KR102324223B1 - Pharmaceutical composition for preventing or treating inflammatory diseases comprising saccharin, acesulfame K or mixture thereof - Google Patents

Abstract

The present invention relates to a composition for preventing or treating inflammatory diseases containing saccharin, acesulfame potassium or a mixture thereof as an active ingredient, wherein the saccharin or acesulfame potassium (acesulfame K) ) was confirmed to have an effect of suppressing the expression of inflammatory cytokines that cause inflammation, and in particular, as it was confirmed to effectively inhibit the secretion of IL-1β, which is a major cause of chronic sinusitis, the saccharin, aceseol Palm potassium (acesulfame K) or a mixture thereof may serve as a therapeutic agent for an inflammatory disease caused by IL-1β.

Description

TECHNICAL FIELD [0001] Pharmaceutical composition for preventing or treating inflammatory diseases comprising saccharin, acesulfame potassium or a mixture thereof as an active ingredient;

The present invention relates to a composition for preventing or treating inflammatory diseases containing saccharin, acesulfame potassium or a mixture thereof as an active ingredient.

Inflammation is an important part of the body's immune response, a useful defense response to injury or damage, and is designed to suppress the detrimental effects of injury. However, excessive acute or chronic inflammation can cause serious disorders such as arthritis, asthma, colitis, Parkinson's disease, Alzheimer's disease or sepsis.

Chronic Rhinosinusitis (CRS) is one of the diseases with a high incidence of about 7% among otolaryngology diseases. It is known as a chronic inflammatory disease of the upper respiratory tract.

The treatment of chronic sinusitis can be divided into drug treatment and surgical treatment, and antibiotics play a very important role in drug treatment. About 70-80% of patients with chronic sinusitis remain symptom-free with medication or surgery, but about 20-30% of patients have poor response to surgery and drug therapy, and recurrence occurs repeatedly even after treatment. In these cases, antibiotic treatment is required for more than 2 to 3 months during drug treatment, complications increase even after reoperation, and it is not easily cured after surgery. Because it is used, there is a problem of side effects due to misuse of drugs.

The molecular mechanism of chronic sinusitis is that a marked increase in eosinophils is observed in the nasal polyp tissue that accompanies chronic sinusitis, and the frequency varies from 30 to 60%. Also, in lymphocytes, CD8 ⁺ T cells are increased, and mast cells and plasma cells are increased. In addition, secretion of IL-4, IL-5, IL-6 and IL-3, which means activation of the type 2 immune response, increases, and these inflammatory mediators interact to contribute to the proliferation and chronicity of costs. is known

According to a report in 2017, it was confirmed that the inflammatory cytokine IL-1β secreted by macrophages is an important mediator in the pathogenesis of CRS. In addition, in an animal model of chronic sinusitis using myeloid immune cell-specific autophagy-deficient mice, it was found that autophagy abnormalities were significantly involved in IL-1β secretion by macrophages, and when IL-1β action was blocked, inflammation was significantly decrease was confirmed.

Recently, as a decrease in LC3 and overexpression of COX-2, which are biomarkers of autophagy, have been observed at the expense of patients with chronic sinusitis, it is necessary to develop a new therapeutic agent that is independent of autophagy dysfunction and can inhibit IL-1β secretion. the current situation.

Republic of Korea Patent Publication No. 10-2015-0129528 (published on November 20, 2015)

The present invention relates to a composition for preventing or treating inflammatory diseases containing saccharin, acesulfame potassium or a mixture thereof as an active ingredient, wherein the composition effectively inhibits IL-1β secretion regardless of autophagy function, thereby treating chronic sinusitis. It can effectively prevent or treat inflammatory diseases, including

The present invention provides a pharmaceutical composition for preventing or treating inflammatory diseases containing saccharin, acesulfame potassium, or a mixture thereof as an active ingredient.

In addition, the present invention provides a health food for preventing or improving inflammatory diseases containing saccharin, acesulfame potassium, or a mixture thereof as an active ingredient.

According to the present invention, it has been confirmed that saccharin or acesulfame potassium exhibits an effect of inhibiting the expression of inflammatory cytokines that cause inflammation, and in particular, it has been confirmed that it effectively inhibits the secretion of IL-1β, a major cause of chronic sinusitis. Accordingly, the saccharin, acesulfame potassium, or a mixture thereof may serve as a therapeutic agent for an inflammatory disease caused by IL-1β.

1 is a result of confirming the anti-inflammatory effect of trehalose in an animal model induced with chronic sinusitis, FIG. 1 (A) is a schematic diagram showing the production process of an animal model of chronic sinusitis, H&E, sirius red and toluidine blue staining results confirming changes in the thickness of nasal tissue and changes in the number of eosinophils and mast cells after intraperitoneal administration (ip) or nasal administration (in) of Oss, Figure 1(C) shows the staining results The graph shown is the result.
2 is a result of confirming the improvement of chronic sinusitis of trehalose in a myeloid immune cell-specific autophagy deficient mouse model (Atg7 ^f/f ; Lyz2-Cre) and a normal mouse model (Atg7 ^{f/f), FIG.} 2(A) is the result of H&E, sirius red and toluidine blue staining confirming the change in the thickness of nasal tissue and the number of eosinophils and mast cells after intraperitoneal administration (ip) or nasal administration (in) of trehalose, respectively, FIG. 2 (B) is a graph result showing the 2(A) staining result, and FIG. 2(C) is the thickness change and IL-1β expression level after intraperitoneal administration (ip) or nasal administration (in) of trehalose. It is the confirmed immunofluorescence analysis result, and FIG. 2(D) is the result of confirming the TNF-α, IL-1β and IL-6 mRNA expression levels after trehalose administration.
Figure 3 is the result of confirming the inflammatory response inhibitory effect of various saccharin, trehalose, saccharin, acesurfame K in macrophages collected from thioglycolate-treated myeloid immune cell-specific autophagy-deficient mice or normal mice abdominal cavity , sucralose, glucose, sucrose, maltose, and lactose were treated, and the expression levels of TNF-α, IL-1β, IL-6 and IL-10 were confirmed.
Figure 4 shows TNF-α, IL-1β, IL-6 and IL-10 mRNA expression levels after treatment with trehalose or saccharin in macrophages collected from the abdominal cavity of mice lacking myeloid immune cell-specific autophagy function or normal mice. is the result of checking
5 is a result of confirming the inflammation improvement effect after saccharin treatment in an animal model of chronic sinusitis. It is a staining result, and FIG. 5(B) is a graph result showing the staining result.
6 is a result of confirming the inflammation improvement effect after saccharin treatment in an animal model of psoriasis. FIG. 6 (A) is a schematic diagram showing the psoriasis animal model production and saccharin treatment process, and FIG. 6 (B) is the ear of the animal model after saccharin treatment. It is the graph result confirming the thickness change.

Hereinafter, the present invention will be described in more detail.

The present invention can provide a pharmaceutical composition for preventing or treating inflammatory diseases containing saccharin, acesulfame potassium, or a mixture thereof as an active ingredient.

The saccharin, acesulfame potassium, or a mixture thereof may inhibit IL-1β secretion.

The inflammatory disease may be induced by IL-1β secreted from macrophages.

The inflammatory disease may be selected from the group consisting of chronic sinusitis, bronchitis, hepatitis, ulcerative colitis, asthma, psoriasis, arthritis, dermatitis, gingivitis, keratitis, atopic dermatitis, gout, and chronic obstructive pulmonary disease (COPD).

In one embodiment of the present invention, the pharmaceutical composition containing saccharin, acesulfame potassium or a mixture thereof as an active ingredient can be prepared by injection, granule, powder, tablet, pill, capsule, suppository, gel, Any one formulation selected from the group consisting of suspensions, emulsions, drops or solutions may be used.

In another embodiment of the present invention, the pharmaceutical composition containing saccharin, acesulfame potassium or a mixture thereof as an active ingredient is a suitable carrier, excipient, disintegrant, sweetening agent, coating agent, and swelling agent commonly used in the manufacture of pharmaceutical compositions. , lubricants, lubricants, flavoring agents, antioxidants, buffers, bacteriostats, diluents, dispersants, surfactants, binders and lubricants may further include one or more additives selected from the group consisting of.

Specifically, carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline Cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil can be used. Solid preparations for oral administration include tablets, pills, powders, granules, and capsules. agent and the like, and such a solid preparation may be prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, and the like in the composition. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Liquid formulations for oral use include suspensions, solutions, emulsions, syrups, and the like, and various excipients such as wetting agents, sweeteners, fragrances, and preservatives in addition to commonly used simple diluents such as water and liquid paraffin may be included. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base material for the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like can be used.

According to an embodiment of the present invention, the pharmaceutical composition is administered intravenously, intraarterially, intraperitoneally, intramuscularly, intraarterially, intraperitoneally, intrasternally, transdermally, intranasally, inhalation, topical, rectal, oral, intraocular or intradermal The route may be administered to a subject in a conventional manner.

The preferred dosage of saccharin, acesulfame potassium or a mixture thereof may vary depending on the condition and weight of the subject, the type and extent of the disease, the drug form, the route and duration of administration, and may be appropriately selected by those skilled in the art. According to an embodiment of the present invention, although not limited thereto, the daily dose may be 0.01 to 200 mg/kg, specifically 0.1 to 200 mg/kg, and more specifically 0.1 to 100 mg/kg. Administration may be administered once a day or may be administered in several divided doses, thereby not limiting the scope of the present invention.

In the present invention, the 'subject' may be a mammal including a human, but is not limited to these examples.

The present invention can provide a health food for preventing or improving inflammatory diseases containing saccharin, acesulfame potassium, or a mixture thereof as an active ingredient.

The health food is used together with other foods or food additives other than the saccharin, acesulfame potassium (acesulfame K) or mixtures thereof, and may be appropriately used according to a conventional method. The mixed amount of the active ingredient may be suitably determined according to the intended use thereof, for example, prophylactic, health or therapeutic treatment.

The effective dose of the compound contained in the health food may be used according to the effective dose of the therapeutic agent, but in the case of long-term intake for health and hygiene or health control, it may be less than or equal to the above range, It is clear that the ingredient can be used in an amount above the above range because there is no problem in terms of safety.

The type of health food is not particularly limited, and examples include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes.

Hereinafter, to help the understanding of the present invention, examples will be described in detail. However, the following examples are merely illustrative of the contents of the present invention, and the scope of the present invention is not limited to the following examples. The embodiments of the present invention are provided to more completely explain the present invention to those of ordinary skill in the art.

<Reference example> Reagent

Aspergillus oryzae protease and trehalose, saccharin, acesulfame potassium, sucralose, glucose, sucrose, maltose, lactose ( lactose) and ATP were purchased from Sigma. Ovalbumin was purchased from Worthington Biochemical, and LPS from InvivoGen was used.

The following experimental examples are intended to provide experimental examples commonly applied to each embodiment according to the present invention.

<Experimental Example 1> Myeloid immune cell-specific autophagy-deficient mouse production

To construct myeloid cell-specific Atg7 deficient Atg7 ^f/f ; Lyz2-Cre mice, Lyz2-Cre mice (stock number 4781; Jackson Laboratories, Bar Harbor, Me) and Atg7 ^f/f mice (Masaaki) Komatsu (provided by Niigata University, Niigata, Japan) was bred Litter mates were used for all experiments, and all methods for mouse experiments were provided by IACUC of Asan Life Science Research Institute.

<Experimental Example 2> Chronic rhinosinusitis (CRS) animal model production

To induce a chronic sinusitis (CRS) mouse model, 0.7 U of Aspergillus oryzae protease per mouse and 75 μg of ovalbumin (Sigma-Aldrich, St Louis, Mo) were mixed in the nose of mice 3 times a week for 5 weeks (total 15). times) was injected. To confirm the therapeutic effect of trehalose and saccharin, 50 mg/kg of trehalose (stock 50 mg/mL) and 1 mg/kg of saccharin (stock 1 mg/mL) per mouse In an amount of kg, it was pre-injected into the nasal cavity 5 hours before CRS induction.

After induction of the CRS model, a section of mouse nasal tissue was used for staining, and H&E staining was provided by the tissue staining room of the Asan Life Science Research Institute. Eosinophil infiltration was confirmed using Sirius red staining, and mast cell infiltration was confirmed by toluidine blue staining.

<Experimental Example 3> Immunofluorescence and confocal analysis

To detect macrophages and IL-1β-secreting cells in the nasal tissues of CRS-induced mice, tissue slides were prepared with CD68 (1:100, FA-11; AbD Serotec) and IL-1β (1: 100, 3A6). , Cell Signaling, Danvers, Mass) was incubated with a primary antibody for 1 hour. Afterwards, secondary antibodies to each primary antibody were Alexa Fluor 488-conjugated goat anti-rat F(ab′)2 (1:250; Jackson ImmunoResearch) and Alexa Fluor 488-conjugated goat anti-mouse F(ab′). 2 (1:250; incubated with Jackson ImmunoResearch for 30 minutes. All antibodies were diluted in PBS containing 1% BSA and 1% goat serum. After all incubation, slides were washed 3 times with PBS. , A cover-glass was mounted with ProLong Gold anti-fade reagent (Molecular Probes, Eugene, Ore) Tissue slides were imaged with an LSM 710 laser scanning confocal microscope (Carl Zeiss, Oberkochen, Germany).

<Experimental Example 4> Macrophage isolation and culture

3 mL of 3.85% thioglycolate (difco) was injected into the abdominal cavity of 8-10 week old mice, and after 3 days, the peritoneal was washed with 10 mL of Dulbecco's modified Eagle medium (DMEM) to obtain peritoneal macrophage cells. ) was obtained. The cells were spread on a plastic tissue culture plate and cultured at 37° C. for 3 hours. Cells that did not adhere were washed with fresh DMEM, and only adherent peritoneal macrophages were used for the experiment.

<Experimental Example 5> Enzyme-linked immunoprecipitation assay (ELISA)

Abdominal macrophages prepared by the method described above were pretreated with trehalose, saccharin, acesulfame potassium, sucralose, glucose, sucrose, maltose and lactose (sigma) at the concentrations shown in the figure for 15 minutes, followed by LPS (InvivoGen) 10ng/ml was treated and cultured for 8 hours. When treated with ATP, incubated for 5 hours and incubated with 5 mM ATP for the last 1 hour, and each cytokine was measured with the R&D duoset ELISA kit using the supernatant.

First, a capture antibody was coated on a 96-well plate at room temperature, washed three times with a washing buffer the next day, and blocked with a diluent buffer for 1 hour. After repeating the washing process, the prepared sample and standard sample were placed in each well and incubated at room temperature for 2 hours. After the incubation was completed, the washing process was repeated once again, and the detection antibody was added to each well and incubated again at room temperature for 2 hours.

After washing again, streptavidin-HRP was put into each well and incubated for 20 minutes in dark conditions and at room temperature. After the last washing, a substrate solution was added, incubated for about 20 minutes in a dark place, and the reaction was stopped with a stop solution. The concentration of cytokines was calculated by measuring the optical density of each well using a microplate reader (VICTOR) set at 450 nm.

<Experimental Example 6> Quantitative real-time RT-PCR analysis

Abdominal macrophages prepared by the method described above were pretreated with 100 mM trehalose and 500 μM saccharin at the indicated concentrations for 15 minutes, then treated with LPS (InvivoGen) 10ng/ml and cultured for 1 hour. After extracting the mRNA with Trizol, cDNA was synthesized using the reverTraAce qPCR kit (TOYOBO, Japan), mixed with a primer and SYBR Green real time PCR master MIX (TOYOBO), and qPCR with a Roche light cycler 480 (Roche) was performed. The expression level of each gene was calculated by comparing it with the expression level of the control group after normalizing it with the expression level of GAPDH. The primer sequences corresponding to each gene used in the experiment are as follows.

1. TNF-a - Forward: 5’- CCT GTA GCC CAC GTC GTA GC -3’

- Reverse: 5’- TTG ACC TCA GCG CTG AGT TG -3’

2. IL-1b - Forward: 5’- CAT CGG CAT TTT GAA CGA GGT -3’

- Reverse: 5'- CAT GCA GTA GAC ATG GCA GAA -3'

3. IL-6 - Forward: 5’- TGG AGT CAC AGA AGG AGT GGC TAA G -3’

- Reverse: 5’- TCT GAC CAC AGT GAG GAA TGT CCA C -3’

4. IL-10 - Forward: 5’- GTG AAG ACT TTC TTT CAA ACA AAG -3’

- Reverse: 5’- CTG CTC CAC TGC CTT GCT CTT ATT -3’

5. IL-18 - Forward: 5’- TGG AGA CCT GGA ATC AGA CAA C -3’

- Reverse: 5’- TAT TTT CAG GTG GAT CCA TTT C -3’

6. TGF-b - Forward: 5’- CCC GAA GCG GAC TAC TAT GC -3’

- Reverse: 5’- CAT AGA TGG CGT TGT TGC GG -3’

7. GAPDH - Forward: 5’- AGT ATG ATG ACA TCA AGA AGG -3’

- Reverse: 5'- ATG GTA TTC AAG AGA GTA GGG -3'

<Example 1> Confirmation of the effect of trehalose in an animal model of chronic sinusitis

1. Confirmation of the inhibitory effect of trehalose in chronic sinusitis

The effect of trehalose was confirmed in an animal model of chronic sinusitis.

To induce a chronic sinusitis (CRS) mouse model, 0.7U of Aspergillus oryzae protease per mouse and 75ug of ovalbumin (Sigma-Aldrich, St Louis, Mo) were mixed in the nose of the mouse 3 times a week for 5 weeks (total 15 times) ) was injected. In order to confirm the therapeutic effect of trehalose, 50 mg/kg of trehalose (stock 50 mg/mL) per mouse was administered to the nasal cavity 5 hours prior to CRS induction in the same manner as in FIG. 1(A). injected.

After induction of the CRS model, mouse nasal tissue was made into sections and used for staining, H&E staining was performed, Eosinophil infiltration was confirmed using Sirius red staining, and mast cell infiltration Silver was confirmed by staining with toluidine blue.

As a result, as shown in FIGS. 1 (B) and 1 (C), it was confirmed that the thickness of the nasal tissue and the number of eosinophils and mast cells, which are major mediators of chronic sinusitis, decreased.

From the above results, it was confirmed that trehalose had an excellent effect on alleviating inflammation, and it was confirmed that trehalose treatment was more effective than intraperitoneal administration.

2. Confirmation of the inhibitory effect of trehalose on IL-1β production

In an animal model of chronic sinusitis, the inhibitory effect of trehalose on IL-1β production and thus the treatment effect of chronic sinusitis were confirmed.

To detect macrophages and IL-1β-secreting cells in nasal tissue after chronic sinusitis was induced in myeloid immune cell-specific ^{autophagy-deficient Atg7 f/f} ;Lyz2-Cre mice and normal Atg7 ^{f/f mice.} Tissue slides were incubated with primary antibodies against CD68 (1:100, FA-11; AbD Serotec) and IL-1β (1:100, 3A6, Cell Signaling, Danvers, Mass) for 1 hour. Afterwards, secondary antibodies to each primary antibody were Alexa Fluor 488-conjugated goat anti-rat F(ab′)2 (1:250; Jackson ImmunoResearch) and Alexa Fluor 488-conjugated goat anti-mouse F(ab′). 2 (1:250; incubated with Jackson ImmunoResearch for 30 minutes. All antibodies were diluted in PBS containing 1% BSA and 1% goat serum. After all incubation, slides were washed 3 times with PBS. , A cover-glass was mounted with ProLong Gold anti-fade reagent (Molecular Probes, Eugene, Ore) Tissue slides were imaged with an LSM 710 laser scanning confocal microscope (Carl Zeiss, Oberkochen, Germany).

As a result, it was confirmed that trehalose relieved chronic sinusitis regardless of autophagy dysfunction in an animal model of chronic sinusitis using a mouse lacking myeloid immune cell-specific autophagy function, as shown in FIG. 2 . In addition, it was confirmed that the secretion of IL-1β by macrophages, which is a major mediator of chronic sinusitis, was inhibited during treatment with trehalose.

From the above results, it was confirmed that trehalose exhibits an effect of alleviating chronic sinusitis through inhibition of IL-1β production.

<Example 2> Confirmation of the inhibitory effect of inflammatory cytokine expression by treatment with various saccharides

In vitro experiments were performed to confirm the effect of various saccharides on cytokine secretion of macrophages.

First, 3 mL of 3.85% thioglycolate (difco) was injected into the abdominal cavity of an 8-10 week-old mouse, and 3 days later, the peritoneal was washed with 10 mL of Dulbecco's modified Eagle medium (DMEM) and peritoneal macrophages ( peritoneal macrophages) were obtained. The cells were spread on a plastic tissue culture plate and cultured at 37° C. for 3 hours. Cells that did not adhere were washed with fresh DMEM, and only adherent peritoneal macrophages were used for the experiment.

After pretreatment with trehalose, saccharin, acesulfame potassium, sucralose, glucose, sucrose, maltose and lactose (sigma) to the abdominal macrophages at the concentrations shown in the figure for 15 minutes, LPS (InvivoGen) 10ng/ ml and incubated for 8 hours. When treated with ATP, incubated for 5 hours and incubated with 5 mM ATP for the last 1 hour, and each cytokine was measured with the R&D duoset ELISA kit using the supernatant.

As a result of confirming the expression level of inflammatory cytokines in peritoneal macrophages treated with each synthetic sweetener by the above process, TNF-a and IL-10 among various inflammatory cytokines as shown in FIG. On the other hand, it was confirmed that the secretion of IL-1β and IL-6 was significantly decreased.

In addition, it was confirmed that saccharin exhibits the same effect as trehalose among various saccharides, and it was confirmed that acesulfame potassium also exhibited an excellent effect on the inhibition of IL-1β expression.

<Example 3> Confirmation of the inhibitory effect of saccharin on the expression of proinflammatory cytokines

The effect of trehalose and saccharin confirmed in the previous experiment on the expression of pro-inflammatory cytokines was confirmed.

Abdominal macrophages prepared in the same manner as in Example 3 were pretreated with 100 mM trehalose and 500 μM saccharin at the indicated concentrations for 15 minutes, then treated with LPS (InvivoGen) 10ng/ml and cultured for 1 hour. mRNA was extracted and qPCR was performed.

As a result, as shown in FIG. 4, it was confirmed that the cytokine expression inhibitory effect by saccharin was not only at the trehalose and protein levels, but also at the gene expression level, and saccharin was also found in myeloid immune cell-specific autophagy-deficient mice. It was confirmed that the inhibitory effect on the expression of cytokines regardless of autophagy dysfunction.

<Example 4> Confirmation of the inhibitory effect of saccharin on chronic sinusitis

In an animal model of chronic sinusitis, it was confirmed whether saccharin could also have an anti-inflammatory effect.

Saccharin (stock 1mg/mL) 1mg/kg per mouse was injected into the nasal cavity 5 hours before chronic sinusitis induction into the previously prepared chronic sinusitis animal model.

After induction of the CRS model, mouse nasal tissue was prepared in sections and H&E staining was performed. Eosinophil infiltration was confirmed using Sirius red staining, and mast cell infiltration was toluidine blue (toluidine blue). toluidine blue) staining.

As a result, as shown in FIG. 5 , there was a change in the thickness of the nasal tissue of the saccharin-treated experimental animal and a decrease in the number of eosinophils and mast cells, which are major mediators of chronic sinusitis, and it was confirmed that saccharin also exhibited an excellent effect on alleviating inflammation.

From the above results, it was confirmed that the secretion of IL-1β, a major factor in inducing chronic sinusitis in macrophages, was inhibited by treatment with saccharin and acesulfame potassium. It has been confirmed that it can be developed as a therapeutic agent that exhibits excellent therapeutic effects.

<Example 5> Confirmation of psoriasis inhibitory effect of saccharin

The psoriasis improvement effect of saccharin was confirmed in an animal model of psoriasis.

In order to induce a psoriasis animal model in the same manner as in FIG. 6A, Aldara cream (including imiquimod 5%, Dong-A ST) was applied to each ear of each mouse at a rate of 5 mg daily for 8 days, confirming the anti-inflammatory effect of saccharin To do this, a saccharin solution (1mg/ml in tertiary distilled water) was wetted with gauze 1 hour before Aldara cream was applied and fixed with a band-aid so that it was sufficiently absorbed into the ear skin.

By confirming the change in ear thickness, the induction of inflammation and the effect of saccharin were confirmed.

The thickness measurement was performed once every two days before saccharin solution treatment, and the change in ear thickness was confirmed by subtracting the original thickness before inducing the psoriasis animal model from the thickness measured on each day.

swelling = Δthickness (thickness measured on each day - thickness measured on day 0)

As a result, as shown in FIG. 6B , it was confirmed that the ear thickness change of the saccharin solution treatment group (IMQ-sacch) compared to the control group (IMQ-veh) was very small in the animal model induced by Aldara cream for 8 days.

From the above results, it was confirmed that saccharin relieves inflammation in the psoriasis model and has a therapeutic effect on psoriasis.

As described above in detail a specific part of the content of the present invention, for those of ordinary skill in the art, it is clear that this specific description is only a preferred embodiment, and the scope of the present invention is not limited thereby. something to do. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

<110> THE ASAN FOUNDATION & University of Ulsan Foundation For Industry Cooperation <120> Pharmaceutical composition for preventing or treating inflammatory diseases comprising saccharin, acesulfame K or mixture <130> ADP-2020-0169 <160> 14 <170> KoPatentIn 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TNF-a Forward primer <400> 1 cctgtagccc acgtcgtagc 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TNF-a Reverse primer <400> 2 ttgacctcag cgctgagttg 20 <210> 3 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> IL-1b Forward primer <400> 3 catcggcatt ttgaacgagg t 21 <210> 4 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> IL-1b Reverse primer <400> 4 catgcagtag acatggcaga a 21 <210> 5 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> IL-6 Forward primer <400> 5 tggagtcaca gaaggagtgg ctaag 25 <210> 6 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> IL-6 Reverse primer <400> 6 tctgaccaca gtgaggaatg tccac 25 <210> 7 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> IL-10 Forward primer <400> 7 gtgaagactt tctttcaaac aaag 24 <210> 8 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> IL-10 Reverse primer <400> 8 ctgctccact gccttgctct tatt 24 <210> 9 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> IL-18 Forward primer <400> 9 tggagacctg gaatcagaca ac 22 <210> 10 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> IL-18 Reverse primer <400> 10 tattttcagg tggatccatt tc 22 <210> 11 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TGF-b Forward primer <400> 11 cccgaagcgg actactatgc 20 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TGF-b Reverse primer <400> 12 catagatggc gttgttgcgg 20 <210> 13 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> GAPDH Forward primer <400> 13 agtatgatga catcaagaag g 21 <210> 14 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> GAPDH Reverse primer <400> 14 atggtattca agagagtagg g 21

Claims (5)

Contains saccharin, acesulfame potassium or a mixture thereof as an active ingredient, chronic sinusitis, bronchitis, hepatitis, ulcerative colitis, asthma, psoriasis, arthritis, dermatitis, gingivitis, keratitis, atopic dermatitis, gout And a pharmaceutical composition for preventing or treating inflammatory diseases selected from the group consisting of chronic obstructive pulmonary disease (COPD). The pharmaceutical composition for preventing or treating inflammatory diseases according to claim 1, wherein the saccharin, acesulfame potassium, or a mixture thereof inhibits IL-1β secretion. The pharmaceutical composition for preventing or treating an inflammatory disease according to claim 1, wherein the inflammatory disease is induced by IL-1β secreted from macrophages. delete Contains saccharin, acesulfame potassium or a mixture thereof as an active ingredient, chronic sinusitis, bronchitis, hepatitis, ulcerative colitis, asthma, psoriasis, arthritis, dermatitis, gingivitis, keratitis, atopic dermatitis, gout And chronic obstructive pulmonary disease (COPD) inflammatory disease prevention or improvement health food selected from the group consisting of.

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