JP2004107333A - Cxcr4 antagonist and use of the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new anticancer and anti-chronic rheumatoid arthritis agent. <P>SOLUTION: This preventing and/or treating agent of cancer and chronic rheumatoid arthritis contains a peptide, its amide, its ester or its salt, having a CXCR4-antagonizing activity. The new anticancer and anti-chronic rheumatoid arthritis agent is the new peptide, its amide, its ester or its salt, having the CXCR4-antagonizing activity. <P>COPYRIGHT: (C)2004,JPO

Description

The present invention relates to a compound having CXCR4 antagonistic activity, and a preventive and / or therapeutic agent for cancer and rheumatoid arthritis containing the compound.

Many hormones and neurotransmitters regulate biological functions through specific receptors present on cell membranes. Many of these receptors perform intracellular signal transduction through activation of a conjugated guanine nucleotide-binding protein (hereinafter sometimes abbreviated as G protein). Further, since these receptors have a common structure having seven cell transmembrane regions, they are collectively referred to as G protein-coupled receptors or seven transmembrane receptors (7TMR).
As one of such G protein-coupled receptor proteins, a human receptor protein encoded by the CXCR4 gene [Non-Patent Document 1] is known.
Also, CXCL12 / SDF-1 [Non-Patent Document 2] is known as a physiologically active peptide that functions as a ligand for CXCR4.
Fujii [Patent Document 1] discloses peptidic compounds having an antagonistic effect on CXCR4, and describes that those compounds have anti-HIV activity.

転 移 Cancer metastasis is an important factor in determining a patient's life expectancy. It has been reported that the expression of CXCR4 is enhanced in breast cancer and the like, and that the expression of CXCL12 / SDF-1α, which is its ligand, is enhanced in organs to which metastases (lymph node, lung, liver and bone) [ Non-Patent Document 3]. In rheumatoid arthritis, infiltration of CD4-positive T cells into joint cavity fluid affects the progress of the disease state. It has been reported that CXCR4 gene expression is enhanced in CD4-positive memory T cells in joint cavity fluid of rheumatoid arthritis patients, and CXCL12 / SDF-1α gene expression is enhanced in joint synovial tissues [Non-patent literature] 4].

WO 02/20561 pamphlet (March 14, 2003) Journal of biological chemistry, vol. 273, p. 4754 (1998) Science, Vol. 261, pp. 600-603 (1993) Nature, Volume 410, pp. 50-56 (2001) Journal of Immunology, Vol. 165, pp. 6590-98 (2000)

The object of the present invention is to provide a novel means for preventing and / or treating cancer and rheumatoid arthritis using a compound having a CXCR4 antagonistic action. The present invention also provides novel compounds having prophylactic and / or therapeutic activity for cancer and rheumatoid arthritis, in particular, various oligopeptides having a common structure.

The present inventors have conducted intensive studies to solve the above problems, and as a result, a compound having a CXCR4 antagonistic activity, which has been considered to be effective as a chemotherapeutic agent for AIDS, has been developed for cancer including metastasis and rheumatoid arthritis. The present inventors have found that the present invention is effective for prevention and / or treatment, and as a result of further research, have completed the present invention.

That is, the present invention
(1) The following formula (Ia)
1 2 3 4 5 6 7 8 9 10 11 12 13 14
A1-A2-A3-Cys-Tyr-A4-A5-A6-A7-A8-A9-A10-Cys-A11 (Ia)
(Where
A1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is deleted;
A2 represents an arginine or glutamic acid residue when A1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus, and when A1 is deleted Represents an arginine or glutamic acid residue which may be derivatized at the N-terminus;
A3 represents an aromatic amino acid residue;
A4, A5 and A9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
A6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
A7 represents a proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
A8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
A10 represents citrulline, glutamic acid, arginine or lysine residue;
A11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, and the cysteine residues at positions 4 and 13 may be linked by a disulfide bond, and the amino acid may be in L-form or D-form. Which comprises a peptide or a salt thereof, a prophylactic and / or therapeutic agent for cancer or rheumatoid arthritis,
(2) In the above formula (Ia),
A1 is an arginine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is deleted;
A2 represents an arginine or glutamic acid residue when A1 is an arginine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus; Represents an arginine or glutamic acid residue which may be derivatized at the terminus;
A4 represents arginine, citrulline, alanine or glutamic acid residue;
A5 represents arginine, citrulline, alanine, lysine or glutamic acid residue;
A6 represents a lysine, alanine, citrulline or glutamic acid residue;
A7 represents a proline or alanine residue;
A8 represents a tyrosine, alanine or glutamic acid residue;
A9 represents an arginine, citrulline or glutamic acid residue;
A10 represents citrulline or glutamic acid residue;
A11 represents an arginine or glutamic acid residue which may be derivatized at the C-terminal, the prophylactic and / or therapeutic agent according to (1),
(3) The following formula (Ib)
1 2 3 4 5 6 7 8 9 10 11 12 13 14
B1-B2-B3-Cys-Tyr-B4-B5-B6-B7-B8-B9-B10-Cys-B11 (Ib)
(Where
B1 is a glutamic acid residue which may be derivatized at the N-terminus or is missing;
B2 represents an arginine or glutamic acid residue when B1 is a glutamic acid residue which may be derivatized at the N-terminus, and is derivatized at the N-terminus when B1 is deleted. Represents an arginine or glutamic acid residue which may be present;
B3 represents an aromatic amino acid residue;
B4, B5 and B9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
B6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
B7 represents proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
B8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
B10 represents citrulline, glutamic acid, arginine or lysine residue;
B11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, and the cysteine residues at the 4-position and the 13-position may be linked by a disulfide bond. Or a salt thereof,
(4) The peptide or a salt thereof according to (3), wherein B1 is a glutamic acid residue which may be derivatized at the N-terminus,
(5) The following formula (Ic)
1 2 3 4 5 6 7 8 9 10 11 12 13 14
C1-C2-C3-Cys-Tyr-C4-C5-C6-C7-C8-C9-C10-Cys-C11 (Ic)
(Where
C1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is missing;
C2 represents a glutamic acid residue if C1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus, and if C1 is deleted, Represents a glutamic acid residue which may be derivatized at the N-terminus;
C3 represents an aromatic amino acid residue;
C4, C5 and C9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
C6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
C7 represents a proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
C8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
C10 represents citrulline, glutamic acid, arginine or lysine residue;
C11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, and the cysteine residues at the 4-position and the 13-position may be linked by a disulfide bond. Or a salt thereof,
(6) The following formula (Id)
1 2 3 4 5 6 7 8 9 10 11 12 13 14
D1-D2-D3-Cys-Tyr-D4-D5-D6-D7-D8-D9-D10-Cys-D11 (Id)
(Where
D1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is deleted;
D2 represents an arginine or glutamic acid residue when D1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus, and when D1 is deleted Represents an arginine or glutamic acid residue which may be derivatized at the N-terminus;
D3 represents an aromatic amino acid residue;
D4 represents a glutamic acid residue;
D5 and D9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
D6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
D7 represents a proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
D8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
D10 represents a citrulline, glutamic acid, arginine or lysine residue;
D11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, and the cysteine residues at the 4-position and the 13-position may be linked by a disulfide bond. Or a salt thereof,
(7) The following formula (Ie)
1 2 3 4 5 6 7 8 9 10 11 12 13 14
E1-E2-E3-Cys-Tyr-E4-E5-E6-E7-E8-E9-E10-Cys-E11 (Ie)
(Where
E1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or has been deleted;
E2 represents an arginine or glutamic acid residue when E1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus; Represents an arginine or glutamic acid residue which may be derivatized at the N-terminus;
E3 represents an aromatic amino acid residue;
E4 and E9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
E5 represents an arginine or glutamic acid residue;
E6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
E7 represents a proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
E8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
E10 represents a citrulline, glutamic acid, arginine or lysine residue;
E11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, and the cysteine residues at the 4-position and the 13-position may be linked by a disulfide bond. Or a salt thereof,
(8) The peptide or salt thereof according to (7), wherein E5 represents a glutamic acid residue,
(9) The following formula (If)
1 2 3 4 5 6 7 8 9 10 11 12 13 14
F1-F2-F3-Cys-Tyr-F4-F5-F6-F7-F8-F9-F10-Cys-F11 (If)
(Where
F1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is missing;
F2 represents an arginine or glutamic acid residue when F1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus; Represents an arginine or glutamic acid residue which may be derivatized at the N-terminus;
F3 represents an aromatic amino acid residue;
F4, F5 and F9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
F6 represents a glutamic acid residue;
F7 represents proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
F8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
F10 represents citrulline, glutamic acid, arginine or lysine residue;
F11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, and the cysteine residues at positions 4 and 13 may be linked by a disulfide bond, and the amino acid may be in L-form or D-form. Or a salt thereof,
(10) The following formula (Ig)
1 2 3 4 5 6 7 8 9 10 11 12 13 14
G1-G2-G3-Cys-Tyr-G4-G5-G6-G7-G8-G9-G10-Cys-G11 (Ig)
(Where
G1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is deleted;
G2 represents an arginine or glutamic acid residue when G1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus; Represents an arginine or glutamic acid residue which may be derivatized at the N-terminus;
G3 represents an aromatic amino acid residue;
G4, G5 and G9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
G6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
G7 represents a proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
G8 represents a glutamic acid residue;
G10 represents a citrulline, glutamic acid, arginine or lysine residue;
G11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, and the cysteine residues at the 4-position and the 13-position may be linked by a disulfide bond. Or a salt thereof,
(11) The following formula (Ih)
1 2 3 4 5 6 7 8 9 10 11 12 13 14
H1-H2-H3-Cys-Tyr-H4-H5-H6-H7-H8-H9-H10-Cys-H11 (Ih)
(Where
H1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is missing;
H2 represents an arginine or glutamic acid residue when H1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus; Represents an arginine or glutamic acid residue which may be derivatized at the N-terminus;
H3 represents an aromatic amino acid residue;
H4 and H5 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
H6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
H7 represents a proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
H8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
H9 represents a glutamic acid residue;
H10 represents a citrulline, glutamic acid, arginine or lysine residue;
H11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, and the cysteine residues at the 4-position and the 13-position may be linked by a disulfide bond. Or a salt thereof,
(12) The following formula (Ii)
1 2 3 4 5 6 7 8 9 10 11 12 13 14
I1-I2-I3-Cys-Tyr-I4-I5-I6-I7-I8-I9-I10-Cys-I11 (Ii)
(Where
I1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is missing;
I2 represents an arginine or glutamic acid residue when I1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus; Represents an arginine or glutamic acid residue which may be derivatized at the N-terminus;
I3 represents an aromatic amino acid residue;
I4, I5 and I9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
I6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
17 represents a proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
I8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
I10 represents glutamic acid, arginine or lysine residue;
I11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, and the cysteine residues at the 4-position and the 13-position may be linked by a disulfide bond. Or a salt thereof,
(13) The following formula (Ij)
1 2 3 4 5 6 7 8 9 10 11 12 13 14
J1-J2-J3-Cys-Tyr-J4-J5-J6-J7-J8-J9-J10-Cys-J11 (Ij)
(Where
J1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is deleted;
J2 represents an arginine or glutamic acid residue when J1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus; Represents an arginine or glutamic acid residue which may be derivatized at the N-terminus;
J3 represents an aromatic amino acid residue;
J4, J5 and J9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
J6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
J7 represents a proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
J8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
J10 represents citrulline, glutamic acid, arginine or lysine residue;
J11 represents a glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, and the cysteine residues at the 4-position and the 13-position may be linked by a disulfide bond. Or a salt thereof,
(14) The peptide of any of the following (1) to (58) or a salt thereof:
(1) Ac-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH;
(2) Ac-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-OH;
(3) Ac-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-OH;
(4) Ac-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Cit-Cit-Cys-Arg-OH;
(5) Ac-Cit-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH;
(6) Ac-Cit-Arg-Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-OH;
(7) Ac-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Cit-Cit-Cys-Arg-OH;
(8) Ac-Cit-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Cit-Cit-Cys-Arg-OH;
(9) Ac-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(10) Ac-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Cit-Cit-Cys-Arg-NH 2;
(11) Ac-Cit-Arg -Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(12) Ac-Cit-Arg -Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(13) Ac-Arg-Arg -Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Cit-Cit-Cys-Arg-NH 2;
(14) Ac-Cit-Arg -Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Cit-Cit-Cys-Arg-NH 2;
(15) H-DGlu-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH;
(16) H-Arg-Glu-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH;
(17) H-Arg-Arg-Nal-Cys-Tyr-Glu-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH;
(18) H-Arg-Arg-Nal-Cys-Tyr-Arg-Glu-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH;
(19) H-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-OH;
(20) H-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Glu-Cit-Cys-Arg-OH;
(21) H-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Glu-OH;
(22) H-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(23) H-Arg-Arg -Nal-Cys-Tyr-DGlu-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(24) H-Arg-Arg -Nal-Cys-Tyr-DGlu-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(25) H-DGlu-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(26) H-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-DGlu-Arg-Cit-Cys-Arg-NH 2;
(27) H-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-DGlu-Cys-Arg-NH 2;
(28) Ac-DGlu-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(29) Ac-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-DGlu-Arg-Cit-Cys-Arg-NH 2;
(30) Ac-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-DGlu-Cys-Arg-NH 2;
(31) Ac-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(32) guanyl-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(33) TMguanyl-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(34) TMguanyl-Arg-Nal -Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(35) 4F-benzoyl-Arg -Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(36) 2F-benzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂ ;
(37) APA-Arg-Nal -Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(38) desamino-R-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(39) guanyl-Arg-Nal -Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(40) succinyl-Arg-Nal -Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(41) glutaryl-Arg-Nal -Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(42) deaminoTMG-APA-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(43) nelfinabiryl-succinyl-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂ ;
(44) AZT-glutaryl-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
_{(45) R-CH 2 -Arg} -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(46) H-Arg-Nal -Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(47) TMguanyl-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(48) ACA-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(49) ACA-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH;
(50) H-Arg-Arg -Nal-Cys-Tyr-Cit-Arg-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(51) Ac-Arg-Arg -Nal-Cys-Tyr-Cit-Arg-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(52) Ac-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(53) Ac-Arg-Arg -Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(54) 4F-benzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(55) 4F-benzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NHMe
(56) 4F-benzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NHEt
(57) 4F-benzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NHiPr
(58) 4F-benzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-tyramine
(In each sequence, the symbol to the left of the N-terminal amino acid indicates that the amino group is derivatized or not, H indicates that it is not derivatized, Ac indicates an acetyl group, and guanyl indicates a guanyl group. Succinyl is a succinyl group, glutaryl is a glutaryl group, TMguanyl is a tetramethylguanyl group, 2F-beozoyl is a 2-fluorobenzoyl group, 4F-benzoyl is a 4-fluorobenzoyl group, and APA is a 5-amino group. A pentanoyl group, ACA is a 6-aminohexanoyl group, desamino-R is a 2-desamino-arginyl group, deaminoTMG-APA is the following formula (II),

nelfinabiryl-succinyl has the following formula (III),

AZT-glutaryl has the following formula (IV):

R-CH ₂ has the following formula (V)

Arg represents an L-arginine residue, Nal represents an L-3- (2-naphthyl) alanine residue, Cys represents an L-cysteine residue, Tyr represents an L-tyrosine residue, and Cit represents L-tyrosine residue. -Citrulline residue, Lys is L-lysine residue, DLys is D-lysine residue, Pro is L-proline residue, DCit is D-citrulline residue, DGlu is D-glutamic acid residue. , Glu each represents an L-glutamic acid residue, two cysteine residues are linked by an intramolecular disulfide bond, and the symbol to the right of the C-terminal amino acid means that the carboxyl group is derivatized or not. Shows that OH is not derivatized, NH ₂ is an amino group, NMe is a methylamino group, NEt is an ethylamino group, NiPr is an isopropylamino group, and tyramine is a p-hydroxyphenylethylamino group. Amidated with Door are shown, respectively),
(15) a medicine comprising the peptide or the salt thereof according to any of (3) to (14);
(16) the medicament according to (15), which is a CXCR4 antagonist;
(17) the medicament according to (15), which is a prophylactic and / or therapeutic agent for cancer or rheumatoid arthritis;
(18) the medicament according to (17), wherein the cancer type is breast cancer or pancreatic cancer;
(19) a method for preventing or treating cancer or rheumatoid arthritis, which comprises administering to a mammal an effective amount of the peptide according to any of (3) to (14) or a salt thereof;
(20) Use of the peptide or the salt thereof according to any of (3) to (14) for producing a prophylactic and / or therapeutic agent for cancer or rheumatoid arthritis,
(21) The following formula (Ia) for mammals
1 2 3 4 5 6 7 8 9 10 11 12 13 14
A1-A2-A3-Cys-Tyr-A4-A5-A6-A7-A8-A9-A10-Cys-A11 (Ia)
(Where
A1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is deleted;
A2 represents an arginine or glutamic acid residue when A1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus, and when A1 is deleted Represents an arginine or glutamic acid residue which may be derivatized at the N-terminus;
A3 represents an aromatic amino acid residue;
A4, A5 and A9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
A6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
A7 represents a proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
A8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
A10 represents citrulline, glutamic acid, arginine or lysine residue;
A11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, and the cysteine residues at positions 4 and 13 may be linked by a disulfide bond, and the amino acid may be in L-form or D-form. Or a method for preventing or treating cancer or rheumatoid arthritis, and (22) prevention and / or treatment of cancer or rheumatoid arthritis. Formula (Ia) for producing a therapeutic agent
1 2 3 4 5 6 7 8 9 10 11 12 13 14
A1-A2-A3-Cys-Tyr-A4-A5-A6-A7-A8-A9-A10-Cys-A11 (Ia)
(Where
A1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is deleted;
A2 represents an arginine or glutamic acid residue when A1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus, and when A1 is deleted Represents an arginine or glutamic acid residue which may be derivatized at the N-terminus;
A3 represents an aromatic amino acid residue;
A4, A5 and A9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
A6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
A7 represents a proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
A8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
A10 represents citrulline, glutamic acid, arginine or lysine residue;
A11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, and the cysteine residues at positions 4 and 13 may be linked by a disulfide bond, and the amino acid may be in L-form or D-form. May be used) or a salt thereof,
And so on.

ペ プ チ ド The peptidic compound of the present invention has a strong CXCR4 antagonistic effect, and exhibits a therapeutic effect on cancer and rheumatoid arthritis by inhibiting the binding between CXCR4 and CXCL12 / SDF-1.

The peptide described herein has the N-terminus (amino terminus) at the left end and the C-terminus (carboxyl terminus) at the right end according to the convention of peptide labeling.
In the present specification and the drawings, when amino acids and the like are represented by abbreviations, the abbreviations are based on the abbreviations by IUPAC-IUB Commision on Biochemical Nomenclature or abbreviations commonly used in the art, and examples thereof are described below. In addition, when an amino acid has an optical isomer, the L-form is indicated unless otherwise specified.
Gly or G: glycine Ala or A: alanine Val or V: valine Leu or L: leucine Ile or I: isoleucine Ser or S: serine Thr or T: threonine Cys or C: cysteine Met or M: methionine Glu or E: glutamine Asp or D: Aspartic acid Lys or K: Lysine Arg or R: Arginine His or H: Histidine Phe or F: Phenylalanine Tyr or Y: Tyrosine Trp or W: Tryptophan Pro or P: Proline Asn or N: Asparagine Gln Glutamine pGlu: pyroglutamic acid Nal: 3- (2-naphthyl) alanine Cit: citrulline DLys: D-lysine DCi : D-citrulline DGIu: D-glutamic acid Me: methyl group Et: ethyl group Bu: butyl group Ph: phenyl group

Further, substituents, protecting groups and reagents frequently used in the present specification are represented by the following symbols.
BHA: benzhydrylamine pMBHA: p-methylbenzhydrylamine Tos: p-toluenesulfonyl CHO: formyl HONB: N-hydroxy-5-norbornene over 2,3-dicarboximide OcHex: Cyclohexyl ester Bzl: benzyl Cl ₂ -Bzl: Dichlorobenzyl Bom: benzyloxymethyl Z: benzyloxycarbonyl Br-Z: 2-bromobenzyloxycarbonyl Boc: t-butyloxycarbonyl DCM: dichloromethane HOBt: 1-hydroxybenztriazole DCC: N, N'-dicyclohexylcarbodiimide TFA: Trifluoroacetic acid DIEA: diisopropylethylamine Fmoc: N-9-fluorenylmethoxycarbonyl DNP: dinitrophenyl Bum: tertiary Butoxymethyl Trt: Trityl Ac: Acetyl guanyl: Guanyl succinyl: Succinyl glutaryl: Glutaryl TM guanyl: Tetramethylguanyl 2F-bezoyl: 2-Fluorobenzoyl 4F-benzoyl: 4-Fluorobenzoyl APA: 5-Aminopentanoyl ACA: 6- Aminohexanoyl desamino-R: 2-desamino-arginyl deaminoTMG-APA: Formula (II) below

nelfinabilyl-succinyl: the following formula (III)

AZT-glutaryl: the following formula (IV)

R-CH ₂ : the following formula (V)

"H-" in the N-terminal amino acid of the peptide indicates that the terminal amino group is not derivatized, and "-OH" in the C-terminal amino acid indicates that the terminal carboxyl group is not derivatized.

The present invention provides a prophylactic and / or therapeutic agent for cancer and rheumatoid arthritis containing a compound having a CXCR4 antagonistic action. The “compound having a CXCR4 antagonistic effect” is a compound having an anticancer effect (eg, a migration inhibitory effect, an invasion inhibitory effect, and an anti-metastatic effect) by antagonistically inhibiting the binding between CXCR4 and its physiological ligand, CXCL12 / SDF-1α. Etc.) or an anti-rheumatoid arthritis effect (for example, a migration inhibitory effect), and more specifically, the following formula (Ia)
1 2 3 4 5 6 7 8 9 10 11 12 13 14
A1-A2-A3-Cys-Tyr-A4-A5-A6-A7-A8-A9-A10-Cys-A11 (Ia)
(Where
A1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is deleted;
A2 represents an arginine or glutamic acid residue when A1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus, and when A1 is deleted Represents an arginine or glutamic acid residue which may be derivatized at the N-terminus;
A3 represents an aromatic amino acid residue;
A4, A5 and A9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
A6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
A7 represents a proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
A8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
A10 represents citrulline, glutamic acid, arginine or lysine residue;
A11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, and the cysteine residues at the 4-position and the 13-position may be connected by a disulfide bond. May be)
Or an amide, an ester or a salt thereof (hereinafter may be collectively referred to as “peptide of the present invention”).

A1 represented by the above formula (Ia) represents an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue (which may be L-form or D-form) which may be derivatized at the N-terminus. Represents or deleted, preferably an arginine, citrulline, alanine or D-glutamic acid residue which may be derivatized at the N-terminus. "N-terminal derivatized" includes, for example, a formyl group; an acyl group, for example, a C _2-6 alkanoyl group such as an acetyl group, a propionyl group, a butyryl group, a pentanoyl group, and a hexanoyl group; a benzoyl group; Benzoyl group (Example: 2-fluorobenzoyl, 3-fluorobenzoyl group, 4-fluorobenzoyl group, 2-bromobenzoyl group, 3-bromobenzoyl group, 4-bromobenzoyl group, 2-nitrobenzoyl group, 3-nitrobenzoyl Arylcarbonyl groups such as nitrosyl group, 4-nitrobenzoyl group), succinyl group, glutaryl group; nicotinyl group; isonicotinyl group; alkylsulfonyl group (eg: methanesulfonyl group, ethanesulfonyl group, propanesulfonyl group, camphorsulfonyl group); aryl Sulfonyl group (Example: p-toluenesulfonyl group, 4 -Fluorobenzenesulfonyl group, mesitylenesulfonyl group, 4-aminobenzenesulfonyl group, dansyl group, 4-bromobenzenesulfonyl group) and the like, but are not limited thereto. Alternatively, the N-terminal amino group may be deleted.

A2 represented by the above formula (Ia) is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue (L-form or D-form) in which A1 may be derivatized at the N-terminus. ) Represents arginine or glutamic acid residue (which may be L-form or D-form), and when A1 is deleted, it may be derivatized at the N-terminus. Represents a good arginine or glutamic acid residue (which may be L-form or D-form), and preferably A1 is an arginine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus. In this case, it represents an arginine or glutamic acid residue, and when A1 is deleted, it represents an arginine or glutamic acid residue which may be derivatized at the N-terminus. . "N-terminal derivatized" includes, but is not limited to, those described above for A1.

A3 represented by the above formula (Ia) is an aromatic amino acid residue (for example, phenylalanine, tryptophan, 3- (2-naphthyl) alanine, tyrosine, 4-fluorophenylalanine, 3- (1-naphthyl) alanine) (L And phenylalanine, tryptophan and 3- (2-naphthyl) alanine.

A4 represented by the above formula (Ia) represents arginine, lysine, ornithine, citrulline, alanine or a glutamic acid residue (which may be L-form or D-form), preferably arginine, citrulline, alanine Or L- or D-glutamic acid residue.

A5 represented by the above formula (Ia) represents an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue, and preferably represents an arginine, citrulline, alanine, lysine or glutamic acid residue.

A6 represented by the above formula (Ia) represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or a glutamic acid residue (which may be L-form or D-form), and is preferably D -Represents a lysine, D-alanine, D-citrulline or D-glutamic acid residue.

A7 represented by the above formula (Ia) represents a proline or alanine residue (which may be L-form or D-form), and preferably represents a proline or alanine residue.

A8 represented by the above formula (Ia) represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or a glutamic acid residue (which may be L-form or D-form), and is preferably tyrosine, alanine or Represents a D-glutamic acid residue.

A9 represented by the above formula (Ia) represents arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue (which may be L-form or D-form), and is preferably arginine, citrulline or glutamic acid. Represents a residue.

A10 represented by the above formula (Ia) represents a citrulline, glutamic acid, arginine or lysine residue (which may be L-form or D-form), and preferably represents citrulline or D-glutamic acid residue. .

A11 represented by the above formula (Ia) represents an arginine, glutamic acid, lysine or citrulline residue (which may be L-form or D-form) which may be derivatized at the C-terminus, and is preferably , An arginine or glutamic acid residue which may be derivatized at the C-terminus. Examples of “C-terminal derivatization” include amidation (—NH ₂ , —NHR, —NRR ′), esterification (—COOR), and the like. Here, as R and R ′ in the amide and ester, for example, a C _1-6 alkyl group such as methyl, ethyl, n-propyl, isopropyl or n-butyl, for example, a C _3-8 cycloalkyl such as cyclopentyl and cyclohexyl Groups, for example, C _6-12 aryl groups such as phenyl and α-naphthyl, for example, phenyl-C _1-2 alkyl groups such as benzyl and phenethyl or α-naphthyl-C _1-2 alkyl groups such as α-naphthylmethyl In addition to the C _7-14 aralkyl group, a pivaloyloxymethyl group commonly used as an oral ester is used.

When the peptide of the present invention has a carboxyl group (or carboxylate) other than at the C-terminus, those in which the carboxyl group is amidated or esterified are also included in the peptide of the present invention. As the amide and ester in this case, for example, the C-terminal amide and ester exemplified for the above-mentioned A11 are used in the same manner. Further, in the peptide of the present invention, a substituent (for example, -OH, -SH, an amino group, an imidazole group, an indole group, a guanidino group, etc.) on a side chain of an amino acid in the molecule in the peptide described above is suitable. Those protected with a protecting group (for example, a C _1-6 acyl group such as a C _2-6 alkanoyl group such as a formyl group or an acetyl group), or a complex peptide such as a so-called glycopeptide having a sugar chain bonded thereto are also included. included.

塩 Examples of the salt of the peptide of the present invention include a physiologically acceptable salt with an acid or a base, and a physiologically acceptable acid addition salt is particularly preferable. Such salts include, for example, salts with inorganic acids (eg, hydrochloric acid, phosphoric acid, hydrobromic acid, sulfuric acid) or organic acids (eg, acetic acid, formic acid, propionic acid, fumaric acid, maleic acid, succinic acid) Acid, tartaric acid, citric acid, malic acid, oxalic acid, benzoic acid, methanesulfonic acid, benzenesulfonic acid) and the like.

Further, the peptide of the present invention is a novel peptide when any one of A1 to A11 represented by the above formula (Ia) is as follows:
(I) when A1 is a glutamic acid residue or is deleted (that is, in the case of the above formula (Ib));
(Ii) when any one of A2, A4, A6, A8 and A9 is a glutamic acid residue (that is, any of the above formulas (Ic) to (Ig));
(Iii) when A5 is an arginine or glutamic acid residue (that is, in the case of the above formula (Ih));
(Iv) when A10 is a glutamic acid, arginine or lysine residue (that is, in the case of the above formula (Ii));
(V) When A11 is a glutamic acid, lysine or citrulline residue (that is, in the case of the above formula (Ij)).
The amino acid residue may be in L-form or D-form.

Preferable examples of the novel peptide of the present invention include peptides having the following amino acid sequences (1) to (58) (in each sequence, two cysteine residues are linked by disulfide bonds).
(1) Ac-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(AcTC14003)
(2) Ac-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(AcTC14005)
(3) Ac-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(AcTC14011)
(4) Ac-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Cit-Cit-Cys-Arg-OH
(AcTC14013)
(5) Ac-Cit-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(AcTC14015)
(6) Ac-Cit-Arg-Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(AcTC14017)
(7) Ac-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Cit-Cit-Cys-Arg-OH
(AcTC14019)
(8) Ac-Cit-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Cit-Cit-Cys-Arg-OH
(AcTC14021)
(9) Ac-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(AcTC14012)
(10) Ac-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Cit-Cit-Cys-Arg-NH ₂
(AcTC14014)
(11) Ac-Cit-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(AcTC14016)
(12) Ac-Cit-Arg-Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(AcTC14018)
(13) Ac-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Cit-Cit-Cys-Arg-NH ₂
(AcTC14020)
(14) Ac-Cit-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Cit-Cit-Cys-Arg-NH ₂
(AcTC14022)
(15) H-DGlu-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(TE14001)
(16) H-Arg-Glu-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(TE14002)
(17) H-Arg-Arg-Nal-Cys-Tyr-Glu-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(TE14003)
(18) H-Arg-Arg-Nal-Cys-Tyr-Arg-Glu-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(TE14004)
(19) H-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(TE14005)
(20) H-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Glu-Cit-Cys-Arg-OH
(TE14006)
(21) H-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Glu-OH
(TE14007)
(22) H-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TE14011)
(23) H-Arg-Arg-Nal-Cys-Tyr-DGlu-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TE14012)
(24) H-Arg-Arg-Nal-Cys-Tyr-DGlu-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TE14013)
(25) H-DGlu-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TE14014)
(26) H-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-DGlu-Arg-Cit-Cys-Arg-NH ₂
(TE14015)
(27) H-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-DGlu-Cys-Arg-NH ₂
(TE14016)
(28) Ac-DGlu-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(AcTE14014)
(29) Ac-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-DGlu-Arg-Cit-Cys-Arg-NH ₂
(AcTE14015)
(30) Ac-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-DGlu-Cys-Arg-NH ₂
(AcTE14016)
(31) Ac-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF1: AcTE14011)
(32) guanyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF2: guanyl-TE14011)
(33) TMguanyl-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2
(TF3: TMguanyl-TE14011)
(34) TMguanyl-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF4: TMguanyl-TE14011 (2-14))
(35) 4F-benzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF5: 4F-benzoyl-TE14011)
(36) 2F-benzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF6: 2F-benzoyl-TE14011)
(37) APA-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF7: APA-TE14011 (2-14))
(38) desamino-R-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF8: desamino-R-TE14011 (2-14))
(39) guanyl-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF9: guanyl-TE14011 (2-14))
(40) succinyl-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF10: succinyl-TE14011 (2-14))
(41) glutaryl-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF11: glutaryl-TE14011 (2-14))
(42) deaminoTMG-APA-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2
(TF12: deaminoTMG-APA-TE14011 (2-14))
(43) nelfinabiryl-succinyl-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF13: nelfinabiryl-succinyl-TE14011 (2-14))
(44) AZT-glutaryl-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2
(TF14: AZT-glutaryl-TE14011 (2-14))
(45) R-CH ₂ -Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF15: R-CH ₂ NH-RTE14011 (2-14))
(46) H-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF17: TE14011 (2-14))
(47) TMguanyl-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2
(TF18: TMguanyl-TC14012)
(48) ACA-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF19: ACA-TC14012)
(49) ACA-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(TF20: ACA-T140)
(50) H-Arg-Arg -Nal-Cys-Tyr-Cit-Arg-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2
(TZ14011)
(51) Ac-Arg-Arg-Nal-Cys-Tyr-Cit-Arg-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(AcTZ14011)
(52) Ac-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(AcTN14003)
(53) Ac-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(AcTN14005)
(54) 4F-benzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(4F-benzoyl-TN14003)
(55) 4F-benzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NHMe
(4F-benzoyl-TN14011-Me)
(56) 4F-benzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NHEt
(4F-benzoyl-TN14011-Et)
(57) 4F-benzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NHiPr
(4F-benzoyl-TN14011-iPr)
(58) 4F-benzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-tyramine
(4F-benzoyl-TN14011-tyramine)

The peptide of the present invention also includes a peptide containing an amino acid sequence substantially identical to the amino acid sequence of any of the above-mentioned peptides, or an amide, ester or salt thereof. As used herein, the term “substantially identical amino acid sequence” refers to the activity of a peptide (for example, a ligand-receptor binding inhibitory activity) or the anticancer activity of a peptide (for example, a migration inhibitory activity, an invasion inhibitory activity, and an anti-metastatic activity). Etc.) or anti-rheumatic effects (eg, migration inhibitory effects) and the like are homogenous in nature [accordingly, to a certain extent (eg, about 0.01 to 100 times, preferably about 0.5 to 20 times, More preferably about 0.5 to 2 times). Therefore, one or more amino acids may be mutated in the amino acid sequence represented by any of the above formulas (Ia) to (Ij) and (1) to (58) as long as the above properties are maintained. .
That is, in the present invention, a significant (significant) change in the physiological properties or chemical properties of the original (unmutated) peptide (= significantly different even in nature, or significantly different in quantity even if it is homogeneous) Peptide (mutant peptide) obtained as a result of a mutation in the amino acid sequence such as substitution, deletion or insertion (addition) that does not result in such a mutation). The amino acid sequence of the variant peptide is considered to be substantially identical to the amino acid sequence of the original (unmutated) peptide.
In general, mutations such as amino acid substitutions, deletions or insertions (additions) in a peptide sequence often do not result in large (significant) changes in the physiological or chemical properties of the peptide. This is a well-known fact. Examples of the substitution include a substitution of a certain amino acid with another amino acid having similar properties (characteristics). Generally, when substitution is performed between amino acids having strong similarity in properties. It is considered that the smaller the substitution, the smaller the change in properties of the original peptide before substitution. Amino acids are classified into the following classes based on similarity in their properties, for example: (i) non-polar (hydrophobic) amino acids (eg, alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan, methionine, etc.) (Ii) polar (neutral) amino acids (eg, glycine, serine, threonine, cysteine, tyrosine, asparagine, glutamine, etc.); (iii) positively charged (basic) amino acids (eg, arginine, lysine, histidine) And (iv) are classified as negatively charged (acidic) amino acids (eg, aspartic acid, glutamic acid, etc.), so that amino acid substitutions within each class are conservative (ie, “ Substitutions that result in "substantially identical" amino acid sequences.

In other words, "substantially identical amino acid sequences"
(I) one or more, preferably one to ten, and more preferably one to five in the amino acid sequence represented by any of the above formulas (Ia) to (Ij) and (1) to (58). An amino acid sequence in which not more than one amino acid has been replaced by another amino acid,
(Ii) 1 to 7 amino acids, preferably 1 to 5 amino acids, more preferably 1 to 5 amino acids in the amino acid sequence represented by any of the above formulas (Ia) to (Ij) and (1) to (58). An amino acid sequence in which at least 3 and no more than 3 amino acids have been deleted,
(Iii) The amino acid sequence represented by any one of the formulas (Ia) to (Ij) and (1) to (58) has 1 to 15 amino acids, preferably 1 to 10 amino acids, more preferably 1 amino acid The amino acid sequence having (inserted) at least 5 or less amino acids, or (iv) a constituent amino acid (particularly, its side chain) in a peptide having the amino acid sequence of (i), (ii) or (iii) above. Peptides containing modifications, or amides, esters or salts thereof, may be included.
The peptide of the present invention is obtained by subjecting the amino acid sequence (i) to (iv) to substitution, deletion, insertion (addition), modification, or the like intentionally or accidentally, to thereby obtain a stable peptide against heat or protease. Alternatively, the peptide can be converted to a highly active peptide having an increased inhibitory activity. The peptide of the present invention also includes these mutant peptides or amides, esters or salts thereof.

Further, as the peptide of the present invention, in addition to the peptide consisting of the amino acid sequence represented by any one of the above formulas (Ia) to (Ij) and (1) to (58), the amino acid sequence and the amino acid sequence of about 50 to 99. It contains an amino acid sequence having a homology of 9% (preferably 70 to 99.9%, more preferably 80 to 99.9%, and still more preferably 90 to 99.9%), and contains the above-mentioned formulas (Ia) to (Ia). A peptide having substantially the same activity as the peptide having the amino acid sequence represented by any one of Ij) and (1) to (58), an amide thereof, an ester thereof, or a salt thereof. Examples of the activity include an inhibitory activity of the peptide, such as a binding inhibitory activity between a ligand and a receptor and a signal transduction inhibitory activity. The expression “substantially the same” as the inhibitory activity indicates that the properties such as the ligand binding inhibitory activity to the receptor are the same. Therefore, the ligand binding inhibitory activity on the receptor may be insignificantly strong or weak, and the difference in molecular weight is not a problem.

Examples of the amide, ester or salt of the peptide having the amino acid sequence represented by any one of the above formulas (1) to (58) include those exemplified for the peptide represented by the above general formula (Ia). . Preferably, in the peptide having the amino acid sequence represented by any of the above formulas (1) to (58), the carboxyl group of the C-terminal amino acid residue is desirably amidated.

The peptides of the present invention, including the peptides containing the amino acid sequences represented by the above formulas (1) to (58), can be produced according to peptide synthesis methods known per se. As a method for synthesizing a peptide, for example, any of a solid phase synthesis method and a liquid phase synthesis method may be used. That is, a partial peptide or amino acid that can constitute a peptide is condensed with the remaining portion, and when the product has a protecting group, the protecting group is eliminated to produce the target peptide. Known methods for condensation and elimination of protecting groups include, for example, the methods described in 1) to 5) below.
1) M. Bodanszky and MA Ondetti, Peptide Synthesis, Interscience Publishers, New York (1966)
2) Schroeder and Luebke, The Peptide, Academic Press, New York (1965)
3) Nobuo Izumiya et al., Basics and Experiments of Peptide Synthesis, Maruzen Co., Ltd. (1975)
4) Haruaki Yajima and Shunpei Sakakibara, Laboratory of Biochemistry 1, Protein Chemistry IV, 205, (1977)
5) Supervised by Haruaki Yajima, Development of Continuing Drugs Volume 14 Peptide Synthesis Hirokawa Shoten

As a specific peptide synthesis method, for example, the following method can be mentioned.
Usually, a commercially available resin for polypeptide synthesis can be used. Examples of such a resin include chloromethyl resin, hydroxymethyl resin, benzhydrylamine resin, aminomethyl resin, 4-benzyloxybenzyl alcohol resin, 4-methylbenzhydrylamine resin, PAM resin, and 4-hydroxymethylmethyl. Phenylacetamidomethyl resin, polyacrylamide resin, 4- (2 ′, 4′-dimethoxyphenyl-hydroxymethyl) phenoxy resin, 4- (2 ′, 4′-dimethoxyphenyl-Fmocaminoethyl) phenoxy resin, and the like. it can. Using such a resin, an amino acid having an α-amino group and a side chain functional group appropriately protected is condensed on the resin according to the sequence of the target polypeptide according to various condensation methods known per se. At the end of the reaction, the polypeptide is cleaved from the resin, and at the same time, various protecting groups are removed. Further, an intramolecular disulfide bond formation reaction is carried out in a highly diluted solution to obtain a target polypeptide or an amide thereof. For the condensation of the protected amino acids described above, various activating reagents that can be used for polypeptide synthesis can be used, and carbodiimides are particularly preferable. As the carbodiimides, DCC, N, N′-diisopropylcarbodiimide, N-ethyl-N ′-(3-dimethylaminoprolyl) carbodiimide and the like are used. For these activations, the protected amino acid was directly added to the resin together with a racemization inhibitor additive (eg, HOBt, HOOBt), or the protected amino acid was previously activated as a symmetric anhydride or HOBt ester or HOOBt ester. Later it can be added to the resin.

溶媒 The solvent used for activating the protected amino acid or condensing with the resin may be appropriately selected from solvents known to be usable for the polypeptide condensation reaction. For example, acid amides such as N, N-dimethylformamide, N, N-dimethylacetamide, N-methylpyrrolidone, halogenated hydrocarbons such as methylene chloride and chloroform, alcohols such as trifluoroethanol, and dimethyl sulfoxide. Sulfoxides, ethers such as pyridine, dioxane, and tetrahydrofuran, nitriles such as acetonitrile and propionitrile, esters such as methyl acetate and ethyl acetate, and appropriate mixtures thereof are used. The reaction temperature is appropriately selected from a range known to be usable for a polypeptide bond formation reaction, and is usually appropriately selected from a range of about -20 ° C to 50 ° C. The activated amino acid derivative is usually used in a 1.5 to 4-fold excess. As a result of the test using the ninhydrin reaction, if the condensation is insufficient, sufficient condensation can be performed by repeating the condensation reaction without removing the protecting group. When a sufficient condensation cannot be obtained by repeating the reaction, the unreacted amino acid can be acetylated using acetic anhydride or acetylimidazole.

Examples of the protecting group for the amino group of the starting material include Z, Boc, tertiary pentyloxycarbonyl, isobornyloxycarbonyl, 4-methoxybenzyloxycarbonyl, Cl-Z, Br-Z, adamantyloxycarbonyl, trifluoroacetyl , Phthaloyl, formyl, 2-nitrophenylsulfenyl, diphenylphosphinothioyl, Fmoc and the like. The carboxyl group may be, for example, a linear, branched or cyclic alkyl ester such as an alkyl esterified ester (eg, methyl, ethyl, propyl, butyl, tertiary butyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 2-adamantyl). ), Aralkyl esterification (eg, benzyl ester, 4-nitrobenzyl ester, 4-methoxybenzyl ester, 4-chlorobenzyl ester, benzhydryl esterification), phenacyl esterification, benzyloxycarbonyl hydrazide, tertiary butoxy It can be protected by carbonyl hydrazide, trityl hydrazide and the like. The hydroxyl group of serine can be protected, for example, by esterification or etherification. As a group suitable for this esterification, for example, a lower alkanoyl group such as an acetyl group, an aroyl group such as a benzoyl group, and a group derived from carbonic acid such as a benzyloxycarbonyl group and an ethoxycarbonyl group are used. Further, groups suitable for etherification include, for example, a benzyl group, a tetrahydropyranyl group, a t-butyl group and the like. As the protecting group for the phenolic hydroxyl group of tyrosine, for example, Bzl, Cl ₂ -Bzl, 2-nitrobenzyl, Br-Z, tert-butyl and the like are used. As the protecting group for imidazole of histidine, for example, Tos, 4-methoxy-2,3,6-trimethylbenzenesulfonyl, DNP, benzyloxymethyl, Bum, Boc, Trt, Fmoc and the like are used.

Examples of activated carboxyl groups of the raw materials include, for example, corresponding acid anhydrides, azides, active esters [alcohols (eg, pentachlorophenol, 2,4,5-trichlorophenol, 2,4-dinitrophenol, Cyanomethyl alcohol, paranitrophenol, HONB, N-hydroxysuccinimide, N-hydroxyphthalimide, esters with HOBt), and the like. As the activated amino group of the raw material, for example, a corresponding phosphoric amide is used. As a method for removing (eliminating) the protecting group, for example, catalytic reduction in a hydrogen stream in the presence of a catalyst such as Pd-black or Pd-carbon, or hydrogen fluoride anhydride, methanesulfonic acid, trifluoromethane or the like can be used. Acid treatment with dichloromethane, trifluoroacetic acid or a mixture thereof, base treatment with diisopropylethylamine, triethylamine, piperidine, piperazine, etc., reduction with sodium in liquid ammonia, and the like are also used. The elimination reaction by the above acid treatment is generally performed at a temperature of about -20 ° C to 40 ° C. In the acid treatment, for example, anisole, phenol, thioanisole, metacresol, paracresol, dimethylsulfide, 1,4 -Addition of a cation scavenger such as butanedithiol, 1,2-ethanedithiol and the like is effective. In addition, the 2,4-dinitrophenyl group used as the imidazole protecting group of histidine is removed by thiophenol treatment, and the formyl group used as the indole protecting group of tryptophan is 1,2-ethanedithiol, 1,4-butane described above. In addition to deprotection by acid treatment in the presence of dithiol or the like, it is also removed by alkali treatment with dilute sodium hydroxide solution, dilute ammonia or the like.

保護 The protection of the functional group which should not be involved in the reaction of the raw material, the protective group, the elimination of the protective group, the activation of the functional group involved in the reaction, and the like can be appropriately selected from known groups or known means. As another method for obtaining an amide form of a polypeptide, for example, first, after protecting the α-carboxyl group of the carboxy terminal amino acid by amidation, the peptide chain is extended to a desired chain length on the amino group side, and A polypeptide in which only the protecting group for the α-amino group at the N-terminus of the peptide chain has been removed and a polypeptide in which only the protecting group for the carboxyl group at the C-terminus has been removed are prepared. Condensation in Details of the condensation reaction are the same as described above. After purifying the protected polypeptide obtained by the condensation, all the protecting groups are removed by the above-mentioned method to obtain a desired crude polypeptide. The crude polypeptide is purified by various known purification means, and the main fraction is freeze-dried to obtain an amide of the desired polypeptide. In order to obtain an ester of the polypeptide, for example, after condensing the α-carboxyl group of the carboxy terminal amino acid with a desired alcohol to form an amino acid ester, the ester of the desired polypeptide can be obtained in the same manner as the amide of the polypeptide. You can get the body.

ペ プ チ ド After the reaction, the peptide of the present invention can be purified and isolated by a combination of ordinary purification methods, for example, solvent extraction, distillation, column chromatography, liquid chromatography, recrystallization and the like. When the peptide obtained by the above method is a free form, it can be converted into an appropriate salt by a known method, and when obtained by a salt, it can be converted into a free form by a known method. it can.

Preferably, the novel peptide of the present invention having the amino acid sequences represented by the above formulas (1) to (58) can be produced by the method described in Examples below or a method analogous thereto. Further, the peptide of the present invention can be produced by the method described in WO02 / 20561 pamphlet or a method analogous thereto.

ペ プ チ ド When the peptide of the present invention is used as a pharmaceutical or veterinary drug, it may be performed according to a conventional method. For example, tablets, capsules, elixirs, microcapsules, and the like, which are sugar-coated as necessary, or sterile solution or suspension with water or other pharmaceutically acceptable liquids It can be used parenterally in the form of injections. For example, the peptide of the present invention may be mixed with physiologically acceptable carriers, flavors, excipients, vehicles, preservatives, stabilizers, binders, and the like in a unit dosage form required for generally accepted pharmaceutical practice. Can be manufactured by The amount of the active ingredient in these preparations is such that an appropriate dose in the specified range can be obtained.

Examples of additives that can be incorporated into tablets, capsules and the like include binders such as gelatin, corn starch, tragacanth gum, gum arabic, excipients such as crystalline cellulose, corn starch, gelatin, alginic acid and the like. Swelling agents, lubricants such as magnesium stearate, sweetening agents such as sucrose, lactose or saccharin, flavoring agents such as peppermint, reddish oil or cherry and the like are used. When the preparation unit form is a capsule, a liquid carrier such as oil and fat can be further contained in the above-mentioned type of material. Sterile compositions for injection can be formulated according to normal pharmaceutical practice such as dissolving or suspending the active substance in vehicles such as water for injection, and naturally occurring vegetable oils such as sesame oil, coconut oil and the like. .
Examples of aqueous liquids for injection include physiological saline, isotonic solutions containing glucose and other adjuvants (eg, D-sorbitol, D-mannitol, sodium chloride, etc.), and suitable solubilizing agents. For example, alcohols (eg, ethanol), polyalcohols (eg, propylene glycol, polyethylene glycol), non-ionic surfactants (eg, Polysorbate 80 ^™ , HCO-50) and the like may be used in combination. The oily liquid includes sesame oil, soybean oil and the like, and may be used in combination with solubilizers such as benzyl benzoate and benzyl alcohol. In addition, buffers (eg, phosphate buffer, sodium acetate buffer), soothing agents (eg, benzalkonium chloride, procaine hydrochloride, etc.), stabilizers (eg, human serum albumin, polyethylene glycol, etc.), storage Agents (eg, benzyl alcohol, phenol, etc.), antioxidants and the like. The prepared injection is usually aseptically filled in a suitable ampoule.
Since the preparation obtained in this way is safe and has low toxicity, it can be used in humans and mammals (eg, mouse, rat, guinea pig, rabbit, sheep, pig, cow, cat, dog, monkey, baboon, chimpanzee, etc.). Can be administered.
The dosage of the peptide of the present invention may vary depending on symptoms, but when administered orally, it is usually about 0.1 to 1000 mg, preferably about 1.0 to 500 mg, more preferably about 1.0 to 500 mg per 60 kg body weight at a time. It is about 1.0 to 200 mg. In the case of parenteral administration, a single dose for a body weight of 60 kg varies depending on an administration subject, a symptom, an administration method and the like. For example, in the case of an injection, it is usually about 0.01 to 300 mg at a time. The dose, preferably about 0.1 to 200 mg, more preferably about 0.1 to 100 mg may be administered by intravenous injection. In the case of other animals, the amount can be administered in terms of the weight per 60 kg.

The peptide of the present invention has an anticancer effect, that is, a cancer cell motility suppressing effect and a cancer metastasis inhibiting effect. That is, since the peptide of the present invention has a cancer metastasis inhibitory action, as will be apparent from the examples described later, it can be used as a prophylactic and therapeutic drug relating to cancer, particularly to cancer metastasis. Therefore, the peptide of the present invention, as an anticancer drug, oral cancer, pharyngeal cancer, lip cancer, tongue cancer, gingival cancer, nasopharyngeal cancer, esophageal cancer, gastric cancer, small intestine cancer, colon cancer including colon cancer, liver cancer, Gallbladder cancer, pancreatic cancer, nasal cavity cancer, lung cancer, osteosarcoma, soft tissue cancer, skin cancer, melanoma, breast cancer, uterine cancer, ovarian cancer, prostate cancer, testicular cancer, penis cancer, bladder cancer, kidney cancer, brain tumor, thyroid gland It is useful as an agent for improving, preventing and treating cancer, lymphoma, leukemia and the like. Further, the peptide of the present invention has an anti-chronic rheumatoid action, that is, a T cell motility suppressing action. That is, since the peptide of the present invention has a T cell motility inhibitory effect, as will be apparent from Examples described later, it can be used as a preventive and therapeutic drug for rheumatoid arthritis. Therefore, the peptide of the present invention is useful as an agent for improving, preventing and treating rheumatoid arthritis.
It is known that compounds having CXCR4 antagonistic activity have antiviral activity. Therefore, it will be obvious to those skilled in the art that the peptide of the present invention can also be used as a prophylactic and therapeutic agent for viral infections (eg, AIDS, SARS, etc.).

For example, when the peptide of the present invention is used as an anticancer drug, other anticancer drugs (eg, chemotherapeutic drugs, immunotherapeutic drugs, or drugs that inhibit the action of cell growth factors and their receptors), etc. (Abbreviated as drug).
Although the peptide of the present invention exhibits excellent anticancer activity even when used as a single agent, the effect can be further enhanced by using it in combination with one or several of the above concomitant drugs (multiple drug combination). Can be.

Examples of the "chemotherapeutic agent" include an alkylating agent, an antimetabolite, an anticancer antibiotic, and a plant-derived anticancer agent.
Examples of the "alkylating agent" include, for example, nitrogen mustard, nitrogen mustard hydrochloride-N-oxide, chlorambutyl, cyclophosphamide, ifosfamide, thiotepa, carbone, improsulfan tosylate, busulfan, nimustine hydrochloride, mitobronitol, mellon Phalan, Dacarbazine, Ranimustine, Estramustine sodium phosphate, Triethylenemelamine, Carmustine, Lomustine, Streptozocin, Pipobroman, Etogluside, Artretamine, Ambamustine, Dibrospidium hydrochloride, Fotemustine, Prednistin, Bumitepa, Ribomustine, Temozolomid , Trofosfamide, dinostatin stimaramar, carbone, adzeresin, systemistin, viseresi , Platinum complexes (carboplatin, cisplatin, miboplatin, nedaplatin, such as oxaliplatin), and the like.

Examples of “antimetabolites” include, for example, mercaptopurine, 6-mercaptopurine riboside, thioinosine, methotrexate, enositabine, cytarabine, cytarabine octophosphate, ancitabine hydrochloride, 5-FU drugs (eg, fluorouracil, tegafur, UFT, Doxyfluridine, carmofur, galocitabine, mititefur), aminopterin, leucovorin calcium, tabloid, butosin, folinate calcium, levofolinate calcium, cladribine, emitefur, fludarabine, gemcitabine, hydroxycarbamide, pentostatin, pyritreximite, idoxiurizin, idoxiurizine, idoxiurizine, idoxiuridine , Ambamustine, gemcitabine and the like.

Examples of the "anticancer antibiotic" include anthracycline anticancer drugs (doxorubicin hydrochloride, daunorubicin hydrochloride, aclarubicin hydrochloride, pirarubicin hydrochloride, epirubicin hydrochloride, etc.), actinomycin D, actinomycin C, mitomycin C, chromomycin A3 Bleomycin hydrochloride, bleomycin sulfate, peplomycin sulfate, neocarzinostatin, mythramycin, sarcomycin, cartinophilin, mitotane, sorbicin hydrochloride, mitoxantrone hydrochloride, idarubicin hydrochloride and the like.

Examples of the “plant-derived anticancer agent” include, for example, vinca alkaloid anticancer drugs (vinblastine sulfate, vincristine sulfate, vindesine sulfate, vinorelbine, etc.), taxane anticancer drugs (paclitaxel, docetaxel, etc.), etoposide, etoposide phosphate, teniposide, Vinorelbine and the like.
The “cell growth factor” in the “drug that inhibits the action of cell growth factor and its receptor” may be any substance that promotes cell growth, and usually has a molecular weight of 20,000. Among the following peptides, there may be mentioned factors which exert an effect at a low concentration by binding to a receptor. Specifically, (1) EGF (epidermal growth factor) or a substance having substantially the same activity as that [ (Eg, EGF, hallegulin (HER2 ligand), etc.), (2) insulin or a substance having substantially the same activity as that (eg, insulin, IGF (insulin-like growth factor) -1, IGF-2, etc.), ( 3) FGF (fibroblast growth factor) or a substance having substantially the same activity as that [eg, acidic FGF, basic FGF, KGF (keratinocyte growth factor), FGF-10, etc.] (4) Other cell growth factors [eg, CSF (colony stimulating factor), EPO (erythropoietin), IL-2 (interleukin-2), NGF (nerve growth factor), PDGF (platelet-derived growth factor), TGFβ ( transforming growth factor β), HGF (hepatocyte growth factor), VEGF (vascular endothelial growth factor) and the like].

The "cell growth factor receptor" may be any receptor as long as it has the ability to bind to the above-mentioned cell growth factor, and specifically, EGF receptor, hallegulin receptor (HER2) , Insulin receptor, IGF receptor, FGF receptor-1 or FGF receptor-2, HGF receptor (c-met), VEGF receptor, SCF receptor (c-kit) and the like.
Examples of the “drug that inhibits the action of cell growth factor” include Herceptin (HER2 antibody), GLEEVEC (c-met, c-kit, abl inhibitor), Iressa (EGF receptor inhibitor) and the like.

の 他 In addition to the above agents, topoisomerase I inhibitors (eg, irinotecan, topotecan, etc.), topoisomerase II inhibitors (eg, sobuzoxane, etc.), angiogenesis inhibitors and the like can also be used.

When the peptide of the present invention is used as a prophylactic and / or therapeutic agent for rheumatoid arthritis, it can be used in combination with a prophylactic and / or therapeutic agent for other joint diseases. Examples of the concomitant drug include anti-inflammatory steroids (eg, prednisolone, hydrocortisone, methylprednisolone, dexamethasone, betamethasone, etc.), non-steroidal anti-inflammatory analgesics (eg, indomethacin, diclofenac, loxoprofen, ibuprofen, aspirin, piroxicam) , Sulindac, etc.) or hyaluronic acid preparations (eg, sodium hyaluronate), COX-II inhibitors and the like.
Although the peptide of the present invention exhibits excellent anti-rheumatic rheumatoid action even when used as a single agent, the effect is further enhanced by using it in combination with one or several of the above concomitant drugs (multiple drug combination). Can be done.

When the peptide of the present invention is used in combination with the concomitant drug, the administration time of the peptide of the present invention and the concomitant drug is not limited, and the peptide of the present invention and the concomitant drug may be simultaneously administered to a subject to be administered. It may be administered with a time lag. The dose of the concomitant drug may be in accordance with the dose clinically used, and can be appropriately selected depending on the administration subject, administration route, disease, combination, and the like.

投 与 The administration form of the peptide of the present invention and the concomitant drug is not particularly limited, as long as the polypeptide of the present invention or a salt thereof and the concomitant drug are combined at the time of administration. Such administration forms include, for example, (1) administration of a single preparation obtained by simultaneously preparing the peptide of the present invention and the concomitant drug, and (2) separate preparation of the peptide of the present invention and the concomitant drug. (2) Simultaneous administration of the two preparations obtained in the same administration route by the same administration route, (3) a time difference in the same administration route of the two preparations obtained by separately preparing the peptide of the present invention and the concomitant drug. (4) Simultaneous administration of two types of preparations obtained by separately formulating the peptide of the present invention and the concomitant drug by different administration routes, (5) Separate preparation of the peptide of the present invention and the concomitant drug And administration of the two preparations obtained in different times with different administration routes (for example, administration in the order of the peptide of the present invention → concomitant drug or administration in the reverse order) and the like. Hereinafter, these administration forms are collectively abbreviated as the concomitant drug of the present invention.

The concomitant drug of the present invention has low toxicity. For example, the peptide of the present invention and / or the concomitant drug is mixed with a pharmacologically acceptable carrier according to a method known per se, and a pharmaceutical composition such as a tablet (sugar-coated tablet) is prepared. , Film-coated tablets), powders, granules, capsules, (including soft capsules), liquids, injections, suppositories, sustained-release preparations, etc., orally or parenterally (eg, topical, rectal, intravenous) Administration etc.) can be safely administered. Injectables can be administered intravenously, intramuscularly, subcutaneously, intraorganically, intranasally, intradermally, instilled, intracerebrally, rectally, intravaginally and intraperitoneally, inside tumors, proximal to tumors, etc. Can be administered.

As the pharmacologically acceptable carrier that may be used for the production of the concomitant drug of the present invention, the same carriers as those used in the above-mentioned pharmaceutical composition of the present invention can be used.
The compounding ratio of the peptide of the present invention to the concomitant drug in the concomitant drug of the present invention can be appropriately selected depending on the administration subject, administration route, disease and the like.
The content of the concomitant drug in the concomitant drug of the present invention varies depending on the form of the preparation, but is usually about 0.01 to 100% by weight, preferably about 0.1 to 50% by weight, more preferably about 0.1 to 50% by weight, based on the whole preparation. It is about 0.5 to 20% by weight.
The content of additives such as carriers in the concomitant drug of the present invention varies depending on the form of the preparation, but is usually about 1 to 99.99% by weight, preferably about 10 to 90% by weight, based on the whole preparation. .

The present invention will be described in more detail with reference to examples below, but these examples do not limit the scope of the present invention.

<Production Example 1: Production of polypeptide TC14003>
H-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(TC14003)
1. Synthesis of TC14003 protected polypeptide resin Fmoc-Cys (Trt) corresponding to the 13th position after removing the Fmoc group from the alcoholic resin Fmoc-Arg (Pbf) -Alko resin introduced with arginine at the 14th position with 20% piperidine / DMF -OH (2.5 eq) was added, and a condensation reaction was performed in DMF by the DIPCDI-HOBt method. The progress of the condensation reaction was determined by the ninhydrin test of Kaiser, E. et al. (Anal. Biochem., 34: 595 (1970)).

2.Introduction of amino acids at positions 12 to 1 Similarly, Cit, Arg (Pbf), Tyr (t-Bu), Pro, DLys (Boc), Lys (Boc), Cit, Tyr (t-Bu) ), Cys (Trt), Nal, Arg (Pbf) and Arg (Pbf) residues were introduced into the resin to obtain a protecting group-protected polypeptide resin.

3. Deprotection Group, Separation and Purification of Polypeptide from Resin The protecting group protected polypeptide resin was treated with 20% piperidine / DMF to remove the Fmoc group, and then the resin was subjected to 1M TMSBr-thioanisole / TFA (trifluoro The reaction was performed at 25 ° C. for 2 hours in an acetic acid system (in the presence of m-cresol (100 eq) and ethanedithiol (300 eq)). The resin was filtered off from the reaction mixture, washed twice with TFA, the combined filtrate and washings were concentrated under reduced pressure, water-cooled dry ether was added to the residue, and the resulting precipitate was centrifuged and centrifuged and decanted. Separated from The obtained residue was washed with cold ether, dissolved in 1N acetic acid, and diluted with distilled water.

4. Cyclization by Air Oxidation The diluted aqueous solution of the above polypeptide was adjusted to pH 7.5 with concentrated aqueous ammonia, and cyclized by air oxidation by aeration. The aqueous solution was purified by large-scale preparative HPLC (Cosmozil 5C18 AR-II column: acetonitrile water) and gel chromatography (Sephadex G-15, eluent: 0.1N AcOH) to obtain a single peak polypeptide. Lyophilized. Purity was confirmed by HPLC.

<Production Example 2: Production of TC14005>
H-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(TC14005)
TC14005 was produced in the same manner as in Production Example 1. However, at the introduction of the 12th to 1st amino acids, DCit was used in place of DLys (Boc) in position 8, and Arg (Pbf) was used in place of Cit in position 6.

<Production Example 3: Production of TC14011>
H-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(TC14011)
TC14011 was produced in the same manner as in Production Example 1. However, DCit was used instead of DLys (Boc) at position 8 at the introduction of amino acids at positions 12 to 1.

<Production Example 4: Production of TC14013>
H-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Cit-Cit-Cys-Arg-OH
(TC14013)
TC14013 was produced in the same manner as in Production Example 1. However, Cit was used instead of Arg (Pbf) at position 11 at the introduction of amino acids at positions 12 to 1.

<Production Example 5: Production of TC14015>
H-Cit-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(TC14015)
TC14015 was produced in the same manner as in Production Example 1. However, Cit was used instead of Arg (Pbf) at position 1 at the introduction of amino acids at positions 12 to 1.

<Production Example 6: Production of TC14017>
H-Cit-Arg-Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(TC14017)
TC14017 was produced in the same manner as in Production Example 1. However, at the introduction of the 12th to 1st amino acids, use DCit instead of DLys (Boc) at position 8, Arg (Pbf) instead of Cit at position 6, and Cit instead of Arg (Pbf) at position 1. Was.

<Production Example 7: Production of TC14019>
H-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Cit-Cit-Cys-Arg-OH
(TC14019)
TC14019 was produced in the same manner as in Production Example 1. However, at the introduction of the 12th to 1st amino acids, use Cit in place of Arg (Pbf) in position 11, DCit in place of DLys (Boc) in position 8, and Arg (Pbf) in place of Cit in position 6. Was.

<Production Example 8: Production of TC14021>
H-Cit-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Cit-Cit-Cys-Arg-OH
(TC14021)
TC14021 was produced in the same manner as in Production Example 1. However, at the introduction of the 12th to 1st amino acids, use Cit instead of Arg (Pbf) at position 11, use Arg (Pbf) instead of Cit at position 6, and use Cit instead of Arg (Pbf) at position 1. Was.

<Production Example 9: Production of TC14012>
H-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TC14012)
1. Synthesis of TC14012 protected polypeptide resin After removing the Fmoc group from Fmoc-Rink amide resin with 20% piperidine / DMF, add Fmoc-Arg (Pbf) -OH (2.5 eq) corresponding to position 14 in DMF, DIPCDI-HOBt A condensation reaction was performed by the method. The progress of the condensation reaction was determined by the ninhydrin test of Kaiser, E. et al. (Anal. Biochem., 34: 595 (1970)).

2. Introduction of amino acids at positions 12 to 1 Similarly, Cys (Trt), Cit, Arg (Pbf), Tyr (t-Bu), Pro, DCit, Lys (Boc), Cit, Tyr (tyr) -Bu), Cys (Trt), Nal, Arg (Pbf) and Arg (Pbf) residues were introduced into a Rink amide resin to obtain a protecting group-protected polypeptide resin.

3. Deprotection Group, Separation and Purification of Polypeptide from Resin The protecting group-protected polypeptide resin was treated with 20% piperidine / DMF to remove the Fmoc group, and then the resin was subjected to 1 M TMSBr-thioanisole / TFA (trifluoro The reaction was performed at 25 ° C. for 3 hours in an acetic acid system (in the presence of m-cresol (100 eq) and ethanedithiol (300 eq)). The resin was filtered off from the reaction mixture, washed twice with TFA, the combined filtrate and washings were concentrated under reduced pressure, water-cooled dry ether was added to the residue, and the resulting precipitate was centrifuged and centrifuged and decanted. Separated from The obtained residue was washed with cold ether, dissolved in 1N acetic acid, and diluted with distilled water.

4. Cyclization by Air Oxidation The diluted aqueous solution of the above polypeptide was adjusted to pH 7.5 with concentrated aqueous ammonia, and cyclized by air oxidation by aeration. This aqueous solution was purified by large-scale preparative HPLC (Cosmozil 5C18 AR-II column: acetonitrile water) and gel chromatography (Sephadex G-15, eluent: 0.1N AcOH) to obtain a single peak polypeptide. Lyophilized. Purity was confirmed by HPLC.

<Production Example 10: Production of TC14014>
H-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Cit-Cit-Cys-Arg-NH ₂
(TC14014)
TC14014 was produced in the same manner as in Production Example 9. However, Cit was used instead of Arg (Pbf) at position 11 and DLys (Boc) was used instead of DCit at position 8 at the introduction of amino acids at positions 12 to 1.

<Production Example 11: Production of TC14016>
H-Cit-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TC14016)
TC14016 was produced in the same manner as in Production Example 9. However, at the introduction of the 12th to 1st amino acids, DLys (Boc) was used instead of DCit at position 8, and Cit was used instead of Arg (Pbf) at position 1.

<Production Example 12: Production of TC14018>
H-Cit-Arg-Nal- Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2
(TC14018)
TC14018 was produced in the same manner as in Production Example 9. However, Arg (Pbf) was used instead of Cit at position 6 and Cit was used instead of Arg (Pbf) at position 1 at the introduction of amino acids at positions 12 to 1.

<Production Example 13: Production of TC14020>
H-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Cit-Cit-Cys-Arg-NH ₂
(TC14020)
AcTC14020 was produced in the same manner as in Production Example 9. However, Cit was used instead of Arg (Pbf) at position 11 and Arg (Pbf) was used instead of Cit at position 6 at the introduction of amino acids at positions 12 to 1.

<Production Example 14: Production of TC14022>
H-Cit-Arg-Nal- Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Cit-Cit-Cys-Arg-NH 2
(TC14022)
TC14022 was produced in the same manner as in Production Example 9. However, at the introduction of the 12th to 1st amino acids, Cit instead of Arg (Pbf) at position 11, DLys (Boc) instead of DCit at position 8, Arg (Pbf), 1 instead of Cit at position 6 Cit was used instead of Arg (Pbf).

<Production Example 15: Production of TA14001, TA14005 to TA14009, TC14001, and TC14004>
H-Ala-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(TA14001)
H-Arg-Arg-Nal-Cys-Tyr-Ala-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(TA14005)
H-Arg-Arg-Nal-Cys-Tyr-Arg-Ala-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(TA14006)
H-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DAla-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(TA14007)
H-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Ala-Tyr-Arg-Cit-Cys-Arg-OH
(TA14008)
H-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Ala-Arg-Cit-Cys-Arg-OH
(TA14009)
H-Cit-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(TC14001)
H-Arg-Arg-Nal- Cys-Tyr-Arg-Cit-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2
(TC14004)
By changing amino acids to be introduced in the same manner as in Production Example 1 or 9, TA14001, TA14005 to TA14009, TC14001, and TC14004 shown above can be produced.

<Production Example 16: Production of AcTC14003, AcTC14005, AcTC14011 to AcTC14022>
Ac-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(AcTC14003)
Ac-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(AcTC14005)
Ac-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(AcTC14011)
Ac-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Cit-Cit-Cys-Arg-OH
(AcTC14013)
Ac-Cit-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(AcTC14015)
Ac-Cit-Arg-Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(AcTC14017)
Ac-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Cit-Cit-Cys-Arg-OH
(AcTC14019)
Ac-Cit-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Cit-Cit-Cys-Arg-OH
(AcTC14021)
Ac-Arg-Arg-Nal- Cys-Tyr-Cit-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2
(AcTC14012)
Ac-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Cit-Cit-Cys-Arg-NH ₂
(AcTC14014)
Ac-Cit-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(AcTC14016)
Ac-Cit-Arg-Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(AcTC14018)
Ac-Arg-Arg-Nal- Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Cit-Cit-Cys-Arg-NH 2
(AcTC14020)
Ac-Cit-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Cit-Cit-Cys-Arg-NH ₂
(AcTC14022)
By the same method as in Production Examples 1 to 14, TC14003, TC14005, and acetylated products of TC14011 to TC14022 were produced. However, the protecting group-protected polypeptide resin was treated with 20% piperidine / DMF to remove the Fmoc group, acetic anhydride (100 eq) -s lysine (100 eq) / DMF treatment, and then acetylated with the resin. TMSBr-thioanisole / TFA (trifluoroacetic acid) system (in the presence of m-cresol (100 eq) and ethanedithiol (300 eq)) at 25 ° C for 2 hours (when the C-terminal is a carboxylic acid compound) or 3 hours ( (C-terminal is an amide form).

<Production Example 17: Production of polypeptide TE14005>
H-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(TE14005)
1. Synthesis of TE14005-Protected Polypeptide Resin After removing the Fmoc group from 270 mg (0.2 mmol) of Fmoc-Arg (Pbf) -Alko resin (0.74 mmol / g) introduced with arginine at the 14th position using 20% piperidine / DMF And Fmoc-Cys (Trt) -OH (2.5 eq) corresponding to position 12 was added, and a condensation reaction was performed in DMF by the DIPCDI-HOBt method. The progress of the condensation reaction was determined by the ninhydrin test of Kaiser, E. et al. (Anal. Biochem., 34: 595 (1970)).

2.Introduction of amino acids at positions 12 to 1 In the same manner, Cit, Arg (Pbf), Tyr (t-Bu), Pro, DGlu (0-t-Bu), Lys (Boc), Arg (Pbf ), Tyr (t-Bu), Cys (Trt), Nal, Arg (Pbf) and Arg (Pbf) residues were introduced into the resin to obtain a protecting group-protected polypeptide resin.

3. Deprotection Group, Separation and Purification of Polypeptide from Resin The protecting group-protected polypeptide resin was treated with 20% piperidine / DMF to remove the Fmoc group, and then 200 mg of resin with 1 M TMSBr-thioanisole / TFA ( The reaction was performed at 25 ° C. for 2 hours with 10 mL of trifluoroacetic acid) (in the presence of m-cresol (100 eq) and ethanedithiol (300 eq)). The resin was filtered off from the reaction mixture, washed twice with 1 mL of TFA, the combined filtrate and washings were concentrated under reduced pressure, 30 mL of water-cooled dry ether was added to the residue, and the resulting precipitate was centrifuged. Separated from the supernatant by decantation. The obtained residue was washed with cold ether, dissolved in 1N acetic acid (50 mL), and diluted with distilled water to 250 mL.

4. Cyclization by Air Oxidation The diluted aqueous solution of the above polypeptide was adjusted to pH 7.5 with concentrated aqueous ammonia, and cyclized by air oxidation by aeration. This aqueous solution was purified by large-scale preparative HPLC (Cosmozil 5C18 AR-II column: acetonitrile-water) and gel chromatography (Sephadex G-15, eluent: 0.1 N AcOH) to obtain a single peak polypeptide. Lyophilized. Purity was confirmed by HPLC.
Yield 24.1 mg (7 AcOH salt) (21.3%)
[α] _D ^23.6 =-5.36 (c 1.12, H ₂ O)
Ion spray mass spectrum (IS-MS): C ₈₉ H ₁₃₆ N ₃₂ 0 ₂₀ S ₂
Calculated: 2038.38 Observed: 2038
(Triple stage quadrupole mass spectrometer API-IIIE (Sciex))

<Production Example 18: Production of polypeptide TE14001>
H-DGlu-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(TE14001)
TE14001 was produced in the same manner as in Production Example 17. However, DLys (Boc) was used instead of DGlu (Ot-Bu) at position 8 and DGlu (Ot-Bu) was used instead of Arg (Pbf) at position 1 at the introduction of amino acids at positions 12 to 1.

<Production Example 19: Production of polypeptide TE14002>
H-Arg-Glu-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(TE14002)
TE14002 was produced in the same manner as in Production Example 17. However, DLys (Boc) was used instead of DGlu (Ot-Bu) at position 8, and Glu (Ot-Bu) was used instead of Arg (Pbf) at position 12 when the amino acids at positions 12 to 1 were introduced.

<Production Example 20: Production of polypeptide TE14003>
H-Arg-Arg-Nal-Cys-Tyr-Glu-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(TE14003)
TE14003 was produced in the same manner as in Production Example 17. However, DLys (Boc) was used instead of DGlu (Ot-Bu) at position 8, and Glu (Ot-Bu) instead of Arg (Pbf) at position 6 at the introduction of amino acids at positions 12 to 1.

<Production Example 21: Production of polypeptide TE14004>
H-Arg-Arg-Nal-Cys-Tyr-Arg-Glu-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(TE14004)
TE14004 was produced in the same manner as in Production Example 17. However, DLys (Boc) was used instead of DGlu (Ot-Bu) at position 8, and Glu (Ot-Bu) was used instead of Lys (Boc) at position 7 at the introduction of amino acids at positions 12 to 1.

<Production Example 22: Production of polypeptide TE14006>
H-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Glu-Cit-Cys-Arg-OH
(TE14006)
TE14006 was produced in the same manner as in Production Example 17. However, at the introduction of the 12th to 1st amino acids, DLys (Boc) was used instead of DGlu (Ot-Bu) at position 8, and Glu (Ot-Bu) was used instead of Arg (Pbf) at position 11.

<Production Example 23: Production of polypeptide TE14007>
H-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Glu-OH
(TE14007)
TE14007 was produced in the same manner as in Production Example 17. However, instead of the Fmoc-Arg (Pbf) -Alko resin introduced with the first 14-position arginine, the Fmoc-Glu (Ot-Bu) -Alko resin obtained by introducing the first 14-position glutamic acid was further used. At the introduction of amino acids from position 1 to position 1, DLys (Boc) was used instead of DGlu (Ot-Bu) at position 8.

<Production Example 24: Production of polypeptide TE14011>
H-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TE14011)
1. Synthesis of TE14011 protected polypeptide resin After removing the Fmoc group from Fmoc-Rink amide resin with 20% piperidine / DMF, add Fmoc-Arg (Pbf) -OH (2.5 eq) corresponding to position 14 in DMF, DIPCDI-HOBt A condensation reaction was performed by the method. The progress of the condensation reaction was determined by the ninhydrin test of Kaiser, E. et al. (Anal. Biochem., 34: 595 (1970)).

2. Introduction of 12th to 1st amino acids In the same manner, Cys (Trt), Cit, Arg (Pbf), Tyr (t-Bu), Pro, DGlu (Ot-Bu), Lys (Boc), Cit, Tyr (t-Bu), Cys (Trt), Nal, Arg (Pbf) and Arg (Pbf) residues were introduced into a Rink amide resin to obtain a protecting group-protected polypeptide resin.

3. Deprotection Group, Separation and Purification of Polypeptide from Resin The protecting group-protected polypeptide resin was treated with 20% piperidine / DMF to remove the Fmoc group, and then the resin was subjected to 1 M TMSBr-thioanisole / TFA (trifluoro The reaction was performed at 25 ° C. for 3 hours in an acetic acid system (in the presence of m-cresol (100 eq) and ethanedithiol (300 eq)). The resin was filtered off from the reaction mixture, washed twice with TFA, the combined filtrate and washings were concentrated under reduced pressure, water-cooled dry ether was added to the residue, and the resulting precipitate was centrifuged and centrifuged and decanted. Separated from The obtained residue was washed with cold ether, dissolved in 1N acetic acid, and diluted with distilled water.

4. Cyclization by Air Oxidation The diluted aqueous solution of the above polypeptide was adjusted to pH 7.5 with concentrated aqueous ammonia, and cyclized by air oxidation by aeration. This aqueous solution was purified by large-scale preparative HPLC (Cosmozil 5C18 AR-II column: acetonitrile-water) and gel chromatography (Sephadex G-15, eluent: 0.1 N AcOH) to obtain a single peak polypeptide, Lyophilized. Purity was confirmed by HPLC.

<Production Example 25: Production of polypeptide TE14012>
H-Arg-Arg-Nal-Cys-Tyr-DGlu-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TE14012)
TE14012 was produced in the same manner as in Production Example 24. However, at the introduction of amino acids at positions 12 to 1, DCit was used in place of DGlu (Ot-Bu) at position 8, and DGlu (Ot-Bu) was used instead of Cit at position 6.

<Production Example 26: Production of polypeptide TE14013>
H-Arg-Arg-Nal-Cys-Tyr-DGlu-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TE14013)
TE14013 was produced in the same manner as in Production Example 24. However, DGlu (Ot-Bu) was used instead of Cit at position 6 at the introduction of amino acids at positions 12 to 1.

<Production Example 27: Production of polypeptide TE14014>
H-DGlu-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TE14014)
TE14014 was produced in the same manner as in Production Example 24. However, DGlu (Ot-Bu) was used instead of Arg (Pbf) at position 1 at the introduction of amino acids at positions 12 to 1.

<Production Example 28: Production of polypeptide TE14015>
H-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-DGlu-Arg-Cit-Cys-Arg-NH ₂
(TE14015)
TE14015 was produced in the same manner as in Production Example 24. However, DGlu (Ot-Bu) was used instead of Tyr (t-Bu) at position 10 at the introduction of amino acids at positions 12 to 1.

<Production Example 29: Production of polypeptide TE14016>
H-Arg-Arg-Nal- Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-DGlu-Cys-Arg-NH 2
(TE14016)
TE14016 was produced in the same manner as in Production Example 24. However, DGlu (Ot-Bu) was used instead of Cit at position 12 at the introduction of amino acids at positions 12 to 1.

<Production Example 30: Production of polypeptides AcTE14014 to AcTE14016>
Ac-DGlu-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(AcTE14014)
Ac-Arg-Arg-Nal- Cys-Tyr-Cit-Lys-DGlu-Pro-DGlu-Arg-Cit-Cys-Arg-NH 2
(AcTE14015)
Ac-Arg-Arg-Nal- Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-DGlu-Cys-Arg-NH 2
(AcTE14016)
In the same manner as in Production Examples 27 to 29, acetylated products of TE14014 to TE14016 were produced. However, the protecting group-protected polypeptide resin was treated with 20% piperidine / DMF to remove the Fmoc group, acetic anhydride (100 eq) -s lysine (100 eq) / DMF treatment, and then acetylated with the resin. The reaction was performed at 25 ° C. for 3 hours in a TMSBr-thioanisole / TFA (trifluoroacetic acid) system (in the presence of m-cresol (100 eq) and ethanedithiol (300 eq)).

<Production Example 31: Production of polypeptide TF1>
Ac-Arg-Arg-Nal- Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2
(TF1: AcTE14011)
TF1 was produced in the same manner as in Production Example 24. However, the protecting group-protected polypeptide resin was treated with 20% piperidine / DMF to remove the Fmoc group, acetic anhydride (100 eq) -s lysine (100 eq) / DMF treatment, and then acetylated with the resin. The reaction was performed at 25 ° C. for 3 hours in a TMSBr-thioanisole / TFA (trifluoroacetic acid) system (in the presence of m-cresol (100 eq) and ethanedithiol (300 eq)).

<Production Example 32: Production of polypeptide TF2>
guanyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF2: guanyl-TE14011)
TF2 was produced in the same manner as in Production Example 24. However, for the protecting group-protected polypeptide resin, the Fmoc group was removed by treatment with 20% piperidine / DMF, and guanyl was treated with 1H-pyrazole-1-carboxamidine (5 eq)-, N-diisopropylethylamine (10 eq) / DMF. The resin was then reacted with a 1 M TMSBr-thioanisole / TFA (trifluoroacetic acid) system (in the presence of m-cresol (100 eq) and ethanedithiol (300 eq)) at 25 ° C for 3 hours.

<Production Example 33: Production of polypeptide TF3>
TMguanyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF3: TMguanyl-TE14011)
TF3 was produced in the same manner as in Production Example 24. However, the protecting group-protected polypeptide resin was treated with 20% piperidine / DMF to remove the Fmoc group and give 2- (1H-benzotriazol-1-yl) -1,1,3,3-tetramethyluronium tetra Tetramethylguanylation by fluoroborate (5 eq) / DMF treatment, and then 1M TMSBr-thioanisole / TFA (trifluoroacetic acid) system (m-cresol (100 eq), ethanedithiol (300 eq)) ) At 25 ° C for 3 hours.

<Production Example 34: Production of polypeptide TF4>
TMguanyl-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF4: TMguanyl-TE14011 (2-14))
TF4 was produced in the same manner as in Production Example 24. However, arginine at position 1 was not condensed.

<Production Example 35: Production of polypeptide TF5>
4F-benzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF5: 4F-benzoyl-TE14011)
TF5 was produced in the same manner as in Production Example 24. However, for the protecting group-protected polypeptide resin, the Fmoc group was removed by treatment with 20% piperidine / DMF, 4-fluorobenzoic acid (2.5 eq) was condensed by the DIPCDI-HOBt method, and then 1 M TMSBr- The reaction was carried out at 25 ° C. for 3 hours in a thioanisole / TFA (trifluoroacetic acid) system (in the presence of m-cresol (100 eq) and ethanedithiol (300 eq)).

<Production Example 36: Production of polypeptide TF6>
2F-benzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂ (TF6: 2F-benzoyl-TE14011)
TF6 was produced in the same manner as in Production Example 24. However, for the protecting group-protected polypeptide resin, the Fmoc group was removed by treatment with 20% piperidine / DMF, 2-fluorobenzoic acid (2.5 eq) was condensed by the DIPCDI-HOBt method, and then 1 M TMSBr- The reaction was carried out at 25 ° C. for 3 hours in a thioanisole / TFA (trifluoroacetic acid) system (in the presence of m-cresol (100 eq) and ethanedithiol (300 eq)).

<Production Example 37: Production of polypeptide TF7>
APA-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF7: APA-TE14011 (2-14))
TF7 was produced in the same manner as in Production Example 24. However, 5-aminopentanoic acid (protected Fmoc) was introduced instead of Arg (Pbf) at position 1 at the introduction of amino acids at positions 12 to 1.

<Production Example 38: Production of polypeptide TF8>
desamino-R-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF8: desamino-R-TE14011 (2-14))
TF8 was produced in the same manner as in Production Example 24. However, at the introduction of the 12th to 1st amino acids, 5-aminopentanoic acid (protected Fmoc) was introduced instead of Arg (Pbf) at the 1st position, and further, the protecting group-protected polypeptide resin had 20 Fmoc group was removed by treatment with 1% piperidine / DMF, guanylated by treatment with 1H-pyrazole-1-carboxamidine (5 eq)-, N-diisopropylethylamine (10 eq) / DMF, and then 1M TMSBr-thioanisole was applied to the resin. The reaction was carried out at 25 ° C. for 3 hours in a / TFA (trifluoroacetic acid) system (in the presence of m-cresol (100 eq) and ethanedithiol (300 eq)).

<Production Example 39: Production of polypeptide TF9>
guanyl-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF9: guanyl-TE14011 (2-14))
TF9 was produced using the same method as in Production Example 32. However, arginine at position 1 was not condensed.

<Production Example 40: Production of polypeptide TF10>
succinyl-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF10: succinyl-TE14011 (2-14))
TF10 was produced in the same manner as in Production Example 24. However, arginine at position 1 was not condensed. The protecting group-protected polypeptide resin was treated with 20% piperidine / DMF to remove the Fmoc group, and treated with succinic anhydride (5 eq) / pyridine for hemisuccinylation. The reaction was carried out at 25 ° C. for 3 hours in a (trifluoroacetic acid) system (in the presence of m-cresol (100 eq) and ethanedithiol (300 eq)).

<Production Example 41: Production of polypeptide TF11>
glutaryl-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF11: glutaryl-TE14011 (2-14))
TF11 was produced in the same manner as in Production Example 40. However, the protecting group-protected polypeptide resin was treated with 20% piperidine / DMF to remove the Fmoc group, and treated with glutaric anhydride (5 eq) / pyridine for hemisuccinylation. The reaction was carried out at 25 ° C. for 3 hours in a (trifluoroacetic acid) system (in the presence of m-cresol (100 eq) and ethanedithiol (300 eq)).

<Production Example 42: Production of polypeptide TF12>
deaminoTMG-APA-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF12: deaminoTMG-APA-TE14011 (2-14))
TF12 was produced in the same manner as in Production Example 24. However, at the introduction of the 12th to 1st amino acids, 5-aminopentanoic acid (protected Fmoc) was introduced instead of Arg (Pbf) at the 1st position, and further, the protecting group-protected polypeptide resin had 20 Fmoc group was removed by treatment with% piperidine / DMF, and 2- (1H-benzotriazol-1-yl) -1,1,3,3-tetramethyluronium tetrafluoroborate (5 eq) / tetramethyl borate was treated with DMF. After guanylation, the resin was reacted with a 1M TMSBr-thioanisole / TFA (trifluoroacetic acid) system (in the presence of m-cresol (100 eq) and ethanedithiol (300 eq)) at 25 ° C. for 3 hours.

<Production Example 43: Production of polypeptide TF15>
R-CH ₂ -Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF15: R-CH ₂ NH-RTE14011)
TF15 was produced in the same manner as in Production Example 24. However, instead of condensing Fmoc-Arg (Pbf) -OH to the 1-position at the introduction of the 12th to 1st amino acids, Fmoc-Arg (Pbf) -H (aldehyde) was condensed by reductive amination ( NaB (CN) H ₃ (3 eq), AcOH (1 eq) / DMF).

<Production Example 44: Production of polypeptide TF17>
H-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂ (TF17)
(TF17: TE14011 (2-14))
TF17 was produced in the same manner as in Production Example 24. However, arginine at position 1 was not condensed.

<Production Example 45: Production of polypeptide TF18>
TMguanyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF18: TMguanyl-TC14012)
TF18 was produced in the same manner as in Production Example 9. However, the protecting group-protected polypeptide resin was treated with 20% piperidine / DMF to remove the Fmoc group and give 2- (1H-benzotriazol-1-yl) -1,1,3,3-tetramethyluronium tetra Tetramethylguanylation by fluoroborate (5 eq) / DMF treatment, and then 1M TMSBr-thioanisole / TFA (trifluoroacetic acid) system (m-cresol (100 eq), ethanedithiol (300 eq)) ) At 25 ° C for 3 hours.

<Production Example 46: Production of polypeptide TF19>
ACA-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TF19: ACA-TC14012)
TF19 was produced in the same manner as in Production Example 9. However, 6-aminohexanoic acid (protected Fmoc) was introduced after Arg (Pbf) at position 1 at the introduction of amino acids at positions 12 to 1.

<Production Example 47: Production of polypeptide TF20>
ACA-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH
(TF20: ACA-T140)
TF20 was produced in the same manner as in Production Example 17. However, at the introduction of amino acids at positions 12 to 1, DLys (Boc) in place of DGlu (Ot-Bu) in position 8, and 6-aminohexanoic acid instead of Arg (Pbf) in position 1 (protected Fmoc) Was used.

<Production Example 48: Production of polypeptide TZ14011>
H-Arg-Arg-Nal- Cys-Tyr-Cit-Arg-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2
(TZ14011)
TZ14011 was produced in the same manner as in Production Example 24. However, DLys (Boc) was used instead of DGlu (Ot-Bu) at position 8 and Arg (Pbf) instead of Lys (Boc) at position 7 when amino acids at positions 12 to 1 were introduced.

<Production Example 49: Production of polypeptide AcTZ14011>
Ac-Arg-Arg-Nal- Cys-Tyr-Cit-Arg-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2
(AcTZ14011)
AcTZ14011 was produced in the same manner as in Production Example 48. However, the protecting group-protected polypeptide resin was treated with 20% piperidine / DMF to remove the Fmoc group, acetic anhydride (100 eq) -s lysine (100 eq) / DMF treatment, and then acetylated with the resin. The reaction was performed at 25 ° C. for 3 hours in a TMSBr-thioanisole / TFA (trifluoroacetic acid) system (in the presence of m-cresol (100 eq) and ethanedithiol (300 eq)).

<Production Example 50: Production of polypeptide TN14003>
H-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(TN14003)
TN14003 was produced in the same manner as in Production Example 9. However, DLys (Boc) was used instead of DCit at position 8 at the introduction of amino acids at positions 12 to 1.

<Production Example 51: Production of polypeptide TN14005>
H-Arg-Arg-Nal- Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2
(TN14005)
TN14005 was produced in the same manner as in Production Example 9. However, Arg (Pbf) was used instead of Cit at position 6 at the introduction of amino acids at positions 12 to 1.

<Production Example 52: Production of polypeptide AcTN14003>
Ac-Arg-Arg-Nal- Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2
(AcTN14003)
AcTN14003 was produced in the same manner as in Production Example 50. However, the protecting group-protected polypeptide resin was treated with 20% piperidine / DMF to remove the Fmoc group, acetic anhydride (100 eq) -s lysine (100 eq) / DMF treatment, and then acetylated with the resin. The reaction was performed at 25 ° C. for 3 hours in a TMSBr-thioanisole / TFA (trifluoroacetic acid) system (in the presence of m-cresol (100 eq) and ethanedithiol (300 eq)).

<Production Example 53: Production of polypeptide AcTN14005>
Ac-Arg-Arg-Nal- Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2
(AcTN14005)
AcTN14005 was produced in the same manner as in Production Example 51. However, the protecting group-protected polypeptide resin was treated with 20% piperidine / DMF to remove the Fmoc group, acetic anhydride (100 eq) -s lysine (100 eq) / DMF treatment, and then acetylated with the resin. The reaction was performed at 25 ° C. for 3 hours in a TMSBr-thioanisole / TFA (trifluoroacetic acid) system (in the presence of m-cresol (100 eq) and ethanedithiol (300 eq)).

<Production Example 54: Production of polypeptide 4F-benzoyl-TN14003>
4F-benzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(4F-benzoyl-TN14003)
1.4 Synthesis of 4F-benzoyl-TN14003 protected polypeptide resin After removing the Fmoc group from 2.94 g (1 mmol) of Fmoc-Rink amide resin (0.34 mmol / g) with 20% piperidine / DMF, Fmoc-corresponding to position 14 was removed. Arg (Pbf) -OH (2.5 eq) was added, and a condensation reaction was performed in DMF by the DIPCDI-HOBt method. The progress of the condensation reaction was determined by the ninhydrin test of Kaiser, E. et al. (Anal. Biochem., 34: 595 (1970)).

2. Introduction of amino acids at positions 13 to 1 Similarly, Cys (Trt), Cit, Arg (Pbf), Tyr (t-Bu), Pro, DLys (Boc), Lys (Boc), Cit, Tyr (t-Bu), Cys (Trt), Nal, Arg (Pbf), Arg (Pbf) residues were introduced into Rink amide resin, and finally 4-fluorobenzoic acid (2.5 eq) at the N-terminus was DIPCDI-HOBt. Condensation was performed by a method to obtain a protecting group-protected polypeptide resin.

3. Deprotection group, separation and purification of polypeptide from resin 1 M TMSBr-thioanisole / TFA (trifluoroacetic acid) system (m-cresol (100 eq), ethanedithiol (300 eq) ) Presence) The reaction was carried out at 270 mL at 25 ° C for 3 hours. The resin was separated from the reaction mixture by filtration, washed twice with 5 mL of TFA, and the combined filtrate and washings were concentrated under reduced pressure. 300 mL of water-cooled dry ether was added to the residue, and the resulting precipitate was decanted. Separated from the supernatant. The obtained residue was washed with cold ether, dissolved in 500 mL of 1N acetic acid, and diluted to 2.5 L with distilled water.

4. Cyclization by Air Oxidation The diluted aqueous solution of the above polypeptide was adjusted to pH 7.5 with concentrated aqueous ammonia, and cyclized by air oxidation by aeration. This aqueous solution was purified by large-scale preparative HPLC (Cosmozil 5C18 AR-II column: acetonitrile water) to obtain a single peak polypeptide, which was lyophilized. Purity was confirmed by HPLC.
Yield 551.5 mg (6 TFA salt) (19.4%)
[α] _D ^28.6 =-10.25 (c 0.39, H ₂ O)
Ion spray mass spectrum (IS-MS): C ₉₇ H ₁₄₄ FN ₃₃ 0 ₁₉ S ₂
Calculated: 2159.52 Observed: 2161
(Triple stage quadrupole mass spectrometer API-IIIE (Sciex))

<Production Example 55: Production of polypeptide 4F-benzoyl-TE14011-Me>
4-fluorobenzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NHMe
(4F-benzoyl-TE14011-Me)
1. Synthesis of 4F-benzoyl-TE14011-Me protected polypeptide resin To 4-sulfamylbutyryl AM NovaGel resin, add Fmoc-Arg (Pbf) -OH (4 eq) corresponding to position 14 and add PyBOP (3 eq) in CHCl _3. The condensation reaction was carried out at -0 ° C by the -DIPEA (6 eq) method (this condensation reaction was carried out twice). After removing the Fmoc group from the resin with 20% piperidine / DMF, Fmoc-Cys (Trt) -OH (2.5 eq) corresponding to the 13-position was added, and a condensation reaction was performed in DMF by the DIPCDI-HOBt method. The progress of the condensation reaction was determined by the ninhydrin test of Kaiser, E. et al. (Anal. Biochem., 34: 595 (1970)). Hereinafter, similarly, Cit, Arg (Pbf), Tyr (t-Bu), Pro, DGlu (Ot-Bu), Lys (Boc), Cit, Tyr (t- Bu), Cys (Trt), Nal, Arg (Pbf), Arg (Pbf) residue introduced into sulfamylbutyryl resin, 4-fluorobenzoic acid (2.5 eq) at the end N-terminal condensed by DIPCDI-HOBt method, protection A group-protected polypeptide resin was obtained.

2. C-terminal alkylamidation, deprotection, separation and purification of the polypeptide from the resin. The protected group-protected polypeptide resin was treated with ICH ₂ CN (40 eq), DIPEA (10 eq) / NMP (48 hours) for cyanomethylation. Then, methylamine (excess) in THF / DMF was allowed to act on the resin to separate the C-terminal methylamidated protecting group-protected polypeptide from the resin. Next, the protecting group-protected polypeptide was reacted in a 1 M thioanisole / TFA (trifluoroacetic acid) system (in the presence of m-cresol (100 eq) and ethanedithiol (300 eq)) at 25 ° C. for 3 hours. The reaction solution was concentrated under reduced pressure, water-cooled dry ether was added to the residue, and the resulting precipitate was separated from the supernatant by centrifugation and decantation. The obtained residue was washed with cold ether, dissolved in 1N acetic acid, and diluted with distilled water.

3. Cyclization by Air Oxidation The diluted aqueous solution of the above polypeptide was adjusted to pH 7.5 with concentrated aqueous ammonia, and cyclized by air oxidation by aeration. This aqueous solution was purified by large-scale preparative HPLC (Cosmozil 5C18 AR-II column: acetonitrile water) and gel chromatography (Sephadex G-15, eluent: 0.1N AcOH) to obtain a single peak polypeptide. Lyophilized. Purity was confirmed by HPLC.

<Production Example 56: Production of polypeptide 4F-benzoyl-TE14011-Et>
4-fluorobenzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NHEt
(4F-benzoyl-TE14011-Et)
4F-benzoyl-TE14011-Et was produced in the same manner as in Production Example 55. However, ethylamine was used instead of methylamine.

<Production Example 57: Production of polypeptide: 4F-benzoyl-TE14011-iPr>
4-fluorobenzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NHiPr
(4F-benzoyl-TE14011-iPr)
4F-benzoyl-TE14011-iPr was produced in the same manner as in Production Example 55. However, isopropylamine was used instead of methylamine.

<Production Example 58: Production of polypeptide 4F-benzoyl-TE14011-tyramine>
4-fluorobenzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-tyramine
(4F-benzoyl-TE14011-tyramine)
4F-benzoyl-TE14011-tyramine was produced in the same manner as in Production Example 55. However, tyramine (p-hydroxyphenylethylamine) was used instead of methylamine.

<Test Example 1: Inhibitory activity of polypeptide of the present invention on binding of CXCL12 to CXCR4 receptor>
50 μL of Jurkat human T-cell leukemia cells prepared at ⁶ × 10 ⁶ cells / mL in a buffer (Dulbecco's PBS solution (pH 7.0) containing 0.5% BSA and 20 mM HEPES) in an assay plate, diluted with the buffer, test compounds (Table 1: Each compound used was synthesized in the form of an acetate) and 25 μL of 200 pM ¹²⁵ I-CXCL12 solution were respectively dispensed, and allowed to react at room temperature for 1 hour to carry out a binding reaction. After the reaction, the reaction solution was subjected to suction filtration through a 96-well GF / C filter plate, and the radioactivity of each well was measured by a top count. The inhibitory activity of each test compound was determined assuming that the radioactivity when no test compound was added was 100% and the radioactivity when 100 nM of unlabeled CXCL12 was added was 0%. The results are shown in Table 1 below.

結果 The results in Table 1 show that this compound has strong binding inhibitory activity.

<Test Example 2: Inhibitory activity of TE-14005 on breast cancer cell migration induced by CXCL12>
A transwell filter (polycarbonate filter, 8 μm diameter, Costar) was treated in a 10 μg / mL fibronectin solution at 37 ° C. for 6 hours, and then air-dried. 600 μL / well of DMEM (Gibco BRL) containing 100 nM of CXCL12 (R & D Systems) and buffer A (0.1% bovine serum albumin, 12 mM HEPES) containing a test substance was added to the lower chamber of the transwell. Substance and human breast cancer MDA-MB-231 cells (purchased from American Tissue Culture Collection), 100 μL / well of buffer A containing 2 × 10 ⁶ cells / mL were added, and cultured at 37 ° C. for 15 hours in a 5% CO ₂ incubator. The upper surface of the filter was wiped off to remove the cells, the cells on the lower surface of the filter were fixed and stained with a 25% methanol solution containing 0.5% crystal violet (Wako Pure Chemical), washed with distilled water, and air-dried. Disconnect, 0.1 M sodium citrate / 50 An ethanol solution was added to dissolve the crystal violet, and the absorbance was measured at 550 nm, and the results are shown in Fig. 1. The negative control shows the migration without CXCL12, and MDA-MB-231 by adding CXCL12. The cell migration was enhanced (positive control) and the CXCL12-induced migration of MDA-MB-231 cells was inhibited by an antagonist, TE14005, up to 10 nM.

<Test Example 3: Inhibitory activity of TC14012 and TN14003 on binding of CXCL12 to CXCR4 receptor>
50 μL of Jurkat human T-cell leukemia cells prepared at ⁶ × 10 ⁶ cells / mL in a buffer (Dulbecco's PBS solution (pH 7.0) containing 0.5% BSA and 20 mM HEPES) in an assay plate, diluted with the buffer, test compounds (Table 2: Each compound used was synthesized in the form of acetate) and 25 μL of 200 pM ¹²⁵ I-CXCL12 solution were respectively dispensed, and allowed to react at room temperature for 1 hour to perform a binding reaction. After the reaction, the reaction solution was subjected to suction filtration through a 96-well GF / C filter plate, and the radioactivity of each well was measured by a top count. The inhibitory activity of each test compound was determined assuming that the radioactivity when no test compound was added was 100% and the radioactivity when 100 nM of unlabeled CXCL12 was added was 0%. The results are shown in Table 2 below.

結果 The results in Table 2 show that this compound has strong binding inhibitory activity.

<Test Example 4: Inhibitory activity of TC-14012 on breast cancer cell migration induced by CXCL12>
A transwell filter (polycarbonate filter, 8 μm diameter, Costar) was treated in a 10 μg / mL fibronectin solution at 37 ° C. for 6 hours, and then air-dried. 600 μL / well of DMEM (Gibco BRL) containing 100 nM of CXCL12 (R & D Systems) and buffer A (0.1% bovine serum albumin, 12 mM HEPES) containing a test substance was added to the lower chamber of the transwell. Substance and human breast cancer MDA-MB-231 cells (purchased from American Tissue Culture Collection), 100 μL / well of buffer A containing 2 × 10 ⁶ cells / mL were added, and cultured at 37 ° C. for 15 hours in a 5% CO ₂ incubator. The upper surface of the filter was wiped off to remove the cells, the cells on the lower surface of the filter were fixed and stained with a 25% methanol solution containing 0.5% crystal violet (Wako Pure Chemical), washed with distilled water, and air-dried. Disconnect, 0.1 M sodium citrate / 50 An ethanol solution was added to dissolve crystal violet, and the absorbance was measured at 550 nm. The addition of CXCL12 enhanced the migration of MDA-MB-231 cells, and the antagonist, TC-14012, inhibited the migration of MDA-MB-231 cells induced by CXCL12 to 10 nM.

<Test Example 5: Inhibitory activity of 4Fbenzoyl-TN-14003 on T cell-derived leukemia cell migration induced by CXCL12>
In a lower chamber of a transwell (polycarbonate filter, 8 μm diameter, Costar), 600 μL of DMEM (GibcoBRL) containing 30 nM of CXCL12 (R & D Systems) and buffer A (0.1% bovine serum albumin, 12 mM HEPES) containing a test substance was placed in the lower chamber. A test substance and 100 μL / well of buffer A containing 2 × 10 ⁶ cells / mL were added to the upper chamber of the test substance and SUP-T1 cells derived from human T cells (purchased from American Tissue Culture Collection). After culturing in a 5% CO ₂ incubator for 4 hours, the number of cells that had migrated to the lower chamber was measured using a Coulter counter, and the results are shown in Fig. 3. The negative control indicates migratory activity without the addition of CXCL12. Control is migration when CXCL12 was added. The addition of CXCL12 enhanced the migration of SUP-T1 cells.The antagonistic activity of SUP-T1 cells induced by CXCL12 was inhibited by 4Fbenzoyl-TN-14003 up to 10 nM. 4Fbenzoyl-TN-14003 is considered to be useful as an inhibitor of rheumatoid arthritis by inhibiting T cell motility at a low concentration.

<Test Example 6: Inhibitory activity of 4F-benzoyl-TN-14003 on breast cancer cell migration induced by CXCL12>
A transwell filter (polycarbonate filter, 8 μm diameter, Costar) was treated in a 10 μg / mL fibronectin solution at 37 ° C. overnight, and then air-dried. 600 μL / well of DMEM (GibcoBRL) containing buffer A (0.1% bovine serum albumin, 12 mM HEPES) containing 100 nM of CXCL12 (R & D Systems) and 4F-benzoyl-TN-14003 was added to the lower chamber of the transwell. 100 μL / well of buffer F containing 4 Fbenzoyl-TN-14003 and human breast cancer MDA-MB-231 cells (purchased from American Tissue Culture Collection), 2 × 10 ⁶ cells / mL was added to the chamber at 37 ° C. in a 5% CO ₂ incubator. After culturing for 15 hours, the upper surface of the filter is wiped off, the cells are removed, the cells on the lower surface of the filter are fixed and stained with a 25% methanol solution containing 0.5% crystal violet (Wako Pure Chemical), and washed with distilled water. Air dried. The filter part was cut off, 0.1 M sodium citrate / 50% ethanol solution was added to dissolve the crystal violet, and the absorbance was measured at 550 nm.The results are shown in Fig. 4. The negative control is the migratory property without adding CXCL12. The positive control indicates the migration when CXCL12 was added.The addition of CXCL12 enhanced the migration of MDA-MB-231 cells.The migration of MDA-MB-231 cells induced by this CXCL12 The sex antagonist 4Fbenzoyl-TN-14003 inhibited up to 100 nM.

<Test Example 7: Anti-metastatic activity of 4Fbenzoyl-TN-14003>
10 ⁶ MDA-MB-231 human breast cancer cells were transplanted through the tail vein to 5 week old female CB-17 SCID mice (CLEA Japan). 4Fbenzoyl-TN-14003 was adjusted to 80 mg / mL with physiological saline, and sealed in a sustained-release osmotic pump (Alzet Pump, Alza, for two-week sustained release) (the dose was 18.2 mg each). / kg / day), and was mounted subcutaneously on the back of the mouse on the day before transplantation. Further, 14 days after transplantation, an Alzet pump containing the same dose of drug was additionally installed. The control group was additionally equipped with an Alzette pump infused with physiological saline. Twenty-eight days after the transplantation, the mice were dissected, about 2 mL of a 0.2% Evans blue solution was injected from the trachea, and the lungs were stained. The lungs were removed, immersed in Bouin's solution, stained and fixed. The metastatic foci (the part stained in yellow) was visually observed to determine whether there was clear anti-metastatic activity. FIG. 5 shows the results. In the control group lung (A), a portion stained yellow was uniformly observed, and metastasis to the lung was observed. On the other hand, in the 4Fbenzoyl-TN-14003-administered group (B), a case in which the rate of yellow staining was small was observed. In comparison, it was observed that metastasis was suppressed in the 4Fbenzoyl-TN-14003 administration group.

<Test Example 8: Inhibitory activity of 4Fbenzoyl-TN14003 on migration of human T cell line (Jurkat) and mouse splenocytes induced by CXCL12>
1. Effect on migration reaction of human T cell line RPMI-1640 and fetal calf serum (FCS) from BioWhittaker, penicillin-streptomycin solution, RPMI-1640 (without phenol red) and HEPES from Invitrogen, BSA from Sigma, human SDF-1α (CXCL12) was purchased from Genzyme. The human T lymphocyte cell line Jurkat was purchased from ATCC and cultured in RPMI-1640 10% FCS. 4F-benzoyl-TN14003 was dissolved in PBS and used for the experiment.
A migration reaction was performed using a 24-well Transwell (Costar, polycarbonate membrane, pore size 5 μm). 600 μL of SDF-1α (final concentration 1 ng / mL) was added to the lower layer of the Transwell, 5 × 10 ⁵ cells (200 μL) were added to the insert, and the mixture was reacted at 37 ° C. for 4 hours. Cells were pre-incubated with drug for 30 minutes at 37 ° C. The migration reaction was performed in RPMI-1640 medium containing 20 mmol / L HEPES and 0.5% BSA. Cells migrated to the lower layer were collected, and the number of cells was counted using a Coulter Counter. The inhibition rate (%) for the migration reaction due to the drug at each concentration was determined by the following formula, and the IC ₅₀ value was calculated from the inhibition rate.

As a result, as shown in FIG. 6, the Jurkat cells showed strong cell migration reactivity to SDF-1α. 4F-benzoyl-TN14003 inhibited this reaction in a concentration-dependent manner, and its IC ₅₀ value was 0.65 nmol / L.

2. Effect of mouse spleen cells on migration reaction Spleens were collected from BALB / c mice (male, Charles River Japan), used as a single cell suspension, and erythrocytes were disrupted to prepare spleen cells.
SDF-1α (Peprotech, final concentration: 100 ng / mL) was added to the lower layer of Transwell (pore size: 5 μm), and 1 × 10 ⁶ / well cells (100 μL) were added to the insert, and reacted at 37 ° C. for 2.5 hours. Cells were pre-incubated with drug for 30 minutes at 37 ° C. The migration reaction was performed in RPMI-1640 medium containing 20 mmol / L HEPES and 0.5% BSA. The number of cells that migrated to the lower layer was counted using a Coulter Counter. In the same manner as in the previous section, the inhibition rate of the migration reaction by each concentration of the drug and the IC ₅₀ value of the inhibitory action were calculated.
As a result, as shown in FIG. 7, 4F-benzoyl-TN14003 showed a concentration-dependent inhibitory effect on the migration reaction of mouse spleen cells to SDF-1α. Its IC ₅₀ value was 0.54 nmol / L, showing the same inhibitory effect as that of human cells. This confirmed that there was almost no species difference between the present peptide and human.

<Test Example 9: Effect of 4Fbenzoyl-TN14003 on mouse delayed type hypersensitivity reaction (DTH)>
Sheep preserved blood was purchased from Japan Biological Materials Center. Sheep preserved blood was washed twice with physiological saline, suspended in physiological saline, and used as sheep red blood cells (SRBC). Since the OD 541 nm value of oxyhemoglobin when hemolyzing 1.0 × 10 ⁹ cells / mL of the SRBC suspension with 14 times the volume of distilled water is said to correspond to approximately 0.700, the SRBC density was adjusted accordingly.
BALB / c mice (male, 6 weeks old, Charles River Japan) were sensitized by subcutaneously administering 2 × 10 ⁷ cells / 50 μl of SRBC to the left hind footpad. Five days later, 10 ⁸ cells / 50 μl of SRBC was subcutaneously administered to the right hind footpad to induce a DTH reaction. Immediately before and 24 hours after antigen induction, the thickness of the right hind footpad was measured using a digital micrometer (Mitutoyo CD-15B), and the increase in footpad thickness (mm) due to swelling was used as an index of the DTH response.
4F-benzoyl-TN14003 was dissolved in PBS and continuously administered using an Alzet osmotic pump (Alza, 0.5 μL / hr for 7 days). The osmotic pump was implanted subcutaneously in the back under ether anesthesia the day before sensitization. A pump injected with PBS as a control was similarly implanted. 4F-benzoyl-TN14003 was administered at doses of 4.8, 24 and 120 μg / day.
Data were expressed as mean ± standard error (n = 7). Performed by the Williams test, and p ≦ 0.025 was considered significant.
As a result, as shown in FIG. 8, 4F-benzoyl-TN14003 (4.8, 24 and 120 μg / day) dose-dependently and significantly suppressed footpad edema, and the inhibition rates were 9, 31 and 51%, respectively. Met. This suggested that CXCR4 plays an important role in cell-mediated immunity such as DTH reaction.

<Test Example 10: Therapeutic effect of 4Fbenzoyl-TN14003 on mouse collagen arthritis>
FK-506 was purified by a method known per se (Kino T. et al., J. Antibiot., 1987 40 (9): 1249-55). Methotrexate was purchased from Wako Pure Chemical Industries, indomethacin from Sigma, bovine type II collagen from Collagen Technology Workshop, Freund's complete adjuvant (FCA) from Difco, and anti-mouse IgG2a antibody from Zymed.
Bovine type II collagen was dissolved at a concentration of 2 mg / mL with a 0.05 mol / L acetic acid solution, and an emulsion was prepared with an equal amount of FCA. DBA / 1JN mice (male, 6 weeks old, Charles River Japan) were sensitized by inoculating 50 μL of the emulsion into the skin of the ridge. Booster immunization was performed similarly 21 days later. Body weight and hind limb thickness measurements and arthritis scoring were performed for two weeks after the boost. The arthritis score was scored on a scale of 0-3 on each limb and evaluated as a total (out of 12) (0, normal; 1, mild or single finger swelling; 2, moderate or multiple finger swelling) 3, severe swelling). Limbs and serum were collected two weeks after the boost.
After coating and blocking the immunoplate with bovine type II collagen (10 μg / mL PBS solution), 100 μL of a 1000-fold diluted mouse serum was added and left at room temperature for 2 hours. After washing, an anti-mouse IgG2a antibody (1000-fold dilution) was added. After washing, TMB was added, and after standing at room temperature for 30 minutes, an equal amount of H ₂ SO ₄ was added, and A450 nm was measured.
Indomethacin (1 mg / kg), methotrexate (3 mg / kg) and FK-506 (10 mg / kg) were suspended in 0.5% methylcellulose and orally administered daily for 2 weeks from the day of booster immunization in a volume of 0.1 mL / 10 g body weight. did. The control group was orally administered the same volume of a 0.5% methylcellulose solution. 4F-benzoyl-TN14003 was dissolved in PBS and continuously administered using an Alzet osmotic pump (Alza, 0.5 μL / hr for 2 weeks). The osmotic pump was implanted subcutaneously on the back of the mouse under ether anesthesia the day before the booster immunization. A pump injected with PBS as a control was similarly implanted. Each drug was evaluated based on the value 2 weeks after the booster immunization.
Data were expressed as mean ± standard error (n = 8-12). Comparison between the two groups was performed by Student's t-test, and p ≦ 0.05 was considered significant. Multiple comparison was performed by Dunnett's test, and p ≦ 0.05 was considered significant.
As a result, Indomethacin (1 mg / kg, po), methotrexate (3 mg / kg, po) and FK-506 (10 mg / kg, po) all significantly suppressed heel swelling of the hind limbs, A significant or clear inhibitory effect was exhibited (FIG. 9).
4F-benzoyl-TN14003 (120 μg / day) showed a significant inhibitory effect on heel swelling of the hind limbs, arthritis score and weight loss. Further, the anti-type II collagen IgG2a antibody tended to suppress the increase in the antibody titer (FIG. 10). These inhibitory effects were equal to or higher than the above-mentioned existing drugs.

Since the peptide compound of the present invention having CXCR4 antagonistic activity inhibits the binding between CXCR4 and CXCL12 / SDF-1α, cancer types expressing CXCR4, for example, oral cancer, pharyngeal cancer, lip cancer, tongue Cancer, gingival, nasopharyngeal, esophageal, gastric, small intestine, colon, including colon, liver, gallbladder, pancreatic, nasal, lung, osteosarcoma, soft tissue, skin, melanoma Can suppress the migration reaction of cancer cells such as breast cancer, uterine cancer, ovarian cancer, prostate cancer, testicular cancer, penis cancer, bladder cancer, kidney cancer, brain tumor, thyroid cancer, lymphoma, leukemia, etc. It is useful as a prophylactic and / or therapeutic agent. Further, the peptidic compound of the present invention can suppress the immune cell migration reaction induced by CXCL12 / SDF-1α, and is useful as a preventive and / or therapeutic agent for rheumatoid arthritis.

It is a figure which shows the inhibitory activity of TE-14005 with respect to the breast cancer cell migration induced by CXCL12. The vertical axis indicates cell migration (absorbance at OD 550 nm). From the left, CXCL12 was not added (negative control), CXCL12 was added (positive control), CXCL12 and TE-14005 were added at 10 nM, CXCL12 and TE-14005 were added at 100 nM, CXCL12 and TE-14005 were added at 1 μM, CXCL12 and TE- 14005 shows the results when 10 μM was added (mean value + standard deviation, n = 2). * Indicates the significance of each TE-14005 addition group with the positive control group (Williams test, p ≦ 0.025). It is a figure which shows the inhibitory activity of TC-14012 with respect to the breast cancer cell migration induced by CXCL12. The vertical axis indicates cell migration (absorbance at OD 550 nm). From the left, without CXCL12 (negative control), with CXCL12 (positive control), with 1 nM CXCL12 and TC-14012, with 10 nM CXCL12 and TC-14012, with 100 nM CXCL12 and TC-14012, CXCL12 and TC- 14012 shows the results when 1 μM was added (mean value + standard deviation, n = 2). * Indicates the significance of each TC-14012-added group to the positive control group (Williams test, p ≦ 0.025). It is a figure which shows the inhibitory activity of 4Fbenzoyl-TN-14003 on the migration of T cell-derived leukemia cells induced by CXCL12. The vertical axis indicates cell migration (cell number). From the left, CXCL12 was not added (negative control), CXCL12 was added (positive control), CXCL12 and 4Fbenzoyl-TN-14003 were added at 1 nM, CXCL12 and 4Fbenzoyl-TN-14003 were added at 10 nM, and CXCL12 and 4Fbenzoyl-TN-14003 were added. The results at the time of addition of 1 μM of CXCL12 and 4Fbenzoyl-TN-14003 are shown (mean value + standard deviation, n = 2). * Indicates the significance of each 4Fbenzoyl-TN-14003 addition group with the positive control group (Williams test, p ≦ 0.025). FIG. 2 shows the inhibitory activity of 4Fbenzoyl-TN-14003 on breast cancer cell migration induced by CXCL12. The vertical axis indicates cell migration (absorbance at OD 550 nm). From left, when CXCL12 was not added (negative control), when CXCL12 was added (positive control), when CXCL12 and 4Fbenzoyl-TN-14003 were added at 10 nM, when CXCL12 and 4Fbenzoyl-TN-14003 were added at 100 nM, and when CXCL12 and 4Fbenzoyl-TN-14003 were added. The results at the time of addition of 10 μM of CXCL12 and 4Fbenzoyl-TN-14003 are shown (mean value + standard deviation, n = 2). * Indicates the significance of each 4Fbenzoyl-TN-14003 addition group with the positive control group (Williams test, p ≦ 0.025). It is a lung tissue staining image showing the lung metastasis inhibitory activity of 4Fbenzoyl-TN-14003 in a mouse transplanted with human breast cancer cells. A is an image of a lung of a control group (group administered with saline), and B is an image of a lung of a group administered with 4Fbenzoyl-TN-14003. It is a figure which shows the inhibitory activity of 4Fbenzoyl-TN-14003 on the migration of Jurkat cells induced by SDF-1α (CXCL12). The vertical axis indicates cell migration (ratio of moving cells to input). From left, when SDF-1α is not added, when SDF-1α is added, when SDF-1α and 4Fbenzoyl-TN-14003 are added at 10 pM, when SDF-1α and 4Fbenzoyl-TN-14003 are added at 100 pM, SDF-1α and 4Fbenzoyl-TN-14003 are added. The results when 1 nM is added, when SDF-1α and 4Fbenzoyl-TN-14003 are added at 10 nM, and when SDF-1α and 4Fbenzoyl-TN-14003 are added at 100 nM are shown. It is a figure which shows the inhibitory activity of 4Fbenzoyl-TN-14003 on the migration of mouse spleen cells induced by SDF-1α (CXCL12). The vertical axis indicates cell migration (ratio of moving cells to input). From left, when SDF-1α is not added, when SDF-1α is added, when SDF-1α and 4Fbenzoyl-TN-14003 are added at 10 pM, when SDF-1α and 4Fbenzoyl-TN-14003 are added at 100 pM, SDF-1α and 4Fbenzoyl-TN-14003 are added. The results when 1 nM is added, when SDF-1α and 4Fbenzoyl-TN-14003 are added at 10 nM, and when SDF-1α and 4Fbenzoyl-TN-14003 are added at 100 nM are shown. It is a figure which shows the inhibitory effect of 4Fbenzoyl-TN-14003 on mouse DTH response induced by SRBC. The vertical axis indicates the increase in footpad thickness due to swelling (mean ± standard error, n = 7). From the left, the results of the PBS administration (control) group, the 4Fbenzoyl-TN-14003 4.8 μg / day administration group, the 4Fbenzoyl-TN-14003 24 μg / day administration group, and the 4Fbenzoyl-TN-14003 120 μg / day administration group are shown. * Indicates P ≦ 0.025 (comparison with PBS administration group; Williams test). It is a figure which shows the effect of the existing drug on mouse collagen arthritis. A is a change in body weight [vertical axis: change in body weight (g) (mean ± standard error, n = 8), abscissa: days after booster immunization], B is a change in disease incidence [vertical axis: disease incidence ( %) (N = 8), horizontal axis: days after booster immunity], C: fluctuation of arthritis score [vertical axis: arthritis score (mean ± standard error, n = 8), horizontal axis: days after booster immunization ] And D show the variation of heel thickness [vertical axis: heel thickness (mm) (mean ± standard error, n = 8), horizontal axis: days after booster immunization] (□: normal mouse group, Δ: No drug administration (control) group, Δ: indomethacin administration group, 群: methotrexate administration group, ●: FK506 administration group). E is the effect of each drug on hind limb swelling 2 weeks after booster immunization [vertical axis: hind limb weight (mg) (mean ± standard error, n = 8)]; F is anti-bovine 2 weeks after booster immunization Shows the effect of each drug on type II collagen IgG2a antibody titer [vertical axis: antibody titer (A450) (mean ± standard error, n = 8)], respectively. [From left: normal mouse group, drug-free (control) group] , Indomethacin (IND) administration group, methotrexate (MTX) administration group, and FK506 administration group]. ## is P ≦ 0.01 (comparison with normal mouse group; t test), * and ** are P ≦ 0.05 and P ≦ 0.01 (comparison with drug non-administration group; Dunnett test), respectively. Is shown. It is a figure which shows the effect of 4Fbenzoyl-TN-14003 on mouse collagen arthritis. A: variation in body weight [vertical axis: variation in body weight (g) (mean ± standard error), horizontal axis: days after booster immunization], B: variation in incidence [vertical axis: incidence (%), horizontal Axis: days after booster immunity], C: fluctuation of arthritis score [vertical axis: arthritis score (mean ± standard error), horizontal axis: days after booster immunity], D: fluctuation of heel thickness [vertical axis: heel] Thickness (mm) (mean ± standard error), horizontal axis: days after booster immunization] [□: normal mouse group (n = 8), Δ: drug-free (control) group (n = 12) , Δ: 4Fbenzoyl-TN-14003 administration group (n = 11)]. E is the effect of 4Fbenzoyl-TN-14003 on hind limb swelling 2 weeks after booster immunization [vertical axis: hind limb weight (mg) (mean ± standard error)], and F is anti-bovine II 2 weeks after booster immunization. Shows the effect of 4Fbenzoyl-TN-14003 on type I collagen IgG2a antibody titer [vertical axis: antibody titer (A450) (mean ± standard error)] [Normal mouse group (n = 8) from left; Control) group (n = 12) and 4Fbenzoyl-TN-14003 administration group (n = 11) are shown]. ## indicates P ≦ 0.01 (comparison with normal mouse group; t test), ** indicates P ≦ 0.01 (comparison with drug non-administration group; t test).

Claims (22)

The following formula (Ia)
1 2 3 4 5 6 7 8 9 10 11 12 13 14
A1-A2-A3-Cys-Tyr-A4-A5-A6-A7-A8-A9-A10-Cys-A11 (Ia)
(Where
A1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is deleted;
A2 represents an arginine or glutamic acid residue when A1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus, and when A1 is deleted Represents an arginine or glutamic acid residue which may be derivatized at the N-terminus;
A3 represents an aromatic amino acid residue;
A4, A5 and A9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
A6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
A7 represents a proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
A8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
A10 represents citrulline, glutamic acid, arginine or lysine residue;
A11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, the cysteine residues at the 4-position and the 13-position may be connected by a disulfide bond, and the amino acid may be L-form or D-form. Or a salt thereof, for preventing and / or treating cancer or rheumatoid arthritis. In the above formula (Ia),
A1 is an arginine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is deleted;
A2 represents an arginine or glutamic acid residue when A1 is an arginine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus; Represents an arginine or glutamic acid residue which may be derivatized at the terminus;
A4 represents arginine, citrulline, alanine or glutamic acid residue;
A5 represents arginine, citrulline, alanine, lysine or glutamic acid residue;
A6 represents a lysine, alanine, citrulline or glutamic acid residue;
A7 represents a proline or alanine residue;
A8 represents a tyrosine, alanine or glutamic acid residue;
A9 represents an arginine, citrulline or glutamic acid residue;
A10 represents citrulline or glutamic acid residue;
The prophylactic and / or therapeutic agent according to claim 1, wherein A11 represents an arginine or glutamic acid residue which may be derivatized at the C-terminus. The following formula (Ib)
1 2 3 4 5 6 7 8 9 10 11 12 13 14
B1-B2-B3-Cys-Tyr-B4-B5-B6-B7-B8-B9-B10-Cys-B11 (Ib)
(Where
B1 is a glutamic acid residue which may be derivatized at the N-terminus or is missing;
B2 represents an arginine or glutamic acid residue when B1 is a glutamic acid residue which may be derivatized at the N-terminus, and is derivatized at the N-terminus when B1 is deleted. Represents an arginine or glutamic acid residue which may be present;
B3 represents an aromatic amino acid residue;
B4, B5 and B9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
B6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
B7 represents proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
B8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
B10 represents citrulline, glutamic acid, arginine or lysine residue;
B11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, and the cysteine residues at the 4-position and the 13-position may be connected by a disulfide bond. Or a salt thereof. The peptide or a salt thereof according to claim 3, wherein B1 is a glutamic acid residue which may be derivatized at the N-terminus. The following formula (Ic)
1 2 3 4 5 6 7 8 9 10 11 12 13 14
C1-C2-C3-Cys-Tyr-C4-C5-C6-C7-C8-C9-C10-Cys-C11 (Ic)
(Where
C1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is missing;
C2 represents a glutamic acid residue if C1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus, and if C1 is deleted, Represents a glutamic acid residue which may be derivatized at the N-terminus;
C3 represents an aromatic amino acid residue;
C4, C5 and C9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
C6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
C7 represents a proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
C8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
C10 represents citrulline, glutamic acid, arginine or lysine residue;
C11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, and the cysteine residues at the 4-position and the 13-position may be connected by a disulfide bond. Or a salt thereof. The following formula (Id)
1 2 3 4 5 6 7 8 9 10 11 12 13 14
D1-D2-D3-Cys-Tyr-D4-D5-D6-D7-D8-D9-D10-Cys-D11 (Id)
(Where
D1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is deleted;
D2 represents an arginine or glutamic acid residue when D1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus, and when D1 is deleted Represents an arginine or glutamic acid residue which may be derivatized at the N-terminus;
D3 represents an aromatic amino acid residue;
D4 represents a glutamic acid residue;
D5 and D9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
D6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
D7 represents a proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
D8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
D10 represents a citrulline, glutamic acid, arginine or lysine residue;
D11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, and the cysteine residues at the 4-position and the 13-position may be connected by a disulfide bond. Or a salt thereof. The following formula (Ie)
1 2 3 4 5 6 7 8 9 10 11 12 13 14
E1-E2-E3-Cys-Tyr-E4-E5-E6-E7-E8-E9-E10-Cys-E11 (Ie)
(Where
E1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or has been deleted;
E2 represents an arginine or glutamic acid residue when E1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus; Represents an arginine or glutamic acid residue which may be derivatized at the N-terminus;
E3 represents an aromatic amino acid residue;
E4 and E9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
E5 represents an arginine or glutamic acid residue;
E6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
E7 represents a proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
E8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
E10 represents a citrulline, glutamic acid, arginine or lysine residue;
E11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, and the cysteine residues at the 4-position and the 13-position may be connected by a disulfide bond. Or a salt thereof. The peptide of claim 7, wherein ΔE5 represents a glutamic acid residue, or a salt thereof. The following formula (If)
1 2 3 4 5 6 7 8 9 10 11 12 13 14
F1-F2-F3-Cys-Tyr-F4-F5-F6-F7-F8-F9-F10-Cys-F11 (If)
(Where
F1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is missing;
F2 represents an arginine or glutamic acid residue when F1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus; Represents an arginine or glutamic acid residue which may be derivatized at the N-terminus;
F3 represents an aromatic amino acid residue;
F4, F5 and F9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
F6 represents a glutamic acid residue;
F7 represents proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
F8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
F10 represents citrulline, glutamic acid, arginine or lysine residue;
F11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, and the cysteine residues at the 4-position and the 13-position may be connected by a disulfide bond. Or a salt thereof. The following formula (Ig)
1 2 3 4 5 6 7 8 9 10 11 12 13 14
G1-G2-G3-Cys-Tyr-G4-G5-G6-G7-G8-G9-G10-Cys-G11 (Ig)
(Where
G1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is deleted;
G2 represents an arginine or glutamic acid residue when G1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus; Represents an arginine or glutamic acid residue which may be derivatized at the N-terminus;
G3 represents an aromatic amino acid residue;
G4, G5 and G9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
G6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
G7 represents a proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
G8 represents a glutamic acid residue;
G10 represents a citrulline, glutamic acid, arginine or lysine residue;
G11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, and the cysteine residues at the 4-position and the 13-position may be connected by a disulfide bond. Or a salt thereof. The following formula (Ih)
1 2 3 4 5 6 7 8 9 10 11 12 13 14
H1-H2-H3-Cys-Tyr-H4-H5-H6-H7-H8-H9-H10-Cys-H11 (Ih)
(Where
H1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is missing;
H2 represents an arginine or glutamic acid residue when H1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus; Represents an arginine or glutamic acid residue which may be derivatized at the N-terminus;
H3 represents an aromatic amino acid residue;
H4 and H5 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
H6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
H7 represents a proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
H8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
H9 represents a glutamic acid residue;
H10 represents a citrulline, glutamic acid, arginine or lysine residue;
H11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, and the cysteine residues at the 4-position and the 13-position may be connected by a disulfide bond. Or a salt thereof. The following formula (Ii)
1 2 3 4 5 6 7 8 9 10 11 12 13 14
I1-I2-I3-Cys-Tyr-I4-I5-I6-I7-I8-I9-I10-Cys-I11 (Ii)
(Where
I1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is missing;
I2 represents an arginine or glutamic acid residue when I1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus; Represents an arginine or glutamic acid residue which may be derivatized at the N-terminus;
I3 represents an aromatic amino acid residue;
I4, I5 and I9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
I6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
17 represents a proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
I8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
I10 represents glutamic acid, arginine or lysine residue;
I11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, and the cysteine residues at the 4-position and the 13-position may be connected by a disulfide bond. Or a salt thereof. The following formula (Ij)
1 2 3 4 5 6 7 8 9 10 11 12 13 14
J1-J2-J3-Cys-Tyr-J4-J5-J6-J7-J8-J9-J10-Cys-J11 (Ij)
(Where
J1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is deleted;
J2 represents an arginine or glutamic acid residue when J1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus; Represents an arginine or glutamic acid residue which may be derivatized at the N-terminus;
J3 represents an aromatic amino acid residue;
J4, J5 and J9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
J6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
J7 represents a proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
J8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
J10 represents citrulline, glutamic acid, arginine or lysine residue;
J11 represents a glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, and the cysteine residues at the 4-position and the 13-position may be connected by a disulfide bond. Or a salt thereof. Any of the following peptides (1) to (58) or a salt thereof:
(1) Ac-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH;
(2) Ac-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-OH;
(3) Ac-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-OH;
(4) Ac-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Cit-Cit-Cys-Arg-OH;
(5) Ac-Cit-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH;
(6) Ac-Cit-Arg-Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-OH;
(7) Ac-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Cit-Cit-Cys-Arg-OH;
(8) Ac-Cit-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Cit-Cit-Cys-Arg-OH;
(9) Ac-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(10) Ac-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Cit-Cit-Cys-Arg-NH 2;
(11) Ac-Cit-Arg -Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(12) Ac-Cit-Arg -Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(13) Ac-Arg-Arg -Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Cit-Cit-Cys-Arg-NH 2;
(14) Ac-Cit-Arg -Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Cit-Cit-Cys-Arg-NH 2;
(15) H-DGlu-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH;
(16) H-Arg-Glu-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH;
(17) H-Arg-Arg-Nal-Cys-Tyr-Glu-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH;
(18) H-Arg-Arg-Nal-Cys-Tyr-Arg-Glu-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH;
(19) H-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-OH;
(20) H-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Glu-Cit-Cys-Arg-OH;
(21) H-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Glu-OH;
(22) H-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(23) H-Arg-Arg -Nal-Cys-Tyr-DGlu-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(24) H-Arg-Arg -Nal-Cys-Tyr-DGlu-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(25) H-DGlu-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(26) H-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-DGlu-Arg-Cit-Cys-Arg-NH 2;
(27) H-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-DGlu-Cys-Arg-NH 2;
(28) Ac-DGlu-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(29) Ac-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-DGlu-Arg-Cit-Cys-Arg-NH 2;
(30) Ac-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-DGlu-Cys-Arg-NH 2;
(31) Ac-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(32) guanyl-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(33) TMguanyl-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(34) TMguanyl-Arg-Nal -Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(35) 4F-benzoyl-Arg -Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(36) 2F-benzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂ ;
(37) APA-Arg-Nal -Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(38) desamino-R-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(39) guanyl-Arg-Nal -Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(40) succinyl-Arg-Nal -Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(41) glutaryl-Arg-Nal -Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(42) deaminoTMG-APA-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(43) nelfinabiryl-succinyl-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂ ;
(44) AZT-glutaryl-Arg -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
_{(45) R-CH 2 -Arg} -Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(46) H-Arg-Nal -Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(47) TMguanyl-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(48) ACA-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(49) ACA-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH;
(50) H-Arg-Arg -Nal-Cys-Tyr-Cit-Arg-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(51) Ac-Arg-Arg -Nal-Cys-Tyr-Cit-Arg-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(52) Ac-Arg-Arg -Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(53) Ac-Arg-Arg -Nal-Cys-Tyr-Arg-Lys-DCit-Pro-Tyr-Arg-Cit-Cys-Arg-NH 2;
(54) 4F-benzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-NH ₂
(55) 4F-benzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NHMe
(56) 4F-benzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NHEt
(57) 4F-benzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-NHiPr
(58) 4F-benzoyl-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DGlu-Pro-Tyr-Arg-Cit-Cys-Arg-tyramine
(In each sequence, the symbol to the left of the N-terminal amino acid indicates that the amino group is derivatized or not, H indicates that it is not derivatized, Ac indicates an acetyl group, and guanyl indicates a guanyl group. Succinyl is a succinyl group, glutaryl is a glutaryl group, TMguanyl is a tetramethylguanyl group, 2F-beozoyl is a 2-fluorobenzoyl group, 4F-benzoyl is a 4-fluorobenzoyl group, and APA is a 5-amino group. A pentanoyl group, ACA is a 6-aminohexanoyl group, desamino-R is a 2-desamino-arginyl group, deaminoTMG-APA is the following formula (II),

nelfinabiryl-succinyl has the following formula (III),

AZT-glutaryl has the following formula (IV):

R-CH ₂ has the following formula (V)

Arg represents an L-arginine residue, Nal represents an L-3- (2-naphthyl) alanine residue, Cys represents an L-cysteine residue, Tyr represents an L-tyrosine residue, and Cit represents L-tyrosine residue. -Citrulline residue, Lys is L-lysine residue, DLys is D-lysine residue, Pro is L-proline residue, DCit is D-citrulline residue, DGlu is D-glutamic acid residue. , Glu each represents an L-glutamic acid residue, two cysteine residues are linked by an intramolecular disulfide bond, and the symbol to the right of the C-terminal amino acid means that the carboxyl group is derivatized or not. Shows that OH is not derivatized, NH ₂ is an amino group, NMe is a methylamino group, NEt is an ethylamino group, NiPr is an isopropylamino group, and tyramine is a p-hydroxyphenylethylamino group. Amidated with Door are shown, respectively). A pharmaceutical comprising the peptide according to any one of claims 3 to 14 or a salt thereof. 16. The medicament according to claim 15, which is a CXCR4 antagonist. The medicament according to claim 15, which is an agent for preventing and / or treating cancer or rheumatoid arthritis. 18. The medicament according to claim 17, wherein the cancer type is breast cancer or pancreatic cancer. 方法 A method for preventing or treating cancer or rheumatoid arthritis, which comprises administering to a mammal an effective amount of the peptide according to any one of claims 3 to 14 or a salt thereof. 使用 Use of the peptide or a salt thereof according to any one of claims 3 to 14 for producing a prophylactic and / or therapeutic agent for cancer or rheumatoid arthritis. For mammals, the following formula (Ia)
1 2 3 4 5 6 7 8 9 10 11 12 13 14
A1-A2-A3-Cys-Tyr-A4-A5-A6-A7-A8-A9-A10-Cys-A11 (Ia)
(Where
A1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is deleted;
A2 represents an arginine or glutamic acid residue when A1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus, and when A1 is deleted Represents an arginine or glutamic acid residue which may be derivatized at the N-terminus;
A3 represents an aromatic amino acid residue;
A4, A5 and A9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
A6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
A7 represents a proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
A8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
A10 represents citrulline, glutamic acid, arginine or lysine residue;
A11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, the cysteine residues at the 4-position and the 13-position may be connected by a disulfide bond, and the amino acid may be L-form or D-form. Wherein the peptide or a salt thereof is administered in an effective amount. Formula (Ia) for producing a prophylactic and / or therapeutic agent for cancer or rheumatoid arthritis
1 2 3 4 5 6 7 8 9 10 11 12 13 14
A1-A2-A3-Cys-Tyr-A4-A5-A6-A7-A8-A9-A10-Cys-A11 (Ia)
(Where
A1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus or is deleted;
A2 represents an arginine or glutamic acid residue when A1 is an arginine, lysine, ornithine, citrulline, alanine or glutamic acid residue which may be derivatized at the N-terminus, and when A1 is deleted Represents an arginine or glutamic acid residue which may be derivatized at the N-terminus;
A3 represents an aromatic amino acid residue;
A4, A5 and A9 independently represent arginine, lysine, ornithine, citrulline, alanine or glutamic acid residues;
A6 represents proline, glycine, ornithine, lysine, alanine, citrulline, arginine or glutamic acid residue;
A7 represents a proline, glycine, ornithine, lysine, alanine, citrulline or arginine residue;
A8 represents tyrosine, phenylalanine, alanine, naphthylalanine, citrulline or glutamic acid residue;
A10 represents citrulline, glutamic acid, arginine or lysine residue;
A11 represents an arginine, glutamic acid, lysine or citrulline residue which may be derivatized at the C-terminus;
In the above formula, Cys represents a cysteine residue, Tyr represents a tyrosine residue, the cysteine residues at the 4-position and the 13-position may be connected by a disulfide bond, and the amino acid may be L-form or D-form. Or a salt thereof.

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