JPH09194367A - Tocopherol-containing synthesis suppressant for protein belonging to hsp60 family - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a tocopherol-containing synthesis suppressant, comprising tocopherol or its derivative as an active ingredient, capable of specifically suppressing the synthesis of proteins belonging to the heat shock protein (HSP)60 family and useful for preventing and treating autoimmune diseases, e.g. insulin- dependent diabetes mellitus (IDDM). SOLUTION: This tocopherol-containing synthesis suppressant for a heat shock protein having 57-68 Dal molecular weight comprises tocopherol, preferably α-tocopherol [3,4-dihydro-2,5,7,8-tetramethyl-2-(4,8,12-trimethyltridecyl)-2H-1- benzopyran-6-ol], i.e., vitamin E or a derivative thereof as an active ingredient. For example, the suppressant comprises the active ingredient in an amount of 0.1-80wt.% and the daily dose thereof for an adult is about 1mg to 10g divided in about one to four portions.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a heat shock protein having a molecular weight of 57 kilodalton (kD) to 68 kD, which contains tocopherol or a derivative thereof, particularly α-tocopherol (that is, vitamin E) as an active ingredient. Group (hereinafter referred to as HSP60 family)
The present invention relates to a protein synthesis inhibitor belonging to. The synthesis inhibitor of the protein belonging to the HSP60 family according to the present invention is particularly considered to be an autoimmune disease in which the protein belonging to the HSP60 family is involved in the onset by suppressing the intra-tissue synthesis of the protein belonging to the HSP60 family. For example, it is possible to effectively improve the physiological condition of a patient suffering from a disease such as type I diabetes and rheumatoid arthritis, and to effectively treat an autoimmune disease such as type I diabetes and rheumatoid arthritis.

[0002]

2. Description of the Related Art In recent years, autoimmune diseases have become a major problem. An autoimmune disease is a disease in which the immune system, which normally should not attack the components that make up your body, reacts with your own tissue and destroys it.
Includes type I diabetes and rheumatoid arthritis. For example, in recent years in Japan, with the development of economy, society, and culture, as well as the improvement of living standards and changes in lifestyles, the number of diabetic patients has increased significantly, and the condition has become more severe and complicated. Advances in diabetology have improved patient prognosis, but have increased frequent retinopathies, nephropathy, and neuropathy, as well as accelerated arterial stiffness, which has severely impaired health and social activities . Among diabetes, type I diabetes (insulin-dependent diabetes mellitus; IDD)
The incidence of M) has increased several fold in many countries in the last few decades, with 1% of currently living humans becoming I by the age of 70.
It is expected that type 2 diabetes will occur.

[0003] Type I diabetes is a disease in which only β cells of the pancreatic islets of Langerhans, which are insulin-producing cells, are autoimmunely destroyed, resulting in an insulin deficiency state, and is an organ-specific autoimmune disease (Atkinson, MA et al .:
"Sci. Am.", 263 : 42-49, 1990; Todd JA .: "Immunol.
Today ", 11 : 122-129, 1990). The autoimmune process that causes type I diabetes is very strictly limited to the pancreas,
Often develops before becoming an adult. When type I diabetes clinically develops, there is inflammation of the islets (isletitis), and most of the β cells that produce insulin are specifically lost (Atkinson, MA, et al .: "Sci. A
m. ", 263 : 62-67, 1990). The clinical manifestation of diabetes only appears after the majority of β-cells (probably 90% or more) have been destroyed to the extent that they cannot regenerate, and the survival of the patient is out of insulin. The autoimmune response is already irreversibly damaged by the time it can be detected by clinical diagnosis, and many do not show significant subjective symptoms, etc. Type I diabetes has many problems.

[0004] Rheumatoid arthritis is a chronic inflammatory disease in which the synovium of the joint is the main lesion. The site of the lesion is sometimes not limited to the synovium of the joint, and it is not uncommon for it to extend to the whole body. The inflammation that initially occurs in the synovium of the joint eventually causes synovial proliferation, further destruction of cartilage and bone, and destruction of joint tissue. As a result, the patient is not only severely restricted socially and at home, but also has a considerable financial burden. The number of patients with rheumatoid arthritis accounts for 0.1-0.3% of the population. This is an example of confirming rheumatoid arthritis, and it is expected that the number of patients will increase to about 10 times that of rheumatoid arthritis, including suspected cases and rheumatoid arthritis-related diseases.

On the other hand, heat shock proteins (heat shock proteins)
protein; also called HSP, stress protein)
Cells with some stress, such as heat, heavy metals, drugs,
A group of proteins expressed in cells by stimulation with amino acid analogs or hypoxia (low oxygen concentration). Heat shock proteins are ubiquitous in nature and are produced by higher animals, including bacteria, yeast, plants, insects, and humans.

[0006] HSPs are of various types,
Due to the molecular weight, the HSP90 family (for example, 9
0 kD or 110 kD HSP, etc.), HSP70 family (eg, 70-73 kD HSP, etc.), HS
P60 family (eg, 57-68 kD HSP etc.), small molecule HSP family (eg, 20 kD, 2
5-28 kD or 47 kD HSP). In the present specification, an HSP having a specific molecular weight is indicated by an HSP and a numeral described immediately after the HSP. For example, an HSP having a molecular weight of 60 kD is referred to as “HSP60”. As described above, there are many types of HSPs, which differ not only in molecular weight but also in structure, function, or property. In addition to the response to stress, some of these proteins are constitutively synthesized and, under normal circumstances, regulate protein folding, unfolding, protein subunit assembly, and protein membrane trafficking. It has been shown to play an essential physiological role. These functions as heat shock proteins are called molecular chaperones.

[0007] One of the attentions regarding the etiology of autoimmune diseases is molecular homology. In other words, when a self-antigen has a common antigenicity with a foreign antigen such as a microorganism, antibodies and sensitized lymphocytes produced by microbial infection attack their own tissues by cross-reaction, resulting in the development of an autoimmune disease. (Atkinson, MA et al .: "Sci. A
m. ", 263 : 42-49, 1990; Shinha, AA et al .:" Scie
nce ", 248 : 1380-1388, 1990). For example, bacterial HSP
Proteins belonging to 60 families include tuberculosis, leprosy,
It is a major antigen such as syphilis, veterans' disease, or Lyme disease (Young, RA et al .: "Cell", 59 : 5-8, 1989),
In addition, proteins belonging to the HSP60 family of bacteria have strong immunogenicity and have molecular homology with their own (host human) proteins, so that they are not affected by infectious disease-triggered molecular homology. It is believed that autoimmune diseases develop.

For example, an antibody (64 kD) that reacts with a 64 kD protein detected in the blood of diabetic patients and their families
D autoantibodies) are β-cell specific, and often appear immediately before diabetes is diagnosed. That is, pancreatic islet cell antigen, which is considered to be the cause of onset in type I diabetes, is a glycoprotein having a molecular weight of 64 kD (Baekkeskov, S. e.
t al .: "Nature", 298 : 167-169, 1982). 64
Antibodies against the kD protein are not limited to human diabetes (Atkinson, MA et al .: "Lancet", 335 : 1357-136).
0, 1990), BB rats (Baekkeskov, S. et al .: "Scie
nce ", 224 : 1348-1350, 1984) and NOD mice (Atkins
on, MA et al .: "Diabetes", 37 : 1587-1590, 1988)
As described above, it is also detected in a type I diabetes model animal that naturally develops type I diabetes and exhibits many characteristics of human type I diabetes. The 64 kD protein of pancreatic β-cell in type I diabetes may be a heat shock protein because it is induced by cytokines and heat stimulation.

The 64 kD protein of the pancreatic Langerhans islet β-cell in NOD mouse, a model animal of type I diabetes, is derived from H. tuberculosis (Mycobacterium tuberculosis).
It has been shown to be an autoantigen that is immunologically cross-reactive with antibodies to proteins belonging to the SP60 family. As described above, the immunological crossover between the protein belonging to the HSP60 family and the 64 kD protein autoantigen was observed.
The D protein may be a member of the HSP60 family, and it has been suggested that an autoimmune mechanism that crosses an epitope of a protein belonging to the HSP60 family is involved in the development of type I diabetes. When a T lymphocyte clone having specificity for a protein belonging to the HSP60 family of Mycobacterium tuberculosis is transferred, it causes Langerhans insulitis and hyperglycemia in young NOD mice. In addition, when a protein belonging to the HSP60 family of Mycobacterium tuberculosis is injected into NOD mice by an immunogenic administration method, that is, by injection into a NOD mouse together with an adjuvant, diabetes can be developed early (Elias, D. et al .: "Proc. Natl. Acad. . Sci.
USA ", 87 : 1576-1580, 1990). HS of mycobacteria
These facts that the immune response of animals to proteins belonging to the P60 family cause type I diabetes are due to the attack by the immune system on antigens that cross-react with antibodies to proteins belonging to the HSP60 family of mycobacteria, damaging β cells. It is shown that.

It is known that proteins belonging to the HSP60 family are related to rat adjuvant arthritis, which is an animal model for rheumatoid arthritis, and to human rheumatoid arthritis. For example, in the case of rheumatoid arthritis, among the heat shock proteins that are bacterial cell proteins, proteins belonging to the HSP60 family have been found to have molecular homology to proteoglycans present in articular cartilage. The involvement of protein-reactive T lymphocytes belonging to the HSP60 family has been shown in adjuvant arthritis in rats ("C
urr. Top. Microbiol. Immunol. ", 145 : 27-83, 198.
9). The disease is transferred to irradiated immunologically naive rats by transferring a clone of T lymphocytes reactive to proteins belonging to the HSP60 family of M. tuberculosis. ("Science", 219 : 56-5
8, 1983; "Nature", 331 : 171-173, 1988). The T lymphocytes also show cross-reactivity with joint proteoglycans ("Proc. Natl. Acad. Sci. USA", 82 : 5117-512).
0, 1985). Regulatory T lymphocytes induced by proteins belonging to the HSP60 family are also recognized in arthritis due to streptococcus and pristine. Thus, adjuvant arthritis appears to be an autoimmune disease caused by protein T lymphocytes belonging to the anti-HSP60 family. Also, in human juvenile rheumatoid arthritis, HS
Involvement of protein-reactive T lymphocytes belonging to the P60 family is considered.

In addition, T lymphocytes that specifically react with proteins belonging to the HSP60 family derived from mycobacteria have been extracted from the synovial fluid of patients with rheumatoid arthritis ("Lancet", II : 478-). 480, 1988; "Na
ture ", 339 : 226, 1989;" Annu. Rev. Immunol. ", 1
1 : 637, 1993). Thus, the mycobacterial HS
Proteins that show cross-reactivity with proteins belonging to the P60 family are found at high concentrations in the cartilage / pannus junction of rheumatoid arthritis, whereas they are found in normal tissues and tissues that exhibit chronic inflammation due to other diseases. No ("Scand. J. Immunol.", 31 : 283-288, 1990). Furthermore, antibodies against proteins belonging to the HSP60 family are detected in human and rat rheumatoid arthritis (Kaufmann, SHE, et al .: "Immunol. Today", 11 :
129-136, 1990), it is possible that the etiology of rheumatoid arthritis is autoimmunity against self antigens similar in structure to proteins belonging to the HSP60 family of mycobacteria. Thus, the presence of an immune response to proteins belonging to the HSP60 family is associated with both rat and human arthritis.

[0012]

SUMMARY OF THE INVENTION In view of the above circumstances, the present inventors have been able to effectively improve the physiological condition of patients with autoimmune diseases such as type I diabetes and rheumatoid arthritis, In order to develop a method that can effectively treat immune diseases, various studies have been made on compounds that exhibit a synthesis inhibitory effect on proteins belonging to the HSP60 family. As a result, the inventors
Surprisingly, it was found that tocopherol or a derivative thereof, particularly α-tocopherol (that is, vitamin E) specifically inhibits the synthesis of proteins belonging to the HSP60 family in cells of tissues showing pathological conditions. That is, it was found that the administration of tocopherol or a derivative thereof suppresses the intracellular synthesis of proteins belonging to the HSP60 family, and thus can treat autoimmune diseases such as type I diabetes and rheumatoid arthritis. It was. The present invention is based on such findings, and has as its object to provide a synthesis inhibitor of a protein belonging to the HSP60 family, which can effectively treat autoimmune diseases such as type I diabetes and rheumatoid arthritis. .

[0013]

Therefore, the present invention is characterized by containing tocopherol or a derivative thereof, particularly α-tocopherol (that is, vitamin E) as an active ingredient, and having a molecular weight of 57 kilodaltons to 68 kilodaltons. Between heat shock proteins (ie,
HSP60 family protein). In the present specification, the “HSP60 family” has a molecular weight of 57 kD to 68 kD as described above.
Mean heat shock proteins. In addition, HSP6
Examples of proteins belonging to family 0 include H
SP60 (ie, a heat shock protein with a molecular weight of 60 kD), HSP58 (ie, a heat shock protein with a molecular weight of 58 kD), HSP65 (ie, a heat shock protein with a molecular weight of 65 kD), or GroEL
(Ie, a prokaryote, for example, a heat shock protein having a molecular weight of about 64 kD such as Escherichia coli).

[0014]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention will be described in detail below. The synthesis inhibitor of the present invention contains tocopherol or its derivative as an active ingredient. In the present specification, tocopherol includes, for example, α-tocopherol and β-
It means tocopherol, γ-tocopherol, δ-tocopherol, or the like, and in the present invention, tocopherol homologues thereof can be used alone or in a mixture thereof. As tocopherol, especially α-
Tocopherol [3,4-dihydro-2,5,7,8-
Most preferred is tetramethyl-2- (4,8,12-trimethyltridecyl) -2H-1-benzopyran-6-ol], that is, vitamin E.

As the tocopherol derivative, for example,
Tocopherol derivatives originally contained in natural products,
Examples thereof include a tocopherol derivative converted by a chemical treatment during extraction or fractionation, and a chemically modified tocopherol derivative. Examples of the tocopherol derivative originally contained in the natural product include a tocotrienol derivative, that is, α-tocotrienol, β-tocotrienol, γ-tocotrienol, δ-tocotrienol, and the like. Examples of the tocopherol derivative converted by a chemical treatment during extraction or fractionation or the chemically modified tocopherol derivative include a tocopherol ester derivative or a tocotrienol ester derivative.

Specific examples of the tocopherol ester derivative include, for example, a linear or branched alkyl monocarboxylic acid or dicarboxylic acid (preferably having 1 carbon atom).
To straight chain or branched alkyl monocarboxylic acid or dicarboxylic acid), straight chain or branched alkenyl monocarboxylic acid or dicarboxylic acid (preferably straight chain or branched chain having 2 to 30 carbon atoms) Alkenyl monocarboxylic acid or dicarboxylic acid), aryl monocarboxylic acid or dicarboxylic acid having 6 to 30 carbon atoms in the aryl moiety (for example, benzoic acid), or nitrogen atom 1
Alternatively, a 5- or 6-membered heterocyclic monocarboxylic acid or dicarboxylic acid containing two (eg, nicotinic acid)
And an ester compound of tocopherol, such as tocopherol acetate, tocopherol succinate, or tocopherol nicotinate. The tocopherol or its derivative has stereoisomers,
Any of their pure stereoisomers or mixtures thereof can be used in the present invention. The tocopherol or its derivative contained in the synthetic inhibitor of the present invention can be prepared by chemical synthesis or by extraction from a natural product and purification. Alternatively, a commercially available product may be used.

The synthetic inhibitor of the present invention comprises an animal, preferably a mammal (particularly, a tocopherol or a derivative thereof, alone or preferably together with a conventional carrier which is pharmaceutically or veterinarily acceptable. Can be administered to humans). The dosage form is not particularly limited and includes, for example, powders, fine granules, granules, tablets, capsules, suspensions, emulsions, syrups, extracts,
Or, oral preparations such as pills, parenteral preparations such as injections, solutions for external use, ointments, suppositories, creams for topical administration, and eye drops can be mentioned. These oral preparations include, for example, gelatin, sodium alginate, starch, corn starch, sucrose, lactose, glucose, mannitol, carboxymethylcellulose, dextrin, polyvinylpyrrolidone, crystalline cellulose, soy lecithin, sucrose, fatty acid esters, talc, stearic acid. Excipients such as magnesium, polyethylene glycol, magnesium silicate, anhydrous silicic acid, or synthetic aluminum silicate, binders, disintegrants, surfactants, lubricants, glidants, diluents, preservatives, coloring It can be produced according to a conventional method using an agent, a flavor, a flavoring agent, a stabilizer, a humectant, a preservative, an antioxidant, or the like.

For example, in the case of producing soft capsules, vitamin E is dissolved in vegetable oil or synthetic oil while heating, if necessary, and gelatin produced by a conventional method,
A soft capsule preparation can be produced by a soft capsule filling machine using a coating composition obtained by mixing and melting glycerin and / or a mildewproofing agent. In the case of producing a hard capsule, for example, vitamin E is heated and adsorbed on an excipient such as soft anhydrous silicic acid, and if necessary, crystalline cellulose and / or corn starch or the like is added, mixture·
After granulation, the sieved product can be produced by filling hard capsules using a capsule filling machine. In addition, when producing an oral liquid preparation, for example,
It can be produced by emulsifying or solubilizing by a conventional method using a synthetic surfactant such as polyethylene hardened castor oil or a natural surfactant such as lecithin.

Examples of the parenteral administration method include injection (subcutaneous, intravenous, etc.) and rectal administration. Of these, injections are most preferably used. For example, in the preparation of injectables, in addition to tocopherol or its derivative as an active ingredient, for example, physiological saline, water-soluble solvent such as Ringer's solution, vegetable oil, or water-insoluble solvent such as fatty acid ester, glucose, or sodium chloride. Any isotonic agent, solubilizing agent, stabilizer, preservative, suspending agent, emulsifying agent or the like can be used. Further, the synthetic inhibitor of the present invention may be administered using a sustained-release preparation technique using a sustained-release polymer or the like. For example, the synthetic inhibitors of the present invention can be incorporated into a pellet of ethylene vinyl acetate polymer and the pellet can be surgically implanted into the tissue to be treated. Although the synthesis inhibitor of the present invention is not limited to this, 0.01 to 99% by weight of tocopherol or its derivative, preferably 0.1 to 8%
It can be contained in an amount of 0% by weight.

The dose of the synthetic inhibitor of the present invention varies depending on the type of disease, age, sex, weight of patient, degree of symptom, administration method, etc., and is not particularly limited, but the amount of α-tocopherol As usual 1m per adult
About g to 10 g are orally or parenterally administered in about 1 to 4 times a day. Furthermore, the use is not limited to pharmaceuticals, and various uses, for example, functional foods and health foods can be given in the form of food and drink.

[0021]

As described above, tocopherol or its derivative contained in the synthetic inhibitor of the present invention, especially α-tocopherol (that is, vitamin E) is used in intracellular HSP60.
Since it has an action of specifically suppressing the synthesis of proteins belonging to the family, administration of the tocopherol or its derivative specifically reduces the biosynthesis of proteins belonging to the HSP60 family in cells. Therefore, the tocopherol or its derivative can be used for prevention and treatment of autoimmune diseases associated with the onset of proteins belonging to the HSP60 family, such as type I diabetes and rheumatoid arthritis.

[0022]

EXAMPLES The present invention will be described below in more detail with reference to examples, but these examples do not limit the scope of the present invention. Example 1: Measurement of HSP expression level in cultured human cancer cells (1) Culture of cultured human cancer cells Breast cancer cell line MCF7 (ATCC HTB 22)
The cells were cultured in RPMI1640 medium containing 0% inactivated fetal bovine serum and 10 ⁻⁸ Mβ-estradiol at 37 ° C. under 5% carbon dioxide conditions except for heat shock treatment.

(2) α-Tocophenol Treatment and Heat Shock Treatment D-α-tocopherol (Funakoshi, Catalog No. ICN 81-1521) was added to the medium of the breast cancer cell line MCF7 2 days after seeding so that the final concentration was 20 μM. -47) is added, and 24
Cultured for hours. Thereafter, the cells were subjected to a heat shock treatment at 45 ° C. for 15 minutes, and then cultured at 37 ° C. overnight. The control test was carried out in the same manner as above except that α-tocopherol was not added.

(3) Measurement of HSP expression level in cultured human cancer cells Each cell treated in (2) above was homogenized by the following method, and the HSP expression level was measured by Western blotting. That is, the cells treated in the above (2) were treated with phosphate buffered saline [composition: KCl = 0.2 g /
1, KH ₂ PO ₄ = 0.2 g / l, NaCl = 8 g /
1, Na ₂ HPO ₄ (anhydrous) = 1.15 g / l;
PBS (-)), and then lysed buffer (lysis buffer) [1.0% NP-4
0, 0.15 M sodium chloride, 50 mM Tris-HC
1 (pH 8.0), 1 mM of 5 mM-EDTA, 2 mM-N-ethylmaleimide, 2 mM phenylmethylsulfonyl fluoride, 2 μg / ml leupeptin and 2 μg / ml pepstatin], and allowed to stand on ice for 20 minutes.
Thereafter, centrifugation was performed at 12,000 rpm at 4 ° C. for 20 minutes. 10 μl of the supernatant after centrifugation was added to PBS (−) 790
μl, add protein assay stain (Dye Reag
ent Concentrate: Bio-Rad, Catalog No. 500-00
06) 200 μl was added. After standing at room temperature for 5 minutes, the protein was quantified by measuring the absorbance at 595 nm.

Using the sample for which protein was quantified, L
aemmli buffer system (Laemmli, NK, "Natur
e ", 283 : pp. 249-256, 1970), lysates containing equal amounts of protein were subjected to SDS polyacrylamide gel electrophoresis. After electrophoresis, blotting and subsequent blocking were performed. Using a transfer device (Trans-Blot Electrophoretic Transfer Cell: Bio-Rad, Catalog No. 170-3946), adhere the gel to a 0.45 μm nitrocellulose membrane (Schleicher & Schuell, Catalog No. 401196) at room temperature and 100 V at room temperature. A tris glycine buffer (Tris Gly Running and Bloom) containing 0.025 M Tris and 0.192 M glycine and adjusted to pH 8.5 was used as a blotting buffer.
tting Buffer; Enprotech, Massachusetts, USA
A buffer prepared by adding methyl alcohol to catalog number SA100034) to 20% was used. After blotting, the nitrocellulose membrane was added to a 10% skim milk (Snow Brand Milk Products) -PBS (-) solution at room temperature for 30 minutes.
Incubate to block non-specific binding.

After blocking, anti-human HSP60 mouse monoclonal antibody (Stre
ssGen, Victoria, BC, Canada, Cat.No. SPA-8
06), a primary antibody reaction was performed. This anti-human HSP
60 murine monoclonal antibody is an antibody to human HSP60 prepared was prepared as an immunogen by recombinant DNA methods using E. coli ( "J. Exp. Med." 175
, 1805-1810, 1992), mammalian HSP60 (primate HS)
P60, mouse HSP60, rat HSP60, and hamster HSP60) ("J. Exp. M
ed. " 175 , 1805-1810, 1992). This anti-human HSP60
The epitope recognized by the mouse monoclonal antibody is located in the amino acid sequence consisting of amino acid residues 383 to 447 of the human HSP60 amino acid sequence ("J. Exp. Med." 175 , 1805-1810, 1992). ). After the primary antibody reaction, the solution was exchanged with PBS (-) for 5 minutes each, and the washing was performed twice with a slow rocking shaker, and PBS (-)-0.1% Tween 20 (Bio-Rad, Catalog No.). 170-6531) The solution was replaced for 15 minutes, and the solution was washed four times. Finally, the cells were washed twice with PBS (-) for 5 minutes each.

After washing, the peroxidase-labeled goat anti-mouse IgG antibody (CAPPEL, Catalog No. 55550) was added to the 2
% For 2 hours using 5 ml of an antibody solution prepared by diluting 5000-fold with a PBS (-) solution containing 5% skim milk.
The next antibody reaction was performed. After the completion of the reaction, the nitrocellulose membrane was washed twice with a PBS (-) solution for 5 minutes while changing the solution twice, and further with a PBS (-)-0.1% Tween20 solution for 15 minutes for 5 times. Performed by slow rocking shaker. Finally PBS (-)
The solution was washed twice for 5 minutes each. Extra PBS
(-) After removing the solution, the western blotting detection reagent (ACL Western blotting detection reagent; Ame
rsham, catalog number RPN2106), sprinkle onto the nitrocellulose membrane, incubate for 1 minute, remove excess detection reagent, wrap the nitrocellulose membrane in wrap, and cover the reaction surface with X-ray film (Kodak X-OMAT, AR, (Catalog No. 165 1454), exposed, developed and HSP6
The existence of 0 was examined.

In a control test, ie breast cancer cell line MCF7 without the addition of α-tocopherol, a molecular weight of about 6 was obtained.
One band of 0 kD was detected. The molecular weight was determined by binding to the anti-human HSP60 mouse monoclonal antibody and molecular weight markers (egg white ovalbumin and bovine serum albumin). The expression of HSP60 was suppressed in the breast cancer cell line MCF7 to which α-tocopherol was added. That is, α-tocopherol is
It can be concluded that it has the activity of a synthetic inhibitor that suppresses the expression of 60.

[0029]

INDUSTRIAL APPLICABILITY As described above in detail, tocopherol or its derivative, particularly α-tocopherol (that is, vitamin E) has the activity of a synthetic inhibitor that suppresses the expression of intracellular proteins belonging to the HSP60 family. Therefore, tocopherol or its derivatives, especially α-
By administering tocopherol (that is, vitamin E), for example, the physiological condition of patients with autoimmune diseases (for example, type I diabetes, rheumatoid arthritis, etc.) in which the proteins belonging to the HSP60 family are involved in the onset is effectively improved. Therefore, the illness can be effectively treated.

Claims (2)

[Claims] 1. A heat shock protein synthesis inhibitor having a molecular weight of 57 kilodaltons to 68 kilodaltons, which contains tocopherol or a derivative thereof as an active ingredient. 2. The molecular weight of 57 kilodaltons to 6 according to claim 1, wherein the tocopherol is α-tocopherol.
Inhibitor of heat shock protein synthesis up to 8 kilodaltons.