KR20100099849A - Composition for treatment of atopic dermatitis comprising immunosuppressant and inhibitors of transglutaminase 2 - Google Patents

Abstract

PURPOSE: A composition containing immunosuppressant and transglutaminase 2(TG2) inhibitor for treating atopic dermatitis is provided to reduce dose of the immunosuppressant. CONSTITUTION: A composition for treating atopic dermatitis contains an immunosuppressant and transglutaminase 1 inhibitor. The immunosuppressant is cyclosporine. The TG2 inhibitor is an antibody to TG2 and antisense molecule to the gene of TG2. The antisense molecule is ribozyme, RNAi or antisense nucleic acid. The transglutaminase 2 inhibitor is glucosamine. The immunosuppressant and TG2 inhibitor is mixed in a ratio of 0.3-0.4:1.

Description

Composition for Treatment of Atopic Dermatitis Comprising Immunosuppressant and Inhibitors of Transglutaminase 2}

The present invention relates to a composition for treating atopic dermatitis containing an immunosuppressive agent and an inhibitor of transglutaminase 2.

Atopic dematitis is a common skin disorder with unexplained strong pruritus, combined with dry skin, erythema and inflammation (Lancet. 1998; 351: 1715-1721). Atopic dermatitis is a chronic disease that causes great discomfort in long-term life, mainly in infants and young children. It occurs in 10 to 15% of all infants and infants, with the incidence in the atopy group being 60% in infants less than 1 year and 85% in infants less than 5 years (Lancet. 1998; 351: 1715-1721). 40% of children with atopic dermatitis gradually improved with growth, but the incidence rate of adults was about 1 to 3% of all adults due to pollution and environmental pollution caused by rapid industrialization (Immunol. Allergy. Cli. North Am. 2002; 22: 1-24), and the incidence rate is increased two to three times as before the industrialization (Lancet. 2003; 361: 151-160). For this reason, many researchers are interested in reducing the clinical symptoms of atopic dermatitis.

The clinical symptoms of atopic dermatitis may vary from person to person, but the most common symptoms are first, severe itching, second, the patient's skin remains dry, and third, dry skin. Due to the loss of the defense function of the skin surface, it is easy to invade the skin of irritants from the outside, and the invasion causes the skin to reject the inflammatory reaction (Lancet. 1998; 351: 1715-1721, J. Pediatr). Health Care. 2002; 16; 143-145.

Many studies have been conducted on the cause and treatment of atopic dermatitis until recently, but due to the complexity of the disease itself and conflicting research data, no clear theory has yet been established on the exact etiology and effective treatment. Therefore, therapeutic agents for atopic dermatitis, which have been developed to date, include topical anti-inflammatory agents such as steroids, tacrolimus, pimecrolimus, immunosuppressive agents such as antihistamines and cyclosporin. In addition, hypotensive moisturizers and photochemotherapy with ultraviolet rays A have been used as clinical supplements. However, these therapies or therapies help to control the symptoms to an appropriate level rather than the underlying treatment, and so far do not fully meet the needs of patients with atopic dermatitis.

In order to meet these demands, the present inventors have developed a therapeutic agent for atopic dermatitis, which contains glucosamine as an active ingredient, a transglutaminase inhibitor, which is a commercially available natural substance, which has been recognized for its safety (Korea Patent Publication No. 2008- 0035997). However, glucosamine was only effective in some subjects, and was poor in patients with severe atopic dermatitis.

Meanwhile, immunosuppressive agents such as cyclosporin have been used in patients with severe atopic dermatitis who do not know the existing treatments. Immunosuppressants have excellent effects, but have disadvantages such as renal toxicity, gingival thickening, hirsutism, neurotoxicity, hypertension, hyperlipidemia, hyperglycemia, hyperkalemia, hyperuricemia, hemolytic uremic syndrome, and infection. In addition, despite the difficulty in long-term administration due to the side effects of the immunosuppressive agent has a problem of reducing to the original state when the administration is stopped.

The present inventors have attempted to develop a treatment that can gradually replace it while being used in combination with an immunosuppressive agent to solve the above problems.

In other words, while reducing the dose of immunosuppressant, while trying to develop a combination therapy that can show the same or better therapeutic effect than the immunosuppressant, the immunosuppressive agent and the inhibitor of transglutaminase 2 are administered in combination. While reducing the dose of, the therapeutic effect was confirmed to be higher than when the immunosuppressive agent was administered alone, and the present invention was completed.

Accordingly, the present invention is to provide a composition for treating atopic dermatitis containing an immunosuppressant and an inhibitor of transglutaminase 2.

"Immune inhibitor", which is an active ingredient of the composition according to the present invention, refers to a drug that suppresses the immune response of a living body. The immunosuppressive agents include glucocorticoid, cyclosporine, tacrolimus (FK506), pimecrolimus, ISA (TX) 247, calcineurin inhibitors, rapamycin (rapycin) Rapamycin), Type IV PDE inhibitors (Type IV PDE inhibitors), Mycophenolate Mofetil (Mycophenolate Mofetil) and the like can be exemplified, but not limited thereto, all known immunosuppressive agents can be used. One type of immunosuppressant may be used alone, or two or more types of immunosuppressive agents may be used in combination. In the present invention, it is particularly preferable to use cyclosporin.

Another active ingredient of the composition according to the invention is an "inhibitor of transglutaminase 2 (TG2)". Transglutaminase is an enzyme (Methods in Enzymol. 1985; 113: 358-375) that catalyzes the binding of epsilon gamma glutamilysine (ε-γ-glutamyl-lysine) to glutamine and lysine groups of different proteins. Was considered an important factor in preventing and healing damaged tissues (FASEB J. 1999; 13: 1787-1795). However, abnormally activated transglutaminase contributes to the development of various diseases such as cerebral disease, arteriosclerosis, inflammatory diseases, autoimmune diseases and fibrosis (Neurochem. Int. 2002; 40: 85-103) .

Inhibitors of TG2 in the present invention may be an inhibitor of TG2 activity or an inhibitor of expression of TG2.

The inhibitor of activity of TG2 in the present invention may preferably be an anti-TG2 antibody capable of neutralizing the activity of TG2. In the present invention, the anti-TG2 antibody may be a polyclonal antibody or a monoclonal antibody. Antibodies of the invention can be prepared by conventional methods well known in the art of immunology using TG2 protein as an antigen.

Another inhibitor of TG2 activity is a compound that inhibits TG2 activity, for example cystamine, putrescine, monodansylcadaverine, w-dibenzylaminoacylamine ( w-dibenzylaminoalkylamine), 3-halo-4,5-dihydroisoxazole, 2-[(2-oxopropyl) thio] imidazolium derivative (2-[(2 -oxopropyl) thio] imidazolium derivatives). In addition, epicalocatechin gallate or chlorogenic acid may be used.

In the present invention, "inhibition of expression of gene (TG2)" includes inhibition of gene transcription and translation into protein. In addition, not only the gene expression is completely stopped, but also the expression is reduced.

Antisense molecules are most commonly used as a method of inhibiting gene expression. Antisense molecules inhibit the expression of target genes by inhibiting transcriptional initiation by triple chain formation, transcriptional inhibition by hybridization at sites where local open loop structure is formed by RNA polymerase, and in RNA where synthesis is in progress. Inhibition of transcription by hybrid formation, inhibition of splicing by hybridization at the junction of introns and exons, inhibition of splicing by hybridization at the site of splicosomal formation, transition from nucleus to cytoplasm by hybridization with mRNA Inhibition of translation initiation by hybridization at the site of translation initiation factor binding. They inhibit the expression of target genes by inhibiting transcription, splicing or translation processes.

The antisense molecule used in the present invention may inhibit the expression of the target gene by any of the above actions. Representative antisense molecules include triplets, ribozymes, RNAi, or antisense nucleic acids.

In the present invention, it is particularly preferable to use glucosamine as an inhibitor of TG2. "Glucosamine" is a breakdown product of chitosan, a major component of sea crab and shrimp shells. The main components of sea crab or shrimp shell are chitin and chitosan, chitin is composed of 2-acetamido-2-deoxy-beta-D-glucose (N-acetylglucosamine), and chitosan is a polysaccharide that has undergone deacetylation of chitin. Poly (beta- (1,4) -glucosamine). Glucosamine has the structure of Formula 1 below.

In the present invention, a glucosamine derivative may be used instead of or in combination with glucosamine. In the present invention, the term "glucosamine derivative" refers to a hydrogen having a hydroxy group of glucosamine substituted with an acyl or alkyl compound, and having a structure of Formula 2 below.

Wherein R is an acyl group having 2 to 18 carbon atoms or straight or branched alkyl having 1 to 5 carbon atoms, preferably acetyl propionyl, butyryl, pentanoyl, hexanoyl, heptanoyl, octanoyl, nonanoyl, deca Acyl groups such as noyl, undecanoyl, lauryl, tridecanoyl, myristyl, pentadecanoyl, palmitoyl, margaryl or stearyl groups, or methyl, ethyl, propyl, butyl, pentyl, isopropyl, iso Alkyl groups, such as a butyl or a secbutyl group, may be sufficient.

In addition, glucosamine or derivatives thereof used in the present invention may be included in the pharmaceutical composition of the present invention in the form of a pharmaceutically acceptable salt according to the use thereof. Such salt forms include, but are not limited to, for example, sulfates of glucosamine or glucosamine derivatives, hydrochlorides of glucosamine and glucosamine derivatives, malate of glucosamine or glucosamine derivatives, and the like. Preferred salt forms are sulfates of glucosamine or glucosamine derivatives.

In the composition of the present invention, the immunosuppressant and the inhibitor of TG2 are preferably mixed at a ratio of 0.3-0.4 to 1. The more severe the severity of atopic dermatitis, the higher the content of the immunosuppressive agent, and the lower the severity by treatment, the lower the content of the immunosuppressive agent is desirable.

The compositions of the present invention can be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral formulations, sterile injectable solutions, suppositories, and transdermal formulations according to conventional methods. Carriers, excipients and diluents that may be included in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, Microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. If necessary, it is formulated with diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants and the like.

The composition of the present invention is preferably formulated into a solid preparation for oral administration. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, which include at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin and the like in the active ingredient. Formulated by mixing. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used.

In addition, the composition of the present invention is preferably formulated into a liquid formulation for oral administration. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups, and the like. In addition to conventional inert diluents (e.g., purified water, ethanol, liquid paraffin), various excipients may be used. Wetting agents, sweetening agents, fragrances, preservatives and the like can be included.

In addition, the compositions of the present invention may be formulated into a preparation for parenteral administration. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As a sterile aqueous solution, suitable buffering solutions such as Hanks' solution, Ringer's solution, or physically buffered saline can be used. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, and olive oil. Same vegetable oils, injectable esters such as ethyloleate, and the like can be used. If necessary, preservatives, stabilizers, wetting or emulsifying agents, salts for controlling osmotic pressure and / or buffers may also be used. Meanwhile, in the case of suppositories, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin, and the like, which are conventional bases thereof, may be used.

When administered to a human patient, the total daily usage of the compositions of the present invention can be determined by treatment within the medical judgment. The specific therapeutically effective amount for a particular patient depends on the type and severity of the reactions to be achieved, whether other agents are used in some cases, the age, body weight, general health, sex, diet, time of administration, route of administration, composition of the patient. It is desirable to apply differently depending on various factors including secretion rate, duration of treatment, and similar factors well known in the medical field.

In combination with an immunosuppressant and an inhibitor of transglutaminase 2 according to the present invention, a therapeutic effect superior to the administration of the immunosuppressant alone can be achieved while reducing the dose of the immunosuppressive agent which is difficult to administer for a long time due to side effects.

Hereinafter, the examples are only for illustrating the present invention in more detail, and the scope of the present invention is not limited by these examples in accordance with the gist of the present invention, those skilled in the art. Will be self-evident.

Example 1: Subject Selection

Severe atopic dermatitis patients requiring cyclosporin administration were selected as subjects, and two weeks of the specified dose of cyclosporin alone, two weeks in combination with glucosamine sulfate (500 mg), two weeks alone, and two weeks in combination. In this example, 100 mg (Cs1), 150 mg (Cs1.5), or 200 mg (Cs2), which is less than half the usual dosage of cyclosporin, was used.

SCORAD index was measured, photographed, and blood collected to measure changes in cytokine expression such as IL-4, IL-5, IFN-β and IFN-γ.

Example 2-1: Clinical Effect

Shown as the SCORAD index. This index sets the area occupied by the lesion to 0-100% according to the rule of nine principles, with 1/5 of the area and severity of erythema, edema / papulation, and exudation / oozing. / crusts), scratches, lichenification, skin dryness, divided into six categories, severity divided by 0-3, summed (0-18) x 7/2, and called objective SCORAD. In addition, itching and sleep disturbances during awakening of subjective symptoms are calculated as a sum (0-103) on each analog scale from 0 to 10 (European Task Force on Atopic Dermatitis, 1993). However, when the actual patients were asked to evaluate the symptoms of itching and sleep disorders on a scale of 0-10, they experienced many cases that were difficult to do properly, and they scored 0-83 points including the objective SCORAD index except the actual subjective scale. (Kunz et al., Dermatology 1997; 195: 10-19), this example evaluated only the objective SCORAD index.

SCORAD for one patient is shown in FIG. 1A. As shown in FIG. 1A, there was little change in SCORAD during the glucosamine alone administration period (November 10 to November 24). However, the combination of cyclosporin and glucosamine resulted in a sharp decrease in SCORAD from 72.5 to 51.5. Elevated SCDRAD after drug discontinuation (December 9) was lowered by cyclosporine alone (December 18) and subsequent co-administration (December 30). Subsequent administration of cyclosporine alone (January 9) again raised SCORAD, which had been lowered by previous treatment.

SCORAD for the other patients is shown in Figure 1b. As shown in FIG. 1B, there was little change in SCORAD during the period of cyclosporine alone (November 17-December 1). These results support previous reports that some individuals do not hear cyclosporin well. During the combined dose of cyclosporine and glucosamine (Dec 1–Dec 15), SCORAD dropped sharply from 41 to 34. Cyclosporine alone (December 15) was administered to increase SCORAD, and SCORAD was lowered by combination (December 30).

That is, the administration of glucosamine alone showed insufficient treatment effect, and the administration of cyclosporine alone showed an increase in SCORAD, that is, a decrease in treatment effect. In contrast, the combined administration of cyclosporin and glucosamine was superior to the administration of glucosamine or cyclosporin alone, and the dose of cyclosporine at the time of the combined administration was less than half of the usual cyclosporin use dose.

Example 2-2: Photographing

Photographs were taken at intervals of 4 weeks with the ruler next to the lesion at a distance of 30 cm from the lesion. The person in charge of photography was taken by the specialty in charge of the subject, and the education for standardizing photography was conducted. The patient was photographed at a certain distance from a certain posture, and the illumination at the location of photography was kept constant. Photographs were taken each time with the same composition, making it easier to compare before and after.

In Figure 2, the left side of the photo before using the combination therapy erythema and swelling of the lesion is severe, both the lower extremity due to the overall erythema and edema, the patient complained of pain and scratches due to itching was obvious. In the middle picture, two cycles of combination therapy were taken. Many lesions were lost, and erythema and edema of each lesion were significantly reduced. The photo on the right shows a slight deterioration of the lesion after a combination change alone, with red spots recurring in some skin areas and a slight increase in scratches.

Example 2-3: Vital Signs

Vital signs, ie blood pressure and pulse rate, were measured after 3 minutes of stability in a sitting position.

The blood pressure and pulse rate of the subjects were in the normal range.

Example 2-4 Analysis of Cytokine Expression

At the time of dosing and when the dosing method was changed, that is, every two weeks, blood was collected and the cytokines such as IL-4, IL-5, IFN-β and IFN-γ were measured by RT-PCR.

As shown in FIG. 3, the combined amounts of glucosamine or cyclosporine alone decreased cytokine levels such as IL-4, IL-5, IFN-β, IFN-γ, and the like. In particular, the expression level of IL-4 was significantly decreased by the co-administration, and in the case of patient 1, the expression level of IL-5 was also markedly decreased.

Figure 1 shows the SCORAD index calculated each time the dosage method is changed.

2 is a photograph of the affected part of the patient at a designated time.

Figure 3 shows the amount of cytokine expression of IL-4, IL-5, IFN-β, IFN-γ, etc. measured every time the dosage method is changed.

Claims (8)

A composition for treating atopic dermatitis, comprising an immunosuppressant and an inhibitor of transglutaminase 2. The method of claim 1, The immunosuppressive agent is a composition for treating atopic dermatitis, characterized in that cyclosporin. The method of claim 1, The inhibitor of transglutaminase 2 is a composition for treating atopic dermatitis, characterized in that the antibody to transglutaminase 2. The method of claim 1, The inhibitor of transglutaminase 2 is a composition for treating atopic dermatitis, characterized in that the antisense molecule for the gene of transglutaminase 2. The method of claim 4, wherein The antisense molecule for the gene of the transglutaminase 2 is a composition for treating atopic dermatitis, characterized in that the triple agent, ribozyme, RNAi or antisense nucleic acid. The method of claim 1, The inhibitor of transglutaminase 2 is a composition for treating atopic dermatitis, characterized in that the glucosamine. The method of claim 6, The glucosamine is a composition for treating atopic dermatitis, characterized in that glucosamine sulfate. The method of claim 1, Immunosuppressant and inhibitor of transglutaminase 2 is a composition for the treatment of atopic dermatitis, characterized in that mixed in a 0.3-0.4 to 1 ratio.

Images