond, CH in the structure represented by formula (4) according to claim 1 2 The point of attachment to the moiety can be the C ring or Z 1 It may also be a ring having the following structure:

2. The peptide molecule of claim 1 or a pharmaceutically acceptable salt thereof.

4. R 1 is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, a monocyclic cycloalkyl group having 3 to 7 carbon atoms, or an aromatic hydrocarbon group or bicyclic heteroaromatic group having a cyclic structure represented by the following formula (6-1): (wherein ring A1 is a ring in which the structure represented by formula (6-1) is a 9- to 10-membered bicyclic aromatic ring, or is absent, The atoms constituting the A1 ring include at least one carbon atom and may include one or two heteroatoms selected from an oxygen atom, a sulfur atom, and a nitrogen atom; X according to claim 1 1 or in the side chain of the structure represented by formula (2) of claim 1 2 The point of attachment to the moiety may be at the A1 ring or the benzene ring. R 2 is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, or an aromatic hydrocarbon group or bicyclic heteroaromatic group having a cyclic structure represented by the following formula (7-1): (wherein ring B1 is a ring in which the structure represented by formula (7-1) is a 9- to 10-membered bicyclic aromatic ring, or is absent, The atoms constituting ring B1 include at least one carbon atom and may include one or two heteroatoms selected from an oxygen atom, a sulfur atom, and a nitrogen atom; CH in the structure represented by formula (3) according to claim 1 2 The point of attachment to the moiety may be at the B1 ring or the benzene ring. and R 3 is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, a monocyclic cycloalkyl group having 3 to 7 carbon atoms, or an aromatic hydrocarbon group or bicyclic heteroaromatic group having a cyclic structure moiety represented by the following formula (8-1): (wherein ring C1 is a ring in which the structure represented by formula (8-1) is a 9- to 10-membered bicyclic aromatic ring, or is absent; The atoms constituting the C1 ring include at least one carbon atom and may include one or two heteroatoms selected from an oxygen atom, a sulfur atom, and a nitrogen atom; CH in the structure represented by formula (4) according to claim 1 2 The point of attachment to the moiety may be at the C1 ring or the benzene ring.

2. The peptide molecule of claim 1 or a pharmaceutically acceptable salt thereof.

5. It contains an amino acid sequence represented by formula (1), and X 1 2. The peptide molecule or a pharmaceutically acceptable salt thereof according to claim 1, wherein is an alanine residue, a phenylalanine residue, a tryptophan residue, a cyclohexylalanine residue, or a 2-naphthylalanine residue.

6. X 2 2. The peptide molecule of claim 1, or a pharmaceutically acceptable salt thereof, wherein is an alanine residue, a tryptophan residue, or a homophenylalanine residue.

7. X 3 The peptide molecule or a pharmaceutically acceptable salt thereof according to claim 1, wherein is an alanine residue, a tryptophan residue, a cyclohexylalanine residue, or a 2-naphthylalanine residue.

8. X 4 is not a methionine residue, or a pharmaceutically acceptable salt thereof.

9. R 4 is a hydrogen atom, or -R 5 -Z-R 6 Or -Z-R 7 represents the structure shown in R 5 is an alkylene group having 1 to 6 carbon atoms, and R 6 is an alkyl group having 1 to 6 carbon atoms, or R 5 and R 6 form a 4- to 7-membered heterocyclic ring together with Z to which they are attached, R 7 is an alkyl group having 1 to 6 carbon atoms; 9. The peptide molecule of claim 8, or a pharmaceutically acceptable salt thereof.

10. R 4 But, -R 5 -Z-R 6 represents the structure shown in R 5 is an alkylene group having 1 to 3 carbon atoms, and R 6 is an alkyl group having 1 to 3 carbon atoms, or R 5 and R 6 form a 5- or 6-membered heterocyclic ring together with Z to which they are attached.

9. The peptide molecule of claim 8, or a pharmaceutically acceptable salt thereof.

11. R 5 is an alkylene group having 1 to 3 carbon atoms, and R 6 is an alkyl group having 1 to 3 carbon atoms, Z is an oxygen atom; 11. The peptide molecule of claim 10, or a pharmaceutically acceptable salt thereof.

12. X 4 is an O-methylhomoserine residue, or a pharmaceutically acceptable salt thereof.

13. R 1 is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, a monocyclic cycloalkyl group having 3 to 7 carbon atoms, or an aromatic hydrocarbon group or bicyclic heteroaromatic group having a cyclic structure represented by the following formula (6-1): (wherein ring A1 is a ring in which the structure represented by formula (6-1) is a 9- to 10-membered bicyclic aromatic ring, or is absent, The atoms constituting the A1 ring include at least one carbon atom and may include one or two heteroatoms selected from an oxygen atom, a sulfur atom, and a nitrogen atom; X according to claim 1 1 or in the side chain of the structure represented by formula (2) of claim 1 2 The point of attachment to the moiety may be at the A1 ring or the benzene ring. R 2 is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, or an aromatic hydrocarbon group or bicyclic heteroaromatic group having a cyclic structure represented by the following formula (7-1): (wherein ring B1 is a ring in which the structure represented by formula (7-1) is a 9- to 10-membered bicyclic aromatic ring, or is absent, The atoms constituting ring B1 include at least one carbon atom and may include one or two heteroatoms selected from an oxygen atom, a sulfur atom, and a nitrogen atom; CH in the structure represented by formula (3) according to claim 1 2 The point of attachment to the moiety may be at the B1 ring or the benzene ring. R 3 is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, a monocyclic cycloalkyl group having 3 to 7 carbon atoms, or an aromatic hydrocarbon group or bicyclic heteroaromatic group having a cyclic structure moiety represented by the following formula (8-1): (wherein ring C1 is a ring in which the structure represented by formula (8-1) is a 9- to 10-membered bicyclic aromatic ring, or is absent; The atoms constituting the C1 ring include at least one carbon atom and may include one or two heteroatoms selected from an oxygen atom, a sulfur atom, and a nitrogen atom; CH in the structure represented by formula (4) according to claim 1 2 The point of attachment to the moiety may be at the C1 ring or the benzene ring. R 4 But, -R 5 -Z-R 6 represents the structure shown in R 5 is an alkylene group having 1 to 3 carbon atoms, R 6 is an alkyl group having 1 to 3 carbon atoms, and Z is an oxygen atom; 2. The peptide molecule of claim 1 or a pharmaceutically acceptable salt thereof.

14. R 5 and R 6 and Z are taken together with the Z to which they are attached to form a 5- to 6-membered heteroaromatic ring, or a pharmaceutically acceptable salt thereof.

15. It contains an amino acid sequence represented by formula (1), and X 1 is not a phenylalanine residue, or a pharmaceutically acceptable salt thereof.

16. 2. The peptide molecule or pharmaceutically acceptable salt thereof according to claim 1, wherein the number of amino acid residues is 11 or 12.

17. An agent for inhibiting the formation of extracellular traps in leukocytes, comprising the peptide molecule according to any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof.

18. A pharmaceutical composition for treating or preventing a disease associated with extracellular trap formation in leukocytes, comprising the peptide molecule according to any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof.

19. A pharmaceutical composition for treating or preventing acute kidney injury, comprising the peptide molecule according to any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof.

20. 20. The pharmaceutical composition of claim 19, wherein the acute kidney injury is post-rhabdomyolytic acute kidney injury.

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