KR20230019795A - A composition for treating hypertrophic scar and keloid comprising benzhydrylthio acetamide compounds as an active ingredient - Google Patents

Abstract

The present invention relates to a composition for treating hypertrophic scars and keloids, comprising a benzhydryl thioacetamide compound as an active ingredient. The benzhydryl thioacetamide compound inhibits cell proliferation and an increase in intracellular Ca^(2+) concentration, in fibroblasts, keratinocytes, and vascular endothelial cells that are involved in skin fibrosis, and inhibits the synthesis of collagen, which is the extracellular matrix, and thus has an effect of effectively treating intractable hypertrophic scars and keloids.

Description

A composition for treating hypertrophic scar and keloid comprising benzhydrylthio acetamide compounds as an active ingredient}

The present invention relates to a composition for the treatment of hypertrophic scars and keloids containing a benzhydryl thioacetamide compound as an active ingredient. It has the effect of effectively treating intractable hypertrophic scars and keloids by suppressing the increase in intracellular Ca ²⁺ concentration and cell proliferation in keratinocytes and vascular endothelial cells and suppressing the synthesis of collagen, which is an extracellular matrix.

In general, the process of wound healing in the human body goes through three stages: inflammation, proliferation, and reorganization. First, in the inflammatory stage, the wound is hemostasis, and immune cells, including white blood cells and neutrophils, remove harmful substances and damaged tissues from the outside. In the next proliferation step, vascular endothelial cells migrate to the injured area to form new blood vessels in the damaged tissue, and extracellular tissue-forming proteins such as collagen and elastin are produced, while fibroblasts form granulation tissue.

In the final reconstitution step, proliferated epithelial cells mature and fibrous tissue is formed, increasing adhesion and elasticity between the newly synthesized tissue and the proliferated cells. This wound healing process may take several months to several years depending on the severity of the wound. And in the case of damage to the depth of the dermis layer, collagen proliferates in the dermis layer that maintains the skin's tension, leaving a 'scar', a healing mark on the skin.

If the fibrous tissue is excessively dense during the process of healing a wound, a 'hypertrophic scar' or 'keloid' can be created in which the wound area grows non-growthly. Hypertrophic scars (also called 'hypertrophic scars') do not go beyond the wound area and tend to disappear gradually over time, but keloids grow wider than the wound area over time and invade normal skin. there is

Hypertrophic scars and keloids, which are types of skin fibrosis, occur when cell movement is excessively activated during wound healing, or tissue cells and capillaries are abnormally proliferated and collagen is excessively accumulated. Statistically, it is so common that about 40-70% of surgical patients develop hypertrophic scars or keloids within one year, but a treatment that can effectively suppress them has not yet been developed.

Efforts to develop therapeutic agents capable of effectively inhibiting hypertrophic scars, keloids, or skin fibrosis have been continuously attempted in the past. For example, in Registered Patent No. 10-2026138 (September 23, 2019), a pharmaceutical composition for inhibiting or treating keloid and hypertrophic scar formation containing an expression inhibitor of CRIF1 (CR6 interacting factor 1) gene as an active ingredient is is introduced The CRIF1 is a kind of protein present in mitochondria, and the active ingredient includes an antisense nucleotide or siRNA that specifically binds to the CRIF1 gene.

In addition, in Registered Patent No. 10-2206017 (January 15, 2021), a pharmaceutical composition for preventing or treating skin fibrosis disease containing an inhibitor of expression or activity of PARP1 (poly ADP-ribose polymerase 1) as an active ingredient has been initiated. As the active ingredient, rucaparib used as a treatment for ovarian cancer is exemplified.

In Registered Patent No. 10-2232072 (March 19, 2021), a composition for preventing or treating skin fibrosis comprising a compound represented by the following [Formula A] or a pharmaceutically acceptable salt thereof as an active ingredient is is introduced

[Formula A]

(In Formula A, R1 and R2 are each independently C1-C3 alkyl.)

The [Formula A] compound inhibits the expression of HMGB1 (High-mobility group box 1), collagen I, collagen III, fibronectin and elastin in fibroblasts. In addition, a representative compound among the [Formula A] compounds is ethyl pyruvate modified from pyruvic acid.

On the other hand, in Registered Patent No. 10-1947277 (February 01, 2019), hypertrophic scar formation containing TRAM-34 (1-[(2-chlorophenyl)diphenylmethyl]-1H-pyrazole), a K _Ca 3.1 channel blocker A pharmaceutical composition for the prevention or treatment of is disclosed. The K _Ca 3.1 channel is a type of potassium (K ⁺ ) channel specifically distributed in fibroblasts, and is mainly involved in the activity of fibroblasts.

And in Registered Patent No. 10-1414831 (June 26, 2014), modafinil and its derivatives, which are known to treat narcolepsy, have the effect of suppressing the K _Ca 3.1 channel current, and K _Ca 3.1 channel mediated diseases , that is, it has been reported that it can be used as a treatment for sickle cell anemia, autoimmune diseases, cancer diseases, traumatic brain damage, neurodegenerative diseases, and secretory diarrhea.

In addition, the present inventors registered patent No. 10-2277739 (July 09, 2021) and the corresponding International Publication WO2020 / 055166A1 (March 19, 2020) containing a benzhydryl thioacetamide compound as an active ingredient A composition for treating liver fibrosis or lung fibrosis has been proposed. The benzhydryl thioacetamide compound includes modafinil and has the effect of inhibiting the expression of K _Ca 2.3 channel protein in cell membranes.

Registered Patent No. 10-2026138 (September 23, 2019) Registered Patent No. 10-2206017 (January 15, 2021) Registered Patent No. 10-2232072 (March 19, 2021) Registered Patent No. 10-1414831 (June 26, 2014) Registered Patent No. 10-2277739 (July 09, 2021) International Publication WO 2020/055166A1 (March 19, 2020)

When most cells are stimulated, the concentration of Ca ²⁺ within the cell increases and its function is activated. Ca ²⁺ acts as a secondary messenger in intracellular signal transduction and plays a very important role in cell function control. In the process of skin fibrosis, intracellular Ca ²⁺ plays a very important role in regulating the activation of fibrosis-related cells, the formation of myofibroblasts, and the production and secretion of extracellular matrix. Therefore, inhibiting the increase in intracellular Ca ²⁺ concentration can effectively inhibit skin fibrosis, especially the formation of hypertrophic scars and keloids.

Accordingly, an object of the present invention is to effectively treat the generation of hypertrophic scars and keloids by inhibiting the increase in intracellular Ca ²⁺ concentration and cell proliferation in skin fibrosis-related cells, and further inhibiting the synthesis of collagen, an extracellular matrix. It is to provide a new pharmaceutical composition that can be.

Another object of the present invention is to provide a new pharmaceutical composition that can increase the therapeutic efficacy for hypertrophic scars and keloids by co-administered with TRAM-34, etc., which has the effect of blocking the K _Ca 3.1 channel, as a conventionally developed hypertrophic scar treatment agent. is to do

For reference, the meaning of the term 'treatment' in this specification includes both suppression of the development of a disease, disease or symptom, and alleviation or elimination.

The composition for treating hypertrophic scars and keloids according to the present invention is characterized in that it contains a benzhydryl thioacetamide compound represented by the following [Formula 1] or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula 1]

[In Formula 1, X ₁ to X ₁₀ are hydrogen (H), fluorine (F), or chlorine (Cl), which may be all the same or different; Y is sulfur (S) or sulfoxide (S=O), * denotes a chiral body; R ₁ is hydrogen, hydroxyl group, methyl group, ethyl group, propyl group, isopropyl group, vinyl group, acryl group, butyl group, t-butyl group, isobutyl group, furan group, hydrofuran group, cyclopropane group, oxirane group , Which means any one of a cyclic phenyl group.]

The benzhydryl thioacetamide compound represented by [Formula 1] inhibits an increase in Ca ²⁺ concentration and cell proliferation in fibroblasts, keratinocytes, and vascular endothelial cells, and further inhibits the synthesis of collagen, an extracellular matrix. It is characterized by having an inhibitory effect.

The benzhydryl thioacetamide compound of [Formula 1] according to the present invention has the effect of effectively treating intractable hypertrophic scars and keloids, and, if necessary, combined administration with a conventional hypertrophic scar treatment agent having a potassium channel inhibitory effect It is also expected to be used as a treatment adjuvant or a cosmetic composition.

1 is a graph measuring the inhibitory effect of intracellular Ca ²⁺ increase of CBM-N1 to N10 compounds according to the present invention in fibroblasts;
Figure 2 is a graph measuring the inhibitory effect of intracellular Ca ²⁺ increase of CBM-N1, N3, N5 compounds according to the present invention in keratinocytes and vascular endothelial cells;
Figure 3 is a graph measuring the inhibitory effect of intracellular Ca ²⁺ increase on La ³⁺ , a substance that blocks intracellular Ca ²⁺ influx, in fibroblasts;
Figure 4 is a graph measuring the cell proliferation inhibitory effect of CBM-N1 ~ N5, N8 ~ N10 compounds according to the present invention on fibroblasts;
Figure 5 is a graph measuring the cell proliferation inhibitory effect on CBM-N1 ~ N10 compounds and La ³⁺ according to the present invention in keratinocytes;
Figure 6 is a graph measuring the fibrosis inhibitory effect on CBM-N1 ~ N8 compounds according to the present invention in fibroblasts;
7 is a graph measuring the fibrosis inhibitory effect of the CBM-N1 to N7 compounds according to the present invention in fibroblasts;
8 is a photograph and graph analyzing the effects of CBM-N1, N3, N5, and N9 compounds by observing hypertrophic scars with a skin magnifying glass in a hypertrophic scar model using rabbit ears;
Figure 9 is a photomicrograph and graph analyzing the effects of CBM-N1, N3, N5, and N9 compounds by measuring the epidermal thickness of hypertrophic scars in a hypertrophic scar model using rabbit ears;
Figure 10 is a photomicrograph and graph analyzing the effects of CBM-N1, N3, N5, and N9 compounds by measuring the scar height index (SEI) in a hypertrophic scar model using rabbit ears;
11 is a photomicrograph and a graph analyzing the effects of CBM-N1, N3, N5, and N9 compounds by measuring collagen density in a hypertrophic scar model using rabbit ears.

Hereinafter, the present invention will be described in detail with reference to the accompanying drawings. However, even if the configuration is essential for carrying out the present invention, detailed descriptions of matters that are introduced in the prior art or can be easily implemented by a person skilled in the art from known technologies will be omitted.

The benzhydryl thioacetamide compound of [Formula 1] according to the present invention specifically includes the following CBM-N1 to CBM-N10 compounds. Here, CBM-N1 to N10 are code names given by the present inventors, and each chemical name is as follows.

1) CBM-N1; 2-(Benzhydrylsulfinyl)acetamide

2) CBM-N2; 2-(Benzhydrylthio)-N-[(tetrahydrofuran-2-yl)methyl]acetamide

3) CBM-N3; 2-(Benzhydrylthio)-N-phenylacetamide

4) CBM-N4; 2-(Benzhydrylsulfinyl)-N-methylacetamide

5) CBM-N5; 2-(Benzhydrylsulfinyl)-N-[(tetrahydrofuran-2-yl)methyl]acetamide

6) CBM-N6; 2-(Benzhydrylthio)-N-methylacetamide

7) CBM-N7; 2-[bis(2-fluorophenyl)methanesulfinyl]acetamide

8) CBM-N8; 2-[bis(3-fluorophenyl)methanesulfinyl]acetamide

9) CBM-N9; 2-[bis(4-fluorophenyl)methanesulfinyl]acetamide

10) CBM-N10; 2-[bis(4-chlorophenyl)methanesulfinyl]acetamide

The CBM-N1 compound is known under the general name of 'modafinil', and the CBM-N9 compound is known under the general name of 'lauflumide'. In addition, all of the CBM-N1 to N10 compounds exhibit almost similar pharmacological mechanisms in the human body.

Various methods for preparing the CBM-N1 compound are known, and commercially available raw materials may be purchased and used. In addition, methods for preparing the CBM-N2 to N6 compounds are known in Korean Patent Registration No. 10-1345860 and the like, and methods for preparing the CBM-N7 to N9 compounds are known in International Publication WO 2020/055166A1.

The CBM-10 compound is a novel compound and can be prepared by the same method as the CBM-9 compound. However, bis(4-chlorophenyl)methanol is used instead of bis(4-fluorophenyl)methanol as a starting material. ¹ H NMR (DMSO-d6) of the CBM-10 compound prepared by this method was 7.3-7.7 ppm (m, 8H), 5.4 ppm (s, 1H), 3.45 ppm (d, 1H), 3.2 ppm (d, 1H).

In addition to the CBM-N1 to N10 compounds, 2-(benzhydrylsulfinyl)-N-hydroxyacetamide known under the general name adrafinil is modafinil as a precursor of the CBM-N1 compound, that is, modafinil. It has the same pharmacological action as (CNS Drug Reviews Vol5, No.3 193-212, 1999).

The [Formula 1] compound inhibits an increase in intracellular Ca ²⁺ concentration in fibroblasts, keratinocytes, and vascular endothelial cells involved in skin fibrosis. Intracellular Ca ²⁺ promotes the formation of hypertrophic scars or keloids by being involved in the activation of fibrosis-related cells, the formation of myofibroblasts, and the production and secretion of extracellular matrix.

In addition, the [Formula 1] compound inhibits the function of keratinocytes that play an auxiliary role in the skin fibrosis process. Keratinocytes secrete various growth factors and stimulate fibroblasts to produce growth factors. It also stimulates the formation of myofibroblasts, contributing to the formation of hypertrophic scars and keloids.

In addition, the [Formula 1] compound inhibits the function of vascular endothelial cells that play an auxiliary role in the skin fibrosis process. Vascular endothelial cells contribute to the growth of fibroblasts by regenerating blood vessels and facilitating blood supply necessary for the growth of fibroblasts.

The pharmaceutical composition according to the present invention includes a pharmaceutically acceptable salt of the compound [Formula 1]. Here, the term 'pharmaceutically acceptable salt' includes conventional metal salts, salts with organic bases, salts with inorganic acids, salts with organic acids, salts with basic or acidic amino acids, and the like.

The pharmaceutical composition according to the present invention includes both solvates and hydrates of the compound of Formula 1, and may also include all possible stereoisomers. The solvates, hydrates and stereoisomers can be prepared using conventional methods. In addition, it includes a crystalline form or an amorphous form of the compound [Formula 1], and when prepared in a crystalline form, it may be optionally hydrated or solvated.

The pharmaceutical composition according to the present invention may be prepared in various formulations by itself or by mixing with pharmaceutically acceptable carriers, excipients, diluents, etc., by methods known in the pharmaceutical field. For example, oral formulations such as powders, granules, tablets, capsules, syrups, and emulsions, or formulations such as external preparations or injections may be prepared.

In particular, considering that the use of the composition according to the present invention is the treatment of hypertrophic scars and keloids generated in skin wounds, it is more preferable that it is in the form of an external preparation for skin or an injection that can be administered locally. Examples of the external skin preparation include solutions, ointments, creams, lotions, sprays, patches, gels, aerosols, and the like.

Compositions according to the present invention can be administered in pharmaceutically effective doses. This effective dosage takes into consideration all factors including the type, size and depth of wound, age and sex of the patient, activity of the drug, sensitivity to the drug, administration time, route of administration and excretion rate, duration of treatment, drugs used concurrently, etc. can be determined by

A preferred daily effective dose of the [Formula 1] compound may be 0.2 to 20 mg/kg, specifically 0.5 to 10 mg/kg, and may be administered once or twice a day, but is not limited thereto.

Hereinafter, the treatment efficacy of the benzhydryl thioacetamide compound according to the present invention for hypertrophic scars and keloids was tested in the following manner.

[ In vitro test using fibroblasts, keratinocytes and vascular endothelial cells ]

1) Test method

Fibroblast NIH-3T3 (CRL-2795; American Type Culture Collection, VA, USA) and vascular endothelial cells of mouse aorta were cultured in Dulbecco's Modified Eagle Medium (Hyclone, Logan, UT), and MV2 medium (PromoCell HaCaT keratinocytes were cultured at HaCaT GmbH.

The fibroblasts were exposed to TGF-β (5ng/ml), a fibrosis-inducing substance, and the keratinocytes were exposed to ATP (10 μM), respectively, to increase the intracellular Ca ²⁺ concentration, that is, [Ca ²⁺ ] _i .

When [Ca ²⁺ ] _i reached a stable state, the CBM-N1 to N10 compounds of the present invention were administered and the effect on [Ca ²⁺ ] _i was observed.

Meanwhile, the measurement method of [Ca ²⁺ ] _i is as follows. First, cells were cultured on cover glass in a cell culture plate (12 well), and Fura-2/AM (2 μM), a Ca ²⁺ sensitive dye, was added to the culture medium, and Fura-2/AM was incubated for 25 minutes at 37°C. penetrated into these cells.

Cells were alternately illuminated with 360 and 380 nm light sources at 50 Hz using a photometric Ca ²⁺ measuring device (Photon Technology International Inc) in an inverted microscope (DM IRB, Leica, Germany). Fluorescence emitted from the cells was measured at 510 nm, and the intracellular Ca ²⁺ concentration was calculated from the result. In addition, La ³⁺ (10 μM) was used to block the inflow of Ca ²⁺ through various Ca ²⁺ channels.

The fibroblasts and keratinocytes were exposed to TGF-β (5ng/ml), TGF-β+CBM-N1~N10, and TGF-β+La ³⁺ for 24 hours, and then cells were analyzed by MTT assay or CCK-8 assay method. The effect on proliferation was analyzed.

In addition, the fibroblasts were exposed to TGF-β (5ng/ml) and TGF-β+CBM-N1-N7 for 24 hours, and fibrosis markers α-smooth muscle actin (α-SMA), collagen 1α (Col1α), The expression level of collagen 3α (Col3α) was analyzed by Western blot or PCR.

2) Test results

Attached Figure 1 shows the increase in intracellular Ca ²⁺ concentration ([Ca ²⁺ ] _i ) by TGF-β (5ng/ml) in fibroblasts and the effect of CBM-N1 to N10 on it (mean ± SE , n = 5. *** < 0.001).

Attached Figure 2 shows the increase in [Ca ²⁺ ] _i by ATP (10 μM) in keratinocytes (A and B) and vascular endothelial cells (C and D) and the effect of CBM-N1, N3, and N5 on it (expressed as mean ± SE, n = 5. *** < 0.001).

Attached Figure 3 shows the increase in [Ca ²⁺ ] _i by TGF-β (5ng/ml) in fibroblasts and the effect of La ³⁺ (10 μM) on it (expressed as mean ± SE, n = 5 *** < 0.001). The La ³⁺ (10 μM) is known to block the influx of Ca ²⁺ into cells through various Ca ²⁺ channels.

Attached Figure 4 shows the effect of CBM-N1-N5, N8-N10 compounds in blocking relative proliferation by TGF-β (5ng/ml, 24-hour exposure) in fibroblasts (mean±SE). marked, n = 5. * < 0.05, ** < 0.01). The cell proliferation was measured by MTT assay.

Attached Figure 5 shows the effect of CBM-N1 to N10 compounds to block cell proliferation by TGF _-β (5ng/ml, 24-hour exposure) in keratinocytes (expressed as mean ± SE, n = 5. * ** < 0.001). The cell proliferation was measured by CCK-8 assay.

Attached Figure 6 shows that the CBM-N1 compound (1 μM) in fibroblasts exposed to TGF-β (5 ng/ml) for 24 hours reduced α-smooth muscle actin (α-SMA), collagen 3α (Col3α), and collagen 1α (Col1α). It shows the effect on the expression of (expressed as mean ± SE, n = 5. ** < 0.01). Expression of the α-SMA, Col3α, and Col1α proteins was measured by Western blotting.

Attached Figure 7 shows the effect of CBM-N1 to N7 compounds (1 μM) on the expression of α-SMA and Col1α in fibroblasts exposed to TGF-β (5 ng/ml) for 24 hours (expressed as mean±SE). , n=5. ** < 0.01). Expression of α-SMA and Col1α mRNA was measured by RT-PCR.

3) Evaluation and analysis

As shown in FIGS. 1, 2, 4, and 5, the CBM-N1 to N10 compounds of the present invention are intracellular by TGF-β or ATP in fibroblasts, keratinocytes, and vascular endothelial cells, which are fibrosis-related cells. It has been shown to effectively suppress the increase in Ca ²⁺ concentration and cell proliferation.

In addition, as shown in FIGS. 3 and 5 , it was confirmed that the increase in cell proliferation was suppressed when the influx of Ca ²⁺ into cells was blocked using La ³⁺ . These results indicate that the CBM-N1-N10 compounds of the present invention inhibit the influx of Ca ²⁺ into cells and thereby inhibit cell proliferation.

And as shown in Figures 6 and 7, the CBM-N1 ~ N7 compounds suppressed the increase in the expression of α-SMA, Col1α or Col3α by TGF-β, a fibrotic substance. It is known that intracellular Ca ²⁺ increase in fibroblasts plays an important role in the formation of extracellular matrix such as α-SMA and collagen. Therefore, the suppression of the expression of α-SMA, Col1α or Col3α by the CBM-N1-N7 compounds of the present invention is presumed to be the result of inhibition of Ca ²⁺ increase.

For reference, the most important step in the fibrosis process is the formation of myofibroblasts, and the fibrosis process occurs only when myofibroblasts are formed. Myofibroblasts are formed through activation of fibroblasts or epithelial mesenchymal transition of vascular endothelial cells and smooth muscle cells.

α-SMA is a marker of myofibroblasts, and an increase in α-SMA expression indicates that myofibroblasts are generated. When myofibroblasts are generated, collagen production increases. Therefore, the increase in α-SMA expression and collagen formation means the formation of myofibroblasts, which are a very important basis for the fibrotic process.

The test results as described above mean that the benzhydryl thioacetamide compound of [Formula 1] according to the present invention inhibits the activation of fibroblasts and consequently has the effect of inhibiting skin fibrosis.

[ In vivo test using rabbit hypertrophic scar model ]

1) Preparing the test model

Eight female New Zealand White Rabbits of 2.6-3 kg were anesthetized, and the epidermis, dermis, and perichondrium of rabbit ear skin were circularly removed using an 8 mm biopsy punch to create wounds. Three such wounds were created per rabbit's ears. The surgical site of the rabbit was disinfected, and hypertrophic scars were induced through natural healing for 3 weeks.

After 3 weeks of natural healing, complete epithelialization of the scar area and generation of hypertrophic scars were confirmed, and treatment was started for each test group. The test groups are normal group (normal tissue), control group (untreated wound, hypertrophic scar tissue), test group 1 (solvent application), test group 2 (CBM-N1 10%), test group 3 (CBM-N3 10%) %), test group 4 (CBM-N 10%), and test group 5 (CBM-N 10%).

For each test group, 0.5mL of the drug was applied and treated every day for 2 weeks (14 days), and tissue necropsy was performed. The biopsied tissue was stored in a 10% formalin solution to prepare a tissue slide. The effectiveness of hypertrophic scar treatment was confirmed through skin magnification and histological evaluation.

2) Visual evaluation

The skin tissues of the control and test groups were observed using a skin magnifying glass on the treatment start and treatment end days (14th day of treatment), and scores were given for visual evaluation according to the degree of hypertrophic scar formation as shown in [Table 1]. .

score Degree of formation of hypertrophic scars 0 normal scar One Unrecognizable low height and slight hard scars 2 Slightly convex and hard scar 3 A hard scar that is convex and severely felt

As shown in FIG. 8, there was no significant difference in the visual evaluation scores between the control group and each test group at the time point (Day 0) when hypertrophic scars were formed by natural healing for 3 weeks after the wound. This means that hypertrophic scars were similarly produced in each test group.

However, at the end of the 2-week treatment (Day 14), the evaluation scores were significantly lower in the test groups treated with CBM-N1, N3, N5, and N9 compounds than in the control group and the solvent treatment group (Vehicle). . A low score means that the size of the hypertrophic scar has decreased to be close to that of a normal scar.

3) Analysis of epidermis thickness

A paraffin block of the fixed rabbit ear tissue was prepared and sectioned at a thickness of 3 μm to prepare a tissue slide. After the hydration process, staining was performed with Hematoxylin and Eosin solutions. Stained tissue slides were photographed under a microscope at 200 magnification, and epidermal thickness was measured using ZEISS ZEN Microscope Software.

As shown in FIG. 9, the control group (Control) and the solvent treatment group (Vehicle) with hypertrophic scars had a significant increase in epidermal thickness compared to normal skin (Normal) (p<0.05), but CBM-N1 and N3 , N5, and N9 treated groups showed an average thinner epidermal thickness compared to the control and solvent treated groups.

4) Scar Elevation Index Analysis

The fixed rabbit ear tissue was stained with Hematoxylin and Eosin solutions in the same way as the epidermal thickness analysis, and the stained tissue slides were photographed at 50x magnification. Using ZEISS ZEN Microscope Software, the length from epidermis to cartilage was measured for the scar area and normal skin, and the scar height index (SEI) was calculated from the results. Here, the scar height index (SEI) is a value obtained by dividing the ‘length from the epidermis to the cartilage at the thickest point in the scar area’ by the ‘length from the epidermis to the cartilage in the normal skin area’.

As shown in FIG. 10, the control group (Control) and the solvent treatment group (Vehicle) with hypertrophic scars had significantly increased SEI values compared to normal skin (Normal) (p<0.05), but CBM-N1, N3, In the N5 and N9 treatment groups, average lower SEI values were observed than the control group and the solvent treatment group.

5) Collagen density analysis

A paraffin block was prepared using the fixed rabbit ear tissue, and a tissue slide was prepared by sectioning at a thickness of 3 μm. After the hydration process, dyeing and washing were performed with Biebrich Scarlet-Acid Fuchsin solution for 5 minutes, and then dyed with Phosphotungstic / Phosphomolybdic acid for 5 minutes.

Subsequently, staining was performed with aniline blue and acetic acid solution, and the stained tissue slides were photographed at 400x magnification. The ratio of the area occupied by collagen, that is, the area stained in blue among the total area in the photographed image, was analyzed by Image J (The National Institutes of Health, USA).

As shown in FIG. 11, the control group (Control) and the solvent treatment group (Vehicle) with hypertrophic scars had a significant decrease in collagen density compared to normal skin (Normal), and the CBM-N1, N3, N5, and N9 treatment groups A lower collagen density was observed on average compared to the control and solvent-treated groups.

6) Evaluation and Analysis

As shown in FIG. 8, as a result of treating the hypertrophic scar formed while the rabbit ear wound was naturally healed for 3 weeks with the CBM-N1, N3, N5, and N9 compounds, the size of the hypertrophic scar was significantly larger than that of the control group or the solvent-treated group. decreased. In addition, in the test results of FIGS. 9, 10, and 11, the epidermal thickness, scar height index, and collagen density were relatively decreased in the test groups treated with the CBM-N1, N3, N5, and N9 compounds.

These test results mean that the CBM-N1, N3, N5, and N9 compounds of the present invention reduce collagen density and epidermal thickness and consequently have the effect of suppressing the generation of hypertrophic scars.

[ conclusion ]

The CBM-N1 to N10 compounds representing the [Formula 1] compounds of the present invention inhibit the increase in intracellular Ca ²⁺ concentration to a similar level in all fibroblasts, keratinocytes, and vascular endothelial cells, which are fibrosis-related cells, and furthermore, cells It was confirmed that there is an effect of inhibiting the proliferation of α-SMA and the synthesis of collagen, an extracellular matrix. In addition, it was confirmed that the CBM-N1, N3, N5, and N9 compounds have an effect of inhibiting the generation of hypertrophic scars caused by wounds in a hypertrophic scar model in rabbits.

Although there is a limitation that the same efficacy test could not be conducted for all [Formula 1] compounds in realistic conditions, according to pharmacokinetic experience, other [Formula 1] compounds excluded from the test are the same as the CBM-N1 to N10 compounds. or expected to exhibit similar pharmacological actions. Therefore, it can be said that all of the [Formula 1] benzhydryl thioacetamide compounds according to the present invention have an effect of effectively inhibiting or treating the formation of hypertrophic scars and keloids.

Claims (7)

A composition for the treatment of hypertrophic scars and keloids, characterized in that it comprises a benzhydryl thioacetamide compound represented by the following [Formula 1] or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

[In Formula 1, X ₁ to X ₁₀ are hydrogen (H), fluorine (F), or chlorine (Cl), which may be all the same or different; Y is sulfur (S) or sulfoxide (S=O), * denotes a chiral body; R ₁ is hydrogen, hydroxyl group, methyl group, ethyl group, propyl group, isopropyl group, vinyl group, acryl group, butyl group, t-butyl group, isobutyl group, furan group, hydrofuran group, cyclopropane group, oxirane group , Which means any one of a cyclic phenyl group.]

The method of claim 1, wherein the [Formula 1] compound has an effect of inhibiting an increase in Ca ²⁺ concentration and cell proliferation in fibroblasts, keratinocytes, and vascular endothelial cells, and inhibiting the synthesis of collagen, which is an extracellular matrix. Characterized in that it has, a composition for treating hypertrophic scars and keloids.
The method of claim 1, wherein the [Formula 1] compound is 2- (benzhydrylsulfinyl) acetamide; 2-(benzhydrylthio)-N-[(tetrahydrofuran-2-yl)methyl]acetamide; 2-(benzhydrylthio)-N-phenylacetamide; 2-(benzhydrylsulfinyl)-N-methylacetamide; 2-(benzhydrylsulfinyl)-N-[(tetrahydrofuran-2-yl)methyl]acetamide; 2-(benzhydrylthio)-en-methylacetamide; 2-[bis(2-fluorophenyl)methanesulfinyl]acetamide; 2-[bis(3-fluorophenyl)methanesulfinyl]acetamide; 2-[bis(4-fluorophenyl)methanesulfinyl]acetamide; 2-[bis(4-chlorophenyl)methanesulfinyl]acetamide; A composition for treating hypertrophic scars and keloids, characterized in that it is any one selected from 2-(benzhydrylsulfinyl)-N-hydroxyacetamide.
The composition for treating hypertrophic scars and keloids according to claim 1, wherein the [Formula 1] compound is 2-(benzhydrylsulfinyl)acetamide (modafinil).
The composition for treating hypertrophic scars and keloids according to claim 1, wherein the [Formula 1] compound is 2-(benzhydrylsulfinyl)-N-hydroxyacetamide (adrafinil).
The composition for treating hypertrophic scars and keloids according to claim 1, wherein the [Formula 1] compound is 2-[bis(4-fluorophenyl)methanesulfinyl]acetamide (lauflumide).
The composition for treating hypertrophic scars and keloids according to claim 1, wherein the [Formula 1] compound is 2-[bis(4-chlorophenyl)methanesulfinyl]acetamide.

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