JPH059199A - Peptide or its salt - Google Patents

Abstract

PURPOSE:To provide a new peptide (salt) having a specific amino acid sequence, bondable to human interleukin-1 and useful as a treating agent for diseases relating to interleukin-1 such as rheumatism and osteoporosis. CONSTITUTION:The objective peptide (salt) expressed by formula (A is Val, Leu, etc.; B is Arg, Lys, etc.; X 1 and X 2 are single bond, Gly, Ala, peptide fragment, etc.) is produced by (1) bonding an amino acid corresponding to the C-terminal of the peptide to a solid carrier through the alpha-COOH group of the amino acid using a styrene-divinylbenzene copolymer, etc., by a solid-phase synthesis of peptide, (2) bonding the corresponding amino acid or peptide fragment to the above amino acid along the direction of N-terminal of the peptide after protecting the functional groups other than the alpha-COOH group, (3) removing the protecting group of the protected amino group at the bonded terminal, (4) repeating the steps 2 and 3, (5) separating the formed peptide chain from the polymer and (6) removing the protecting group.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to human interleukin-
The present invention relates to a peptide having one binding ability or a salt thereof. The peptide or its salt provided by the present invention inhibits the expression of human IL-1 activity by binding to human inter-leukin-1 (hereinafter, abbreviated as human IL-1), and thus rheumatism and osteoporosis. It is useful for treating diseases associated with IL-1 such as

[0002]

2. Description of the Related Art IL-1 is an important factor of immune response and plays an important role in various inflammatory diseases. It has been suggested that IL-1 is involved in alveolar bone destruction during periodontitis, bone resorption in osteoporosis, and cartilage destruction in rheumatoid arthritis (interleukin 1, protein nucleic acid enzyme, Vol. 33, No. 10).
No., pp. 1728-1741, 1988). Also,
The IL-1 inhibitory active substance found in the culture supernatant of monosite is an antagonist of IL-1 receptor having a molecular weight of about 17,000, and can inhibit various physiological activities of IL-1. Reported [WP Arend et al., Journal of Clinical Investigation (J. Cli
n.Invest.), Vol. 85, pp. 1694-1697, (199
0))].

[0003]

Since the above-mentioned antagonist has a large molecular weight and binds to IL-1 receptor on cells, it has various side effects such as toxicity to the liver and kidney and non-specific immunosuppression. May occur. Therefore, an object of the present invention is to provide a peptide having IL-1 inhibitory activity without binding to the IL-1 receptor.

[0004]

According to the present invention, the above-mentioned object is achieved by the following general formula (I) H-X ↑ 1-Cys-A-B-B-A-B-A-Ser-X ↑ 2. -Y (I) (In the formula, A represents an amino acid residue selected from the group consisting of Val, Leu, Ile and Nle, and B represents Arg, Lys, Gln.
, His and Ser, which represent an amino acid residue selected from the group consisting of a single bond or X ↑ 1 and X ↑ 2, respectively.
Gly, Ala, Val, Arg, Asn, Ser, Phe, Pro, Le
u, Glu, Asp, Lys, Thr, His, Tyr, Nle and
A peptide fragment consisting of 1 to 10 amino acid residues selected from the group consisting of Ile, Y represents a hydroxyl group or an amino group) and having human interleukin-1 binding ability, or a salt thereof, More preferably, the following general formula (II) H-X ↑ 3-D-X ↑ 4-Y (II) (wherein D is the formula (1): Cys Leu Arg Ile Lys Ile Ser
A peptide fragment represented by (SEQ ID NO: 1), formula (2):
Cys Leu Gln Ile Gly Ile Ser (SEQ ID NO: 2) peptide fragment or formula (3): Cys Leu Gln Ile Gl
n Ile Ser (SEQ ID NO: 3) represents a peptide fragment, wherein X ↑ 3 and X ↑ 4 are single bonds or Gly, respectively.
, Ala, Val, Arg, Asn, Ser, Phe, Pro, Leu
, Glu, Asp, Lys, Thr, His, Tyr and Ile represent a peptide fragment consisting of 1 to 10 amino acid residues, and Y represents a hydroxyl group or an amino group) or a peptide thereof. This is accomplished by providing salt.

In the present specification, various amino acid residues are described by the following abbreviations. Ala: L-alanine residue Arg: L-arginine residue Asn: L-asparagine residue Asp: L-aspartic acid residue Cys: L-cysteine residue Gln: L-glutamine residue Glu: L-glutamic acid residue Gly: Glycine residue His: L-histidine residue Ile: L-isoleucine residue Leu: L-leucine residue Lys: L-lysine residue Phe: L-phenylalanine residue Pro: L-proline residue Ser: L -Serine residue Thr: L-threonine residue Trp: L-tryptophan residue Tyr: L-tyrosine residue Val: L-valine residue Nle: L-norleucine residue The amino acid sequence of a peptide is described so that the N-terminal amino acid residue is located on the left and the C-terminal amino acid residue is located on the right.

Representative examples of the peptides provided by the present invention are shown below. (A): Val Val Arg Asn Ser Ser Tyr CysLeu Arg Ile
Lys Ile Ser Ala Lys (SEQ ID NO: 4), (b): Lys Asn Ser Ser Tyr Cys Leu ArgIle Lys Ile
Ser Ala Lys Phe ValGlu (SEQ ID NO: 5), (c): Lys Tyr Cys Leu Arg Ile Lys IleSer Ala Lys
Phe Val Glu Asn GluPro (SEQ ID NO: 6), (d): Tyr Cys Leu Arg Ile Lys Ile SerAla Lys
(SEQ ID NO: 7), (e): Tyr Cys Leu Arg Ile Lys Ile Ser (SEQ ID NO: 8), (f): Cys Leu Arg Ile Lys Ile Ser (SEQ ID NO:
1), (g): Lys Cys Leu Arg Ile Lys Ile Ser (SEQ ID NO: 9), (h): Cys Leu Gln Ile Lys Ile Ser (SEQ ID NO:
2), (i): Lys Cys Leu Gln Ile Gln Ile Ser (SEQ ID NO: 10), (j): Lys Ile Cys Ile Arg Ile Gln IleSer (SEQ ID NO: 11), (k): Lys Xaa Cys Xaa Arg Xaa Gln XaaSer (SEQ ID NO: 12) (where Xaa represents Nle), (l): Lys Ile Cys Ile His Ile Gln IleSer (SEQ ID NO: 13), (m): Lys Ile Cys Leu Arg Ile Gln IleSer (SEQ ID NO: 14), (n): Lys Cys Val Gln Val Gln Val Ser (SEQ ID NO: 15) and (o): Lys Ile Cys Ile Arg Ile Gln IleXaa (SEQ ID NO: 16) (however, Xaa) Is the --COOH group of Ser--
CONH ↓ 2 group is substituted).

The salt of the peptide of the present invention is a physiologically acceptable salt, and examples of the salt include hydrochloric acid, sulfuric acid,
Salts with acids such as phosphoric acid, lactic acid, tartaric acid, maleic acid, fumaric acid, oxalic acid, malic acid, citric acid, oleic acid, palmitic acid; alkali metals or alkaline earth metals such as sodium, potassium, calcium and aluminum. Salts with metal hydroxides or carbonates; salts with triethylamine, benzylamine, diethanolamine, t-butylamine, dicyclohexylamine, arginine and the like can be mentioned.

The peptide of the present invention is prepared by a method usually used in peptide synthesis, for example, a solid phase synthesis method or a liquid phase synthesis method, and the solid phase synthesis method is easy to operate [for example, the Japanese Biochemical Society. Volume "Sequel to Biochemistry Laboratory 2
Protein Chemistry (below) "(May 20, 1987)
Tokyo Kagaku Doujinshi Co., Ltd.), pp. 641-694].

The preparation of the peptide of the present invention by the solid phase synthesis method can be carried out, for example, by using a polymer which is insoluble in a reaction solvent such as a styrene-divinylbenzene copolymer, and the amino acid corresponding to the C-terminal of the desired peptide is Having α-COO
Α-amino group other than α-COOH group which the amino acid or peptide fragment has by binding via the H group, and then toward the amino acid N-terminal direction of the desired peptide The step of condensing and binding the functional group of 1) and the step of removing the protecting group of the amino group forming a peptide bond such as the α-amino group in the bound amino acid or peptide fragment are sequentially repeated. By extending the peptide chain to form a peptide chain corresponding to the desired peptide, then removing the peptide chain from the polymer, and removing the protecting group from the protected functional group. It is carried out by obtaining the peptide and then purifying it. Here, the elimination of the peptide chain from the polymer and the removal of the protective group are preferably carried out simultaneously using hydrogen fluoride, from the viewpoint of suppressing side reactions.
Further, it is effective to purify the obtained peptide by reverse phase liquid chromatography.

The salt of the peptide of the present invention can be prepared by utilizing a usual salt formation reaction.

The peptides of the present invention and salts thereof (hereinafter abbreviated as peptides) have the ability to bind to human IL-1, and therefore IL-without binding to IL-1 receptor. 1 can be inhibited. In addition, the peptides of the present invention have been confirmed to have low toxicity in a toxicity test.

Therefore, the peptides of the present invention are IL-
It is effective in treating diseases such as rheumatism and osteoporosis caused by overproduction of 1.

The peptides of the present invention can bind to and neutralize IL-1 in serum and synovial fluid by administering it to patients suffering from diseases such as rheumatism and osteoporosis. The symptoms of the person can be reduced.

The dose for the effective expression of peptides is usually 0.01 μg / kg to 2 g / kg (adult), preferably 0.01 μg / kg to 200 mg / kg (adult).

The dosage form is preferably a solution obtained by dissolving peptides in a physiologically acceptable solution such as 5% glucose solution or physiological saline, and the solution is various pharmacologically acceptable solutions. It may contain an additive. Examples of the administration method include intravenous administration, subcutaneous administration, intraperitoneal administration, joint administration and the like, and oral administration is also possible by further encapsulating or liposome-forming the peptides.

[0016]

EXAMPLES The present invention will be specifically described below with reference to examples. The present invention is not limited to these examples.

Example 1 Formula (4): Val Val Arg Asn Ser Ser Tyr Cys Leu Arg
The peptide represented by Ile Lys Ile Ser AlaLys (SEQ ID NO: 4) was synthesized by a solid phase synthesis method using an automatic peptide synthesizer. That is, 4- [N ↑ α- (t-butoxycarbonyl) -N ↑ ε- (p-chlorobenzyloxycarbonyl) -L-lysyloxymethyl] -phenylacetamidomethyl group was added at 0.65 mmol / g (resin). Of a styrene-divinylbenzene copolymer [constituent molar ratio of styrene and divinylbenzene: 99: 1] [PAM Lysine, t-Boc-L-Lys (Cl manufactured by Applied Biosystems, USA). -Z)] 0.1 mmol was used to couple amino acids sequentially toward the N-terminus of the peptide. In the coupling reaction, as amino acids, N ↑ α- (t-butoxycarbonyl) -N ↑ ε- (p-chlorobenzyloxycarbonyl) -L-lysine [t-Boc lysine], N, manufactured by Applied Biosystems, USA −
(T-Butoxycarbonyl) -L-leucine [t-Boc leucine], N- (t-butoxycarbonyl) -O- (2
-Bromobenzyloxycarbonyl) -L-tyrosine [t-Boc tyrosine], N- (t-butoxycarbonyl)
-L-isoleucine [t-Boc isoleucine], N- (t
-Butoxycarbonyl) -L-asparagine [t-Boc asparagine], N- (t-butoxycarbonyl) -O-
Benzyl-L-serine [t-Boc serine], N- (t-butoxycarbonyl) -S- (p-methoxybenzyl)-
L-cysteine [t-Boc cysteine], N- (t-butoxycarbonyl) -L-valine [t-Boc valine], N ↑
α- (t-butoxycarbonyl) -N ↑ G- (mesitylene-2-sulfonyl) -L-arginine [t-Boc arginine], N- (t-butoxycarbonyl) -L-alanine [t-Boc alanine] 1 mmol was used for each.

The obtained peptide was subjected to deprotection and desorption from the solid phase by using HF reactor type I manufactured by Peptide Institute Co., Ltd. The crude product was purified by a preparative high performance liquid chromatograph manufactured by Millipore Waters Co. [column: Deltapack C18 47 × 300 mm with preppack 1000 pressure module]. The obtained purified peptide is a high performance liquid chromatograph for analysis LC6A manufactured by Shimadzu Corporation [Column: TSKgel OD manufactured by Toso Corporation.
S-80TM CTR, mobile phase: mixed solvent of acetonitrile and water containing 0.05% by volume of trifluoroacetic acid (acetonitrile concentration was changed from 5% by volume to 50% by volume in 30 minutes)], A single peak was shown at 17.1 min. The molecular weight of the purified peptide determined by the FAB method mass spectrum was 1838 (theoretical value: 1837.2).

Examples 2 to 15 and Comparative Example Formula (5): Lys Asn Ser Ser Tyr Cys Leu Arg Ile Lys
Peptide represented by Ile Ser Ala Lys PheVal Glu (SEQ ID NO: 5) (Example 2), Formula (6): Lys Tyr Cys Le
u Arg Ile Lys Ile Ser Ala Lys Phe Val Glu Asn Glu
Peptide represented by Pro (SEQ ID NO: 6) (Example 3), formula (7): Tyr Cys Leu Arg Ile LysIle Ser Ala
Lys (SEQ ID NO: 7) peptide (Example 4), Formula (8): Tyr Cys Leu Arg Ile Lys Ile Ser
Peptide represented by (SEQ ID NO: 8) (Example 5), formula (9): Cys Leu Arg Ile Lys Ile Ser (SEQ ID NO:
Peptide represented by 1) (Example 6), Formula (10): Ly
s Cys Leu Arg Ile Lys Ile Ser (SEQ ID NO: 9) peptide (Example 7), Formula (11): Cys Leu Gl
n IleLys Ile Ser (SEQ ID NO: 2) peptide (Example 8), Formula (12): Lys Cys Leu Gln Ile Gl
Peptide represented by n Ile Ser (SEQ ID NO: 10) (Example 9), Formula (13): Lys Ile Cys Ile Arg Ile Gln
Peptide represented by Ile Ser (SEQ ID NO: 11) (Example 10), Formula (14): Lys Xaa Cys Xaa Arg Xaa Gln
Xaa Ser (SEQ ID NO: 12) (however, Xaa is
Peptide represented by Nle) (Example 11), Formula (15): Lys IleCys Ile His Ile Gln Ile Ser (SEQ ID NO: 13) Peptide (Example 12), Formula (16): Lys Ile Cys Leu Arg Ile Gln Ile Ser (SEQ ID NO: 14) Peptide (Example 13), Formula (17): Lys Cys Val Gln Val Gln Val Ser (SEQ ID NO: 15) Peptide (Example 14) ), The formula (1
8): Lys Ile Cys Ile Arg Ile Gln Ile Xaa (SEQ ID NO: 16) (where Xaa represents —COH group of Ser by —C
A peptide represented by ONH ↓ 2 group (represented by substitution with an ONH ↓ 2 group) (Example 15) and formula (19): Tyr Ser Leu Glu
The peptide represented by Ile Lys Ile Ser (SEQ ID NO: 17) (comparative example) was synthesized in the same manner as in Example 1.

In Example 2, styrene-containing 4- [N- (t-butoxycarbonyl) -γ-benzyl-L-glutamyloxymethyl] -phenylacetamidomethyl group at a rate of 0.70 mmol / g (resin). Granular resin consisting of divinylbenzene copolymer [molar ratio of styrene and divinylbenzene: 99: 1] [PAM glutamic acid, t-Boc-L-Gl manufactured by Applied Biosystems, USA]
u (OBzl)] 0.1 mmol was used and in Example 3 4-
Add [N- (t-butoxycarbonyl) -L-prolyloxymethyl] -phenylacetamidomethyl group to 0.78
Granular resin consisting of a styrene-divinylbenzene copolymer [molar ratio of styrene and divinylbenzene: 99: 1] having a ratio of mmol / g (resin) [PAM proline, t-Boc manufactured by Applied Biosystems, USA] -L-
Pro] 0.1 mmol was used, and in Example 4, 4- [N ↑
Styrene-divinyl having α- (t-butoxycarbonyl) -N ↑ ε- (p-chlorobenzyloxycarbonyl) -L-lysyloxymethyl] -phenylacetamidomethyl group at a ratio of 0.65 mmol / g (resin). Granular resin composed of benzene copolymer [molar ratio of styrene and divinylbenzene: 99: 1] [PAM lysine, t-Boc-L-Lys (Cl-, manufactured by Applied Biosystems, USA)
Z)] 0.1 mmol is used and 4- [N- (t-butoxycarbonyl) is used in Examples 5 to 14 and Comparative Example.
-O-benzyl-L-seryloxymethyl] -phenylacetamidomethyl group 0.72 mmol / g (resin)
Of a styrene-divinylbenzene copolymer [constituent molar ratio of styrene and divinylbenzene: 99 to 1] [PAM Serine, t-Boc-L-Ser (Bzl, manufactured by Applied Biosystems, USA) )] 0.1 mmol, and in Example 15 a styrene-divinylbenzene copolymer having an α-amino-p-methylbenzyl group of 0.78 mmol / g (resin) [Structure of styrene and divinylbenzene] Molar ratio: 99 to 1] granular resin [Applied Biosystems, USA, p-
Methyl BHA resin] 0.1 mmol was used.

As the amino acid, in addition to those used in Example 1, N- (t-butoxycarbonyl) -L-phenylalanine [t-Boc phenylalanine], N- (t manufactured by Applied Biosystems, Inc., USA. -Butoxycarbonyl) -γ-benzyl-L-glutamic acid [t-Bo
c glutamic acid], N- (t-butoxycarbonyl)-
L-glutamine [t-Boc glutamine], N- (t-butoxycarbonyl) -L-norleucine [t-Boc norleucine], N- (t-butoxycarbonyl) -N ↑ Im-
(2,4-Dinitrophenyl) -L-histidine [t-Bo
c Histidine] was used.

Deprotection and desorption from the solid phase were performed on each of the obtained peptides in the same manner as in Example 1 to purify the crude product. For each purified peptide, Table 1 shows the elution time in an analytical high performance liquid chromatograph and the molecular weight measurement results by FAB method mass spectrum.

[0023]

[Table 1]

Test Example Binding test with human IL-1 1 g of cellulose particles (commercially available from Seikagaku Corporation, CM-Cellulofine CH) in 5 ml of dioxane obtained by distillation in the presence of sodium metal. And 0.05 g of N-hydroxysuccinimide and 0.
1 g was added, and the mixture was shaken and stirred overnight at room temperature. The resulting mixture was mixed with 0.02 mol / l phosphate buffer (pH:
It wash | cleaned by 7.4) and suction-filtered. The particles obtained contain 1 mg of the peptides obtained in Examples 1-15,
It was mixed with 0.02 mol / l phosphate buffer (pH: 7.4) or 1 ml of purified water, and the mixture was stirred overnight at a temperature of 4 ° C. The resulting mixture was suction filtered. The filtrate was subjected to an analytical reversed-phase high-performance liquid chromatograph, but no residual unreacted peptide was observed (immobilization rate of the peptide on the carrier: about 100%). In this way
1 g of each of the adsorbents on which 1 mg of each of the peptides obtained in Examples 1 to 15 was immobilized was obtained.

100 mg of the adsorbent was weighed, and 100 μl of PBS containing 5% bovine serum albumin (10 mM phosphate buffer containing 0.15 M NaCl, pH: 7.4) was used.
l and incubate overnight at 4 ° C, then ↑ 125
About 300 Bq of I-labeled human IL-1β (purchased from Amersham Japan) was added and incubated at room temperature for 3 hours. The reaction mixture obtained was treated with 0.2% Triton X-1.
After washing 5 times with a Tris buffer (pH: 7.4) containing 00, the radioactivity remaining on the adsorbent was measured with a γ counter. The results are shown in Table 2.

[0026]

[Table 2]

As shown in Table 2, the peptide-immobilized adsorbents obtained in Examples 1 to 15 were the adsorbent immobilized with the comparative example and glycine (H-Gly-OH) (control example). A significantly higher radioactivity was observed as compared with the adsorbent on which was immobilized, and the peptides obtained in Examples 1 to 15 were human IL-1.
It was confirmed to have binding ability.

[0028]

INDUSTRIAL APPLICABILITY According to the present invention, peptides are provided which are effective for treating diseases associated with IL-1 such as rheumatism and osteoporosis. The peptides provided by the present invention are
As is clear from the above-mentioned test example, it has the ability to specifically bind to human IL-1, and human IL-1 and human I
Since it inhibits binding to the L-1 receptor, it can be administered to the above-mentioned affected person to neutralize human IL-1 in the body fluid and reduce the symptoms of the affected person.

[0029]

[Sequence list]

SEQ ID NO: 1 Sequence length: 7 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Array Cys Leu Arg Ile Lys Ile Ser 1 5

[0030] SEQ ID NO: 2 Sequence length: 7 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Array Cys Leu Gln Ile Lys Ile Ser 1 5

[0031] SEQ ID NO: 3 Sequence length: 7 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Array Cys Leu Gln Ile Gln Ile Ser 1 5

[0032] SEQ ID NO: 4 Sequence length: 16 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Array Val Val Arg Asn Ser Ser Tyr Cys Leu Arg Ile Lys Ile Ser Ala Lys  1 5 10 15

[0033] SEQ ID NO: 5 Sequence length: 17 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Array Lys Asn Ser Ser Tyr Cys Leu Arg Ile Lys Ile Ser Ala Lys Phe Val  1 5 10 15 Glu

[0034] SEQ ID NO: 6 Sequence length: 17 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Array Lys Tyr Cys Leu Arg Ile Lys Ile Ser Ala Lys Phe Val Glu Asn Glu  1 5 10 15 Pro

[0035] SEQ ID NO: 7 Sequence length: 10 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Array Tyr Cys Leu Arg Ile Lys Ile Ser AlaLys 1 5 10

[0036] SEQ ID NO: 8 Sequence length: 8 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Array Tyr Cys Leu Arg Ile Lys Ile Ser 1 5

[0037] SEQ ID NO: 9 Sequence length: 8 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Array Lys Cys Leu Arg Ile Lys Ile Ser 1 5

[0038] SEQ ID NO: 10 Sequence length: 8 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Array Lys Cys Leu Gln Ile Gln Ile Ser 1 5

[0039] SEQ ID NO: 11 Sequence length: 9 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Array Lys Ile Cys Ile Arg Ile Gln Ile Ser 1 5

[0040] SEQ ID NO: 12 Sequence length: 9 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Sequence features Characteristic symbol: modified site Location: 2, 4, 6, 8 How the characteristics were determined: E Other information: Xaa represents Nle. Array Lys Xaa Cys Xaa Arg Xaa Gln Xaa Ser 1 5

[0041] SEQ ID NO: 13 Sequence length: 9 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Array Lys Ile Cys Ile His Ile Gln Ile Ser 1 5

[0042] SEQ ID NO: 14 Sequence length: 9 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Array Lys Ile Cys Leu Arg Ile Gln Ile Ser 1 5

[0043] SEQ ID NO: 15 Sequence length: 8 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Array Lys Cys Val Gln Val Gln Val Ser 1 5

SEQ ID NO: 16 Sequence length: 9 Sequence type: Amino acid topology −: Linear sequence type: Peptide sequence characteristic symbol: modified site Location: 9 Method for determining characteristic: E Other information: Xaa is -CONH of Ser -COOH group
↓ Indicates the one substituted with two groups. Sequence Lys Ile Cys Ile Arg Ile Gln Ile Xaa 1 5

[0045] SEQ ID NO: 17 Sequence length: 8 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Array Tyr Ser Leu Glu Ile Lys Ile Ser 1 5

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification number Reference number within the agency FI Technical display location A61K 37/02 ADT C07K 99:00 (72) Inventor Chie Fujiwara 1 at 2045 Sakata, Kurashiki, Okayama Prefecture Kuraray Co., Ltd.

Claims (3)

[Claims] 1. A general formula H-X ↑ 1-Cys-ABABABAA-Ser-X ↑ 2-Y (wherein A is selected from the group consisting of Val, Leu, Ile and Nle. B is Arg, Lys, Gln
, His and Ser, which represent an amino acid residue selected from the group consisting of a single bond or X ↑ 1 and X ↑ 2, respectively.
Gly, Ala, Val, Arg, Asn, Ser, Phe, Pro, Le
u, Glu, Asp, Lys, Thr, His, Tyr, Nle and
A peptide or a salt thereof, which represents a peptide fragment consisting of 1 to 10 amino acid residues selected from the group consisting of Ile, Y represents a hydroxyl group or an amino group, and has human interleukin-1 binding ability. 2. The peptide has the general formula H-X ↑ 3-D-X ↑ 4-Y (wherein D is the formula (1): Cys Leu Arg Ile Lys Ile Ser.
A peptide fragment represented by (SEQ ID NO: 1), formula (2):
Cys Leu Gln Ile Gly Ile Ser (SEQ ID NO: 2) peptide fragment or formula (3): Cys Leu Gln Ile Gl
n Ile Ser (SEQ ID NO: 3) represents a peptide fragment, wherein X ↑ 3 and X ↑ 4 are single bonds or Gly, respectively.
, Ala, Val, Arg, Asn, Ser, Phe, Pro, Leu
, Glu, Asp, Lys, Thr, His, Tyr and Ile represent a peptide fragment consisting of 1 to 10 amino acid residues, and Y represents a hydroxyl group or an amino group). 1. The peptide or salt thereof according to 1. 3. The peptide has the following formula (4): Val Val Arg Asn Ser Ser Tyr Cys Leu Arg.
Ile Lys Ile Ser Ala Lys (SEQ ID NO: 4), Formula (5): Lys Asn Ser Ser Tyr Cys Leu Arg Ile Lys
Ile Ser Ala Lys Phe Val Glu (SEQ ID NO: 5), Formula (6): Lys Tyr Cys Leu Arg Ile Lys Ile Ser Ala
Lys Phe Val Glu Asn Glu Pro (SEQ ID NO: 6), Formula (7): Tyr Cys Leu Arg Ile Lys Ile Ser Ala Lys
(SEQ ID NO: 7), Formula (8): Tyr Cys Leu Arg Ile Lys Ile Ser (SEQ ID NO: 8), Formula (9): Cys Leu Arg Ile Lys Ile Ser (SEQ ID NO:
1), Formula (10): Lys Cys Leu Arg Ile Lys Ile Ser (SEQ ID NO: 9), Formula (11): Cys Leu Gln Ile Lys Ile Ser (SEQ ID NO: 2), Formula (12): Lys Cys Leu Gln Ile Gln Ile Ser (SEQ ID NO: 10), Formula (13): Lys Ile Cys Ile Arg Ile Gln Ile Ser
(SEQ ID NO: 11), Formula (14): Lys Xaa Cys Xaa Arg Xaa Gln Xaa Ser
(SEQ ID NO: 12) (however, Xaa represents Nle), Formula (15): Lys Ile Cys Ile His Ile Gln Ile Ser
(SEQ ID NO: 13), Formula (16): Lys Ile Cys Leu Arg Ile Gln Ile Ser
(SEQ ID NO: 14), Formula (17): Lys Cys Val Gln Val Gln Val Ser (SEQ ID NO: 15) or Formula (18): Lys Ile Cys Ile Arg Ile Gln Ile Xaa
(SEQ ID NO: 16) (where Xaa is Ser's --COOH
The peptide according to claim 1 or 2, or a salt thereof, which is a peptide represented by a group in which a group is substituted with a —CONH ↓ 2 group).