Anti-inflammatory compositions comprising N-benzyl-N-methyl decan-1-amine or derivatives thereof as active ingredients

Abstract

The present invention relates to an anti-inflammatory composition comprising N-benzyl-N-methyl decane-1-amine or a derivative thereof as an effective ingredient, which can comprise a compound represented by the following formula 1 as an effective ingredient. [Chemical Formula 1] In the formula 1, R1 is hydrogen, fluorine, an alkyl group having a carbon number of 1 to 5, or an alkoxy group having a carbon number of 1 to 5, and R2 and R3 are each independently an alkyl group having a carbon number of 1 to 5.

Description

Technical Field

[0001] This invention relates to an anti-inflammatory composition comprising N-benzyl-N-methyldecane-1-amine or a derivative thereof as an active ingredient. Background Technology

[0002] Inflammation refers to the pathological state of abscess formation caused by the invasion of external bacteria. The inflammatory response is a biological defense process used to repair and regenerate damage caused by the invasion of bacteria that cause certain organic changes in living cells or tissues. When tissues (cells) are damaged or infected by external infectious agents (bacteria, fungi, viruses, various allergens), various inflammatory mediators and immune cells in local blood vessels and body fluids participate in the process, showing a series of complex physiological reactions such as enzyme activation, secretion of inflammatory mediators, invasion of body fluids, cell migration, and tissue destruction, as well as external symptoms such as erythema, edema, fever, and pain.

[0003] Under normal circumstances, the inflammatory response clears external sources of infection, regenerates damaged tissues, and restores bodily functions. However, if antigens are not cleared or if the inflammatory response is excessive or persistent due to internal substances, it can become the main pathological phenomenon of the disease (allergic diseases, chronic inflammation) and can also become an obstacle in the treatment process such as blood transfusion, drug administration, and organ transplantation.

[0004] To alleviate these inflammatory symptoms, nonsteroidal anti-inflammatory drugs such as ibuprofen and indomethacin are used, while steroidal drugs such as dexamethasone are used, but their use is limited due to reported side effects related to stability.

[0005] Therefore, there is a need to develop a safe anti-inflammatory substance with maximum efficacy and minimal side effects. Summary of the Invention

[0006] One object of the present invention is to provide an anti-inflammatory composition comprising N-benzyl-N-methyldecane-1-amine or a derivative thereof as an active ingredient.

[0007] One object of the present invention is to provide a cosmetic or pharmaceutical composition for improving atopic dermatitis, comprising N-benzyl-N-methyldecane-1-amine or a derivative thereof as an active ingredient.

[0008] One object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of rheumatoid arthritis, comprising N-benzyl-N-methyldecane-1-amine or a derivative thereof as an active ingredient.

[0009] One object of the present invention is to provide a pharmaceutical composition for treating ulcerative colitis comprising N-benzyl-N-methyldecane-1-amine or a derivative thereof as an active ingredient.

[0010] An anti-inflammatory composition according to an embodiment of the present invention may contain a compound represented by Formula 1 as an active ingredient.

[0011] [Chemical Formula 1]

[0012]

[0013] In chemical formula 1, R1 is hydrogen, fluorine, an alkyl group having 1 to 5 carbon atoms, or an alkoxy group having 1 to 5 carbon atoms, and R2 and R3 are each independently an alkyl group having 1 to 5 carbon atoms.

[0014] In one embodiment, the anti-inflammatory composition of the present invention may contain a compound selected from chemical formulas 1-1 to 1-5 as an active ingredient.

[0015] [Chemical Formula 1-1]

[0016]

[0017] [Chemical Formula 1-2]

[0018]

[0019] [Chemical Formulas 1-3]

[0020]

[0021] [Chemical Formulas 1-4]

[0022]

[0023] [Chemical Formulas 1-5]

[0024]

[0025] In one embodiment, the composition can be therapeutic for inflammatory diseases including atopic dermatitis, rheumatoid arthritis, or ulcerative colitis.

[0026] In one embodiment, the composition can inhibit the activity of lipopolysaccharide-induced p38 MAP kinase and JNK, and inhibit inflammation-related signaling pathways linked to p38 MAPK-MK2.

[0027] In one embodiment, the composition can inhibit the production of pro-inflammatory cytokines such as TNF-α, IL-1β or IL-6, and inhibit the activity of NF-κB transcription factors.

[0028] In one embodiment, the composition may have antioxidant effects by increasing the expression of heme oxygenase 1 (HO-1) or nuclear factor erythroid-related factor 2 (Nrf2).

[0029] In one embodiment, the composition can inhibit the production of chemokine CINC-3.

[0030] In one embodiment, the composition can be a variety of oral or parenteral formulations. In the preparation of formulations, diluents or excipients are typically used, such as fillers, supplements, binders, wetting agents, disintegrants, or surfactants. Solid dosage forms for oral administration include tablets, pills, powders, granules, capsules, etc., which are prepared by mixing one or more compounds with one or more excipients, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Liquid pharmaceutical formulations for oral administration include suspensions, solutions, emulsions, syrups, etc. Besides commonly used simple diluents such as water and liquid paraffin, liquid formulations can also contain a variety of excipients, such as wetting agents, sweeteners, flavoring agents, and preservatives.

[0031] Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. For non-aqueous solvents and suspensions, propylene glycol, polyethylene glycol, vegetable oils such as soybean oil and olive oil, and injectable esters such as ethyl oleate can be used. For suppository bases, Witepsol, macrogol, Tween 61, cocoa paper, laurel, and glycerin gelatin can be used.

[0032] The compositions according to the invention can be administered to mammals, such as rats, mice, livestock, and humans, via various routes, such as parenteral and oral routes. All possible routes of administration are contemplated; for example, the compositions can be administered orally, rectally or intravenously, intramuscularly, subcutaneously, intrauterinely, or intradurally.

[0033] On the other hand, as another embodiment, the present invention can provide a cosmetic composition for improving atopic dermatitis comprising a compound represented by Formula 2 as an active ingredient.

[0034] [Chemical Formula 2]

[0035]

[0036] In chemical formula 2, R4 is hydrogen, fluorine, an alkyl group having 1 to 5 carbon atoms, or an alkoxy group having 1 to 5 carbon atoms, and R5 is an alkyl group having 1 to 5 carbon atoms.

[0037] In one embodiment, the cosmetic composition for improving atopic dermatitis may contain a compound selected from compounds represented by chemical formulas 1-1, 1-2, 1-3, and 1-5 as an active ingredient.

[0038] In one embodiment, the cosmetic composition may further comprise one or more ingredients selected from the group consisting of skin moisturizing ingredients, cosmetic formulation ingredients, fragrances, preservatives, and purified water. Here, known substances such as soybean oil and olive oil can be used as the skin moisturizing ingredients, cosmetic formulation ingredients, fragrances, and preservatives.

[0039] In one embodiment, the formulation of the cosmetic composition may be a toner, cream, lotion, serum, gel, essence, mask, powder, topical skin ointment, patch, suspension, spray, or beauty essence.

[0040] In another embodiment, the present invention can provide a pharmaceutical composition for the prevention or treatment of rheumatoid arthritis comprising a compound represented by the chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0041] In one embodiment, the pharmaceutical composition for the prevention or treatment of rheumatoid arthritis may contain a compound represented by chemical formula 1-1 or 1-2 as an active ingredient. Preferably, the compound represented by chemical formula 1 may be a compound represented by chemical formula 1-1.

[0042] In one embodiment, the pharmaceutical composition can be used for oral administration, in which case the compound represented by Formula 1 can be prepared into a unit-dose formulation suitable for oral administration at a dose of 15 to 25 mg / kg (200 μl volume, approximately 500 μg per dose).

[0043] In one embodiment, the pharmaceutical composition can be administered orally once or multiple times, and the dosage for a specific patient can be varied according to the patient's weight, age, sex, health status, diet, administration time, administration method, excretion rate, disease severity, etc.

[0044] In one embodiment, oral administration of the pharmaceutical composition for 3 weeks may not affect the activity of AST and ALT enzymes in a rheumatoid autoimmune disease model induced by type 2 collagen.

[0045] Additionally, as another embodiment of the present invention, examples include pharmaceutical compositions for treating ulcerative colitis comprising a compound represented by the chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0046] In one embodiment, the pharmaceutical composition for treating ulcerative colitis may contain a compound represented by chemical formula 1-1 or chemical formula 1-2 as an active ingredient. Preferably, the compound represented by chemical formula 1 may be a compound represented by chemical formula 1-2.

[0047] In one embodiment, the pharmaceutical composition can be used for enema administration. In this case, the pharmaceutical composition, as an active ingredient for one day, can be directly administered as an enema to patients with ulcerative colitis at a dose of less than 0.4 mg / kg body weight (500 μL volume, each dose being approximately 100 μL).

[0048] In one embodiment, the pharmaceutical composition can be administered via enema once or multiple times. The dosage for a specific patient can be varied based on the patient's weight, age, sex, health status, diet, administration time, administration method, excretion rate, disease severity, etc.

[0049] In one embodiment, the pharmaceutical composition may be administered as a pharmaceutically acceptable salt, alone or in combination with other pharmaceutically active compounds, or as a suitable aggregate. The salt can be any pharmaceutically acceptable salt without particular limitation; for example, hydrochloric acid, sulfuric acid, nitric acid, etc., can be used.

[0050] In one embodiment, enema administration of the pharmaceutical composition can inhibit the production of the chemokine CINC-3. Additionally, enema administration of the pharmaceutical composition can reduce the production of reactive oxygen species by macrophages.

[0051] Invention Effects

[0052] The N-benzyl-N-methyldecane-1-amine or its derivatives according to the present invention exhibit excellent anti-inflammatory effects by inhibiting the inflammatory response of cells induced by lipopolysaccharide (LPS), reducing the production of reactive oxygen species in macrophages, and inhibiting the expression of chemokine CNIC-3, pro-inflammatory cytokines, and inflammatory complex proteins. In particular, it has anti-inflammatory effects on inflammatory diseases such as atopic dermatitis, rheumatoid arthritis, and ulcerative colitis.

[0053] Specifically, when N-benzyl-N-methyldecane-1-amine or its derivatives according to the present invention are applied to the skin, they exhibit excellent anti-inflammatory effects on atopic dermatitis induced by 1,4-dinitrochlorobenzne (DNCB) compounds. In addition, they exhibit excellent anti-inflammatory effects on rheumatoid arthritis induced by type 2 collagen by oral administration and excellent anti-inflammatory effects on ulcerative colitis by enema administration. Attached Figure Description

[0054] Figure 1 via H 3 -NMR shows the synthesis results and structures of N-benzyl-N-methyldecane-1-amine (BMDA) and its derivative (DMMA) synthesized according to an embodiment of the present invention.

[0055] Figure 2 The lipopolysaccharide of N-benzyl-N-methyldecane-1-amine (BMDA) and its derivative (DMMA) according to an embodiment of the present invention demonstrates the inhibitory effect of lipopolysaccharide on the inflammatory response of the mononuclear cell line THP-1.

[0056] Figure 3 These are experimental results demonstrating the anti-inflammatory effect of enema administration of N-benzyl-N-methyldecane-1-amine (BMDA) and its derivatives (DMMA) according to an embodiment of the present invention in a DNBS-induced colitis model.

[0057] Figure 4 and Figure 5 These are experimental results demonstrating the anti-inflammatory effects of oral administration of N-benzyl-N-methyldecyl-1-amine (BMDA) and its derivatives (DMMA) according to an embodiment of the present invention in a rheumatoid arthritis model induced by type 2 collagen.

[0058] Figure 6 The expression of inflammatory cytokines and antioxidant genes HO-1 (heme oxygenase-1) and Nrf2 (nuclear factor erythroid-related factor 2) induced by oral administration of N-benzyl-N-methyldecyl-1-amine (BMDA) and its derivatives (DMMA) according to an embodiment of the present invention were investigated by qRT-PCR and immunoblotting, respectively.

[0059] Figure 7The results show the AST and ALT activity measurements of N-benzyl-N-methyldecane-1-amine (BMDA) and its derivatives (DMMA) administered orally according to an embodiment of the present invention in a rheumatoid arthritis model induced by type 2 collagen.

[0060] Figure 8 The lipopolysaccharide of N-benzyl-N-methyldecane-1-amine (BMDA) derivatives (benzyl-decyl-methyl-amine, benzyl-dodecyl-methyl-amine, decyl-(4-fluoro-benzyl)-methyl-amine) according to an embodiment of the present invention is shown to inhibit the inflammatory response of the mononuclear cell line THP-1.

[0061] Figure 9 The results of an evaluation of the efficacy of a cosmetic composition comprising N-benzyl-N-methyldecane-1-amine (BMDA) and its derivatives according to an embodiment of the present invention for improving atopic dermatitis are shown.

[0062] Figure 10 and Figure 11 It is further support Figure 9 A and Figure 9 Detailed results for B. Detailed Implementation

[0063] The various embodiments and experimental examples of the present invention will now be described in detail. However, the following embodiments are merely some examples of the present invention, and the present invention should not be construed as being limited to these embodiments.

[0064] <Example 1: Confirmation of the structure and synthesis of BMDA (FMJ-G0) and its derivative (FMJ-G4)>

[0065] 1 M N-benzylmethylamine and decanal were mixed in methanol and stirred for 20 minutes. Then, sodium triacetoxyborohydride [NaBH(OAc)3] was added and the mixture was stirred for another hour. Under vacuum, the volatile organic solvent was removed, and impure BMDA (benzyl-decyl-methyl-amine) was extracted with ethyl acetate saturated with sodium bicarbonate. Further purification of BMDA was obtained by column chromatography, followed by H₂ chromatography. 3 -NMR confirmed its structure (see Figure 1A The chemical structure of the synthesized substance is represented by the following formula 1-1.

[0066] [Chemical Formula 1-1]

[0067]

[0068] Additionally, in the same manner, FMJ-G4 (DMMA, Decyl-(4-methoxy-benzyl)-methyl-amine) was synthesized starting from 1-(4-methoxyphenyl)-N-methylmethanamine and decanal, and this structure was also obtained via H... 3 - NMR confirmed (see Figure 1B The chemical structure of the synthesized substance is represented by the following formula 1-2.

[0069] [Chemical Formula 1-2]

[0070]

[0071] <Example 2: Evaluation of the anti-inflammatory efficacy of BMDA (FMJ-G0) and its derivatives (DMMA, FMJ-G4)>

[0072] 1. To evaluate the inhibitory effects of lipopolysaccharide (LPS) induced by BMDA (FMJ-GO) and its derivative (DMMA) on inflammation-related signaling activity and its inhibitory effects on TNF-α and IL-β inflammatory cytokines.

[0073] Human THP-1 mononuclear cell lines (5 × 10^5 cells / well) were cultured in 6-well plates with BMDA (FMJ-GO) and DMMA (FMJ-G4) at concentrations of 4 μM, 2 μM, and 1 μM for 3 hours. Each well was then treated with lipopolysaccharide at a concentration of 1 mg / ml. After 1 hour, the cells were centrifuged, and the supernatant was used to evaluate inflammatory cytokines by ELISA. The recovered THP-1 cells were then placed in cell lysis buffer for Western blotting.

[0074] In addition, under the same conditions, mononuclear THP-1 cell lines treated with lipopolysaccharide (LPS) and BMDA / DMMA ​​were divided into cytoplasmic and nuclear portions, and the distribution of NF-κBP65 protein in the cytoplasm and nucleus was examined by immunoblotting.

[0075] See results Figure 2A and 2B, BMDA (FMJ-G0) and its derivative DMMA (FMJ-G4), low molecular weight compounds inhibited LPS-activated p38 MAPK and JNK activity. Furthermore, they also inhibited inflammation-related signaling pathways linked to p38 MAPK-MK2. These results confirmed that these low molecular weight compounds exhibit concentration-dependent activity.

[0076] On the one hand, the biological significance of this result lies in the fact that, through ELISA, the low-molecular-weight substances of BMDA (FMJ-GO) and its derivative DMMA (FMJ-G4) of this invention inhibited the production of TNF-α and IL-1β inflammatory cytokines. Figure 2 C) indicates that these low molecular weight compounds also inhibited the activity of the NF-κB transcription factor and suppressed NF-κB expression and nuclear translocation. Figure 2 D).

[0077] 2. Evaluation of the anti-inflammatory efficacy of BMDA (FMJ-G0) and its derivatives (DMMA, FMJ-G4) in a DNBS-induced colitis model.

[0078] For experimental colitis, male SD rats (250-260g) were fasted the day before the onset of colitis. A water bottle was provided, allowing them to drink only water. Then, after anesthesia, a rubber cannula was inserted into the anus, and a substance obtained by dissolving 48.0 mg of DNBS in 0.4 mL of 50% ethanol was injected through the cannula. This procedure was repeated for 3 days.

[0079] Starting on day 4 after DNBS insertion, BMDA (FMJ-G0) and its derivatives (DMMA, FMJ-G4) were diluted with PBS and injected 500 μL rectally at a concentration of 200 μg / mL after anesthesia. This procedure was repeated for 5 days. As a control group, DMSO was diluted with PBS at the same ratio and injected for the same 5 days. Then, on day 6, the rats were asphyxiated, their large intestines were removed, and the degree of colitis was assessed according to known methods.

[0080] In addition, a portion of the large intestine was taken out and placed in cell lysis buffer (20mM Tris-HCl [pH 7.5], 150mM NaCl, 1mM EDTA, 1% Triton X-100, 0.5% SDS) for sonication. Then, it was separated by centrifugation, and only the supernatant was collected. After quantifying the protein, it was subjected to immunoblotting.

[0081] Then, a portion of the supernatant prepared for immunoblotting after centrifugation was used to measure the level of chemokine CINC-3 (cytokine-induced neutrophil chemoattractant) by ELISA. Additionally, to determine the activity of macrophages introduced into the distal colon, the titer of MPO (myeloperoxidase) was measured. The distal colon fragment was pulverized in an ultrasonic grinder, centrifuged, and hydrogen peroxide was added to the supernatant for decomposition. The change in absorbance was measured at 460 nm for 1 minute.

[0082] See results Figure 3 A-3B, BMDA (FMJ-G0) and its derivative DMMA (FMJ-G4) low molecular weight substances significantly reduced the inflammatory response induced by DNBS to meaningful levels.

[0083] Compared to the large intestine, treatment with DNBS+PBS showed that the large intestine became significantly shorter and the ulcer crusts were considerably larger. Figure 3 (A-3B). However, it can be seen that the large intestine length of rats treated with DMMA (FMJ-G4) was longer than that of the group treated with DNBS+PBS, and the ulcer-induced crusts were smaller. Specifically, rats treated with the derivative DMMA (FMJ-G4) showed a slightly longer large intestine length than those treated with BMDA (FMJ-G0), and the ulcer crusts were also smaller. The large intestine length of rats treated with BMDA (FMJ-G0) did not differ significantly from that of the group treated with DNBS+PBS, but showed a marked improvement in ulcer severity. However, BMDA (FMJ-G0) and DMMA (FMJ-G4) treatments did not restore the weight loss caused by DNBS treatment. Figure 3 C).

[0084] In addition, the colon of mice administered via enema with low molecular weight BMDA (FMJ-G0) and its derivative DMMA (FMJ-G4) showed lower macrophage peroxidase activity compared to the colon of mice treated with DNBS+PBS. Figure 3 D), and it was observed that the production of the chemokine CNIC-3 introduced into neutrophils was also lower ( Figure 3 E).

[0085] 3. Anti-inflammatory effects of BMDA (FMJ-G0) and its derivatives (DMMA, FMJ-G4) in a type 2 bovine collagen-induced rheumatoid arthritis model.

[0086] Rheumatoid arthritis was induced in 7-week-old male DBA / J1 mice. Type II collagen (2 mg / mL) was completely mixed with CFA (Complete Freund's Adjuvant) at a 1:1 ratio, and 0.1 mL (100 μg) of the mixture was injected intradermally 2 cm from the tail base. Two weeks later, IFA was used instead of CFA, and mixed uniformly with type II collagen at a 1:1 ratio. The same amount and volume as the first injection were injected subcutaneously at the tail base. Three days later, the mice were divided into four groups for a second injection.

[0087] In addition, male mice were orally administered BMDA (FMJ-G0) and its derivatives (DMMA, FMJ-G4) at a concentration of 2.5 mg / mL for 21 days in a volume of 200 μL (20 mg / kg body weight), and then the control group was orally administered soybean oil.

[0088] During the experiment, the dorsum of the forelimbs and hindlimbs was measured three times a week using calipers. The participants were then subjected to asphyxiation to prepare joint sites for immunohistochemical staining and Western blotting. Additionally, the spleen was removed and its size was compared. Serum was prepared by drawing blood from the portal vein. The specific methods are as follows.

[0089] 1) Prepare paraffin sections, including those from the ankle joint of the forelimb. First, decalcify the sections, then stain them with H&E at a thickness of 4 μm.

[0090] 2) Add cell lysis buffer and use an ultrasonic homogenizer to homogenize the tissue to prepare for immunoblotting.

[0091] 3) After adding trisol solution to the joint tissue, the tissue was pulverized with an ultrasonic grinder. After centrifugation, the proteins and DNA precipitated by the trisol solution were separated by centrifugation. Then, total RNA was recovered from the supernatant, and c-DNA was synthesized using oligo-dT primers and reverse transcriptase. Quantitative qRT-PCR and immunoblotting were attempted.

[0092] See results Figure 4 Mice A-4B, compared to the control group treated with soybean oil, showed significantly lower paw swelling when orally administered BMDA (FMJ-G0) and DMMA (FMJ-G4). Oral administration of BMDA (FMJ-G0) and DMMA (FMJ-G4) did not reverse collagen-induced rheumatoid arthritis-induced weight loss. Figure 4 B), but compared with the control group, the spleen length of mice treated with BMDA (FMJ-G0) and DMMA (FMJ-G4) was smaller and the weight was also reduced. Figure 4 C-4D).

[0093] In addition, observe Figure 5 In rats A-5B, severe inflammation and synovitis were observed after oral administration of soybean oil following collagen induction and investigation of joint tissue staining. However, these inflammations and synovitis were significantly reduced upon oral administration of BMDA (FMJ-G0) and DMMA (FMJ-G4). In particular, a significant reduction in inflammation was observed upon oral administration of BMDA (FMJ-G0).

[0094] In addition, when the expression of inflammatory cytokines in the joint region was investigated by qRT-PCR, low levels of the inflammatory cytokines TNF-α, IL-1β, and IL-6 were also observed in the spleen of mice treated with soybean oil. Figure 6 A). Conversely, in the joint sites of animals orally administered BMDA (FMJ-G0) and DMMA (FMJ-G4), the expression of two proteins involved in antioxidant responses, HO-1 and Nrf2, was increased. Figure 6 B).

[0095] In particular, by showing enzyme activities similar to those in the serum of untreated control mice, it was confirmed that oral administration of BMDA (FMJ-G0) and DMMA (FMJ-G4) for 3 weeks did not increase the activities of AST and ALT enzymes, which are indicators of hepatotoxicity (see [link to original text]). Figure 7 ).

[0096] <Example 3: Synthesis of BMDA(FMJ-GO) Derivatives and Evaluation of Their Anti-inflammatory Effects>

[0097] Except for changing the reaction materials in Example 1 above, benzyl-decyl-ethyl-amine (FMJ-G1), benzyl-dodecyl-methyl-amine (FMJ-G2), and decyl-4-fluoro-benzyl-methyl-amine (FMJ-G3) derivatives of FMJ-G0 were synthesized by the same method.

[0098] Specifically, the FMJ-G1 derivative was synthesized by mixing N-benzylethylamine and decanal at a concentration of 1 M; the FMJ-G2 derivative was synthesized by mixing N-benzylmethylamine and dodecanal; and the FMJ-G3 derivative was synthesized by mixing 1-(4-fluorophenyl)-N-methylmethanamine and decanal in methanol, via reductive amination. The synthesized substances were then purified by column chromatography. The chemical structures of the synthesized substances are represented by formulas 1-3 to 1-5. (See...) Figure 8 A)

[0099] [Chemical Formulas 1-3]

[0100]

[0101] [Chemical Formulas 1-4]

[0102]

[0103] [Chemical Formulas 1-5]

[0104]

[0105] Then, THP-1 cell lines were pretreated with FMJ-G0 derivatives FMJ-G1, FMJ-G2, and FMJ-G3 at a concentration of 4 μm for 1 hour, followed by culturing for 4 hours with lipopolysaccharide (1 μg / mL). The cell lines were immediately recovered, and the signaling pathways involved in the synthesis of inflammatory cytokines were identified. The results are presented in [Table data would be inserted here]. Figure 8 B.

[0106] The results confirmed that pretreatment with FMJ-G1, FMJ-G2, and FMJ-G3 inhibited the phosphorylation of p38 MAP kinase, JNK, and MK2 by lipopolysaccharide (LPS), and that FMJ-G0 derivatives could suppress the inflammatory response.

[0107] <Example 4: Preparation of cosmetic compositions containing BMDA (FMJ-GO) and its derivatives and evaluation of their efficacy in improving atopic dermatitis>

[0108] 1. Preparation of cosmetic compositions

[0109] Add 0.3g of olive wax to 2mL of soybean oil (Sigma-Aldrich) and place it in a 15ml glass tube. Then, place the tube in a 75°C water bath and vortex every 10 minutes while boiling for 30 minutes to melt the olive wax. Under the same conditions, place 7.7ml of purified water in a 75°C water bath and heat for 30 minutes. Then, pour 7.5ml of purified water into the tube containing the olive wax and vortex thoroughly. Next, add 30ug (0.3%) of BMDA and its derivatives (FMJ-G0, FMJ-G4, FMJ-G1, FMJ-G2, FMJ-G3) synthesized in Examples 1 and 3, mix thoroughly, and cool on ice to prepare a cosmetic cream.

[0110] 2. Evaluation of efficacy in improving atopic dermatitis

[0111] To induce atopic dermatitis in 5-week-old Balb / C mice, the fur on the backs of the mice was shaved. One day later, a 0.5% solution of DNCB (2,4-dinitrochlorobenzne) was prepared by adding it to a solution of acetone and olive oil (3:1). 150 μL was applied each time for 3 consecutive days. Then, the same concentration was applied twice a week to the shaved backs for two weeks. On the other hand, to alleviate the induced atopic dermatitis, a cosmetic cream containing BMDA (FMJ-G0) and its derivatives (FMJ-G4, FMJ-G1, FMJ-G2, FMJ-G3) of the present invention was applied to the backs of the mice with induced atopic dermatitis for 2 weeks, and changes in skin inflammation were observed.

[0112] As a result, Figure 9 As shown in Figure A, cosmetic creams containing BMDA (FMJ-G0) and DMMA (FMJ-G4) significantly reduced DNCB-induced atopic dermatitis, and regrowth of hair was also observed on the relieved skin. Additionally, from... Figure 9 B. It was observed that in DNCB-induced acute atopic dermatitis, immune cells were activated and the spleen appeared enlarged. However, the spleen size of mice with skin treated with a cosmetic cream containing BMDA (FMJ-G0) and DMMA (FMJ-G4) was similar to that of mice without induced atopic dermatitis. Furthermore, the size of the inguinal lymph nodes was also similar to that of the spleen.

[0113] In addition, under the same conditions, a commercially available steroid ointment (Lidomax) (0.15%) was applied to the skin, and the results confirmed similarities in dermatitis severity, spleen size, and lymph node size to those of the control group (vehicle) which showed similarities to those of the cosmetic cream without the active ingredient. Figure 9 AB).

[0114] in addition, Figure 9 Figure C shows the results of applying a cosmetic cream containing the derivative synthesized in Example 3. Figure 9 C. It was observed that application of FMJ-G1 and FMJ-G3 cosmetic creams significantly alleviated atopic dermatitis. However, as with the control group (Vehicle), treatment with FMJ-G2 cosmetic cream did not prevent the progression of acute dermatitis induced by DNCB. These results are clearly closely related to the size of the spleen and lymphatic fluid. That is, from... Figure 9 D confirmed that the spleen and lymph nodes of mice treated with cosmetic cream containing FMJ-G2 increased in size in the same way as those of mice treated with the control group (Vehicle).

[0115] Figure 10 A and Figure 10 B showed that applying BMDA (FMJ-G0) and DMMA (FMJ-G4) to the skin significantly prevented acute dermatitis caused by DNCB and significantly reduced dermal thickness.

[0116] Figure 10 The results of A and B are as follows: Figure 9 As shown, skin tissue was excised from the back of mice in the group, fixed in 10% formaldehyde for 48 hours, paraffin slides were prepared, and then thinned to a thickness of 4 μm. After preparation of thin slides, hematoxylin-eosin staining was performed. Then, the results of whether the damaged skin tissue could be restored by BMDA (FMJ-G0) and DMMA (FMJ-G4) were investigated using an optical microscope equipped with the Leica application (Leica Microsystems, Swistzerland).

[0117] Treatment with BMDA or DMMA (FMJ-G0 / G4) confirmed that inflammation caused by DNCB thinning of the dermis. The dermis is... Figure 10 In photo A, from top to bottom, the first dark purple layer is the epidermis, the pink layer below it is the dermis, the dark layer below that is the subcutaneous layer, and below that is what is equivalent to muscle.

[0118] Figure 10C shows that toluidine blue staining of BMDA (FMJ-G0) and DMMA (FMJ-G4) prevents mast cells from invading the dermis, thereby inhibiting the inflammatory response in the dermis. Figure 10 C is obtained by using toluidine blue solution to... Figure 10 After staining a portion of the thin section prepared in AB, the results of mast cells invading the dermis were observed using an optical microscope according to the described method. By preventing mast cells from invading the dermis, inflammation in the dermis is protected from mast cells.

[0119] Figure 10 D showed that mice treated with BMDA (FMJ-G0) and DMMA (FMJ-G4) had significantly lower concentrations of IgE in their blood, which is closely related to allergic reactions, compared with the control group. Figure 10 D is in Figure 9 Atopic dermatitis was induced by treating mice with BMDA (FMJ-G0) and DMMA (FMJ-G4). Two weeks later, blood (500 μL) was collected from the hepatic veins of mice in each group and kept at room temperature for 30 minutes. The blood was then centrifuged to separate the serum. The serum was then used to measure the IgE concentration associated with the allergic reaction by ELISA.

[0120] Anti-mouse IgE (Pharmingen, CA, USA) was diluted with PBS (pH 7.4) in 96-well plates, with 100 μL coating each well. The plates were then incubated at 4°C for 12 hours. After washing the plates with PBS containing 0.05% Tween, they were blocked with PBS containing 1% BSA, 5% sucrose, and 0.05% NaN3 for 1 hour. After washing several times with washing buffer, recombinant IgE and serum were added, and the mixture was incubated at 37°C for 2 hours. The plates were then washed again, followed by the addition of biotinylated IgE (Pharmingen, CA, USA), and incubation for another 2 hours. After washing the plates, avidin peroxidase was added, and the mixture was incubated at 37°C for 30 minutes. After washing the plates again, ABTS solution was added as a substrate, and the absorbance was measured at 405 nm using an ELISA reader (Epoch2, BioTek, VT, USA). The results indicate that treatment with BMDA or DMMA (FMJ-G0 / G4) also inhibited IgE production associated with allergic reactions.

[0121] Figure 11 Is it like this? Figure 9 After inducing atopic dermatitis, mice were treated with BMDA (FMJ-G0) and DMMA (FMJ-G4). After two weeks, a portion of skin tissue from each group of mice was placed in a trisol solution and centrifuged using an ultrasonic homogenizer. To separate total RNA, proteins and DNA were precipitated in the trisol solution and centrifuged. Total RNA was then recovered from the supernatant, and c-DNA was synthesized using oligo-dT primers and reverse transcriptase. Quantitative data of TNF-α, IL-1β, IL-6, iNOS, and Cox2 transcription levels were obtained by qRT-PCR.

[0122] In the control group skin treated with DNCB, the amounts of transcripts of inflammatory cytokines TNF-α, IL-1β, and IL-6, as well as proteins iNOS and COX2 that produce inflammatory mediators (NO, PDE2), were dramatically increased. Furthermore, treatment with BMDA (FMJ-G0) and DMMA (FMJ-G4) significantly reduced the amounts of these transcripts. Figure 11 AC).

[0123] When the activity of signaling proteins affecting the transcription of these proteins was investigated by immunoblotting, as supporting the previous results, BMDA (FMJ-G0) and DMMA (FMJ-G4) treatments significantly reduced the phosphorylation levels of inflammation-related JNK, p38MAPK, and NF-κB compared to the control group. Figure 11 D).

[0124] Although the present invention has been described above with reference to preferred embodiments, those skilled in the art will understand that various modifications and changes can be made to the invention without departing from the spirit and scope of the invention as set forth in the claims.

Claims

1. The use of the compound in the preparation of a cosmetic composition for improving atopic dermatitis, characterized in that, The compounds comprise compounds represented by the following chemical formulas 1-1, 1-2, 1-3, and 1-5: [Chemical Formula 1-1] ； [Chemical Formula 1-2] ； [Chemical Formulas 1-3] ； [Chemical Formulas 1-5] 。 2. The use according to claim 1, characterized in that, The cosmetic composition further includes one or more of the following: skin moisturizing ingredients, fragrance, preservatives, and purified water.

3. The use according to claim 1 or 2, characterized in that, The cosmetic composition is formulated as a cream, lotion, gel, powder, topical skin ointment, patch, or spray.

4. The use of the compound in the preparation of pharmaceutical compositions for the prevention or treatment of rheumatoid arthritis, characterized in that, The compound comprises compounds represented by the following chemical formulas 1-1 or 1-2: [Chemical Formula 1-1] ； [Chemical Formula 1-2] 。 5. The use according to claim 4, characterized in that, The compound is represented by the following chemical formula 1-1: [Chemical Formula 1-1] 。 6. The use according to claim 4, characterized in that, The pharmaceutical composition is for oral administration.

7. The use according to claim 6, characterized in that, Formulations prepared from compounds represented by chemical formula 1-1 or formula 1-2 in a unit-dose form are suitable for oral administration at a dose of 15 to 25 mg / kg.

8. The use of the compound in the preparation of a pharmaceutical composition for treating ulcerative colitis, characterized in that, The compound comprises compounds represented by the following chemical formulas 1-1 or 1-2: [Chemical Formula 1-1] ； [Chemical Formula 1-2] 。 9. The use according to claim 8, characterized in that, The compound is represented by the following chemical formulas 1-2: [Chemical Formula 1-2] 。 10. The use according to claim 8, characterized in that, The pharmaceutical composition is used for enema administration.

11. The use according to claim 10, characterized in that, The pharmaceutical composition is administered as an effective ingredient for one day via enema to patients with ulcerative colitis at a dose of less than 0.4 mg / kg.

12. The use according to claim 8, characterized in that, Enema administration of the pharmaceutical composition inhibits the production of chemokine CINC-3.

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