JP2002338600A - Hiv-1 infection-suppressing polypeptide - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a HIV-1 infection-suppressing polypeptide. SOLUTION: Peptides corresponding to an extracellular domain of the N- terminal of CXCR4, CCR5, CCR3 and GPR1 were synthesized, and the influence of the peptides on coreceptor-using HIV-1 infection was studied, and it was found that a peptide of the first to 26th at the N-terminal of GPR1, which has not been considered as a chemokine receptor, extremely suppresses the HIV-1 infection. That is, the present invention provides polypeptides having an amino acid sequence of MEDLEETLFEEFENYSYDLDYYSLES (Sequence No.1), EFENYSYDLDYY (Sequence No.2) or ENYSYDLDYYSLE (Sequence No.3).

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a peptide which suppresses HIV-1 infection, and more particularly, to a peptide which suppresses HIV-1 infection.
The present invention relates to an N-terminal peptide of GPR1, which is a co-receptor of HIV-1, and a pharmaceutical composition comprising the same.

[0002]

BACKGROUND OF THE INVENTION Human acquired immunodeficiency syndrome (AIDS)
Is a disease with a very high incidence and mortality characterized by a severe systemic immunodeficiency state caused by human immunodeficiency virus (HIV) infection. H
IV has a property of infecting cells by using a cell membrane protein called CD4 as a receptor, and thus infects and destroys CD4-positive helper T cells, which are central cells controlling cell-mediated immunity. This leads to a marked decline in cellular immunity,
The result is a systemic immunodeficiency condition. HIV, a pathogenic virus of AIDS, is broadly classified into HIV-1 and HIV-2, which have different serological and genetic properties.
HIV-1 is a virus that is currently the dominant pandemic and is distributed worldwide. In contrast, HIV-2
Is mainly limited to the West African region, and only sporadic cases related to the West African region have been reported in France, the United States, and Belgium. HIV-1 is further roughly divided into three groups: groups M, O and N. this house,
Group M is the most prevalent lineage, but this group is further classified into at least 10 subtypes of subtypes AJ. HIV infects CD4 molecules present on the membranes of helper T cells and macrophages as receptors, and requires cofactors (co-receptors) that cooperate with CD4 molecules to promote viral entry. The HIV-1 co-receptor has long been a mystery, but in 1996, it was identified as the chemokine (inflammatory cytokine) receptors CXCR4 and CCR5 (Feng Y, Broder CC, Kennedy PE , Berger EA,
Science 272 (5263): 809-10 (1996); Alkhatib G, Comba
diere C, Broder CC, Feng Y, Kennedy PE, Murphy PM,
Berger EA. Science 272 (5270): 1955-8 (1996); Deng
H, Liu R, Ellmeier W, Choe S, Unutmaz D, Burkhart
M, Di Marzio P, Marmon S, Sutton RE, Hill CM, Davi
s CB, Peiper SC, Schall TJ, Littman DR, Landau NR.
Nature. 381 (6584): 661-6 (1996); Dragic T, Litwin
V, Allaway GP, Martin SR, Huang Y, Nagashima KA, C
ayanan C, Maddon PJ, Koup RA, Moore JP, Paxton WA.
Nature. 381 (6584): 667-73 (1996)). Those that utilize CXCR4 as a co-receptor were converted to X4 virus (T-trop
ic), those that use CCR5 are converted to R5 virus (M-trop
ic), which uses both X4-R5 virus (dual-t
ropic).

[0003] Soluble CD4, a protein related to the receptor, was found to have an anti-HIV effect.
D4 has been reported to have little antiviral activity in macrophage-tropic HIV strains (Turner S; Tiza
rd R; DeMarinis J; Pepinsky RB; Zullo J; Schooley
R; Fisher R, Proc Natl Acad Sci USA 1992; 89: 1335-
9). The fusion protein of soluble CD4 and the Fc portion of immunoglobulin has a stronger anti-HIV effect than soluble CD4,
The fusion protein is currently undergoing clinical testing, but its effect has not yet been reported. Anti-HIV activities of CXCR4 and CCR5-related peptides have been reported for coreceptor-related synthetic peptides with small amino acids of 10 or less (Konishi K; Ikeda
K; Achiwa K; Hoshino H; Tanaka K, Chem Pharm Bull
(Tokyo) 2000, 48: 308-9, Farzan M; Vasilieva N; Sc
hnitzler CE; Chung S; Robinson J; Gerard NP; Gerar
d C; Choe H; Sodroski J., J Biol Chem 2000, 275: 33
516-21)), its activity is not very strong.

[0004] Table 2000-507596 states that
A non-chemokine agent capable of binding to a chemokine receptor,
By contacting CD4 ⁺ cells in an amount and under conditions that inhibit fusion of HIV-1 to CD4 ⁺ cells, HIV
-1 infection can be inhibited. Furthermore, JP-A-2001-503608 discloses that a polypeptide having the sequence of the extracellular domain of CC5, which is a chemokine receptor, can inhibit HIV-1 infection.
Here, it is reported that the second extracellular domain of CCR5 (ECL2 peptide) has a virus growth inhibitory effect, but reports that it has no HIV growth inhibitory effect in other places.
On the other hand, GPR1 is a orphan receptor that does not belong to typical chemokine receptors such as CCR5 and CXCR4 and has a structure similar to a G protein-coupled transmembrane receptor seven times. , Conventional GPR1-related peptides and HI
There is no report on the relationship between V-1 and infection inhibition.

[0005] All HIV co-receptors are GPCRs.
(G protein couple 7 times transmembrane receptor).
More than 100 GPCRs have been reported,
CR consists of an N-terminal extracellular domain and first, second and third extracellular loops. The present inventors have found that GPCRs that can already be co-receptors include acidic amino acids (D: aspartic acid, E: glutamic acid) in the extracellular region, especially at the N-terminus.
And tylosin (Y) were reported to be high (Shimizu-N;
Soda-Y; Kanbe-K; Liu-HY; Mukai-R; Kitamura-T; Hosh
ino-H J-Virol. 2000; 74: 619-26). It has been experimentally demonstrated in the case of CCR5 that tylosin sulfation of this region is required for efficient HIV transmission (Kuhm
ann-SE; Platt-EJ; Kozak-SL; Kabat-D J-Virol. 200
0; 74: 7005-15, Farzan-M; Mirzabekov-T; Kolchinsk
yP; Wyatt-R; Cayabyab-M; Gerard-NP; Gerard-C; Sod
roski-J; Choe-H Cell. 1999; 96: 667-76). The coreceptor binds to the V3 region of the HIV coat protein gp120. The V3 region contains many basic amino acids,
It is possible that it binds to the co-receptor acidic amino acid / sulfated tylosin. The need for acidic amino acids is not experimentally proven, but is considered to be a common property of GPCRs that are co-receptors.

[0006]

SUMMARY OF THE INVENTION The CD4 molecule is an HIV
-1, HIV-2, and SIV have been identified as receptors, and several molecules belonging to GPCRs have been reported to act as coreceptors for HIV / SIV infection. In the present invention, HIV-1
The purpose of this study was to investigate the effects of co-receptor-related peptides on the invasion process of cells into cells, to clarify the necessity of these factors in the early stages of infection and to use them in the development of antiviral agents. . So first, HIV
CXCR4 and CCR, the main co-receptors of strain -1
5 and CCR3, and further focused on the co-receptor of GPR1 which the present inventors have frequently studied, synthesized a peptide of its N-terminal extracellular domain, and examined the effect on HIV-1 infection. Surprisingly, the infection of most HIV-1 strains was suppressed in the presence of the GPR1 peptide. Therefore, this mechanism was analyzed and further examined as a clue for the development of a new anti-HIV-1 agent.

[0007]

Means for Solving the Problems and Embodiments of the Invention
The entry of V-1 into cells requires the presence of CD4 and mainly a chemokine receptor as a co-receptor. Chemokine receptor is G-protein-coupled7-transmenbra
belongs to ne receptor (GPCR), and many HIV-1
Cells are infected using chemokine receptors such as CCR5, CXCR4 and CCR3 among GPCRs. One of the orphan receptors, GPR1
-1 strain has already been reported (Shimizu-N; Sod
aY; Kanbe-K; Liu-HY; Jinno-A; Kitamura-T; Hoshino
-H, J-Virol. 73 (6): 5231-9 (1999)). These GPC
The extracellular domain of R is the HIV-1 Env protein g
It is expected to have affinity with p120. So CX
Peptides corresponding to the N-terminal extracellular domain of CR4, CCR5, CCR3, and GPR1 were synthesized, and the effect of each co-receptor on HIV-1 infection was examined. As a result, the N-terminal 1-26 peptide of GPR1, which is not considered to be a chemokine receptor, is HI
V-1 was found to significantly suppress infection. on the other hand,
CXCR4, CCR5, or the N-terminal peptide of CCR3 did not affect infection. The results suggested that the HIV-1 particles bind to the GPR1 peptide, that the binding is inhibited by the anti-V3 antibody, that the tylosin sequence in the peptide is required for inhibitory activity, and the like.
As a result, the GPR1 peptide is a new type of anti-HIV
It was found that the drug had a -1 action.

[0008] That is, the present invention relates to MEDLEETLEE.
It is a polypeptide having the amino acid sequence of FENYSYDLDYYSLES (SEQ ID NO: 1). As will be apparent from the examples described later, MEDLEETL
The polypeptide having the amino acid sequence of DLDYYSLESC (SEQ ID NO: 4) showed an infection-suppressing action on HIV-1, but the C-terminal C of this sequence was added for the sake of experimentation. Since it is not considered to be effective, it was found that MEDLETLEFEFENYSYDYLDYYSLE was effective in this sequence.
It is considered that the polypeptide had the amino acid sequence of S (SEQ ID NO: 1). It is considered that the polypeptide having this sequence has a high possibility of binding to HIV due to a large amount of acidic amino acids and tylosin. Further, considering the effective minimum unit of this sequence from the results shown in Table 2 below, when the number of amino acids is about 14 or less, Y, D and E are contained in half or more, and two or more tyrosines and It is believed that the smallest peptide containing four or more acidic amino acids (D or E) caused this infection inhibitory effect. Therefore, E
FENYSYDLDYY (SEQ ID NO: 2) or ENYSY
It is considered that a polypeptide having any of the amino acid sequences of DLDYYSLE (SEQ ID NO: 3) provided the infection inhibitory effect of the present invention.

[0009] The present invention also comprises an amino acid sequence in which one or several amino acids have been deleted, substituted or added in any of the amino acid sequences of SEQ ID NOs: 1 to 3,
It is a protein that has an HIV-1 infection-suppressing action.
Specifically, these amino acid sequences (SEQ ID NOs: 1 to 3)
It is also considered that C, D, E, Y, or some other amino acids or the like are added or substituted, or natural ones having an intermediate size have the same effect. In addition, since the activity of tylosin is increased by sulphation, it is considered that, in addition to the GPR1 peptide, sulphated tylosin (Y) of the candidate peptide has a higher infection-suppressing effect on HIV-1. Can be Still further, these may include immunoglobulins (eg, IgM and I
peptide fused to Fc portion of gG) or CD4
It is considered that proteins also have an HIV-1 infection-suppressing action.

[0010] The synthetic peptide is an established T cell line directional T
-Tropic X4 AIDS virus (X4HIV-
1) Since it efficiently inhibits the infection of macrophage-tropic M-tropic R5 AIDS virus (R5HIV-1), it can be used as an anti-HIV-1 agent. Because it is a synthetic peptide, oral administration does not allow absorption from the intestinal tract.
It is administered sarcastically and subcutaneously. In recent years, patients frequently infected with HIV-1 resistant to combination therapy of anti-HIV drugs, and anti-HIV-1
Although it is difficult for some patients to use, the peptide of the present invention is expected to be effective when administered to such patients.

Further, the present invention provides a pharmaceutical composition comprising an effective amount of the above-mentioned polypeptide for inhibiting infection of HIV-1 into CD4 cells and a pharmaceutically acceptable carrier,
In addition, HIV-1 using this drug composition in combination with other drugs
It is a pharmaceutical composition which suppresses infection of stomach. Here, other drugs include a nucleoside reverse transcriptase inhibitor AZ
T (zidovudine), ddl (didannosine), ddC (zalcitabine), 3TC (lamivudine) and d4T (stavudine), non-nucleoside reverse transcriptase inhibitors nevirapine and delavirdine, and protease inhibitors indinavir, ritonavir; Saquinavir and nelfinavir and the like.

The drug is dissolved in physiological saline, phosphate buffered saline or the like and administered parenterally (intravenous, intramuscular, intradermal, subcutaneous injection). The concentration is 1 in the blood.
10 to 1,000 times at a time so that it is about 0 μg / ml.
0 mg. Stability in blood is unknown, but at most three times a day. If combined with the Fc of the antibody to increase stability, it may be possible to administer large amounts once a few days.

[0013]

The present invention is illustrated below by way of examples, but is not intended to limit the present invention. As the cells, human glioma-derived NP-2 cells were treated with human CD4 and HIV-1 co-receptor genes CCR3, CCR5, CXCR4, GP.
Cell line NP- into which R1 or the like was introduced and stably expressed them.
2 / CD4 / CCR3, NP-2 / CD4 / CCR5,
NP-2 / CD4 / CXCR4, NP-2 / CD4 / G
PR1 was used. C8166 and Molt-4 cells were also used for infection experiments and syncytium formation assay. H
As the IV-1 strain, GUN-1 / WT wild type (W
T) strain is a dual-tropic HIV of M-tropic and T-tropic
-1 and a mutant of GUN-1 / Ser is obtained by introducing a point mutation (Ser) at the P (proline) of the GPGR at the tip of the V3 region to obtain a BT cell such as BT-20 / N or BT-3. HIV-1 (BT-tropic) and also exhibits T-tropic properties. A typical strain of T-tropic virus is IIIB
BaL was used as the strain and the M-tropic strain.

In addition, the synthetic peptides shown in Table 1 were produced by commissioning a manufacturer. GPR1 pep. (Y / A) is obtained by substituting tylosin (Y) contained in a peptide with alanine (A).

[Table 1]

For preparation of the anti-GPR1 peptide antibody, a synthetic peptide consisting of the extracellular N-terminal amino acid of the GPR1 protein was bound to keyhole limpet hemocyanin, mixed with complete Freund's adjuvant, and subcutaneously injected into rabbits (Japanese rabbits). Was inoculated. GPR immune serum
Purified using a column to which one of the synthetic peptides was bound and used. For the infection experiment, the cells were spread on a 24-well plate and cultured at 37 ° C for 1 day, after which the culture solution was removed and the virus solution was inoculated at 37 ° C. Add the medium and add
After culturing for ７7 days, the medium was removed, washed with PBS (−), and then fixed by adding methanol. HIV-1 antigen positive cells
Foci were detected by immunostaining (Foci immunostaining assay, FIA). Alternatively, cells were detached with trypsin to prepare a cell smear, and examined by indirect fluorescent antibody method. One day after infection of cells with HIV-1, HIV synthesized in the cells
-1 DNA was detected using PCR. Primer is g
The globin gene was amplified as a control. Detection of binding between GPR1 peptide and virus particle HI
After incubating the V-1 particles and the peptide, the mixture was subjected to sucrose density gradient centrifugation to examine whether the virus particles and the peptide co-precipitated. The presence of virus particles was detected using a p24 monoclonal antibody, and the presence of a GPR1 peptide was detected using a rabbit anti-GPR1 peptide antibody. Virus / Viral Protein Binding Experiment Virus binding to cells was detected by Western blotting of viral protein p24 adsorbed to the cells. The binding of the viral protein to the peptide and the binding to soluble CD4 (sCD4) were detected based on the principle of the ELISA method.

[0016]HIV-1 sensitivity of coreceptor-related peptides
Effect on dyeing  The synthetic peptides shown in Table 1 were added to the HIV-1 infection system,
The effect of was studied. The result is shown in FIG. With a virus
The GUN-1 / Ser strain using GPR1, T-tr
opic IIIB strain, M-tropic BaL strain and dual-tropic
NP-2 / CD4 / GPR using GUN-1 / WT strain
1, NP-2 / CD4 / CXCR4, NP-2 / CD
Sensitive to 4 / CCR5 and NP-2 / CD4 / CCR3 cells
Dyed. GPR1 peptide is a combination of all
Inhibited dyeing. On the other hand, CXCR4, CCR3, CCR5
N-terminal peptide had no effect on infection.

Table 2 shows the results of the above study. T-tropic
Alternatively, the IC ₅₀ of the dual-tropic virus is _0.5 μ
M, and at a concentration of about 5 μM, 9
0% (IC ₉₀ ) inhibition. M-tropic virus IC
₅₀ was slightly higher at 10 μM, and the N-terminal peptide of CXCR4, CCR3 and CCR5 did not suppress infection even at a concentration of about 50 μM.

[Table 2]

Next, a partial peptide of the GPR1 peptide and a mutant peptide (GPR1 pep. 1-13, GPR1
pep. 8-20, GPR1 pep. 15-27, G
PR1 pep. (Y / A)) was examined for its anti-HIV-1 action. Table 3 shows the results.

[Table 3] From this table, Pep. Pep. 1-1
3, Pep. 8-20, Pep. It can be seen that the peptide in which 15-27 and tylosin were mutated to alanine showed almost no anti-HIV-1 activity using any virus strain. Next, the effects of these peptides on syncytium formation were examined. The result is shown in FIG. HIV-
-Producing Molt-4 / IIIB cells and C816
When these peptides are added to a mixed culture system of 6 cells, G
Only PR1 peptide 1-27 inhibited syncytium formation. This result indicates that the GPR1 peptide inhibits the process of adsorption and invasion of HIV-1.

[0019]GPCR peptide to viral DNA synthesis
Impact of  In the presence of four types of GPCR peptides, the GUN-1 strain was Mo
lt-4 cells were infected and the virus DNA
The growth was detected by PCR. The result is shown in FIG. 20μ
N-terminal peptide of M CXCR4, CCR5, CCR3
PCR band was detected even in the presence of
The intensity of the R band was not different from the control. On the other hand, GPR1
In the presence of peptide, infection can be suppressed even at a concentration of 0.4 μM.
System was granted. This result indicates that the GPR1 peptide is HI
It shows that V-1 inhibits the process up to reverse transcription.
You. When combined with the results in FIG. 2, the adsorption and infiltration of HIV-1
It is thought to hinder the process. GPR1 peptide
It was examined whether the action of was due to the inhibition of cell proliferation. The result
The results are shown in FIG. In the presence of 50 μM GPR1 peptide
Also had no effect on the growth of Molt-4 cells. That is, H
The inhibition of IV-1 infection inhibited cell proliferation.
It is determined that this is not the case.

[0020]Connection between GPR1 peptide and HIV-1 particles
Combination  The HIV-1 particles were concentrated and GPR1 peptide 1-27
Or by using sucrose density gradient centrifugation. That
FIG. 5 shows the results. Mix the virus particles and peptide,
After the reaction, the mixture was overlaid on the sucrose solution, and ultracentrifuged.
Painted. Virus p24 was detected in 8-9 fractions
(FIG. 5A). That is, the GPR1 peptide is a virus particle
It was considered that coprecipitation was detected in the same
available. Coreceptors link HIV-1 and CD4.
After binding, it is believed to bind to virus particles.
GPR1 peptide is a mechanism that has not been assumed until now.
It is judged to act on virions by nism

[0021]GPR1 peptide and virus gp120
Join  GPR1 peptide binds to recombinant gp120 of strain IIIB
did. This binding is due to a monoclonal antibody that reacts to its V3 region.
Partially inhibited by body or rabbit anti-GPR1 peptide antibody
Was. In addition, although the results are not shown, sCD4 and g
Even if p120 is incubated, the GPR1 peptide
Binding to gp120 was not inhibited. These results
Indicates that the X4 virus is independent of HIV-1 cell tropism.
Binding of gp120 to GPR1
The regions to be given are the V3 region of gp120 and the GPR1 peptide.
And binds to gp120 independently of CD4
It indicates that Synthesis of extracellular N-terminal of GPR1
The peptides were T-tropic, dual-tropic and BT-tropic
Rus infection was efficiently inhibited. Against M-tropic virus
Then the activity was rather weak. GPR1 is a core of HIV-1
Function as a receptor and the coreceptor is a virus
Is thought to react with the V3 region. In fact, we
In this experimental system, the monoclonal antibody against the V3 region
Inhibits binding of PR1 peptide to viral gp120
You are getting results. Generally, the V3 region is
CD4 and g are folded inside the coat protein
It is thought that the binding of p120 is triggered and exposed to the outside.
Have been. Our results show that part of the V3 region
This suggests that the face is outside the virus particle.

The N-terminal peptides of CCR5 and CXCR4 did not affect HIV-1 infection. Since the extracellular domains of CCR5 and CXCR4 emerge outside the virus particles only after binding to CD4, their effects may be masked under the assay conditions and may not have been detected. Tylosin present in the N-terminal part of the coreceptor is thought to play an important role in viral infection. Also in this study, when tylosin was replaced with alanine, the anti-HIV-1 action could not be detected. This result also indicates that tylosin is required for virus and coreceptor binding.

[0023]

The synthetic peptide of the N-terminal extracellular domain of GPR1 which can be a co-receptor for HIV-1 infection,
It showed an infection-suppressing effect on a wide range of HIV-1.
This peptide bound to the virus particles. No compound that inhibits infection with these types of substances is known, and may provide a clue to the development of new anti-HIV-1 agents. The polypeptide of the present invention comprises macrophage-directed H
IV-1, dual cell oriented and T cell oriented HIV-1
Strongly inhibits infection. This peptide did not inhibit the adsorption of HIV-1 to cells, but inhibited the entry into cells after adsorption to cells. Comparing the data of the present study with the data of the present experiment, the synthetic peptide shows a strong anti-HIV action against macrophage-tropic viruses. The synthetic peptide of the present invention is a relatively long synthetic peptide derived from the minor coreceptor GPR1, and has strong anti-HIV-1. Compared to soluble CD4, the activity is also stronger for macrophage-tropic strains. The sequence of this peptide is derived from the sequence of the orphan receptor GPR1 whose ligand in the human body is unknown. Since it is a part of the protein originally expressed in the body, it is not strongly antigenic and is expected to be hardly inactivated in the body.

[0024]

[Sequence List] SEQUENCE LISTING <110> Japan Science and Technology Corporation <120> Polypeptide that suppresses HIV-1 infection <130> PS01-1017 <160> 11 <210> 1 <211> 26 <212> PRT < 213> Artificial Sequence <400> 1 Met Glu Asp Leu Glu Glu Thr Leu Phe Glu Glu Phe Glu Asn Tyr Ser 1 5 10 15 Tyr Asp Leu Asp Tyr Tyr Ser Leu Glu Ser 20 25 <210> 2 <211> 12 <212 > PRT <213> Artificial Sequence <400> 2 Glu Phe Glu Asn Tyr Ser Tyr Asp Leu Asp Tyr Tyr 1 5 10 <210> 3 <211> 13 <212> PRT <213> Artificial Sequence <400> 3 Glu Asn Tyr Ser Tyr Asp Leu Asp Tyr Tyr Ser Leu Glu 1 5 10 <210> 4 <211> 27 <212> PRT <213> Artificial Sequence <400> 4 Met Glu Asp Leu Glu Glu Thr Leu Phe Glu Glu Phe Glu Asn Tyr Ser 1 5 10 15 Tyr Asp Leu Asp Tyr Tyr Ser Leu Glu Ser Cys 20 25 <210> 5 <211> 28 <212> PRT <213> Artificial Sequence <400> 5 Met Glu Gly Ile Ser Ile Tyr Thr Ser Asp Asn Tyr Thr Glu Glu Met 1 5 10 15 Gly Ser Gly Asp Tyr Asp Ser Met Lys Glu Pro Cys 20 25 <210> 6 <211> 24 <212> PRT <213> Artific ial Sequence <400> 6 Met Thr Thr Ser Leu Asp Thr Val Glu Thr Phe Gly Thr Thr Ser Tyr 1 5 10 15 Tyr Asp Asp Val Gly Leu Leu Cys 20 <210> 7 <211> 20 <212> PRT <213> Artificial Sequence <400> 7 Met Asp Tyr Gln Val Ser Ser Pro Ile Tyr Asp Ile Asn Tyr Tyr Thr 1 5 10 15 Ser Glu Pro Cys 20 <210> 8 <211> 13 <212> PRT <213> Artificial Sequence <400 > 8 Met Glu Asp Leu Glu Glu Thr Leu Phe Glu Glu Phe Glu 1 5 10 <210> 9 <211> 13 <212> PRT <213> Artificial Sequence <400> 9 Leu Phe Glu Glu Phe Glu Asn Tyr Ser Tyr Asp Leu Asp 1 5 10 <210> 10 <211> 13 <212> PRT <213> Artificial Sequence <400> 10 Tyr Ser Tyr Asp Leu Asp Tyr Tyr Ser Leu Glu Ser Cys 1 5 10 <210> 11 <211> 27 <212> PRT <213> Artificial Sequence <400> 11 Met Glu Asp Leu Glu Glu Thr Leu Phe Glu Glu Phe Glu Asn Ala Ser 1 5 10 15 Ala Asp Leu Asp Ala Ala Ser Leu Glu Ser Cys 20 25

[Brief description of the drawings]

FIG. 1. GPR1 pep. 1-27
FIG.

FIG. 2 is a diagram showing suppression of syncytium formation by a synthetic peptide.

FIG. 3. GP of viral DNA formation after HIV-1 infection.
R1 pep. It is a figure showing suppression by 1-27.

FIG. 4 shows the effect of GPR1 peptide on the growth of Molt-4 cells.

FIG. 5. GPR1 pep. FIG. 2 is a diagram showing the binding of 1-27 to HIV-1 particles.

Continued on the front page (72) Inventor Yasushi Soda 1-43-14 Denon Chofu, Ota-ku, Tokyo (72) Inventor Atsushi Ohgami 21702, Frederick, Maryland, United States of America 107 Alesandra Coat # 207 (72) Inventor Noriaki Shimizu 72-8 Amagawaharamachi, Maebashi-shi, Gunma (72) Inventor Yuji Haraguchi 864-2 Nakaocho, Takasaki-shi, Gunma F-term (reference) 4C084 AA02 AA03 AA07 BA01 BA09 BA18 BA19 BA23 MA01 MA65 MA66 NA14 ZB332 ZC422 4H045 AA10 AA30 BA16 BA18 BA50 CA40 DA50 EA29 FA40 FA73

Claims (8)

[Claims] 1. A polypeptide having the amino acid sequence of SEQ ID NO: 1. 2. A polypeptide having the amino acid sequence of SEQ ID NO: 2 or 3. 3. A polypeptide comprising an amino acid sequence in which one or several amino acids have been deleted, substituted or added in the amino acid sequence of SEQ ID NO: 1, and having an activity of suppressing HIV-1 infection. 4. A polypeptide comprising an amino acid sequence in which one or several amino acids are deleted, substituted or added in any one of the amino acid sequences of SEQ ID NOs: 2 and 3, and has an activity of suppressing HIV-1 infection. 5. The polypeptide according to claim 3, wherein tylosin in the amino acid sequence is sulfated. 6. A pharmaceutical composition comprising the polypeptide according to any one of claims 1 to 5 and a pharmaceutically acceptable carrier in an amount effective to inhibit infection of HIV-1 to CD4 cells. . 7. A pharmaceutical composition for suppressing HIV-1 infection, comprising a combination of the drug according to claim 6 and another drug. 8. An injection comprising the pharmaceutical composition according to claim 6 or 7 for intravenous, intramuscular, sarcastic or subcutaneous administration.