WO2001019389A1

WO2001019389A1 - Remedies for rheumatoid arthritis and fused protein

Info

Publication number: WO2001019389A1
Application number: PCT/JP2000/006180
Authority: WO
Inventors: Toshihiro Nakajima; Kusuki Nishioka
Original assignee: St. Marianna University School Of Medicine; Santen Pharmaceutical Co., Ltd.
Priority date: 1999-09-10
Filing date: 2000-09-11
Publication date: 2001-03-22
Also published as: AU6877100A

Abstract

Remedies for rheumatoid arthritis and articular synovial cell proliferation inhibitors, in particular, remedies for rheumatoid arthritis and articular synovial cell proliferation inhibitors capable of inhibiting the proliferation and differentiation of rheumatoid arthritis articular synovial cells suffering from abnormality in the function regulatory mechanism at the transcription factor expression mechanism level. Namely, remedies for rheumatoid arthritis and articular synovial cell proliferation inhibitors which contain, as the active ingredient, the phosphorylated CREB recognition region of CBP which is a phosphorylated CREB coactivator; and a fused protein which comprises the sequence represented by SEQ ID NO:2 in Sequence Listing, the sequence represented by SEQ ID NO:3 in Sequence Listing and the sequence consisting of the 553- to 679-residues (KIX 10.4) of mouse CBP.

Description

Remedies for rheumatoid arthritis and fusion

The present invention provides a synovial cell proliferation inhibitor capable of introducing a phosphorylated CREB recognition region partial peptide of CBP, which is a phosphorylated CREB coactivator, into the synovial cell nucleus of rheumatoid arthritis, and The present invention relates to a therapeutic agent for rheumatoid arthritis, which comprises a phosphorylated CREB recognition region partial peptide of CBP, which is a phosphorylated CREB coactivator, as an active ingredient. Background art

Cell-level responses to various stimuli such as hormones and growth factors, in particular, the activation or inactivation of transcription of DNA to RNA are important steps directly involved in cell growth and activation. Transcription is the root of life support. Thus, transcriptional regulation is directly involved in life support in this sense. Regulation of DNA transcription is mediated by a group of mainly proteinaceous factors called transcription factors. For example, in the hormone Zc AMP / protein kinase A (PKA) pathway, the binding of hormones to hormone receptors increases adenylate cyclase activity, and PKA is activated when intracellular c AMP levels increase. It translocates into the nucleus and phosphorylates the transcription factor CREB (CRE binding protein), which binds to a transcriptional control site called CRE (cAMP response element) in DNA. Phosphorylated CREB binds to the phosphorylated CREB coactivator, CBP (CREB binding protein), and as a result, transcription by RNA polymerase II is activated.

In recent years, knowledge about the regulation mechanism of transcription factor expression in synovial cell proliferation and activation has been accumulated. For example, it has been reported that CREB is induced when synovial cell proliferation is inhibited by the addition of cAMP analog (Higaki et al., Clinical Immunity, Vol. 27, No. 2, 182-188 ( 1 995)).

Furthermore, stimulation of rheumatoid arthritis synovial cells with interleukin-11, an inflammatory cytokine, increased CREB expression. High expression of CREB was observed in tissues, and strong expression of CREB was observed even when pannus-like tissue, a granulation tissue of synovium, was formed in an in vitro culture system. It is also known that pr-cAMP showed an inhibitory action on synovial cell proliferation (Suzuki et al., History of Medicine, Vol. 182, No. 9, 586-589, 1997, 8. 20).

On the other hand, Nakajima et al. Reported that a partial peptide of amino acids 455-679 in the phosphorylation CREB recognition region called KIX in CBP (mouse) was 〇 £ 8 !: serine 1 in a region called the ID domain. It has been reported that this is a specific recognition region for phosphorylation of 33 residues (mouse) (MoI. Cell. Bio 1., 16: 694-703 (1969)). .

For CBP and its homolog P300 present in adenovirus E1A, Chrivia et al. (Nature, Vol. 365, No. 6449, 855- 859 (1993)), Livingston et al. v., Vol. 8, No. 8, 869-884 (1994)).

It is known that binding to CREB in which serine 133 residues in the KID domain is phosphorylated is carried out in a region called KIX such as 1S CBP and P300. The structure and amino acid sequence of the KIX domain are already known (eg, Cell, Vol. 91, pp. 741-752, December 12, 199 7).

Nakajima et al. Demonstrated that KIX SZB peptide (mouse IX: 553-679 amino acid residues) was treated as a dominant negative by intracellular microinjection to specifically suppress CREB transcriptional activity in He1a cells. (Cell. Biol., Vol. 16, 694-703 (1996)), suggesting a potential treatment for arteriosclerosis based on transcriptional control (Uehara). Note: Research report of the Life Science Foundation, Vol. 10, pp. 294-295, 1996).

Rheumatoid arthritis is a chronic inflammatory disease of which the cause is currently unidentified, with polyarthritis as the main symptom. In rheumatoid arthritis, the disease progresses with inflammatory infiltration of the synovium, proliferation of synovial cells, stratification and angiogenesis. However, the cause of synovial cell hyperfunction, which plays an important role in the pathogenesis of rheumatoid arthritis, It has not been fully elucidated yet, and there are many unclear points regarding abnormalities in the function control mechanism of synovial cells themselves. Summary of the Invention

In view of the above-mentioned situation, the present invention relates to a therapeutic agent for rheumatoid arthritis and an agent for inhibiting the growth of synovial synovial cells, and in particular, a transcription factor expression mechanism for the proliferation and differentiation of rheumatoid arthritis synovial cells exhibiting abnormal function control mechanisms. It is an object of the present invention to provide a therapeutic agent for rheumatoid arthritis and a synovial cell proliferation inhibitor which can be suppressed at a level. The present invention provides a therapeutic agent for rheumatoid arthritis and an agent for inhibiting synovial cell proliferation of a joint, which comprises a phosphorylated CREB recognition region of CBP, which is a phosphorylated CREB coactivator, as an active ingredient.

In a preferred embodiment, the partial peptide of the phosphorylated CREB recognition region of CBP is all or part of the KIX domain of CBP, in particular, derived from human or mouse.

It is a K I X protein.

In the present invention, the phosphorylated CREB recognition region partial peptide of CBP can act as a dominant negative body in the synovial cell nucleus. The present invention is also a therapeutic agent for treating rheumatoid arthritis, which comprises administering a partial peptide of a phosphorylated CREB recognition region of CBP bound to a partial peptide of the antennadia protein.

In a preferred embodiment, a nuclear localization signal is bound to the phosphorylated CREB recognition region partial peptide of CBP bound to the partial peptide of the antenna dia protein.

The present invention further relates to a sequence represented by SEQ ID NO: 2 in the sequence listing, a sequence represented by SEQ ID NO: 3 in the sequence listing, and residues 553 to 679 (KIX 10.4) of mouse CBP. It is also a fusion protein characterized by consisting of a sequence.

The present invention still further relates to a pharmaceutical composition containing the above-mentioned fusion protein as an active ingredient, particularly a therapeutic agent for rheumatoid arthritis.

In a preferred embodiment, the fusion protein has a daltathione-1 S-transferase domain cleavable with thrombin bound thereto. 'Detailed disclosure of the invention

Hereinafter, the present invention will be described in detail.

The therapeutic agent for rheumatoid arthritis and the synovial cell proliferation inhibitor of the present invention comprise, as an active ingredient, a phosphorylated CREB recognition region partial peptide of CBP, which is a phosphorylated CREB coactivator. This active ingredient can be introduced into the nucleus of synovial cells in rheumatoid arthritis to exert an effect of inhibiting the proliferation of articular synovial cells. The partial peptide has an amino acid sequence of a phosphorylated CREB recognition region of CBP, which is a phosphorylated CREB coactivator. Phosphorylation of CBP The CREB recognition region is called KIX. When the serine residue, which is the 133rd amino acid residue (mouse) of CREB, is phosphorylated, the CREB containing this 133 serine residue Is known to bind to a region called the kinase-inducible domain (KID, amino acid residues 101-160 (mouse). SEQ ID NO: 4 in the sequence listing). Cell. Biol., Vol. 16, No. 2, 694—703 (1996)).

The KID domain has two parallel helix regions A and ct B. The amino acid sequence of this region is extremely conserved irrespective of the origin, especially the serine residue located at the Ν-terminal side of the αB helix. Is common to all as far as it is known. This serine residue is, for example, residue 133 in mouse and corresponds to residue 118 in human. Phosphorylation of this serine residue is essential for binding to KIX. The number of this serine residue in the KID domain varies depending on the origin. For convenience, the KID of each origin is a serine residue corresponding to residue 133 in mouse or rat, unless otherwise specified. It is called No. serine residue.

Κ The IX protein has three helical regions α1, hi2 and α3, and two short three. Includes helix regions G1 and G2. These regions include, for example, in the mouse KIX protein, an α1 helix spanning glutamine 579-isoloisin 611, an arginine 623—α2 helix spanning tyrosine 640, Ginine 646—α3 helix over lysine 662, tryptophan 59 1—G1 over histidine 594 and G2 over proline 617—lysine 621.

The arginine residue in helix α1 (arginine 600 in mouse KIX) is considered to be important for stabilizing the KIX protein.

IX This secondary structure of IX protein is commonly provided in various organisms, for example, human and mouse CBP, human and mouse P300, Drosophila CBP, R10E1 1. 1, K03H 1.10, etc. all have the above secondary structure (eg, Cell, Vol. 91, 741-752, 1997). KIX proteins have amino acid sequences that differ depending on the species.However, hydrophobic residues important for interaction with KID are conserved, and all amino acids in the various KIX proteins exemplified above have the same amino acid sequence. However, they belong to the same family. Therefore, in the present invention, as the partial peptide of the phosphorylated CREB recognition region of CBP, various KIX proteins having the above-mentioned characteristics, for example, human and mouse CBP KIX, human and mouse Ρ300 ΚIX, Drosophila CBP KIX, R10E11.1, K03H1.10. The major part of the amino acid sequence of these KIX proteins has the above-mentioned structure common to CBP protein, R10E11.1, and Κ03Η1.10. Amino acid residue numbers of the ΚIX domain in proteins are generally different. The major part of the amino acid sequence of these IX proteins is described, for example, in FIG. 1A of Cell, Vol. 91, 741-752 (1977).

The partial peptide may be human or mouse KIX (amino acid residues 456 to 680; mouse amino acid residues 45 55 to 679) shown in SEQ ID NO: 1 in the sequence listing. Les ,.

Examples of the partial peptide include a part of KIX of human / mouse CBP, for example, residues 554/553 to 679/680 (also referred to as “KIX SZB”), 5 54 553 residues to 6 79/680 residues (KIX 10. No. 4)), residues 577Z5 76 to 679/680 (also referred to as “KIX 5.4”), 577/5 residue 76 to 662/66 1 residue It may be a peptide having a base sequence (also referred to as “KIX 5.6”). Of these, XS / B, KIX 10.4, and X5.4 are preferred. However, for example, residues 456/455 to 598/597 (residues also referred to as “KIX II”), residues 617 to 616 to residues 680/679 (“KIX 13.4)) and residues 5775576 to 649-648 (also referred to as “KIX 5.1.2”). Not used in the present invention. The above-mentioned examples are non-limiting listings of substances that can be used in the present invention. For example, as long as they have a binding property to phosphorylated CREB, for example, the above-mentioned IX protein secondary structure and KID Amino acid substitutions that do not affect hydrophobic interaction are also applicable to the present invention.

As a method for preparing the KIX protein, for example, the gene sequence of the KIX domain corresponding to residues 586 to 679 of mouse CBP is amplified by PCR, inserted into an appropriate expression vector, and introduced into E. coli. Then, the cultured cells are collected, and the supernatant obtained by ultrasonic treatment and centrifugation is applied to an FP LC column to obtain a protein fraction and purified.

Phosphorylation of the CB Ρ CREB recognition domain partial peptide is introduced into the synovial nucleus of the rheumatoid arthritis joint, along with chemicals such as microinjection, electroporation, scrape bite-dating, and calcium phosphate. Although a known method such as a method can be used, in the present invention, it is preferable to use an internalization vector derived from a home domain, which will be described below.

In other words, the Drosophila transcription factor antenna dia is a DNA-binding protein with a home domain, but the home domain (60 amino acid residues) of this antenna dia translocates through the cell membrane. It is known. In recent years, the minimum amino acid sequence required for this antenna media transfer has been determined. This region is a third helix consisting of 16 amino acid residues consisting of residues 43 to 58 of the homeodomain of the antenna media (SEQ ID NO: 2 in the sequence listing). This region (hereinafter also referred to as Ant_Mini) or the home domain of the antenna media containing this region is preferably bound to a physiologically active substance of 10 kDa or less, for example, a protein, a nucleic acid, or another compound. This allows efficient and stable introduction into cells.

Note that the third helix structure itself is not always necessary for exerting the transfer function.For example, even if glutamine 50 is replaced with proline, or isofisine 45, glutamine 50, and lysine 55 are each substituted. Replacement with proline reduces translocation into the nucleus but allows translocation into cells. In addition, the reverse sequence of the 16 amino acid residue sequence represented by SEQ ID NO: 2 in the above sequence listing (residue from arginine at residue 43 to lysine at residue 58), that is, the above amino acid sequence in reverse order from N-terminal to C-terminal In addition, even if the synthetic peptide is arranged from the lysine at residue 58 to the arginine at residue 43, the ability to enter the nucleus is maintained. Furthermore, a peptide in which the above 16 amino acid residue sequence is composed of D amino acids may be used. In addition, even if there is an amino acid substitution in the home domain of the antenna media, for example, in the second helix, the transfer function is not necessarily inhibited (for example, Curr ent Op In io i n i n N e u r

0 b i o 1 o g y, 1 996, 6: 629—634, especially Tables 1 and 2). Therefore, in the present invention, when the phosphorylated CREB recognition region partial peptide of CBP is introduced into the synovial cell nucleus of a joint of rheumatoid arthritis, a partial peptide of antennadia, particularly Ant- Preferably, the fusion protein is introduced by binding to Mini or a home domain containing the same.

It is preferable to include a nuclear localization signal for introduction into the nucleus of the synovial cells. Nuclear localization dignity sign nanore (nucl ear loc aliiz ations

1 gna 1; NLS) is an amino acid sequence that functions in the translocation of a peptide from the cytoplasm through the nuclear pore into the nucleus. Therefore, the fusion protein preferably comprises Ant-Mini or a homeodomain containing the same, a nuclear localization signal, and a partial peptide of the phosphorylated CREB recognition region of CBP. The nuclear localization signal may be, for example, a basic amino acid cluster (SEQ ID NO: 3 in the sequence listing) such as SV40 virus large T antigen, a bipolar signal such as nucleoplasmin, or the like. Conventionally, it has been considered that the maximum number of peptides that can be transferred and introduced by using partial peptides of antenna media is about 100 amino acid residues (Curent Opinionin Neurobiology, 1996, 6: 629). — 6 3 4) In the present invention, by including a nuclear localization signal, even a longer peptide can be introduced into the nucleus.

Examples of the method for preparing the fusion protein include: a) phosphorylation of 〇8? £ B recognition region partial peptide, or, if applicable, phosphorylation of CBP to which a nuclear localization signal is added CREB recognition region A peptide in which a thiol-protected cysteine is linked to a peptide, an internalization vector Ant-Mini or a thiol-protected cysteine at the end of a homeodomain containing the same. To form an S--S bond between the two through deprotection of the thiol.b) Continuously connect the internalization vector with the phosphorylated CR EB recognition region partial peptide of CBP. And synthesizing them into a series of fusion proteins. In addition, the phosphorylated CREB recognition region partial peptide of CBP can bind to the N-terminus of Ant-Mini, but does not exclude binding to the C-terminus.

In the present invention, it is preferable that a daltathione-1 S-transferase (GST) domain, which can be cleaved with thrombin, is bound to the fusion protein. The daltathione-S-transferase domain is a protein with a molecular weight of 260,000 and cannot translocate into cells. Then, the fusion protein can be translocated into cells by cutting off the fusion protein with thrombin. Therefore, the fusion protein can be transferred into cells only in the presence of thrombin, and the introduction of the fusion protein into cells can be controlled. This makes it possible, for example, to establish a drug delivery system that allows a pharmaceutical composition containing the above-mentioned fusion protein as an active ingredient to efficiently act on a target tissue, for example, an affected area in rheumatoid arthritis. , Is advantageous.

In this case, there must be no thrombin cleavage site in the fusion protein. However, the fusion protein, for example, Ant-Mini or a homeodomain containing the same, a nuclear localization signal, and CBP Phosphorylated CREB recognition region Fusion proteins consisting of partial peptides serve this purpose.

The GST fusion protein can be prepared, for example, by incorporating the above fusion protein gene sequence into an expression vector capable of expressing a fusion protein of the target protein and the GST domain, and placing the gene fusion vector in an appropriate host. And a method of expressing the same. As such expression vectors, commercially available expression vectors can be used. For example, pGEX-2 and pGEX-2TK in GSTGene Fusion Vetors series manufactured by Amersham Pharmacia Biotech, Inc .: , PGEX—4T—1, pGEX—4T—2, pGEX—4T—3 and so on.

In the present invention, a phosphorylated CREB recognition region partial peptide of CB, which is a phosphorylated CREB coactivator, is used as a dominant negative body, that is, a peptide that binds to the KID region of phosphorylated CREB competitively with CBP. Rheumatoid arthritis can be introduced into synovial cell nuclei. Since the above phosphorylated CREB recognition region partial peptide can bind to a region called 〇 £ 8!: 1D, it blocks the binding of phosphorylated CREB coactivator, CBP, and transcribes it. It is considered to prevent the start.

Therefore, the fusion protein of the present invention may be able to suppress the proliferation and activation of joint synovial cells in the lesions of patients with rheumatoid arthritis, and is expected to be useful as a therapeutic agent for rheumatoid arthritis. .

For such a therapeutic agent, other well-known components such as a stabilizer can be used as long as the object of the present invention is not hindered. Further, a pharmacologically acceptable carrier can be blended. Further, the method may include a means for transporting the active ingredient to a target tissue. BEST MODE FOR CARRYING OUT THE INVENTION

Hereinafter, the present invention will be described in more detail with reference to Examples and the like, but the present invention is not limited thereto. Example 1 Preparation of KIX fusion protein A gene sequence encoding mouse IX (residues 586-6979) was prepared by the method described in Chrivia et al., Nature 365, 855-8559 (1993). And amplified by the PCR method. Using p GEX-2T (manufactured by Amersham Pharmacia Biotech) as an expression vector, the Ant-Mini gene-NLS gene-KIX gene was incorporated into the vector and expressed in E.co1i. The resulting mixture was purified by a daltathione-Sepharose column to obtain a KIX fusion protein consisting of N-terminal, GST domain, Ant-Mini domain, NLS domain, KIX domain, and C-terminal. Ant—Mini used the amino acid sequence of SEQ ID NO: 2 in the sequence listing, and NLS used the amino acid sequence of SEQ ID NO: 3 in the sequence listing. Example 2 Growth inhibition test of FGF stimulation of synovial cells

Synovial cells were obtained by removing blood cells from synovial tissue obtained with consent at the time of knee surgery in patients with rheumatoid arthritis and then counting the cells on a 96-well cultured plastic plate (100 microliters). at ^{1 X 1 0 4 Zw e 1} 1 conditions were used after 3-5 passages cultured in 2% FCS-DMEM culture medium.

After synovial cells were cultured for 24 hours in 2% FCS, the GST / Ant—Mini / NL S / KIX fusion protein obtained in Example 1 was treated with thrombin to obtain Ant—Mini / NL SZK. The IX fusion protein was added to a concentration of 5 / ig gml, 10 μg / m 125 μg / m I, 50 μg / m 1 and 100 g / m 1 . 30 minutes after the addition of the fusion protein, FGF (fibroblast growth factor) was added. Then, after culturing for 12 hours, the number of synovial cells in each we11 was quantified by the MTT method. The MTT assay was performed after adding MTT and reacting for 6 hours, and then measuring the OD using a plate absorbance meter. As a control, one without the fusion protein was used.

The results are shown in Table 1. The numerical values are relative values when the control is 100. The case where no FGF was added was also shown. When FGF was not added, it was also confirmed that the addition of the fusion protein did not affect the proliferation up to at least 50 μg Zm1. table 1

From the results in Table 1, it can be seen that the fusion protein of the present invention is rapidly introduced into synovial cell nuclei, that it can affect the expression of transcription factors as dominant negative CBP-KIX, and that it is stimulated by FGF. It was found that activation of synovial cells could be effectively suppressed. Industrial applicability

INDUSTRIAL APPLICABILITY As described above, the present invention can provide a novel introduction method capable of rapidly introducing an unprecedented dominant negative CBP-KIX into the synovial cell nucleus of a joint. Proliferation and differentiation of rheumatoid arthritis synovial cells can be suppressed at the level of transcription factor expression mechanism. Also, a therapeutic agent for rheumatoid arthritis and a synovial cell proliferation inhibitor for arthritis can be provided using the dominant negative body as an active ingredient. The pharmaceutical composition containing the fusion peptide of the present invention as an active ingredient can be used as a synovial cell proliferation inhibitor and a therapeutic agent for rheumatoid arthritis.

Claims

The scope of the claims

1. A therapeutic agent for rheumatoid arthritis, comprising a phosphorylated CREB recognition region of CBP, a phosphorylated CREB coactivator, as an active ingredient.

2. An agent for inhibiting synovial cell proliferation, comprising a phosphorylated CREB recognition region of CBP, a phosphorylated CREB coactivator, as an active ingredient.

3. The therapeutic agent for rheumatoid arthritis according to claim 1, wherein the CBP phosphorylated CREB recognition region partial peptide comprises all or a part of the CBP KIX domain.

4. The therapeutic agent for rheumatoid arthritis according to claim 1, wherein the CBP phosphorylated CREB recognition region partial peptide is a human or mouse-derived KIX protein.

5. The phosphorylation of CBP The CREB recognition region partial peptide has a sequence consisting of mouse CBP residues 553 to 679 (KIX 10.4). Rheumatoid arthritis treatment.

6. The therapeutic agent for rheumatoid arthritis according to claim 1, wherein the CBP phosphorylation CREB recognition region partial peptide is a dominant negative body.

7. A therapeutic agent for rheumatoid arthritis, which comprises administering a partial peptide of CBP phosphorylated CREB recognition region bound to a partial peptide of antennadia protein.

8. The therapeutic agent for rheumatoid arthritis according to claim 7, wherein the CBP phosphorylation CREB recognition region is a dominant negative body.

9. The therapeutic agent for rheumatoid arthritis according to claim 7 or 8, wherein the partial peptide of the antennadia protein has an amino acid sequence represented by SEQ ID NO: 2 in the sequence listing.

10. A therapeutic agent for rheumatoid arthritis, which comprises administering a partial peptide of antenna dia protein and phosphorylation of CBP bound to a nuclear localization signal and a partial CREB recognition region peptide.

1 1. The sequence represented by SEQ ID NO: 2 in the sequence listing, the sequence represented by SEQ ID NO: 3 in the sequence listing, and the sequence of residues 553 to 679 (KIX 10.4) of mouse CBP A fusion protein comprising a fusion protein.

1 2. The fusion protein of claim 11 having a daltathione-1 S-transferase domain that can be cleaved with thrombin.

1 3. The sequence represented by SEQ ID NO: 2 in the sequence listing, the sequence represented by SEQ ID NO: 3 in the sequence listing, and the sequence of residues 553 to 679 (KIX 10.4) of mouse CBP A pharmaceutical composition comprising a fusion protein comprising the active ingredient as an active ingredient.

14. The sequence represented by SEQ ID NO: 2 in the sequence listing, the sequence represented by SEQ ID NO: 3 in the sequence listing and the sequence of residues 553 to 679 (KIX 10.4) of mouse CBP, and A pharmaceutical composition comprising, as an active ingredient, a fusion protein comprising a daltathione-S-transferase domain that can be cleaved with thrombin.

15. The composition according to claim 13, which is a therapeutic agent for rheumatoid arthritis.

1 6. Effective amount of phosphorylation CREB phosphorylation of CB P, a coactivator A method for treating rheumatoid arthritis, which comprises administering a REB recognition region as an active ingredient.

1 7. A method for inhibiting the proliferation of articular synovial cells, which comprises administering an effective amount of a phosphorylated CREB recognition region of CBP, which is a phosphorylated CREB coactivator, as an active ingredient.

1 8. The method for treating rheumatoid arthritis according to claim 16, wherein the CBP phosphorylated CREB recognition region partial peptide comprises all or a part of the domain of the 8th to 8th.

1 9. The method for treating rheumatoid arthritis according to claim 16, wherein the CBP phosphorylation CREB recognition region partial peptide is an IX protein derived from human or mouse.

20. The phosphorylated CREB recognition region partial peptide of CBP has a sequence consisting of mouse CBP residues 553 to 679 (KIX 10.4). How to treat rheumatoid arthritis.

21. The method for treating rheumatoid arthritis according to claim 16, wherein the CBP phosphorylated CREB recognition region partial peptide is a dominant negative form.

22. A method for treating rheumatoid arthritis, which comprises administering an effective amount of a phosphorylated CREB recognition region partial peptide of CBP bound to a partial peptide of an antenna protein.

23. The therapeutic agent for rheumatoid arthritis according to claim 22, wherein the phosphorylation of CBP The CREB recognition region is a dominant negative body.

24. The method for treating rheumatoid arthritis according to claim 22 or claim 23, wherein the partial peptide of the antenna dia protein has an amino acid sequence represented by SEQ ID NO: 2 in the sequence listing.

25. A method for treating rheumatoid arthritis, which comprises administering an effective amount of a partial peptide of antennadiadia protein and a partial peptide of CRBP phosphorylated CREB recognition region bound to a nuclear-localized signature.

26. Use of the phosphorylated CREB recognition domain of CBP, a phosphorylated CREB coactivator, for the treatment of rheumatoid arthritis.

27. Use of a phosphorylated CREB recognition domain of CBP, a phosphorylated CREB coactivator, for inhibiting the growth of joint synovial cells.

28. The use of the phosphorylated CREB recognition region of CBP according to claim 26, wherein the partial peptide of the phosphorylated CREB recognition region of CBP contains all or a part of the KIX domain of CBP.

29. Use of the CBP phosphorylated CREB recognition region according to claim 26, wherein the CBP phosphorylated CREB recognition region partial peptide is IX protein derived from human or mouse.

30. The CBP phosphorylation CREB recognition region partial peptide has a sequence consisting of mouse CBP residues 553 to 679 (KIX 10.4). Phosphorylation of CBP Use of CREB recognition region.

31. Use of the phosphorylated CREB recognition region of CBP according to claim 26, wherein the partial peptide of the CBP phosphorylated CREB recognition region is a dominant negative form.

32. Use of CBP phosphorylated CREB recognition domain partial peptides combined with antennae, a partial protein of the deaprotein, for the treatment of rheumatoid arthritis.

33. Use of a phosphorylated CREB recognition region partial peptide of CBP bound to a partial peptide of the antenna vedia protein according to claim 32, wherein the phosphorylated CREB recognition region of CBP is a dominant negative form.

34. The partial peptide of the antenna dia protein according to claim 32 or 33, wherein the partial peptide of the antenna dia protein has an amino acid sequence represented by SEQ ID NO: 2 in the sequence listing. Use of phosphorylated CREB recognition region partial peptides.

35. Use of the antenna, a partial peptide of the diaprotein, and the phosphorylation of CBP in conjunction with a nuclear localization signal, the CREB recognition domain partial peptide, for the treatment of rheumatoid arthritis.

36. As a pharmaceutical composition, a sequence represented by SEQ ID NO: 2 in the sequence listing, a sequence represented by SEQ ID NO: 3 in the sequence listing, and residues 553 to 679 of mouse CBP (KIX 10. Use of a fusion protein comprising the sequence of 4).

37. As a pharmaceutical composition, the sequence represented by SEQ ID NO: 2 in the sequence listing, the sequence represented by SEQ ID NO: 3 in the sequence listing, and residues 553 to 679 of mouse CBP (KIX 10 Use of the sequence of 4), and a fusion protein comprising a daltathione mono-S-transferase domain that can be cleaved with thrombin.

38. Use of the fusion protein according to claim 36 or 37, wherein the pharmaceutical composition is a therapeutic agent for rheumatoid arthritis.