KR101472903B1 - Peptides for Inhibiting an Activity of T Lymphocyte and Uses Thereof - Google Patents

Abstract

The peptides of the present invention can perform the same or similar functions as the functions of native CTLA-4, and have excellent skin permeability according to their small size. The peptides of the present invention can effectively bind to antigen-presenting cell surface proteins (CD80 and CD86) to inhibit the activation of T cells and thereby inhibit the expression of inflammatory cytokines (e.g., IL-2 and IFN-y) . Accordingly, the composition comprising the peptide of the present invention exerts a very excellent effect in preventing, treating or ameliorating a Th1-mediated immune disease. Therefore, the excellent activity and stability of the peptide of the present invention can be very usefully applied to medicines, quasi-drugs, and cosmetics.

Description

T Cell Activity Inhibition Peptides and Uses Thereof < RTI ID = 0.0 >

The present invention relates to novel peptides for inhibiting T cell activity and uses thereof.

The immune response can be activated or inactivated. T cells and B cells are known to be the main cells in this immune response. Much research has been done on the role of these cells. Based on previous studies, it has been shown that the immune response of T cells is mainly related to disease. In the immune response, T cell activity is involved in the activity of cells involved in other immunity. In order for the T cells to be activated, an antigen must be presented from an antigen presenting cell (APC). Antigen presenting cells (APCs) recognize antigens first and present antigen to T cell receptor (TCR) of T cell through MHC (main histocompatibility antigen), and T cell can activate. In addition, activation of T cells requires signaling through TCR as well as additional signaling through surface molecules such as CD28. CD80 (B7-1) and CD86 (B7-2) act as ligands to CD28 and transmit stimulatory signals, which are involved in T cell cytokine production and cell survival.

CTLA-4 (Cytotoxic T lymphocyte-associated antigen 4; CD152) is a member of immunoglobulin (Ig) competitively acting on CD28 and is characterized by high affinity for CD80 and CD86 ligands than CD28. These CTLA-4 interactions are inhibitory signals that inhibit the expression of interleukin-2 (IL-2) and IL-2 receptors in T cells, Inhibit activity and proliferation.

Numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to better understand the state of the art to which the present invention pertains and the content of the present invention.

The present inventors have sought to develop a substance having the same or similar function as that of CTLA-4 and superior in activity and stability to natural CTLA-4 protein. As a result, the present inventors have found that peptides having superior physiological activity (for example, suppression of phosphorylation of JNK and p38 in spleen cells and inhibition of expression of IFN-y and IL-2 produced by T cells) than natural CTLA-4 protein Thereby completing the present invention.

Accordingly, an object of the present invention is to provide a peptide for inhibiting T-cell activity.

It is another object of the present invention to provide a pharmaceutical composition for the prevention or treatment of a Th1-mediated immune disorder.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to one aspect of the present invention, there is provided a peptide for inhibiting T-cell activity comprising an amino acid sequence selected from the group consisting of the amino acid sequences set forth in SEQ ID NOS: 1 and 2.

According to another aspect of the present invention, there is provided a pharmaceutical composition for preventing or treating a Th1-mediated immune disease using the above-mentioned peptide as an active ingredient.

The present inventors have sought to develop a substance having the same or similar function as that of CTLA-4 and superior in activity and stability to natural CTLA-4 protein. As a result, the present inventors synthesized peptides having superior physiological activities (for example, suppression of phosphorylation of JNK and p38 in spleen cells, and suppression of expression of IFN-γ and IL-2 produced by T cells) compared with natural CTLA-4 protein Respectively.

In the present invention, the peptides of the present invention are represented by the following general formulas 1 and 2:

1

Ser-Thr-Gly-Lys-Lys-Ala-Val

Formula 2

Arg-Thr-Glu-Met-Pro-Thr-Leu-Tyr

The present inventors synthesized a number of CTLA-4-derived peptides that exhibit the above-described characteristics based on sites involved in the function of human CTLA (Cytotoxic T lymphocyte-associated antigen) -4 (GenBank Accession No. AAH69566.1) Respectively. In more detail, several regions of CTLA-4 protein, a protein receptor that down-regulates the immune system, are randomly synthesized to first search for binding sites for antigen-presenting cell surface proteins, and then the amino acid sequence of the predicted site To prepare the peptide of the present invention. Receptor-binding predicted sites are prepared to prepare candidate peptides, and the peptides of the present invention are provided by screening peptides having the best inhibitory activity against T cell activation among these rear peptides.

According to some embodiments of the present invention, the Sequence Listing first sequence and Sequence Listing second sequence peptide of the present invention are derived from human CTLA-4, which is illustrated in Table 1.

The peptides of the present invention not only inhibit the activation of T cells by performing the function of native human CTLA-4 (for example, binding to antigen-presenting cell surface proteins), but also inhibit inflammatory cytokines (IL-2 and IFN- Lt; RTI ID = 0.0 > expression. ≪ / RTI >

According to some embodiments of the invention, the peptides of the invention inhibit phosphorylation of JNK (c-Jun N-terminal kinase) and p38 (see Figures 3a and 3b).

According to some embodiments of the invention, the peptides of the invention inhibit the expression of inflammatory cytokines (see Figures 4A and 4B). In Th1-mediated cellular immune responses, when T cells are activated and aged, T cells that prefer cellular immunity produce the factors. Specifically, the peptides of the present invention inhibit the expression of INF (interferon) -γ, IL (interleukin) -2 or TNF (tumor necrosis factor) -α, and more specifically INF- Lt; / RTI > IFN-γ plays a role in activating macrophages, cytotoxic T-lymphocytes (CTLs) and neutrophils, and IL-2 is a cytotoxic T- Promotes lymphocyte reaction.

As used herein, the term "peptide" refers to a linear molecule formed by peptide bonds and amino acid residues joined together. The peptides of the present invention can be prepared according to chemical synthesis methods known in the art, particularly solid-phase synthesis techniques (Merrifield, J. Amer. Chem. Soc. 85: 2149-54 (1963) ; Stewart, et al., Solid Phase Peptide Synthesis , 2nd ed., Pierce Chem. Co .: Rockford, 111 (1984)).

According to the present invention, the peptides of the present invention have very good thermal stability compared to the native CTLA-4 protein (see Figures 5A and 5B). The native CTLA-4 protein (about 30-34 kDa) has difficulties in application such as post-translational modifications (eg, detection in both cytoplasm and membrane fraction) and detection in two different sizes. In addition, it has not only difficulty in production, but also has a low stability in temperature and long-term storage. However, since the peptide of the present invention can be mass-synthesized at a very low cost and is physically and chemically stable at high temperatures, it can prevent the decrease in physiological activity to a maximum extent and has an improved therapeutic effect by increasing the survival time outside the body . Accordingly, the peptide of the present invention can be advantageously applied to products requiring long-term storage such as pharmaceuticals, quasi-drugs and cosmetics.

According to some embodiments of the present invention, the N-terminal or C-terminal of the peptide may be modified with a hydroxy group (-OH), an amino group (-NH ₂ ), an azide (-NHNH ₂ )

According to some embodiments of the present invention, the N-terminal of the peptide is selected from the group consisting of an acetyl group, a fluorenylmethoxycarbonyl group, a formyl group, a palmitoyl group, a myristyl group, a stearyl group and a polyethylene glycol (PEG) The protecting groups selected may be further combined.

The above-mentioned modification of the amino acid serves to greatly improve the stability of the peptide of the present invention. As used herein, the term "stability" refers not only to ' in vivo ' stability, but also to storage stability (e.g., room temperature storage stability). The above-mentioned protecting group acts to protect the peptide of the present invention from attack of a protein cleaving enzyme in vivo.

The present invention also provides a composition for preventing, treating or ameliorating a Th1-mediated immune disorder comprising the above-mentioned peptide as an active ingredient.

As demonstrated in the Examples below, the peptides of the present invention inhibit the phosphorylation of JNK and p38 associated with T cell activation and inhibit the expression or activity of immunological related substances (e.g., INF-y and IL-2) associated therewith (See Figures 3-4).

Since the composition of the present invention contains the aforementioned CTLA-4-related peptide of the present invention as an active ingredient, the description common to both of them is omitted in order to avoid the excessive complexity of the present specification.

According to some embodiments of the invention, the ThI-mediated immune disorders of the invention include transplant rejection, autoimmune diseases and inflammatory diseases.

According to some embodiments of the present invention, the autoimmune disease of the present invention is selected from the group consisting of: lymphocytic leukemia But are not limited to, multiple sclerosis.

According to certain embodiments of the present invention, the inflammatory disease of the present invention may be administered to a patient suffering from a condition selected from the group consisting of asthma, eczema, psoriasis, acne, allergies, rheumatoid arthritis, psoriatic arthritis, atopic dermatitis, Inflammatory skin diseases including dermatitis, chronic sinusitis or seborrheic dermatitis; Inflammatory bowel disease including Crohn's disease or ulcerative colitis; Acute or chronic inflammatory diseases including ankylosing spondylitis, sepsis, pulmonic shock, vasculitis and bursitis; osteoporosis; Osteoarthritis; Type 1 or type 2 diabetes; Solid rock; Atherosclerosis; Viral infection by HIV (human immunodeficiency virus) or influenza virus; And chronic viral infections by Epstein-Barr virus, cytomegalovirus, herpes simplex virus, and the like.

The composition of the present invention comprises (a) a pharmaceutically effective amount of a peptide exhibiting the activity of the above-mentioned CTLA-4 protein of the present invention; And (b) a pharmaceutically acceptable carrier.

As used herein, the term "pharmaceutically effective amount" means an amount sufficient to achieve efficacy or activity of the above-mentioned CTLA-4 related peptide.

The pharmaceutically acceptable carriers to be contained in the pharmaceutical composition of the present invention are those conventionally used in the present invention and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, But are not limited to, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrups, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. It is not. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington ' s Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention can be administered orally or parenterally. In the case of parenteral administration, it can be administered by intravenous injection, subcutaneous injection, muscle injection, intraperitoneal injection, topical administration, transdermal administration and the like.

The appropriate dosage of the pharmaceutical composition of the present invention may vary depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate, . On the other hand, the preferred dosage of the pharmaceutical composition of the present invention is 0.0001-100 占 퐂 / day.

In addition, the pharmaceutical composition of the present invention may be formulated into a unit dosage form by using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by a person having ordinary skill in the art to which the present invention belongs. Or may be manufactured by penetration into a multi-dose container. The formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets, capsules or gels (e.g., hydrogels), and may additionally contain dispersing or stabilizing agents .

In addition, the composition of the present invention comprises (a) a cosmetically effective amount of the peptides of the present invention as described above; And (b) a cosmetically acceptable carrier.

As used herein, the term "a cosmetically effective amount" means an amount sufficient to achieve the cosmetic efficacy of the compositions of the present invention described above.

The cosmetic composition of the present invention may be prepared in any form conventionally produced in the art and may be in the form of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant- , Oil, powder foundation, emulsion foundation, wax foundation and spray, but is not limited thereto. More specifically, it can be manufactured in the form of a soft lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.

When the formulation of the present invention is a paste, cream or gel, an animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier component .

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, / Propane or dimethyl ether.

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

In the case where the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, or ethoxylated glycerol fatty acid esters.

The ingredients contained in the cosmetic composition of the present invention include, in addition to peptides and carrier components as an active ingredient, ingredients commonly used in cosmetic compositions and include conventional ingredients such as antioxidants, stabilizers, solubilizers, vitamins, Phosphorous assistant.

The features and advantages of the present invention are summarized as follows:

(i) The peptides of the present invention can perform the same or similar functions as those of native CTLA-4, and have excellent skin permeability according to their small size.

(ii) The peptides of the present invention inhibit the activation of T cells by effectively binding to antigen-presenting cell surface proteins (CD80 and CD86), thereby inhibiting the expression of inflammatory cytokines (such as IL-2 and IFN-y) can do.

(iii) Thus, the compositions comprising the peptides of the present invention exert very good efficacy in preventing, treating or ameliorating Th1-mediated immune disorders.

(iv) Therefore, the excellent activity and stability of the peptide of the present invention can be very usefully applied to medicines, quasi-drugs and cosmetics.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a graph showing the results of a high-performance liquid chromatography analysis of the peptides of the first and second sequences of the sequence listing prepared by the synthesis example of the present invention. FIG.
FIG. 2A is a result of MTT assay showing the effect of inhibiting the growth of mouse splenocytes by treating the peptide of the first sequence of the sequence listing prepared by the synthesis example of the present invention.
FIG. 2B is a result of MTT assay showing the effect of inhibiting the growth of mouse spleen cells by treating the peptide of the second sequence of the sequence listing prepared by the synthesis example of the present invention.
FIG. 3A is a Western blotting result showing the inhibitory effect of phosphorylation of JNK and p38 treated with the peptide of the first sequence of the sequence listing prepared by the synthesis example of the present invention.
3B is a Western blotting result showing the inhibitory effect of phosphorylation of JNK and p38 treated with the peptide of the second sequence of the sequence listing prepared by the synthesis example of the present invention.
FIG. 4A is a graph showing the effect of reverse transcription-polymerase chain reaction (RT-PCR) on the expression of IFN-.gamma. And IL-2 associated with T cell activation when the peptide of the first sequence of SEQ ID NO: reaction.
FIG. 4B shows the results of RT-PCR showing the inhibitory effect of IFN-y and IL-2 on the T cell activation when the peptide of the second sequence of the sequence listing prepared according to the present invention was treated.
5A is a graph showing the thermal stability of the peptide of the first sequence of the sequence listing prepared by the synthesis example of the present invention.
FIG. 5B shows the thermal stability of the peptide of Sequence Listing No. 2 prepared by the synthesis example of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example

Synthesis Example 1: Synthesis of Ser-Thr-Gly-Lys-Lys-Ala-Val (Sequence Listing 1)

700 mg of chlorotrityl chloride resin (CTL resin, Nova biochem Cat No. 01-64-0021) was placed in a reaction vessel, 10 ml of methylene chloride (MC) was added, and the mixture was stirred for 3 minutes. The solution was removed, and 10 ml of dimethylformamide (DMF) was added thereto. After stirring for 3 minutes, the solvent was removed again. 200 ml of Fmoc-Val-OH (Bachem, Swiss) and 400 mmole of diisopropylethylamine (DIEA) were added to the reactor, and the mixture was stirred well. . The reaction mixture was washed with methanol and DIEA (2: 1), dissolved in DCM, reacted for 10 minutes, and washed with excess DCM / DMF (1: 1). The solution was removed, 10 ml of DMF was added, stirred for 3 minutes, and then the solvent was removed. 10 ml of the deprotection solution (20% Piperidine / DMF) was added to the reaction vessel, stirred for 10 minutes at room temperature, and then the solution was removed. The same amount of the deprotection solution was added and the reaction was maintained for 10 minutes. Then, the solution was removed and the Val-CTL resin was prepared by washing with DMF twice, MC once, and DMF once for 3 minutes each. To the new reactor, 10 ml of DMF solution was added, 200 mmole of Fmoc-Ala-OH (Bachem, Swiss), 200 mmole of HoBt and 200 mmole of Bop were added and stirred well to dissolve. The reactor was charged with 400 mmole DIEA (N, N-Diisopropylethylamine) in two portions and stirred for at least 5 minutes until all solids dissolved. The dissolved amino acid mixture solution was placed in a reaction vessel containing the deprotected resin and allowed to react for 1 hour at room temperature with stirring. The reaction solution was removed, and the mixture was stirred with DMF solution three times for 5 minutes and then removed. A small amount of the reaction resin was taken and the degree of reaction was checked using a Kaiser test (Nihydrin test). The Ala-Val-CTL resin was prepared by the same deprotection reaction twice as described above. After thoroughly washing with DMF and MC and once again performing a Kaiser test, the following amino acid adhesion experiment was performed as described above. Fmoc-Lys (Boc), Fmoc-Lys (Boc), Fmoc-Gly, Fmoc-Thr (tBu) and Fmoc-Ser (tBu) were sequenced according to the selected amino acid sequence. The Fmoc-protecting group was reacted twice with the deprotecting solution for 10 minutes, followed by washing well. Acetic anhydride, DIEA, and HoBt (Hydroxybenzotriazole) were added and acetylation was carried out for 1 hour. The peptidyl resin was washed three times with DMF, MC and methanol, and then nitrogen air was slowly flowed and dried. 30 ml of an aqueous solution [TFA (Trifluoroacetic acid) 95%, distilled water 2.5%, thioanisole 2.5%] was added under reduced pressure under a vacuum of Phosphorus pentoxide and P ₂ O ₅ , and then occasionally shaken at room temperature The reaction was maintained for 2 hours. The resin was filtered and the resin was washed with a small amount of solution and then combined with the mother liquor. The precipitate was collected by distillation using 50 ml of cold ether to induce / separate precipitation, and washed with 2 more cold ether. The mother liquor was removed and sufficiently dried under nitrogen to synthesize 0.069 g of NH ₂ -Ser-Thr-Gly-Lys-Lys-Ala-Val-OH peptide 1 (yield: 89.9%). A molecular weight of 689 (theoretical value: 689.8 Da) was obtained by measurement using a molecular weight meter. Peptide 2 (NH ₂ -Arg-Thr-Glu-Met-Pro-Thr-Leu-Tyr-OH) of Sequence Listing 2 sequence was also synthesized in the same manner as described above. A molecular weight of 1010 (theoretical value: 1,010.2 Da) was obtained by using a molecular weight analyzer (yield: 92.1%).

number Amino acid sequence Analytical value (mass spectrometer) Analytical value Theoretical value First sequence NH ₂ -Ser-Thr-Gly-Lys-Lys-Ala-Val-OH 689 689.8 Second sequence NH ₂ -Arg-Thr-Glu-Met-Pro-Thr-Leu-Tyr-OH 1010 1010.2

Test Example 1: Confirmation of inhibition of cell growth effect using synthetic peptide

(Rizzino, et al. Cancer Res. 48: 1) to analyze the similar effect and inhibitory effect of the growth factors on the peptides of SEQ ID No. 1 and No. 2 synthesized in Synthesis Example 1, 4266 (1988)) using MTT (Sigma, USA) using splenocytes isolated from rat spleen.

Splenocytes isolated from the spleen were cultured in RPMI 1640 culture medium in a 100 mM cell culture dish (Corning, USA). The cultured cell lines were detached from the bottom of the culture container and centrifuged to collect only cell precipitates. The cells were suspended in RPMI 1640 culture medium without FBS, and the cells were plated on a 48-well tissue culture plate at a rate of 3 × 10 ⁷ cells per well, followed by incubation at 37 ° C. and 5% CO ₂ for 24 hours. After 24 hours, the medium was replaced with the same culture medium except for serum, and then the blank samples and the synthetic peptides for standardization were dissolved in 10% distilled water in sterilized condition and then the concentrations of 1 μl / ml, 10 μl / ml and 50 μl / ml For 48 hours under the same conditions as above. After the culture was completed, MTT assay solution was added to the culture supernatant and cultured for 3 hours at 37 ° C and 5% CO ₂ . After 3 hours, the culture solution was completely removed, and the cells were dissolved in 500 μl of DMSO. The absorbance of the cells was measured at 540 nm in ultraviolet light, and the viability of the cells was measured. As shown in Figs. 2A and 2B, the peptides of the present invention effectively inhibited the growth of spleen cells.

Test Example 2: Confirmation of the signal promoting effect of the synthetic peptides CTL4 (SEQ ID No. 1) and CTL8 (SEQ ID No. 2)

Peptides synthesized in Synthesis Example 1 were treated with splenocytes isolated in rat spleen and treated with Con A (concanavalin A, positive control; Sigma, USA) for 1 minute after 1 hour to induce activity on T cells . The cells were obtained and the degree of phosphorylation of JNK and p38, MAPKs involved in CTLA-4 protein, was confirmed by Western blotting using phospho-JNK and phospho-p38 antibody (Cell Signaling, USA) . When pre-treated with the peptides of the present invention, JNK showed a phosphorylation inhibition effect in a peptide treatment concentration-dependent manner (FIGS. 3A and 3B). Furthermore, IFN (interferon) γ and IL-2 were observed by RT-PCR using specific primers. The expression of IFN-y and IL-2, which are expressed by activation of T cells, was inhibited by the peptides of the present invention (FIGS. 4A and 4B).

The results of Test Examples 1 and 2 show that the peptide of the present invention inhibits the phosphorylation of JNK and p38 and induces the decrease of IFN-γ and IL-2 expression by inhibition of T cell activation, I could confirm.

Test Example 3: Thermal stability of prepared peptides

Peptide of Sequence Listing 1st sequence and Sequence Listing 2nd sequence prepared through Synthesis Example 1 and standard growth factor (CTLA-4) purchased from NIBSC (UK) were prepared with a phosphate buffer solution at a concentration of 0.1 mg / ml . The prepared solution was put into a glass vial at 1 ml each and allowed to stand at 37 캜. The solution at 37 ° C was sampled at 0, 5, 10, 20, 30, 50, and 70 days, and the denatured peptides or proteins were removed by centrifugation for each day, and the supernatant was taken and quantified by HPLC 5a and 5b).

Example 1: Preparation of Nano Peptides

50 mg of each of the two peptides obtained in the above Synthesis Example were precisely weighed accurately and dissolved with 500 ml of distilled water. The mixture solution was mixed with 5 g of lecithin, 0.3 ml of sodium oleate, 50 ml of ethanol, and a small amount of oil, and the mixture was adjusted to a total volume of 1 L with distilled water. To prepare a nanosome having a size of about 100 nm. The final concentration of the prepared nanosomes was about 50 ppm, either singly or in combination, for cosmetic use.

Formulation Example 1: Softening Age

The softening water containing the peptide nanosome prepared in Example 1 and having the following composition was prepared according to the general method of producing lotion.

ingredient Content (% by weight) Peptide nanosome 0.001 1,3-butylene glycol 6.0 glycerin 4.0 PEG 1500 1.0 Sodium hyaluronate 1.0 Polysorbate 20 0.5 ethanol 8.0 Preservative, pigment Suitable amount Benzophenone-9 0.05 incense a very small amount Purified water Balance Sum 100

Formulation Example 2: Moisturizing cream

A nutritional cream containing the peptide nanosome prepared in Example 1 and having the following composition was prepared according to a general method for producing a moisturizing cream.

ingredient Content (% by weight) Peptide nanosome 0.001 Meadow Foam Oil 3.0 Cetearyl alcohol 1.5 Stearic acid 1.5 Glyceryl stearate 1.5 Liquid paraffin 10.0 Wax 2.0 Polysorbate 60 0.6 Sorbitan sesquioleate 2.5 Squalane 3.0 1,3-butylene glycol 3.0 glycerin 5.0 Triethanolamine 0.5 Tocopheryl acetate 0.5 Preservative, pigment Suitable amount incense Suitable amount Purified water Balance Sum 100

Formulation Example 3: Moisturizing lotion

A nutritional lotion comprising the peptide nanosome prepared in Example 1 and having the following composition was prepared according to the general method of producing lotion.

ingredient Content (% by weight) Peptide nanosome 0.002 1,3-butylene glycol 4.0 glycerin 4.0 Cetearyl alcohol 0.8 Glyceryl stearate 1.0 Triethanolamine 0.13 Tocopheryl acetate 0.3 Liquid paraffin 5.0 Squalane 3.0 Macadamia nut oil 2.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.5 Carboxyvinyl polymer 1.0 Preservative, pigment Suitable amount incense Suitable amount Purified water Balance Sum 100

Formulation Example 4: Essence

An essence comprising at least one kind of nano-somes of the four kinds of peptide nano-particles prepared in Example 1 and having the following composition was prepared according to a general essence production method.

ingredient Content (% by weight) Peptide nanosome 0.005 glycerin 10.0 1,3-butylene glycol 5.0 PEG 1500 2.0 Allantoin 0.1 DL-Panthenol 0.3 EDTA-2Na 0.02 Hydroxyethylcellulose 0.1 Sodium hyaluronate 8.0 Carboxyvinyl polymer 0.2 Triethanolamine 0.18 Octyldodec-16 0.4 ethanol 6.0 Fragrance, preservative, pigment Suitable amount Purified water Balance Sum 100

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is obvious that the same is by way of illustration and example only and is not to be construed as limiting the scope of the invention. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

<110> Caregen Co. Ltd. <120> Novel Peptides for Inhibiting an Activity of T lymphocytes and          Uses Thereof <130> PN130280 <160> 2 <170> Kopatentin 2.0 <210> 1 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> CTL4 <400> 1 Ser Thr Gly Lys Lys Ala Val   1 5 <210> 2 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> CTL8 <400> 2 Arg Thr Glu Met Pro Thr Leu Tyr   1 5

Claims (11)

A peptide for inhibiting T-cell activity comprising an amino acid sequence selected from the group consisting of the amino acid sequences set forth in SEQ ID NO: 1 and SEQ ID NO: 2.
The peptide according to claim 1, wherein the peptide is derived from human CTLA-4 (Cytotoxic T lymphocyte-associated antigen 4; CD152) protein.
2. The peptide of claim 1, wherein said peptide inhibits phosphorylation of JNK (c-Jun N-terminal kinase) and p38.
2. The peptide of claim 1, wherein the peptide inhibits the expression of an inflammatory cytokine.
5. The peptide of claim 4, wherein said inflammatory cytokine inhibits the expression of interferon-y (IFN-y) or interleukin-2.
delete A pharmaceutical composition for preventing or treating a Th1-mediated immune disease using the peptide of any one of claims 1 to 5 as an active ingredient.
8. The pharmaceutical composition according to claim 7, wherein the Th1-mediated immune disease is transplant rejection, autoimmune disease or inflammatory disease.
9. The method of claim 8, wherein the autoimmune disease is characterized by being a disease selected from the group consisting of Lupus, rheumatoid arthritis, multiple sclerosis, Wegener's granulomatosis, temporal arteritis, cryoglobulinemia, pemphigus vulgaris, thyroiditis or multiple sclerosis &Lt; / RTI &gt;
9. The method of claim 8, wherein the inflammatory disease is selected from the group consisting of asthma, eczema, psoriasis, acne, allergies, rheumatoid arthritis, psoriatic arthritis, atopic dermatitis, atopic dermatitis, allergic dermatitis, chronic sinusitis or seborrheic dermatitis Inflammatory skin diseases comprising; Inflammatory bowel disease including Crohn's disease or ulcerative colitis; Acute or chronic inflammatory diseases including ankylosing spondylitis, sepsis, pulmonic shock, vasculitis and bursitis; osteoporosis; Osteoarthritis; Type 1 or type 2 diabetes; Solid rock; Atherosclerosis; Viral infection by HIV (human immunodeficiency virus) or influenza virus; Or Epstein-Barr virus, a cytomegalovirus, or a herpes simplex virus. A polynucleotide selected from the group consisting of an amino acid sequence represented by SEQ ID No. 1 or 2 and an acetyl group, a fluorenylmethoxycarbonyl group, a formyl group, a palmitoyl group, a myristyl group, a stearyl group and a polyethylene glycol (PEG) A peptide for inhibiting T-cell activation comprising a protective group at N- or C-terminal.

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