A complex composition for preventing or treating hearing loss comprising sarpogrelate and vaccinium myrtillus extract as active ingredients

Abstract

This invention relates to a composite composition for the prevention or treatment of hearing loss, comprising sargrelate and blueberry extract as active ingredients. The composite composition has been shown to effectively inhibit hair cell death caused by reactive oxygen species generated by hydrogen peroxide at low doses and exhibits excellent protective effects on auditory hair cells in zebrafish. Therefore, this composite composition can be effectively used as a medicine or health food for the prevention or treatment of hearing loss.

Description

Technical Field

[0001] This disclosure relates to a composition of a compound formulation for the prevention or treatment of hearing loss, the compound formulation comprising sarpogrelate and Vaccinium myrtillus extract as active ingredients. Background Technology

[0002] Hearing loss or slowing is a common condition affecting approximately 15%-20% of the population, but it has a significant impact on quality of life. In recent years, due to aging and the widespread use of digital devices, the number of people with hearing loss is rapidly and gradually increasing. Because hearing loss is permanent and difficult to cure, it is very important to prevent hearing loss before it occurs.

[0003] The ear is divided into the outer ear (from the tympanic membrane to the ear flap), the middle ear (from the tympanic membrane to the cochlea), and the inner ear (the inside of the cochlea). Hearing loss caused by diseases in the outer and middle ear can be improved once the disease is treated. However, even with treatment, it is difficult to recover hearing loss caused by diseases in the inner ear and auditory nerve system.

[0004] Hearing loss can be classified into age-related hearing loss, childhood hearing loss, Ménière's disease, sudden hearing loss, noise-induced hearing loss, and ototoxic hearing loss. The most common forms are noise-induced hearing loss and ototoxic hearing loss, which are attributed to noise and drugs. In recent years, noise-induced hearing loss has increased dramatically.

[0005] Human auditory organs are affected by noise levels exceeding 75 dBA, equivalent to living in an industrial society where everyone is exposed to harmful noise. Furthermore, the frequent exposure to loud sounds due to headphone use increases the number of people with noise-induced hearing loss across all age groups. Noise-induced hearing loss in youth tends to worsen with age, affecting people of all ages.

[0006] Furthermore, it is known that ototoxic hearing loss occurs due to cell apoptosis induced by reactive oxygen species, which are produced by ototoxic drugs.

[0007] Although studies on the prevention and treatment of noise-induced hearing loss and ototoxic hearing loss have primarily used antioxidants, no drug has yet shown unique preventative effects, and the safety of many drugs remains unproven. Summary of the Invention

[0008] Technical goals

[0009] The purpose of this disclosure is to provide a composition comprising sugrel ester and European blueberry extract as active ingredients, as a pharmaceutical composition for the prevention or treatment of hearing loss caused by reactive oxygen species generated by noise or drug exposure.

[0010] Technical solution

[0011] This disclosure provides a pharmaceutical composition for the prevention or treatment of hearing loss, the pharmaceutical composition comprising sargrelate and blueberry extract as active ingredients.

[0012] Beneficial effects

[0013] According to this disclosure, a composition comprising a composite composition wherein sargrelate and blueberry extract are mixed in a weight ratio of 1:(0.125-8) as active ingredients, effectively inhibiting hair cell death caused by reactive oxygen species generated by hydrogen peroxide at low doses, and exhibiting excellent protective effects on zebrafish auditory hair cells, thereby providing a composition containing sargrelate and blueberry extract as active ingredients for the prevention or treatment of sensorineural hearing loss. Attached Figure Description

[0014] Figure 1 ECs of hair cells obtained as experimental results demonstrating the inhibitory effect of apoptosis in auditory cell lines are shown. 50 The results of the value determination.

[0015] Figure 2 The results of efficacy tests on hair cells in a zebrafish pharmaceutical efficacy model of a composite composition according to the compositional proportions of this disclosure are shown.

[0016] Figure 3 The results of a comparative study show the hearing-improving efficacy of sargrelate, European blueberry extract, and the compound composition of this disclosure in an in vivo mouse model with noise-induced hearing loss. Detailed Implementation

[0017] The disclosure is described in more detail below.

[0018] Since noise and ototoxic drugs generate reactive oxygen species (ROS), and the ROS generated thereby induce apoptosis of auditory cells, leading to noise-induced hearing loss and ototoxic hearing loss, the inventors of this disclosure made this disclosure by discovering that, in the course of research on a more effective treatment for hearing loss caused by ROS, a composite composition containing sugrel ester and blueberry extract in an optimal weight ratio as active ingredients exhibits excellent protective effects on auditory cells exposed to ROS, even at low doses.

[0019] This disclosure may provide a pharmaceutical composition for the prevention or treatment of hearing loss, the pharmaceutical composition comprising sargrelate and blueberry extract as active ingredients.

[0020] More specifically, hearing loss can be selected from the group consisting of noise-induced hearing loss, ototoxic hearing loss, age-related hearing loss, and sudden hearing loss, and as an example, hearing loss can be ototoxic hearing loss induced by hydrogen peroxide or reactive oxygen species (ROS) or noise-induced hearing loss.

[0021] The pharmaceutical composition can be a composite composition comprising sarpogrelate and blueberry extract in a weight ratio of 1:(0.125-8). Preferably, it can be a composite composition in a weight ratio of sarpogrelate and blueberry extract of 1:(2-4), and more preferably, a composite composition in a weight ratio of sarpogrelate and blueberry extract of 1:3.

[0022] The composite composition can exhibit a greater improvement in hearing than a simple increase in the efficacy value of a product composed of a single component such as sarpogrelate or bilberry extract. When the weight ratio of sarpogrelate to bilberry extract is less than 1:0.125, it is difficult to expect a synergistic effect as a composite composition due to the low content of bilberry relative to 300 mg (recommended daily dose) of sarpogrelate. Even when the weight ratio of sarpogrelate to bilberry extract is greater than 1:8, it is still difficult to expect a synergistic effect as a composite composition due to the low content of sarpogrelate relative to 510 mg (recommended daily dose) of bilberry.

[0023] The blueberry extract used in this disclosure is an extract of the fruit / leaf parts of blueberry using water, C1-C4 alcohols, or a mixture thereof as the extraction solvent, and the method for preparing the extract includes: extracting with solid powder of blueberry fruit / leaf parts using 5 to 30 times the extraction solvent by cold precipitation, hot precipitation, and percolation; concentrating the extract; and then processing it into powder by spray drying or freeze drying in the form of adding a soft extract, a soft extract alone, or a soft extract together with an excipient (such as dextran).

[0024] Sargrelate, one of the components disclosed herein, is (-)-4-[1-dimethylamino-3-[2-[2-(3-methoxyphenyl)ethyl]phenoxy]propyl-2-yl]oxy-4-oxobutyric acid, which is a drug having a structure consisting of C 24 H 31NO6 represents the molecular structure and is typically in hydrochloride form. The component sargrelate disclosed herein is not limited to hydrochloride and may include any salt, including free base or hydrochloride.

[0025] [Chemical Formula]

[0026]

[0027] The pharmaceutical composition can inhibit apoptosis of auditory cells and increase the number of hair cells.

[0028] According to exemplary embodiments of the present disclosure, it was observed that the compound extract showed up to 21.5% inhibition of apoptosis in auditory cell lines at the same concentration compared with administration of European blueberry extract alone, and further improved efficacy by up to 22% in comparison with the number of hair cells in zebrafish administered with sargrelate alone.

[0029] Specifically, Example 2, in which saprogallol and blueberry extract were mixed in a 1:3 weight ratio, showed 21.6% p and 28.8% p higher than saprogallol at doses of 50 mpk and 100 mpk under click conditions in 14-day-old mice auditory brainstem response tests, and 19.9% ​​p and 24.5% p higher than blueberry extract at the same doses, respectively, for threshold improvement.

[0030] In an exemplary embodiment of this disclosure, a pharmaceutical composition for the prevention or treatment of hearing loss comprising sargrelate and blueberry extract as active ingredients may be made from any one of the group consisting of injections, granules, powders, tablets, pills, capsules, suppositories, gels, suspensions, emulsions, drops, or liquids prepared according to conventional methods.

[0031] In another exemplary embodiment of this disclosure, the pharmaceutical composition may further comprise one or more suitable additives selected from the group consisting of carriers, excipients, disintegrants, sweeteners, coating agents, swelling agents, lubricants, flavoring agents, antioxidants, buffers, antibacterial agents, diluents, dispersants, surfactants, binders, and friction reducers commonly used in the preparation of pharmaceutical compositions.

[0032] Specifically, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylparaben, propylparaben, talc, magnesium stearate, and mineral oil can be used as carriers, excipients, and diluents for oral administration of solid dosage forms, including tablets, pills, powders, granules, and capsules. These solid dosage forms can be prepared by mixing a composition with at least one excipient, such as starch, calcium carbonate, sucrose, lactose, and gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Orally administered liquid dosage forms include suspensions, solutions, emulsions, and syrups, and can contain various excipients such as wetting agents, sweeteners, flavorings, and preservatives, in addition to commonly used simple diluents (e.g., water and liquid paraffin). Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized formulations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oils (such as olive oil), and injectable esters (such as ethyl oleate) can be used as non-aqueous solvents and suspending agents. Wipedesol, polyethylene glycol (macrogol), Tween 61, cocoa butter, laurin fat, and glycerin gelatin can be used as the base for suppositories.

[0033] According to exemplary embodiments of this disclosure, the pharmaceutical composition is administered to a subject in a conventional manner via intravenous, intra-arterial, intraperitoneal, intramuscular, intrasternal, percutaneous, intranasal, inhalation, local, rectal, oral, intraocular, or intradermal routes.

[0034] Preferred dosages of compositions containing sarpogrelate and blueberry extract as active ingredients may vary depending on the subject's condition and weight, disease type and severity, drug form, route of administration, and duration, and may be appropriately selected by those skilled in the art. According to exemplary embodiments of this disclosure, although not limited thereto, the daily dose may be from 0.01 mg / kg to 200 mg / kg, specifically from 0.1 mg / kg to 200 mg / kg, and more specifically from 0.1 mg / kg to 100 mg / kg. Administration may be given once daily or in multiple separate doses, thus not limiting the scope of this disclosure.

[0035] In exemplary embodiments of this disclosure, the term "subject" as used herein may refer to mammals, including humans, but is not limited to the examples described.

[0036] Example

[0037] In the following, exemplary embodiments will be described in detail to aid in understanding this disclosure. However, the following exemplary embodiments are merely illustrative of the present disclosure, and the scope of this disclosure is not limited to the following exemplary embodiments. Exemplary embodiments of this disclosure are provided to explain this disclosure more completely to those skilled in the art.

[0038] <Examples 1-5> Preparation of a composite composition of European blueberry ethanol extract and sargrelate

[0039] European blueberry extract is primarily extracted as follows: 100g of European blueberry is added to 1L of 90 (v / v)% ethanol aqueous solution, and then stirred at 40°C and 1600rpm for 2 hours. The primary extract is transferred, and 1L of 70 (v / v)% ethanol aqueous solution is added to the residue for secondary extraction, followed by stirring at 40°C and 1600rpm for 2 hours. The secondary extract is then transferred. Next, 1L of 70 (v / v)% ethanol aqueous solution is added to the secondary residue, and the mixture is subjected to tertiary extraction in the same manner.

[0040] After mixing the primary, secondary, and tertiary extracts, the alcohol was evaporated and concentrated at a temperature below 50°C. The concentrate obtained from the above process was diluted to 1.5 ± 5% Brix and centrifuged at 6900 rpm for 6 to 12 minutes.

[0041] The supernatant obtained by centrifugation was hydraulically filtered and purified. Then, 1200 L of water was added to the purified solution for a first wash of 2 hours. 5000 L of 70 (v / v)% ethanol was added to the purified solution after the first wash for a second wash. The wash solution was collected and concentrated.

[0042] The concentrate obtained through the above process was spray-dried to prepare a powdered European blueberry ethanol extract.

[0043] Powdered bilberry ethanol extract was added to saprogrel hydrochloride (manufacturer: Pharmacostech) at the weight ratio shown in Table 1. The bilberry extract and saprogrel were placed in a mixer and mixed at 50 rpm for 5 minutes to prepare the composite composition.

[0044] Table 1

[0045] Example 1 Example 2 Example 3 Example 4 Example 5 European blueberry ethanol extract 10g 30g 80g 10g 10g Sapogrelate 10g 10g 10g 30g 80g Weight of the composite composition 20g 40g 90g 40g 90g

[0046] <Examples 6-10> Preparation of a composite composition of European blueberry ethanol extract and sargrelate

[0047] 100g of blueberries were added to 0.5L of 70 (v / v)% ethanol aqueous solution and stirred at 60°C and 1600rpm for 2 hours for primary extraction. The extract from the primary extraction was transferred, and 0.5L of 70 (v / v)% ethanol aqueous solution was added to the residue, followed by stirring at 60°C and 1600rpm for 2 hours for secondary extraction. The secondary extract was then transferred. Next, 0.5L of 70 (v / v)% ethanol aqueous solution was added to the secondary residue, and the mixture was subjected to tertiary extraction in the same manner.

[0048] After mixing the primary, secondary, and tertiary extracts, the alcohol was evaporated and concentrated at a temperature below 50°C. The concentrate obtained through the above process was diluted to 3 ± 5% Brix and centrifuged at 6900 rpm for 6 to 12 minutes.

[0049] The supernatant obtained by centrifugation was hydraulically filtered and purified. Then, 1200 L of water was added to the purified solution for a first wash of 2 hours. 5000 L of 70 (v / v)% ethanol was added to the purified solution after the first wash for a second wash. The wash solution was collected and concentrated. The resulting concentrate was freeze-dried to prepare a powdered European blueberry ethanol extract.

[0050] Powdered bilberry ethanol extract was added to saprogrel hydrochloride (manufacturer: Pharmacostech) at the weight ratio shown in Table 2. The bilberry extract and saprogrel were placed in a mixer and mixed at 50 rpm for 5 minutes to prepare the composite composition.

[0051] Table 2

[0052] Example 6 Example 7 Example 8 Example 9 Example 10 European blueberry ethanol extract 10g 30g 40g 10g 10g Sapogrelate 10g 10g 10g 20g 50g Weight of the composite composition 20g 40g 50g 30g 60g

[0053] <Examples 11-13> Preparation of a composite composition of European blueberry methanol extract and sargrelate

[0054] The methanol extract of blueberry was prepared as follows: Frozen blueberry pulp was pulverized, and 100 g of the pulverized material was immersed in a 0.5 L to 1 L 70 (v / v)% methanol aqueous solution, followed by extraction at 28 °C to 30 °C. The supernatant was then separated by centrifugation, and citric acid was added to the separated supernatant to remove the resulting precipitate for purification. The remaining extract was then concentrated, diluted with ethanol, and dried at 60 °C for 36 hours to prepare a dry powder.

[0055] The bilberry methanol extract obtained through the above process was added to saprogale hydrochloride (manufacturer: Pharmacostech) at the weight ratio shown in Table 3. The bilberry extract and saprogale were placed in a mixer and mixed at 50 rpm for 5 minutes to prepare the composite composition.

[0056] Table 3

[0057] Example 11 Example 12 Example 13 European blueberry methanol extract 10g 30g 50g Sapogrelate 10g 10g 10g Weight of the composite composition 20g 40g 50g

[0058] <Examples 14-15> Preparation of a composite composition of European blueberry methanol extract and sargrelate

[0059] 100g of blueberries were added to 1L of 70 (v / v)% methanol aqueous solution and stirred at 30°C and 1600 rpm for 2 hours for primary extraction. The extract from the primary extraction was transferred, and 1L of 70 (v / v)% methanol aqueous solution was added to the residue, followed by stirring at 30°C and 1600 rpm for 2 hours for secondary extraction. The secondary extract was then transferred. Next, 1L of 70 (v / v)% methanol aqueous solution was added to the secondary residue, and the mixture was subjected to tertiary extraction in the same manner.

[0060] After mixing the primary, secondary, and tertiary extracts, the alcohol was evaporated and concentrated at a temperature below 50°C. The concentrate obtained through the above process was diluted to 3 ± 5% Brix and centrifuged at 6900 rpm for 6 to 12 minutes.

[0061] The supernatant obtained by centrifugation was hydraulically filtered and purified. Then, 1200 L of water was added to the purified solution for a first wash of 2 hours, followed by a second wash with 5000 L of 70 (v / v)% methanol. The wash solution was collected and concentrated. The resulting concentrate was spray-dried to prepare a powdered European blueberry methanol extract.

[0062] The resulting bilberry methanol extract was added to saprogrel hydrochloride (manufacturer: Pharmacostech) at the weight ratio shown in Table 4. The bilberry extract and saprogrel were placed in a mixer and mixed at 50 rpm for 5 minutes to prepare the composite composition.

[0063] Table 4

[0064] Example 14 Example 15 European blueberry methanol extract 30g 30g Sapogrelate 30g 10g Weight of the composite composition 60g 40g

[0065] <Examples 16-20> Preparation of a composite composition of European blueberry water extract and sargrelate

[0066] European blueberry water extract was obtained by adding 30g of frozen European blueberry pulp to 600mL of water and hot water extraction at 110°C for 3 hours to obtain a hot water extract. The extract was then concentrated using a vacuum concentrator until the solid content was 30%, and then freeze-dried to prepare a dry powder.

[0067] The European blueberry extract obtained through the above process was added to saprogale hydrochloride (manufacturer: Pharmacostech) in the weight ratio shown in Table 5. The European blueberry extract and saprogale were placed in a mixer and mixed at a rate of 50 rpm for 5 minutes to prepare the composite composition.

[0068] Table 5

[0069] Example 16 Example 17 Example 18 Example 19 Example 20 European blueberry water extract 10g 30g 80g 10g 10g Sapogrelate 10g 10g 10g 30g 80g Weight of the composite composition 20g 40g 90g 40g 90g

[0070] <Experimental Example 1> Identification of the inhibitory effect of the composite composition on apoptosis in auditory cell lines

[0071] House Ear Institute Organ of Corti 1 (HEI-OC1) (an auditory hair cell derived from the mouse auditory organ Corti) was cultured in an incubator at 33°C using DMEM (high glucose Dulbecco Eagle medium, Sigma-Aldrich Co., St. Louis, USA) in the presence of 10% CO2 and containing 10% fetal bovine serum (FBS; WELGENE Inc., Gyeongsangbuk-do, South Korea) and 50 U / mL INF-γ (Peprotech Inc., Seoul, South Korea), without the addition of antibiotics.

[0072] To generate reactive oxygen species (ROS) induced by oxidative stress (which occurs during the induction of noise-induced hearing loss), a cell line with noise-induced hearing loss was established by treating with hydrogen peroxide.

[0073] The composite composition from Example 2 was used to treat cell lines with noise-induced hearing loss at concentrations of 0.001 μg / mL, 0.005 μg / mL, 0.01 μg / mL, 0.05 μg / mL, 0.1 μg / mL, and 0.5 μg / mL for 1 hour, followed by treatment with 100 μM hydrogen peroxide for 24 hours to identify the cytotoxic and apoptotic effects of hydrogen peroxide by MTT assay.

[0074] As shown in Table 6 and Figure 1 As shown, the inhibitory effect on apoptosis of auditory cells was identified in the composite composition of Example 2. Compared with sargrelate, the apoptosis-inhibiting effect on auditory cells was found to be significantly superior.

[0075] Table 6

[0076] μg / mL CTL <![CDATA[H2O2]]> 0.001 0.005 0.01 0.05 0.1 0.2 Relative cell viability 100 0.0 53.7 60.7 77.1 89.9 97.3 93.7

[0077] As a result, Figure 1 As shown, the EC50 of the composite composition in Example 2 was found to be 0.015 μg / mL. Furthermore, the EC50s of the examined sargrelate and blueberry extract were found to be 26.43 μg / mL and 0.16 μg / mL, respectively, indicating that the composite composition exhibited higher efficacy than the single formulation.

[0078] Based on the above results, it was determined that the complex composition in Example 2 could exhibit superior apoptosis inhibition effects at low doses compared to each of the individual formulations.

[0079] <Experimental Example 2> Efficacy Test of Hair Cells in a Zebrafish Drug Efficacy Model

[0080] The inner ear structure of zebrafish is similar to that of humans and is transparent, making it easy to observe. At the same time, the lateral line system, which serves as an additional auditory organ, is exposed to the outside. Therefore, changes in auditory hair cells after noise exposure or drug administration can be observed in living organisms, making it a model organism for reactive oxygen species-induced hearing loss.

[0081] To prepare zebrafish larvae, females and males were placed in a 1:1 ratio in a tank equipped with a net to separate the eggs from the adults. The zebrafish embryos were collected after 12 hours.

[0082] After collection, the zebrafish were washed three times with a 0.03% sea salt solution to remove foreign matter and bred in an incubator at 28.5°C with a photoperiod of 14 bright / 10 dark. Six days after fertilization, zebrafish larvae (6 dpf) were placed in 24-well tubs and exposed to 2.5 mM hydrogen peroxide for 3 hours to create a model of hydrogen peroxide-induced hearing loss, and a normal control without hydrogen peroxide treatment was prepared.

[0083] From Examples 1 to 5, zebrafish models with hydrogen peroxide-induced hearing loss were treated with concentrations of 0.1 μg / mL, 0.5 μg / mL, and 1 μg / mL, respectively, and exposed for 12 hours.

[0084] Zebrafish larvae that had been tested for toxicity were anesthetized with 0.02% tricaine, and their auditory hair cells were stained with 0.1% YO-PRO for 30 minutes. The auditory hair cells were then observed and counted using a fluorescence microscope (Olympus 1×70, Olympus, Japan).

[0085] As a result, Figure 2As shown, all treatment groups in Examples 1 to 5 exhibited the effect of inhibiting the cytotoxicity of hydrogen peroxide in zebrafish hair cells.

[0086] In particular, when Figure 3 When the complex composition of Example 2 was administered at a concentration of 0.1 μg / mL, an average of 12.74 hair cells were observed. The highest hair cell protective efficacy of the complex composition of Example 2 was demonstrated by the threshold-improving effects of each of the European blueberry extract and saprogretyl.

[0087] <Experimental Example 3> Comparative Study of Hearing Improvement Efficacy in In Vivo Mouse Models

[0088] The animals used in the experiment were housed in a light environment every 12 hours, with the temperature controlled at 25±2℃. In order to induce noise-induced hearing loss, the mice were exposed to a complex sound of 115dB for 90 minutes to prepare a mouse model of noise-induced hearing loss (NIHL).

[0089] To evaluate the efficacy of the sample in NIHL model mice, auditory brainstem response (ABR) measurements were used to measure auditory thresholds.

[0090] For the composite treatment group in Example 2, the sample was orally administered to NIHL mouse models at the same time daily after 20 hours of noise exposure, while no administration was given to the control group. All mice were measured before the experiment after their ear canals were examined by otoscopy.

[0091] For the stimulus sound, the evaluation was performed by gradually reducing the click sound (wide frequency range) from 80dB by 5dB, and the minimum sound that elicited a response was set as the threshold, with the lowest value being measured over 14 days.

[0092] As a result, such Figure 3 As shown, compared with the control group that did not receive the drug, the hearing threshold was reduced by up to 30% p in the group treated with sargrelate, European blueberry extract and the complex composition in Example 2, and a statistically significant difference was observed.

[0093] Although specific portions of this disclosure have been described in detail above, it will be clear to those skilled in the art that these specific descriptions are merely preferred exemplary embodiments, and the scope of this disclosure is not limited thereto. Therefore, the essential scope of this disclosure will be defined by the appended claims and their equivalents.

Claims

1. A pharmaceutical composition for preventing or treating hearing loss, comprising sarpogrelate and an extract of Vaccinium myrtillus as active ingredients.

2. The pharmaceutical composition according to claim 1, wherein, The extract of Vaccinium myrtillus is obtained by extraction with water, a C1-C4 alcohol or a complex solvent thereof.

3. The pharmaceutical composition according to claim 1 or 2, wherein, The hearing loss is selected from the group consisting of noise-induced hearing loss, ototoxic hearing loss, presbycusis and sudden hearing loss.

4. The pharmaceutical composition according to claim 1 or 2, wherein, The pharmaceutical composition comprises sarpogrelate and the extract of Vaccinium myrtillus in a weight ratio of 1 : (0.125-1 : 8).

5. The pharmaceutical composition according to claim 1 or 2, wherein, The pharmaceutical composition inhibits apoptosis of auditory cells and increases the number of hair cells.

6. Use of the pharmaceutical composition comprising sarpogrelate and the extract of Vaccinium myrtillus as active ingredients according to any one of claims 1-5 in the manufacture of a medicament for preventing or treating hearing loss.

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