KR20230083910A - Composition for preventing or treating hearing loss or tinnitus - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating hearing loss or tinnitus, comprising oxolamine, and a pharmaceutically acceptable salt, solvate or hydrate thereof. The composition containing oxolamine and a pharmaceutically acceptable salt, solvate or hydrate according to the present invention can effectively suppress damage to hair cells caused by the use of antibiotics as well as increased hearing due to hearing loss, and thus can inhibit behaviors caused by tinnitus, so that it is possible to be usefully used as a pharmaceutical composition or health functional food since it is useful for preventing, treating, or alleviating hearing loss or tinnitus.

Description

Composition for preventing or treating hearing loss or tinnitus {Composition for preventing or treating hearing loss or tinnitus}

The present invention relates to a composition for preventing or treating hearing loss or tinnitus, and more preferably to a composition for preventing or treating hearing loss or tinnitus containing oxolamine or a pharmaceutically acceptable salt thereof.

In modern society, due to the increase in noise exposure and the aging of the population, problems related to hearing loss and tinnitus due to cell or nerve damage are emerging more than inflammation-based ear-related diseases.

Hearing loss and tinnitus are so common that most people can distinguish the difference between ear-related diseases caused by inflammation, such as bacterial otitis externa, malignant otitis media, fungal otitis externa, otomycosis, otitis media, or otitis media, and hearing loss and tinnitus. It is one of the most common and widely known diseases.

For the treatment of hearing loss, many specialists and medical practitioners still aim to treat the above diseases with conventional antibiotics or anti-inflammatory drugs based treatments, but these cannot be used as suitable treatments for hearing loss. The use of existing treatments as described above is, firstly, because there is no known treatment for hearing loss, and secondly, because many specialists and people in the pharmaceutical industry are not clearly aware of a treatment for hearing loss. Due to these problems, the problem of not properly performing the treatment of hearing loss continues to occur.

In particular, the use of antibiotics presents a major problem due to ototoxicity. For example, aminoglycoside antibiotics have side effects of ototoxicity and renal toxicity that cause hearing and equilibrium dysfunction in the inner ear, which can occur not only with overdose but also with long-term use in therapeutic doses. Sometimes this happens. Ototoxicity of aminoglycoside antibiotics causes vestibular dysfunction in about 15% of users, hearing loss in 10-30%, and occurs in both ears in the form of sudden severe hearing loss, mainly at high frequencies of 4000 Hz or higher.

Tinnitus is also commonly referred to as tinnitus (the perception of sound in the absence of an external source of acoustic signals). Tinnitus, tinnitus, or tinnitus is the perception of sound in the human ear when there is no corresponding external sound. To put it simply, sounds that do not originate from the outside are heard from the inside. Statistically, about 15 to 20% of adults experience tinnitus of various kinds, of which 4% experience severe tinnitus. In addition, it is said that 70-80% of people with hearing loss experience tinnitus.

Regarding the treatment of tinnitus, most treatments are focused on relieving tinnitus through rehabilitation. For example, a tinnitus masking method in which a sound similar to the patient's tinnitus is heard at a louder volume than the tinnitus from the outside through a device resembling a hearing aid, and a tinnitus masking method that makes the tinnitus inaudible, and a sound that is smaller than the actual tinnitus continuously over a wide frequency range There is a tinnitus retraining therapy that treats tinnitus without accompanying hearing loss by listening to it. To date, there is no treatment for tinnitus approved by the US Food and Drug Administration (FDA).

Under this background, preclinical studies on various drugs have been reported as studies to find substances effective for the prevention and treatment of hearing loss and/or tinnitus, but it has been confirmed that there are limitations to proceeding with clinical trials. In addition, there are currently no drugs approved for the prevention and treatment of hearing loss and/or tinnitus. Moreover, there are few drugs developed for the treatment of hearing loss and/or tinnitus that can be used without toxicity or risk to the human body. That is, unlike ear-related diseases caused by inflammation, a suitable treatment for hearing loss and/or tinnitus has not been developed at all.

An object of the present invention is to provide a pharmaceutical composition for preventing or treating hearing loss or tinnitus comprising oxolamine, a pharmaceutically acceptable salt, solvate or hydrate thereof.

Another object of the present invention is to provide a method for preventing or treating hearing loss or tinnitus by administering oxolamine, a pharmaceutically acceptable salt, solvate or hydrate thereof to a subject in need thereof.

Another object of the present invention is to provide a use of oxolamine, a pharmaceutically acceptable salt, solvate or hydrate thereof for the preparation of a medicament for the prevention or treatment of hearing loss or tinnitus.

Another object of the present invention is to provide a composition comprising oxolamine, a pharmaceutically acceptable salt, solvate or hydrate thereof for use in the prevention or treatment of hearing loss or tinnitus.

Another object of the present invention is to provide a food composition for preventing or improving hearing loss or tinnitus comprising oxolamine, a food-acceptable salt, solvate or hydrate thereof.

Another object of the present invention is to provide a health functional food for preventing or improving hearing loss or tinnitus containing oxolamine, a food-acceptable salt, solvate or hydrate thereof.

Another object of the present invention is to provide a feed composition for preventing or improving hearing loss or tinnitus comprising oxolamine, a food-acceptable salt, solvate or hydrate thereof.

A detailed description of this is as follows. Meanwhile, each description and embodiment disclosed in this application may also be applied to each other description and embodiment. That is, all combinations of various elements disclosed in this application fall within the scope of this application. In addition, the scope of the present application is not to be construed as being limited by the specific descriptions described below.

Pharmaceutical composition for preventing or treating hearing loss or tinnitus containing oxolamine

In order to solve the above problems, one aspect of the present invention provides a pharmaceutical composition for preventing or treating hearing loss or tinnitus comprising oxolamine, a pharmaceutically acceptable salt, solvate or hydrate thereof.

In the present invention, oxolamine may be a compound represented by Formula 1 below.

[Formula 1]

.

.

The oxolamine is known for cough or respiratory anti-inflammatory use.

In the present invention, the oxolamine may be preferably a pharmaceutically acceptable salt of oxolamine, and more preferably oxolamine citrate represented by Formula 2 below.

[Formula 2]

.

.

The present inventors have confirmed that the pharmaceutical composition containing oxolamine has an excellent therapeutic effect on hearing loss by effectively suppressing the increase in hearing threshold and hair cell damage caused by hearing loss.

In the present invention, "hearing loss" means any condition in which hearing is reduced or lost. Hearing loss may include, but is not limited to, conductive hearing loss and sensorineural hearing loss.

The conductive hearing loss is hearing loss caused by an ear disease, and is a hearing loss caused by problems in organs such as the eardrum and ossicles, which are organs that transmit sound. The hearing loss may be due to several causes, such as infections, injuries, inflammation, tumors, and adverse reactions to drugs or other chemicals.

The sensorineural hearing loss is a hearing loss caused by problems in the cochlea, which is an organ that detects sound, the auditory nerve, which transmits sound with electrical energy, and the brain responsible for hearing, which plays a comprehensive role in discriminating and understanding sound. The cause of sensorineural hearing loss may be hearing loss caused by noise, drugs, aging, trauma, or the like, and may be, for example, ototoxic hearing loss. The ototoxic hearing loss is caused by ototoxic drugs such as gentamicin, streptomycin, kanamycin, neomycin, amikacin, tobramycin, netylmycin ( Administration of one or more drugs selected from the group consisting of netilmicin), dibekacin, sisomycin, ribodomycin, cisplatin, carboplatin, and oxaliplatin It may be due to hearing loss.

In addition, the hearing loss may include noise-induced hearing loss, presbyopia, sudden hearing loss, acoustic neuropathy due to diabetes, ototoxic hearing loss, traumatic hearing loss, viral hearing loss, and the like.

If hearing is reduced or lost, it is not limited to the scope of the hearing loss of the present invention and may be included in the range of diseases.

In addition, the present inventors have confirmed that the pharmaceutical composition containing oxolamine has an excellent therapeutic effect on tinnitus by reducing abnormal behavior caused by tinnitus and returning hearing amplitude to normal.

In the present invention, “tinnitus” refers to the perception of sound in the absence of an external source of sound signals, and may include objective tinnitus, subjective tinnitus, peripheral tinnitus, and central tinnitus, but is not limited thereto. Specifically, it may include subjective tinnitus caused by various causes such as noise, drugs, aging, trauma, and viruses, but is not limited thereto.

The objective tinnitus (or objective tinnitus) is tinnitus that can be heard from the outside, and the subjective tinnitus (or subjective tinnitus) means tinnitus that is heard only by the tinnitus patient and cannot be heard from the outside. Tinnitus can also be classified into peripheral tinnitus and central tinnitus based on differences in how it is perceived by affected individuals. Peripheral (or cochlear) tinnitus is presumed to originate from the peripheral nervous system and cochlea, and central tinnitus is presumed to originate from the auditory cortex.

However, if it corresponds to tinnitus, it is not limited to the scope of the tinnitus of the present invention and may be included in the range of diseases.

In the present invention, oxolamine may exist in the form of a pharmaceutically acceptable salt. As the salt, an acid addition salt formed with a pharmaceutically acceptable free acid is useful. As used herein, "pharmaceutically acceptable salt" is a concentration that has an effective effect that is relatively non-toxic and harmless to patients, and any organic or It means inorganic addition salt.

At this time, as the addition salt, inorganic acids such as hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, etc. may be used, and organic acids include citric acid, methanesulfonic acid, p-toluenesulfonic acid, acetic acid, trifluoroacetic acid, maleic acid, succinic acid, Oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, Hydroiodic acid and the like may be used, and preferably, oxolamine citrate salt form may be used using citric acid.

In addition, a pharmaceutically acceptable metal salt may be prepared using a base. The alkali metal salt or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and then evaporating and drying the filtrate. In this case, as the metal salt, it is particularly suitable for preparing sodium, potassium, calcium, magnesium salts or mixed salts thereof, but is not limited thereto.

As used herein, the term "solvate", unless otherwise indicated, refers to a solvate formed from association of one or more solvent molecules in a compound provided herein. The term "solvate" includes hydrates (eg, monohydrate, dihydrate, trihydrate, tetrahydrate, etc.).

The term “hydrate” refers to a compound of the present invention that contains a stoichiometric or non-stoichiometric amount of water bound by non-covalent intermolecular forces. or a salt thereof.

The pharmaceutical composition of the present invention may contain a pharmaceutically acceptable carrier, and may be prepared according to conventional methods, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, oral formulations, external preparations, suppositories, and the like. It can be formulated in the form of a sterile injectable solution.

The pharmaceutically acceptable carrier includes those commonly used in the art, such as lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like. In addition, the pharmaceutical composition of the present invention may include diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, and other pharmaceutically acceptable additives, but is not limited thereto.

When the pharmaceutical composition of the present invention is formulated into oral solid preparations, it includes tablets, pills, powders, granules, capsules, etc., and these solid preparations contain at least one excipient such as starch, calcium carbonate, It may include sucrose or lactose, gelatin, and the like, and include, but are not limited to, lubricants such as magnesium stearate and talc.

When the pharmaceutical composition of the present invention is formulated in liquid form for oral use, it includes suspensions, internal solutions, emulsions, syrups, etc., and diluents such as water and liquid paraffin, wetting agents, sweeteners, aromatics, preservatives, etc., but are not limited thereto.

When the pharmaceutical composition of the present invention is formulated for parenteral use, it includes a sterilized aqueous solution, a non-aqueous solvent, a suspension, an emulsion, a lyophilized preparation, and a suppository, and the non-aqueous solvent and suspension include propylene glycol, polyethylene glycol, and olive vegetable oils such as oils, injectable esters such as ethyl oleate, and the like, but are not limited thereto. As a base material for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin paper, glycerogelatin, etc. may be used, but are not limited thereto.

The composition may be administered singly or in multiple doses in a pharmaceutically effective amount. The term "pharmaceutically effective amount" of the present invention means an amount sufficient to prevent or treat a disease with a reasonable benefit / risk ratio applicable to medical prevention or treatment, and the effective dose level is the severity of the disease, the activity of the drug, Age, weight, health, sex of the patient, sensitivity to the drug of the patient, administration time of the composition of the present invention used, route of administration and excretion rate, treatment period, including drugs used in combination or simultaneous use with the composition of the present invention used factors and other factors well known in the medical field. For example, oxolamine may be administered at a dose of 0.0001 to 100 mg/kg per day, preferably 0.01 to 50 mg/kg, and the administration may be administered once a day or divided into several times.

The pharmaceutical composition of the present invention is administered to mammals such as rats, mice, livestock, and humans by various routes, for example, oral administration, inner ear, abdominal cavity or intravenous, intramuscular, subcutaneous, intrauterine intrathecal or intracerebrovascular injection. It can be.

The pharmaceutical composition of the present invention may include oxolamine in an amount of 0.01 to 95% by weight, preferably 1 to 80% by weight, based on the total weight of the composition.

The pharmaceutical composition for preventing or treating hearing loss or tinnitus of the present invention has an excellent therapeutic effect on hearing loss or tinnitus by reducing abnormal behavior caused by hearing loss or tinnitus.

As used herein, the term "prevention" refers to any action that suppresses hearing loss or tinnitus or delays the onset of hearing loss by administration of the pharmaceutical composition according to the present invention.

As used herein, the term "treatment" refers to all activities that improve or beneficially change symptoms of hearing loss or tinnitus by administration of the pharmaceutical composition according to the present invention.

Method for preventing or treating hearing loss or tinnitus comprising administering oxolamine to a subject in need thereof, and use of oxolamine

Another aspect of the present invention provides a method for preventing or treating hearing loss or tinnitus, comprising administering oxolamine, a pharmaceutically acceptable salt, solvate or hydrate thereof to a subject in need thereof.

In the present invention, the oxolamine may be preferably a pharmaceutically acceptable salt of oxolamine, and more preferably oxolamine citrate.

In the present invention, the terms "oxolamine", "hearing loss" or "tinnitus" are as described above.

The term "subject" of the present invention means any animal that has or may develop hearing loss or tinnitus, and will typically show a beneficial effect by treatment with oxolamine, a pharmaceutically acceptable salt, solvate or hydrate thereof. It may be an animal capable of detecting hearing loss or tinnitus, or an individual that is likely to have such symptoms is included without limitation. As described above, the hearing loss or tinnitus can be effectively prevented or treated by administering the pharmaceutical composition of the present invention to a subject. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent, or in combination with an existing therapeutic agent for hearing loss or tinnitus, and may be administered sequentially or simultaneously with conventional therapeutic agents.

The term "administration" of the present invention means introducing a predetermined substance into a patient by an appropriate method, and the administration route of the composition may be administered through any general route as long as it can reach the target tissue. In addition, the pharmaceutical composition of the present invention may be administered by any device capable of transporting an active substance to a target tissue. For example, oral administration, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration, intranasal administration, intrapulmonary administration, rectal administration, inner ear administration may be used, but is not limited thereto. does not Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories.

Administration of the pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. Considering all of the above factors, it can be administered in an amount that can obtain the maximum effect with the minimum amount without side effects, which can be easily determined by those skilled in the art.

Another aspect of the present invention provides the use of oxolamine, a pharmaceutically acceptable salt, solvate or hydrate thereof for the preparation of a medicament for the prevention or treatment of hearing loss or tinnitus.

Another aspect of the present invention provides a composition comprising oxolamine, a pharmaceutically acceptable salt, solvate or hydrate thereof for use in the prevention or treatment of hearing loss or tinnitus.

Food composition, health functional food and feed composition containing oxolamine

Another aspect of the present invention provides a food composition for preventing or improving hearing loss or tinnitus comprising oxolamine, a pharmaceutically acceptable salt, solvate or hydrate thereof.

Another aspect of the present invention provides a health functional food for preventing or improving hearing loss or tinnitus comprising oxolamine, a pharmaceutically acceptable salt, solvate or hydrate thereof.

In the present invention, the terms "oxolamine", "solvate", "hydrate", "hearing loss" or "tinnitus" are as described above.

In the present invention, oxolamine may exist in the form of a food-acceptable salt. As the salt, an acid addition salt formed by a free acid that is acceptable in food science is useful. As used herein, "food acceptable salt" is a concentration that has an effective effect that is relatively non-toxic and harmless to patients, and any organic or It means inorganic addition salt. Here, the salt is as described above in the pharmaceutical composition.

In the present invention, it is preferable to use oxolamine citrate as oxolamine.

The term of the present invention, "improvement" refers to all activities in which hearing loss or tinnitus is improved or advantageously changed by administration of the composition of the present invention.

The food composition of the present invention may include conventional food additives, and the suitability as the "food additive" is determined according to the general rules of the food additive code approved by the Ministry of Food and Drug Safety and general test methods, etc., unless otherwise specified. It is judged according to the specifications and standards for the item.

Items listed in the "Food Additive Code" include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid, natural additives such as dark pigment, licorice extract, crystalline cellulose, goyang pigment, guar gum, and mixed preparations such as sodium L-glutamate preparations, noodle-added alkali preparations, preservative preparations, and tar color preparations.

In addition, the food composition of the present invention can be prepared and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. for the purpose of preventing and/or improving hearing loss or tinnitus.

The food composition of the present invention can be used as a health functional food. The term “health functional food” refers to food manufactured and processed using raw materials or ingredients having useful functionalities for the human body in accordance with the Health Functional Food Act, and “functional” refers to food that is not related to the structure and function of the human body. It refers to intake for the purpose of obtaining useful effects for health purposes such as regulating nutrients or physiological functions.

Another aspect of the present invention provides a feed composition for preventing or improving hearing loss or tinnitus comprising oxolamine, a food-acceptable salt, solvate or hydrate thereof.

In the present invention, the terms "oxolamine", "solvate", "hydrate", "hearing loss" or "tinnitus" are as described above.

In the present invention, the feed may be fish, bird, or mammal feed, and preferably, the wild habits defined in Article 2, Subparagraph 1 of the Livestock Act and each subparagraph of the Enforcement Rule of the same Act are purified and suitable for breeding. It can be feed for livestock or aquatic organisms that can contribute to farm household income increase. The livestock may be cattle, horses, mules, donkeys, goats, goats, sheep, deer, pigs, rabbits, poultry, etc., and poultry may be chickens, turkeys, ducks, ostriches, geese, quails, etc., preferably chickens, It is not limited thereto as long as it is suitable for breeding and obtaining livestock products. The above "livestock products" are meat, milk, eggs, honey and their processed products, hides (including raw fur), raw wool, and other livestock products produced from livestock, which are defined in Article 2, Subparagraph 3 of the Livestock Act, as determined by the Ordinance of the Ministry of Agriculture and Forestry. means that In addition, it may be a dog, cat, etc., including companion animals, but is not limited thereto.

The term "feed" in the present invention means any natural or artificial diet, meal, etc. or component of said meal, intended for or suitable for eating, ingestion and digestion by an animal. In one aspect, the feed composition containing oxolamine of the present invention may include concentrated feed, roughage and/or special feed.

Concentrated feed includes seed fruits including grains such as wheat, oats, and corn, bran including rice bran, wheat bran, and barley bran as by-products obtained after refining grains, soybeans, fluids, sesame seeds, linseed, and coco palm oil. Fish meal, which is a by-product obtained by extracting starch from sweet potatoes and potatoes, and residual starch, which is the main component of starch residues, fish meal, fish residue, and fish soluble, meat meal, which are concentrated fresh liquids obtained from fish. , animal feed such as blood meal, feather meal, skim milk powder, dried whey obtained by drying whey, which is the balance when cheese is produced from milk and casein is produced from skim milk, yeast, chlorella, and seaweed.

Forage, raw grass feed such as wild grass, grass, and green cutting, turnip for feed, beet for feed, root vegetables such as Lutherbearer, a type of turnip, raw grass, green cut crops, grain, etc. are put into silos. There are silage (silage), which is a stored feed filled and fermented with lactic acid, grass, hay cut and dried, straw from breeding crops, and leaves of leguminous plants. Special feeds include mineral feeds such as oyster shells and rock salt, urea feeds such as urea or its derivative, diureide isobutane, and supplements for ingredients that tend to be insufficient when only natural feed ingredients are mixed, or formulated feeds to improve the storage life of feeds. There are feed additives, which are substances added in small amounts.

The composition comprising oxolamine, a pharmaceutically acceptable salt, solvate or hydrate thereof according to the present invention can effectively suppress damage to hair cells caused by the use of antibiotics as well as increase hearing due to hearing loss, and can cause tinnitus Since it is useful for preventing, treating, or improving hearing loss or tinnitus by suppressing behaviors caused by it, it can be usefully used as a pharmaceutical composition or health functional food.

1 is a diagram showing the inhibitory effect of oxolamine on hair cell damage caused by neomycin treatment. NOR represents a normal control group untreated with neomycin, NM represents a control group treated only with neomycin, and oxolamine citrate represents an experimental group treated with oxolamine at a concentration of 0.1 μM.

Hereinafter, the present invention will be described in more detail through examples. However, these examples are intended to illustrate the present invention, and the scope of the present invention is not limited thereto.

<Example>

Example 1. Effect of oxolamine on ototoxic hearing loss zebrafish model

Zebrafish fry (larvae) 6 days after fertilization were placed in 24 wells, treated with neomycin at a concentration of 2 μM, and exposed for 1 hour to prepare an ototoxic hearing loss model. In addition, an untreated normal control group that was not treated with neomycin was prepared.

The prepared ototoxic deafness zebrafish model was treated with oxolamine citrate at a concentration of 0.1 μM and exposed for 6 hours. As a comparison group, 0.03% sea salt solution was used. The zebrafish fry of the experimental group and the control group, which were treated with oxolamine citrate and sea salt solution in the normal control group and ototoxic hearing loss model, respectively, were anesthetized with 0.02% tricaine, and the hair cells were treated with 0.1% YO-PRO for 30 minutes. After staining, it was directly observed with the naked eye using a fluorescence microscope (Olympus 1×70, Olympus, Japan). Specifically, the number of hair cells observed with the naked eye was counted and graphed, and a fluorescence image was taken and shown in FIG. 1 .

As shown in Figure 1, compared to the normal control group, the number of hair cells was significantly reduced in the control group not treated with oxolamine citrate in the ototoxic hearing loss induced zebrafish model, whereas the ototoxic hearing loss induced zebrafish In the experimental group treated with oxolamine citrate, the number of hair cells was significantly increased. Through this, it was confirmed that treatment with oxolamine citrate was effective in preventing and treating hearing loss by reducing hair cell damage caused by ototoxic hearing loss.

Example 2. Confirmation of improvement effect of hearing loss after exposure to noise

Example 2-1. Confirmation of hearing threshold using click sound, which is a broadband stimulus sound

In order to confirm the effect of oxolamine citrate on hearing threshold after exposure to noise, a hearing threshold measurement experiment using hearing evoked potential was performed. An auditory brainstem response (ABR) measurement method is a method of evaluating a response to sound by measuring the electrical energy when a sound stimulus is transmitted as an electrical signal from the auditory nerve. When sound reaches the auditory nerve via the outer ear, middle ear, and cochlea, the response reflects the condition of the outer ear, middle ear, and cochlea, which reflects the actual sound energy reaching the brain. Hearing threshold refers to the minimum sensory point of barely audible sound, and in the case of a normal mouse, a response is observed even to a small sound of 20 dB on average.

Specifically, the mice to which oxolamine citrate was administered and the untreated control mice were divided and evaluated.

Noise was exposed to 115 dB complex sound for 90 minutes, and oxolamine citrate was orally administered at the same time every day after 24 hours of noise exposure.

Hearing threshold was evaluated before noise exposure and on day 1, day 10, and day 20 after exposure.

For the auditory evoked potential test, mice were anesthetized by intramuscular injection of ketamine (4.57 mg/kg) and xylazine (0.43 mg/kg), and then evaluated while maintaining body temperature at 37±0.5°C. In the auditory evoked potential test, the stimulus was evaluated by gradually lowering the sound by 5 dB from 80 dB with a click sound, which is a broadband stimulus sound, and the lowest sound that responded was set as the threshold.

Example 2-2. Hearing Threshold Verification Using 16 kHz TB Stimulus

In the auditory evoked potential test, the stimulus sound was evaluated as a 16 kHz pure tone, gradually lowering the sound by 5 dB from 80 dB.

Example 3. Evaluation of the protective effect of cochlear hair cells

It was performed in the same manner as in Example 2-1, and the normal group with intact hair cells, the group with damaged hair cells due to exposure to noise (NIHL, Noise-induced hearing loss), and the treatment with oxolamine citrate after exposure to noise One group (NIHL + Oxolamine citrate) was evaluated.

After the end of the threshold evaluation experiment on day 20, the mouse cochlea was separated. The separated cochlea was stored in 4% paraformaldehyde at 4° C. for 12 hours, then treated with 0.1 M EDTA for 5 days, and the organ of Corti was microdissected under a microscope. To confirm hair cells, they were stained with 5 U/mL rhodamine phalloidin and confirmed under a microscope.

Example 4. Evaluation of tinnitus suppression of oxolamine citrate in a zebrafish model of tinnitus induced by salicylic acid

The treatment effect on tinnitus was confirmed using a behavioral response test method that can evaluate tinnitus by inducing tinnitus in zebrafish without physical stress.

Specifically, a control group (NOR) in which adult zebrafish did not induce tinnitus, an experimental group in which tinnitus was induced by exposure to 3 mM salicylic acid for 5 hours (SS), and oxolamine citrate immediately after exposure to 3 mM salicylic acid for 5 hours were treated at 17 It was divided into an experimental group (Oxolamine citrate) exposed for a period of time. At this time, the oxolamine citrate experimental group conducted the experiment by exposing adult zebrafish to concentrations of 10 ug/ml, 100 ug/ml, and 200 ug/ml of a composition containing oxolamine. All experiments use 0.03% sea salt solution as a basic solution, and all experimental groups were adapted to 0.03% sea salt solution for 1 hour immediately before measurement.

To confirm the effect of oxolamine citrate, zebrafish exposed to a composition containing oxolamine citrate were placed in a transparent tube (outer diameter 25 mm, inner diameter 21 mm, total length 60 cm), the passing time and the number of rotations in the transparent tube were analyzed. The water temperature was maintained at 28 ℃ by installing an underwater heater in the water tank.

Example 5. Assessment of Skinner's behavioral response to oxolamine citrate in a rat model of tinnitus induced by salicylic acid

When evaluating Skinner's behavioral response in trained rats, in the case of tinnitus-induced animals, the response to pressing the trigger when there is no cue tone tends to increase. At this time, the induction of tinnitus can be confirmed by the animal mistaking the tinnitus sound as a cue tone and showing the action of pressing the trigger. At this time, a true positive refers to the number of times the trigger is pressed to acquire food when a cue tone is provided, and a false positive refers to the number of times the trigger is pressed when there is no cue tone.

A control group (NOR) in which tinnitus was not induced using oxolamine, an experimental group (SS) in which tinnitus was induced, and an experimental group (SS+Oxolamine citrate) administered with oxolamine citrate were divided into three groups of three each and evaluated.

In the experiment, the SS group and the Oxolamine citrate group used salicylic acid, which is commonly used in tinnitus-induced animal models, to induce tinnitus, and 350 mg/kg was orally administered 3 hours before the test. In the SS group, only salicylic acid was administered orally, and in the SS+Oxolamine citrate group, oxolamine was administered orally 1 hour and 30 minutes before the examination after inducing tinnitus.

The experiment schedule consisted of 1 day of examination before administration (Base), 3 days of examination after concurrent administration with salicylic acid (1 to 3 Days), and 3 days of examination during the tinnitus recovery period (4 to 6 Days). In the case of the SS group, water was administered 1 hour and 30 minutes before the test after salicylic acid was administered 3 hours before the test on the 1st to 3rd days, and only water was administered orally 1 hour 30 minutes before the test on the 4th to 6th day. In the case of the SS+Oxolamine citrate group, on days 1 to 3, salicylic acid was administered 3 hours before the test, and then oxolamine citrate was administered 1 hour and 30 minutes before the test.

For behavioral response analysis, Silence activity ratio (SA ratio) and False positive ratio (FP ratio) were used. The SA ratio is calculated by dividing the number of responses when there is no cue tone by the number of responses during the time when the cue tone is provided, and the FP ratio is calculated as a ratio of the number of responses when there is no cue tone to the total number of responses .

Example 6. Confirmation of hearing amplitude of oxolamine in a tinnitus rat model

Hearing amplitude measurement experiments were performed using auditory evoked potential to confirm the effect of oxolamine on the sense of the presence or absence of sound by measuring hearing amplitude after tinnitus induction.

An auditory brainstem response (ABR) measurement method is a method of evaluating a response to sound by measuring the electrical energy when a sound stimulus is transmitted as an electrical signal from the auditory nerve. When sound reaches the auditory nerve via the outer ear, middle ear, and cochlea, the response reflects the condition of the outer ear, middle ear, and cochlea, which reflects the actual sound energy reaching the brain. Hearing threshold refers to the minimum sensory point of barely audible sound, and in the case of a normal mouse, a response is observed even to a small sound of 20 dB on average.

When measuring hearing amplitude, five amplitudes are usually observed (unit: μV, Wave Ⅰ ~ Wave Ⅴ). Amplitudes 1 to 3 (Wave I to III) are formed by auditory branches extending from the 8th cranial nerve and lower nerves. Here, amplitude 1 is generated from the dendrite of the auditory nerve fiber, amplitude 2 is generated from the cochlear nucleus, and amplitude 3 represents the activity level of the superior olivary complex that receives auditory information from the cochlear nucleus. Also, amplitudes 4 and 5 (Wave IV ~ V) are formed from the upper brainstem and are related to the lateral lemniscus.

The schedule before the click stimulation test and the experimental group were the same as those of Example 5, and the click stimulation test was performed and evaluated on the 5th day of oral administration of oxolamine.

For the auditory evoked potential test, ketamine (11.43 mg/kg) and xylazine (1.08 mg/kg) were injected intramuscularly to anesthetize rats, and then the body temperature was maintained at 37±0.5°C. In the auditory evoked potential test, the stimulus was evaluated by gradually lowering the sound by 5 dB from 90 dB with a click sound, which is a broadband stimulus sound, and the lowest sound that responded was set as the threshold.

Claims (11)

Oxolamine, a pharmaceutical composition for preventing or treating hearing loss or tinnitus comprising a pharmaceutically acceptable salt, solvate or hydrate thereof. The pharmaceutical composition according to claim 1, wherein oxolamine is a compound represented by Formula 1 below:
[Formula 1]

. The pharmaceutical composition according to claim 1, wherein the oxolamine is oxolamine citrate. The pharmaceutical composition according to claim 1, wherein the hearing loss is conductive hearing loss or sensorineural hearing loss. The pharmaceutical composition according to claim 1, wherein the hearing loss is any one selected from the group consisting of noise-induced hearing loss, age-related hearing loss, sudden hearing loss, acoustic neuropathy due to diabetes, ototoxic hearing loss, traumatic hearing loss, and viral hearing loss. The pharmaceutical composition according to claim 1, wherein the tinnitus is any one selected from the group consisting of objective tinnitus, subjective tinnitus, peripheral tinnitus and central tinnitus. A food composition for preventing or improving hearing loss or tinnitus comprising oxolamine, a food-acceptable salt, solvate or hydrate thereof. 8. The pharmaceutical composition according to claim 7, wherein the oxolamine is oxolamine citrate. The food composition according to claim 7, wherein the hearing loss is conductive hearing loss or sensorineural hearing loss. The food composition according to claim 7, wherein the hearing loss is any one selected from the group consisting of noise-induced hearing loss, age-related hearing loss, sudden hearing loss, diabetic acoustic neuropathy, ototoxic hearing loss, traumatic hearing loss, and viral hearing loss. The food composition according to claim 7, wherein the tinnitus is any one selected from the group consisting of objective tinnitus, subjective tinnitus, peripheral tinnitus and central tinnitus.

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