JPH1045603A - TNF-α production inhibitor containing sulfatide as active ingredient - Google Patents

Abstract

(57) [Summary] TNF-α production inhibitor, TNF-α-mediated disease preventive / therapeutic agent, that is, chronic inflammatory disease, acute inflammatory disease, inflammatory disease due to infection, autoimmune disease, Provided is a prophylactic / therapeutic agent for allergic diseases and other TNF-α-mediated diseases. SOLUTION: TNF- containing sulfatide as an active ingredient
α-production inhibitor, TNF-α-mediated disease preventive / therapeutic agent, ie, chronic inflammatory disease, acute inflammatory disease, inflammatory disease due to infection, autoimmune disease, allergic disease, other TNF-α-mediated Prophylactic / therapeutic agent for diseases and the like.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a TNF-α production inhibitor or a TNF-α-mediated disease preventive / therapeutic agent comprising sulfatide as an active ingredient, ie, chronic inflammatory disease, acute inflammatory disease, inflammation due to infection. It is useful as a prophylactic / therapeutic agent for sexual diseases, autoimmune diseases, allergic diseases, and other TNF-α-mediated diseases.

[0002]

2. Description of the Related Art Sulfatide has been historically isolated as a sulfur-containing lipid present in the brain and is a compound having the following structural formula.

[0003]

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[0004] Sulfatide is relatively abundant in brain white matter, is important as a lipid constituting myelin, and is very specifically bound to a basic protein of myelin. It is also thought that it is specifically present in the kidney and is involved in electrolyte ion transport. Recently, sulfatide (I ³ SO ₃ -GalCer) was found to bind to laminin, an intercellular adhesion protein, with a higher affinity than ganglioside (Roberts
DD et al., Proc. Biol. Chem. USA, 82, 1306-1310, 1
985), thrombospondin, an extracellular matrix protein secreted by many cells other than platelets (Roberts
DD et al., J. Biol. Chem., 260, 9405-9411, 1985).
And blood coagulation factor Von Willbrand factor (Roberts
DD et al., J. Biol. Chem., 261, 3306-3309, 1986)
, And these proteins have a higher binding specificity for sulfatide than laminin. Sulfatide also activates the third component of complement, properdin (Hol
t GD et al., J. Biol. Chem., 265, 2852-2855, 199
Antistatin (Holt G. 0), which is contained in certain leech (Mexican leech) and is thought to suppress blood coagulation and cancer transfer.
D. et al., J. Biol. Chem., 264, 12138-12140, 1989)
, A proteinase present in erythrocytes (multicatalytic p
rotenase) (Ohkubo I. et al., Biochem. Biophys. Res.
n., 174, 1133-1140, 1991), malaria circumsporoz
oite protein (Cerami C. et al., Mol. Biochem. Parasito
l., 54, 1-12, 1992) and humman immunodeficiency
virus type I gp120 (Bhat S. et al., AIDS Res.Hum.Retr
oviruses. 9, 175-181, 1993). A common amino acid sequence exists in a plurality of proteins to which these sulfatides bind, and the sequence is estimated to be CSVTCGXGXXXRX (Holt
(GD et al., J. Biol. Chem., 265, 2852-2855, 1990).
.

Recently, it has been reported that sulfatide binds to adhesion molecules L-selectin and P-selectin (Suzuki Y. et al., Biochem. Biophys. Res. Commun., 19
0, 426-434, 1993, Aruffo A. et al., Cell, 67, 35-4.
4, 1991, Imai Y. et al., J. Cell Biol., 111, 1225-1.
232, 1990), suggesting that it is highly expressed in myeloid cells (Aruffo A. et al., Cell, 67, 35-44, 1991).
In addition, sulfatide suppresses liver damage induced by carbon tetrachloride (Kajihara K. et al., Biosci. Biotechenol. Bio.
Chem., 59, 155-157, 1995), cobra venom and IgG.
It has been reported to suppress lung damage caused by immune complexes (Mulligan MS et al., Int. Immunol., 7, 1107-1113.
, 1995), but its mechanism of action has not been clarified.

[0006]

Therefore, the pharmacological action of sulfatide is still largely unknown, and the fact is that no effective drug has been found.

[0007]

Under these circumstances, the present inventors have conducted intensive studies on the pharmacological action of sulfatide, and as a result, have found that sulfatide has an inhibitory action on TNF-α production. Production inhibitor and TN
Agent for preventing and treating F-α-mediated diseases, that is, prevention and treatment of chronic inflammatory diseases, acute inflammatory diseases, inflammatory diseases due to infection, autoimmune diseases, allergic diseases, and other TNF-α-mediated diseases. The present invention was found to be useful as a therapeutic agent and completed the present invention.

That is, the present invention provides (1) a TNF-α production inhibitor containing sulfatide as an active ingredient, (2) a prophylactic / therapeutic agent for TNF-α-mediated disease containing sulfatide as an active ingredient, and (3) TNF-α. The prophylactic / therapeutic agent according to the above (2), wherein the α-mediated disease is a chronic inflammatory disease, (4) TNF-α
The prophylactic / therapeutic agent according to the above (2), wherein the intervening disease is an acute inflammatory disease, (5) the prophylactic / therapeutic agent according to the above (2), wherein the TNF-α-mediated disease is an inflammatory disease due to infection,
(6) the prophylactic / therapeutic agent according to the above (2), wherein the TNF-α-mediated disease is an autoimmune disease; and (7) the prophylactic / therapeutic agent according to the above (2), wherein the TNF-α-mediated disease is an allergic disease.
Therapeutic agent, (8) administration of TNF-α-mediated disease for transplant rejection, graft-versus-host (GvH) disease, osteoporosis, cancer cachexia, disseminated vascular coagulation and trauma, burns, animal and plant components and drugs And (9) a pharmaceutical composition comprising a therapeutically effective amount of sulfatide and a pharmaceutically acceptable excipient.

[0009]

BEST MODE FOR CARRYING OUT THE INVENTION The sulfatide of the present invention is TNF-
Since it has an α production inhibitory action, it is useful as a TNF-α production inhibitor. Further, at present, the following documents are cited as showing the relationship between the TNF-α production inhibitory action and TNF-α-mediated diseases.

[0010] Relationship with Crohn's disease. Van Dullemen et. Al., Gastroenterology, 109, 129-
135, 1995

[0011] Regarding the relationship with contact dermatitis. Piguet et. Al., J. Exp. Med., 173, 673-679, 1991 ・ Relationship with bacterial shock (Septic shock). Exley et.al., Lancet, 335, 1275-1277, 1990

[0012] Regarding the relationship with endotoxemia. Beutler et. Al., Science, 229, 860-871, 1985 ・ Relationship with cerebral malaria. Grau et. Al., Science, 237, 1210-1212, 1987 Grau et. Al., N. Engl. J. Med., 320, 1586-1591,
1989 Kwiatkowski et. Al., QJ Med., 86, 91-98, 1993. Relation to infection.

Chang et. Al., Immunol. Infect. Dis.
, 2, 61-68, 1992 Harvell, J. Immunol., 143, 2894-2899, 1989 Kindler et. Al., Cell, 56, 731-740, 1989 Liew et. Al., Immunology, 69, 570-573. , 1990 Nakane et.al., Infect. Immun., 57, 3331-3337, 19
89 Nakano et.al., J. Immunol., 144, 1935-1941, 19
90 Opal et. Al., J. Infect. Dis., 161, 1148-1152,
1990 ・ About the relationship with AIDS.

Lahdevirt et. Al., Am. J. Med., 85,
289-291, 1988 Tracy, Cancer.Cell, 1, 62-63, 1989 Odeh, J. Intern. Med., 228, 549-556, 1990 Bromberg et.al., J. Immunol., 148, 3412-3417. ,
1992 Wllaurie et.al., AIDS, 6, 1265-1268, 1992 Ayehunie et.al., Clin. Exp. Immunol., 91, 37-42.
, 1993

・ Rheumatoid arthrit
is). Elliot et. Al., Arthririo Rheum., 36, 1681-1690,
1993 ・ Relationship with Demyelinating disease. Probert et. Al., Proc. Natl. Acad. Sic. USA,
92, 11294-11298, 1995

[0016] Relationship with Pulmonary Fibrosis. Piguet et.al., J. Exp.Med., 170, 655-663, 1989 Piguet et.al., Nature, 344, 245-247, 1990

Graft versus host disease
disease: GvHD). Piruet et.al., J. Exp.Med., 170, 655-663, 1987 Holler et.al., Blood, 75, 1011-1016, 1990 Irle et.al., Bone Marrow Transplant., 3, 127,
1988 Symington et.al., Transplantation, 50, 518-521
, 1990 Herve et. Al., Blood, 79, 3362-3368, 1992 Herve et. Al., Immunol. Rev., 129, 31-55, 1992. Regarding the relationship with allograft rejection.

Imagawa et. Al., Transplantation, 5
0, 189-193, 1990 ・ Regarding the relationship with osteoporosis. Ishimi et.al., J. Immunol., 145, 3297-3303, 19
90 MacDonald et. Al., Br. J. Rheumatol., 31, 149-15
5, 1992 ・ Relationship with cancer cachexia (Cancer cachexia).

Sherry et. Al., FASEB J., 3, 1956-196.
2, 1989 Teng et. Al., Proc. Natl. Acad. Sic. USA, 88,
3535-3539, 1991 ・ Disseminated intravascular coagu
About the relationship with thrombus due to lation syndrome (DIC). Tracy et. Al., Surg. Gen. Obstet., 164, 415-422
, 1987 Van et. Al., N. Engl. J. Med., 322, 1622-1629,
1990

Therefore, the sulfatide of the present invention is TN
F-α mediated diseases such as chronic inflammatory disease, acute inflammatory disease, inflammatory disease due to infection, autoimmune disease,
It can be used as an agent for preventing or treating allergic diseases and other TNF-α-mediated diseases.

Here, chronic inflammatory diseases are osteoarthritis,
Psoriatic arthritis, inflammatory skin diseases (psoriasis, eczema dermatitis, seborrheic dermatitis, lichen planus, pemphigus, bullous pemphigoid, epidermolysis bullosa, urticaria, vascular edema, vasculitis, erythema And diseases such as cutaneous eosinophilia, acne, alopecia areata, eosinophilic fasciitis, atherosclerosis, etc., and inflammatory bowel disease (ulcerative colitis, Crohn's disease, etc.) .

Acute inflammatory diseases include diseases such as contact dermatitis, adult respiratory failure syndrome (ARDS), sepsis (including organ damage caused by sepsis), and septic shock.

The inflammatory diseases caused by infection include endotoxin shock, acquired immunodeficiency syndrome (AIDS) cachexia,
In addition, they show diseases such as inflammatory reactions (including fever, pain and organ damage due to epidemic and non-epidemic colds) caused by infection with bacteria, viruses, mycoplasmas and the like.

Autoimmune diseases include rheumatoid arthritis, ankylosing spondylitis, systemic lupus erythematosus, glomerulonephritis (such as nephrotic syndrome (idiopathic nephrotic syndrome, minimal change nephropathy, etc.)), multiple sclerosis, multiple cartilage Inflammation, scleroderma, dermatomyositis, Wegener's granulomatosis, active chronic hepatitis I, primary biliary cirrhosis, myasthenia gravis, idiopathic sprue, Graves' disease, sarcoidosis, Reiter's syndrome, juvenile diabetes (type I diabetes mellitus) ), Autoimmune eye diseases (endocrine eye disorders, non-infectious uveitis, keratitis (dry keratoconjunctivitis, spring keratoconjunctivitis, etc.)), autoimmune blood diseases (hemolytic anemia, irreproducible anemia, Such as idiopathic thrombocytopenia).

Allergic diseases include atopic dermatitis,
It indicates diseases such as asthmatic diseases (bronchial asthma, pediatric asthma, allergic asthma, endogenous asthma, extrinsic asthma, dusty asthma, late asthma, airway irritability, bronchitis, etc.) and allergic rhinitis.

Other TNF-α-mediated diseases include organ or tissue transplantation (eg, heart, kidney, liver, lung,
Bone marrow, cornea, pancreas, islet cells, small intestine, duodenum, limbs,
Allergic and xenografts such as muscle, nerve, fatty marrow, skin, etc.), ie, rejection and graft versus host (GvH) disease, osteoporosis, cancer cachexia,
Diseases such as disseminated vascular coagulation, trauma, burns, inflammatory reactions (including shock) due to animal and plant components (including snake venom) and drug administration, and the like.

[0027] The sulfatide of the present invention may be a commercially available one, or any of those extracted, purified and synthesized from each organ can be used. In particular, the sulfatide used in the present invention can be obtained by the separation / purification method described in the following document (Suzuki Y. et al.,
Biochem. Biophys. Res.Commun., 190, 426-434, 19
93).

The sulfatide of the present invention may be a pharmaceutically acceptable salt, such as a salt with a metal such as sodium, potassium, calcium, magnesium, aluminum and the like. Hydrates, 1/3 hydrates, 2/3 hydrates, monohydrates, 3/2 hydrates, dihydrates and the like are also included.

When the sulfatide of the present invention is used as a TNF-α production inhibitor or a TNF-α-mediated disease preventive / therapeutic agent, it is prepared as a general pharmaceutical preparation. For example, a pharmaceutical composition or a pharmaceutical composition obtained by mixing the sulfatide of the present invention with a pharmaceutically acceptable carrier (excipient, binder, disintegrant, flavoring agent, flavoring agent, emulsifier, diluent, solubilizing agent, etc.) Tablets, pills, powders, granules, capsules, lozenges, syrups, solutions, emulsions, suspensions, injections (solutions,
Suspensions, etc.), suppositories, inhalants, transdermal absorbents, eye drops, eye ointments and the like are formulated in a form suitable for oral or parenteral use.

When a solid preparation is used, additives such as sucrose, lactose, cellulose sugar, D-mannitol,
Maltitol, dextran, starches, agar, alginates, chitins, chitosans, pectins, tran gum, gum arabic, gelatin, collagen, casein, albumin, calcium phosphate, sorbitol, glycine, carboxymethylcellulose, polyvinylpyrrolidone , Hydroxypropylcellulose, hydroxypropylmethylcellulose, glycerin, polyethylene glycol, sodium hydrogencarbonate, magnesium stearate, talc and the like. Further, the tablets can be made into tablets coated with a usual coating, if necessary, such as sugar-coated tablets, enteric-coated tablets, film-coated tablets or two-layer tablets or multilayer tablets.

When a semisolid preparation is used, animal and vegetable fats (eg, olive oil, corn oil, castor oil, etc.), mineral fats (eg, petrolatum, white petrolatum, solid paraffin, etc.), waxes (eg, jojoba oil, carnauba wax, beeswax) And partially or totally synthesized glycerin fatty acid esters (such as lauric acid, myristic acid, and palmitic acid). Examples of these commercially available products include Witepsol (manufactured by Dynamid Nobel), Pharmasol (manufactured by NOF Corporation) and the like.

In the case of a liquid preparation, additives such as sodium chloride, glucose, sorbitol, glycerin, olive oil, propylene glycol, ethyl alcohol and the like can be mentioned. In particular, in the case of an injection, a sterile aqueous solution such as physiological saline, isotonic solution, or oily solution such as sesame oil or soybean oil is used. If necessary, a suitable suspending agent such as sodium carboxymethyl cellulose, a nonionic surfactant, and a solubilizing agent such as benzyl benzoate or benzyl alcohol may be used in combination. Further, in the case of eye drops, an aqueous solution or aqueous solution is used, and particularly, a sterile aqueous solution for injection is used. Buffers (borate buffers, acetate buffers, carbonate buffers and the like are preferable for reducing irritation), isotonic agents, solubilizing agents, preservatives, thickeners, chelates Various additives such as an agent, a pH adjuster (the pH is usually preferably adjusted to about 6 to 8.5), and a fragrance may be appropriately added.

[0033] The amount of the active ingredient in these preparations is 0.1% of the preparation.
It is 1 to 100% by weight, suitably 1 to 50% by weight. The dosage may vary depending on the patient's condition, body weight, age, etc.
About 50 mg, and it is preferable to administer it once or in several divided doses.

[0034]

EXAMPLES The TNF-α production inhibitor and the TNF-α-mediated disease preventive / therapeutic agent of the present invention will now be described more specifically with reference to formulation examples and pharmacological actions. It is not limited.

Formulation Example 1: Tablets 50.0 mg Sulfatide Fine powder cellulose 25.0 mg Lactose 49.5 mg Starch 40.0 mg Talc 5.0 mg Tablets consisting of 0.5 mg magnesium stearate.

If desired, a sugar-coated tablet or a film-coated tablet can be obtained by applying a sugar-coating treatment or a film-coating treatment.

Formulation Example 2: Capsules A capsule consisting of 50.0 mg of sulfatide, 50.0 mg of lactose, 15.0 mg of starch, and 5.0 mg of talc.

Formulation Example 3: Fine Granules Fine granules consisting of 10% sulfatide, 80% lactose, and 10% starch.

Formulation Example 4: Granules Granules comprising 10% of sulfatide, 55% of lactose, 20% of finely powdered cellulose, 20% of starch and 15% of starch.

Formulation Example 5: Suppository A suppository comprising 100 mg of sulfatide, 950 mg of Witepsol H15, and 950 mg of Witepsol E75.

Formulation Example 6: Injection 50.0 mg of sulfatide and 100.0 mg of glucose are dissolved in purified water to prepare a 2 ml injection solution.

Hereinafter, the pharmacological action of the medicament of the present invention will be described with reference to experimental examples.

EXPERIMENTAL EXAMPLE 1: In Vitro TNF-α Production Inhibitory Activity Sulfatide was added to a phosphate buffer (PBS) at 1 mg / m 2.
After adjusting to 1, sonication was performed for 1 minute. In addition, lipopolysaccharide (LPS) was added to PBS
Was adjusted to 1 mg / ml. The human monocytic leukemia cell line THP-1 was transformed with 5% fetal calf serum (fetal calf).
1 in RPMI-1640 medium containing serum (FCS)
It was adjusted to × 10 ⁶ cells / ml and dispensed at 100 μl / well into a 96-well culture plate. Next, after adding sulfatide to various concentrations, 37 ° C, 5%
Incubated under CO ₂ for 15 minutes. After the incubation, LPS was added to a concentration of 10 μg / ml, and the cells were cultured at 37 ° C. and 5% CO ₂ for 6 hours. After completion of the culture, the culture supernatant was recovered, and the amount of TNF-α in the culture supernatant was measured by ELISA. Table 1 shows the results.

[0044]

[Table 1]

From the above experimental examples, in vitro, the human monocyte leukemia cell line THP-1 produces a large amount of TNF-α in the culture supernatant by culturing in the presence of LPS, but 10 μg / ml. The preculture with the sulfatide at the above concentration suppressed the production of TNF-α in the culture supernatant by LPS stimulation of THP-1 cells in a dose-dependent and significant manner.

Experimental Example 2: Serum TNF-α Production Inhibitory Activity PBS and sulfatide were administered to mice (C3H / hen, 9 weeks old, female) via the tail vein, and 30 minutes later PB
LPS adjusted to 10 μg / ml with S.
0.5 ml per 0 g was intraperitoneally administered to the mice.
90 minutes after LPS administration, whole blood was collected from the inferior vena cava of the mouse under ether anesthesia. Serum was obtained by sufficiently coagulating the collected blood at room temperature, leaving it overnight at 4 ° C., and then centrifuging. The amount of TNF-α in serum was determined by ELISA.
It was measured by the method. Table 2 shows the results.

[0047]

[Table 2]

From the above experimental examples, it was found that LP
S increase of TNF-α in serum by 1 and 10 m
A dose of g / kg of sulfatide could significantly suppress.

Experimental Example 3: Endotoxin shock inhibitory activity Mice (C3H / hen, 8 weeks old, female) were administered PBS and sulfatide via the tail vein, and 30 minutes later, P
0.5 ml of LPS adjusted to 1.6 mg / ml with BS
Each mouse was intraperitoneally administered. After LPS administration, survival was calculated by observing every other day for 7 days. Table 3 shows the results.

[0050]

[Table 3]

According to the above experimental examples, sulfatide improved the survival rate in a lethal endotoxin shock model by LPS administration in a dose-dependent manner.

[0052]

According to the above formulation examples and pharmacological experimental examples, the sulfatide of the present invention has a TNF-α production inhibitory effect, and a TNF-α production inhibitor, a TNF-α-mediated disease preventive / therapeutic agent, Prevention of chronic inflammatory disease, acute inflammatory disease, inflammatory disease due to infection, autoimmune disease, allergic disease, and other TNF-α-mediated diseases
Useful as a therapeutic.

Claims (9)

[Claims] 1. TNF-α containing sulfatide as an active ingredient
Production inhibitors. 2. TNF-α containing sulfatide as an active ingredient
Agent for prevention and treatment of mediated diseases. 3. The method according to claim 2, wherein the TNF-α-mediated disease is a chronic inflammatory disease. 4. The method according to claim 2, wherein the TNF-α-mediated disease is an acute inflammatory disease. 5. The preventive / therapeutic agent according to claim 2, wherein the TNF-α-mediated disease is an inflammatory disease caused by infection. 6. The method according to claim 2, wherein the TNF-α-mediated disease is an autoimmune disease. 7. The method according to claim 2, wherein the TNF-α-mediated disease is an allergic disease. 8. The method according to claim 1, wherein the TNF-α-mediated disease is transplant rejection,
Graft versus host (GvH) disease, osteoporosis, cancer cachexia,
3. The preventive / therapeutic agent according to claim 2, wherein the agent is an inflammatory reaction caused by disseminated vascular coagulation and administration of trauma, burns, animal and plant components, and drugs. 9. A pharmaceutical composition comprising a therapeutically effective amount of sulfatide and a pharmaceutically acceptable excipient.