Application of thyroid hormone in preparation of medicine for treating GJB2 gene mutation related congenital deafness

Abstract

The invention provides application of thyroid hormone in preparation of a medicine for treating GJB2 gene mutation related congenital deafness. Experiments prove that injection of the thyroid hormone T3 or T4 can effectively treat congenital deafness and cochlea structure development disorder caused by GJB2 gene mutation, new indications of the thyroid hormone except for hypothyroidism and thyromegaly are expanded, and the application of the thyroid hormone T3 or T4 in treating the congenital deafness caused by GJB2 gene mutation has the advantages that the application of the thyroid hormone T3 or T4 in treating the congenital deafness caused by GJB2 gene mutation has good application prospects. Compared with a traditional treatment scheme, the cochlea dysplasia of the patient can be treated while the hearing of the patient is improved, and a new treatment way is provided for related congenital deafness caused by GJB2 gene mutation.

Description

Technical Field:

[0001] The present invention relates to the field of biomedicine, and particularly to the application of thyroid hormones in the preparation of drugs for treating congenital deafness related to GJB2 gene mutations. Background Art:

[0002] Congenital deafness refers to hearing loss caused by structural or functional disorders of the sound conduction structure, cochlea, auditory pathway, or auditory center due to genetic or environmental factors during maternal pregnancy and childbirth. GJB2 gene mutations are the most common genetic factors leading to congenital deafness, accounting for more than 50% of the causes of hereditary non-syndromic deafness. GJB2 gene mutations result in the failure of the important physiological structure of the cochlea, the Corti tunnel, to open and form, leading to cochlear structural developmental disorders, which is recognized as the main deafness mechanism. The current clinical treatment plan for patients with congenital profound deafness caused by GJB2 gene mutations mainly relies on cochlear implantation, but there are the following limitations: 1. The cost of cochlear implants is high; 2. There are certain risks associated with cochlear implantation surgery; 3. Regular maintenance and repair are required, etc. This brings great burden and inconvenience to patients and there is also a risk of loss. The GJB2 gene therapy programs being developed in the laboratory have only been verified in specific GJB2 gene local mutant mouse models with intact Corti tunnel structures and have not been applied to GJB2 gene widely mutant mouse models with non-open Corti tunnels and cochlear structural developmental disorders. Currently, there is no research report at home and abroad on a drug treatment plan that can effectively treat the non-opening of the cochlear Corti tunnel and cochlear structural malformations caused by widespread GJB2 gene mutations, nor can hearing be restored by saving the cochlear structure.

[0003] Thyroid hormones are hormones secreted by the thyroid gland of the human body or animals, and there are two forms: tetraiodothyronine (T4) and triiodothyronine (T3). Among them, T3 accounts for about 20% of the thyroid hormone content in the blood circulation and is the main form exerting biological activity; T4 accounts for 80% and is converted into T3 in peripheral tissues to exert biological regulatory effects. Thyroid hormones have the functions of promoting the growth and development of organs and tissues and regulating metabolism. Currently, it is mainly used clinically for the treatment of hypothyroidism or goiter, and there is no research indicating the efficacy of thyroid hormones in the treatment of congenital deafness related to GJB2 gene mutations and its cochlear structural developmental disorders. The relationship between the GJB2 gene and thyroid hormones has never been reported in existing research. Summary of the Invention:

[0004] (I) Technical Problems to be Solved

[0005] In view of the deficiencies of the above-mentioned prior art, the purpose of the present invention is to provide the use of thyroid hormones in the preparation of drugs for the treatment of congenital deafness related to GJB2 gene mutations. Through thyroid hormones T3 or T4, congenital deafness and cochlear structural development disorders caused by GJB2 gene mutations can be effectively treated, expanding the new indications of thyroid hormones beyond hypothyroidism and goiter. While improving the patient's hearing, it can treat the patient's cochlear development disorders, providing a new treatment approach for congenital deafness caused by GJB2 gene mutations.

[0006] (II) Technical solution

[0007] To achieve the above object, the present invention adopts the following technical solutions:

[0008] Use of thyroid hormones in the preparation of drugs for the treatment of congenital deafness related to GJB2 gene mutations.

[0009] The thyroid hormone is T3 or T4. T3 is triiodothyronine ≡O-(4-hydroxy-3-iodophenyl)-3,5-diiodo-L-tyrosine, and T4 is levothyroxine ≡O-(4-hydroxy-3,5-diiodophenyl)-3,5-diiodo-L-tyrosine. T3 or T4 in its own form or in the form of sodium salt or hydrate is widely known and can be obtained by synthesis and / or extraction from animal glands (such as pigs, etc.).

[0010] The drug is a water-dispersible agent of thyroid hormone T3 or T4. The preparation method of the water-dispersible agent of T3 is: dissolve T3 powder in 40 mM sodium hydroxide to prepare a liquid with a concentration of 1 μg / μL, and adjust the solution to pH = 7.4 - 7.7; the preparation method of the water-dispersible agent of T4 is: dissolve T4 powder in physiological saline to prepare a liquid with a concentration of 1 μg / μL, and adjust the solution to pH = 7.4 - 7.7.

[0011] The drug is made into an oral dosage form or an injection by adding conventional excipients or in combination with other drugs for treating deafness. The excipients include disintegrants, lubricants, and binders. The other drugs for treating deafness are: dexamethasone, PARP-1 inhibitors, acetylcysteine, gene therapy drugs, etc.

[0012] The drug is used for the treatment of congenital deafness or cochlear structural development disorders caused by GJB2 gene mutations. The administration methods of the drug include subcutaneous administration, gastrointestinal administration, or local cochlear administration.

[0013] (III) Beneficial effects

[0014] The beneficial effects of the present invention are as follows:

[0015] The present invention discovers that congenital deafness and cochlear structural dysplasia caused by GJB2 gene mutation can be effectively treated by thyroid hormone T3 or T4, and proposes the application of thyroid hormone in the preparation of drugs for treating congenital deafness related to GJB2 gene mutation, expanding the new indications of thyroid hormone beyond hypothyroidism and goiter. While improving the patient's hearing, it can treat the cochlear dysplasia of the patient, providing a new treatment approach for congenital deafness caused by GJB2 gene mutation.

[0016] (1) The thyroid hormone and its preparations described in the present invention can treat congenital deafness caused by GJB2 gene mutation on the basis of promoting the opening of the Corti tunnel in the cochlea and rescuing the cochlear structure.

[0017] (2) The thyroid hormone and its preparations described in the present invention can be applied to congenital deafness caused by extensive mutations of the GJB2 gene, not limited to the treatment of deafness caused by targeted mutations or knockouts of the GJB2 gene.

[0018] (3) The thyroid hormone and its preparations are small molecule drugs, which can effectively reach the cochlea through the blood-labyrinth barrier and play a role. Therefore, deafness treatment can be achieved through administration methods such as oral administration or intravenous injection.

[0019] Description of the drawings:

[0020] Figure 1 Picture of GJB2 gene knockout mice;

[0021] Figure 2 Genotype identification results of GJB2 gene knockout mice;

[0022] Figure 3 Hearing thresholds of mice in each group after different treatments;

[0023] Figure 4 Cochlear pathological conditions of mice in each group after different treatments; In the control group, the Corti tunnel in the cochlea was open and formed, and the open Corti tunnel was marked by the yellow curve; No Corti tunnel was observed in the cochlea of GJB2 gene knockout group mice, and the cochlear structure was dysplastic; Corti tunnel could be observed in the cochlea of GJB2 gene knockout group mice treated with T3, and the open Corti tunnel was marked by the yellow curve. After treatment with T3, the cochlear structure of GJB2 gene knockout group mice returned to normal.

[0024] Figure 5To observe the survival of hair cells in mice of different treatment groups, Figure A shows the survival of hair cells in the apical turn of the cochlea of control group mice, and all the hair cells in the apical turn of these mice survived; Figure B shows the survival of hair cells in the apical turn of the cochlea of GJB2 gene knockout group mice, and some of the hair cells in the apical turn of these mice died; Figure C shows the survival of hair cells in the apical turn of the cochlea of GJB2 gene knockout group mice treated with T3, and all the hair cells in the apical turn of these mice survived; Figure D shows the survival of hair cells in the middle turn of the cochlea of control group mice, and all the hair cells in the middle turn of these mice survived; Figure E shows the survival of hair cells in the middle turn of the cochlea of GJB2 gene knockout group mice, and a large number of hair cells in the middle turn of these mice died, and the concentrated area of hair cell death is marked by the green square; Figure F shows the survival of hair cells in the middle turn of the cochlea of GJB2 gene knockout group mice treated with T3, and all the hair cells in the middle turn of these mice survived; Figure G shows the survival of hair cells in the basal turn of the cochlea of control group mice, and all the hair cells in the basal turn of these mice survived; Figure H shows the survival of hair cells in the basal turn of the cochlea of GJB2 gene knockout group mice, and some of the hair cells in the basal turn of these mice died; Figure I shows the survival of hair cells in the basal turn of the cochlea of GJB2 gene knockout group mice treated with T3, and all the hair cells in the basal turn of these mice survived;

[0025] Figure 6 Pathological observation diagram of bilateral cochlear specimens of GJB2 gene knockout group mice treated with T4 Specific implementation method:

[0026] The following further describes the invention content of the present invention in detail in combination with specific implementation methods. It should be understood that the embodiments of the present invention are only used to illustrate the present invention and not to limit the present invention. Without departing from the technical idea of the present invention, various substitutions and changes made according to common technical knowledge and conventional means in the art should be included within the scope of the present invention.

[0027] Example 1. Verification of the efficacy of T3 in the treatment of congenital deafness caused by GJB2 gene mutation

[0028] 1. Experimental materials

[0029] (1) Experimental drugs: Triiodothyronine (the main active form of thyroid hormone), abbreviated as T3. This product is a white powder. Purchased from Sigma Corporation, USA, product number: T2877.

[0030] (2) Other reagents:

[0031] Tamoxifen. Analytical pure, white fine grains. Purchased from Sigma Corporation, USA, product number: T5648.

[0032] Sodium hydroxide (NaOH). Analytical pure, white flakes. Purchased from Sinopharm Chemical Reagent Co., Ltd., product number: 10019718.

[0033] Hydrochloric acid (HCl). Purchased from Sinopharm Chemical Reagent Co., Ltd., product number: 10011028.

[0034] 0.9% normal saline, colorless and transparent liquid. Purchased from Sichuan Kelun Pharmaceutical Co., Ltd.

[0035] Corn oil. Yellow transparent liquid. Purchased from Dalian Meilun Biotechnology Co., Ltd., product number: 8001-30-7.

[0036] Experimental animals:

[0037] The strains of GJB2loxp / loxp mice and Rosa26CreER mice are both of CBA strain, SPF grade. Animal source: Gifted by Professor Xi Lin from Emory University, USA.

[0038] Mouse feeding conditions: Mice are housed in the SPF animal house of the Animal Experiment Center of Huazhong University of Science and Technology. Mice are cage-raised, 3-5 per cage. The feeding environment is kept at a constant temperature and humidity, with a temperature of 22-26 °C and a relative humidity of 40% - 50%. The light and dark durations in the feeding environment are both 12 hours. The mouse feed is cobalt-60 irradiated breeding feed for mice and rats, provided by Spf (Beijing) Biotechnology Co., Ltd. Mice are fed with high-pressure sterilized pure water.

[0039] This example verifies the efficacy of thyroid hormone T3 in the treatment of congenital deafness caused by GJB2 gene mutation. The experimental method is as follows:

[0040] (1) Preparation of drugs:

[0041] Prepare sodium hydroxide solution: Dissolve sodium hydroxide in normal saline to prepare a 40 mM / L sodium hydroxide solution.

[0042] Prepare thyroid hormone T3 solution: Dissolve T3 powder in 40 mM / L sodium hydroxide to prepare a solution with a concentration of 1 μg / μL, and adjust the solution to pH = 7.4 - 7.7 with hydrochloric acid for use.

[0043] Prepare tamoxifen solution: Dissolve tamoxifen powder in corn oil to prepare a 7.5 μg / ml solution. It can be heated in a 55 °C metal bath to accelerate dissolution, and can be stored at -20 °C after preparation.

[0044] (2) Construction of GJB2 gene-wide knockout mice:

[0045] By mating GJB2loxp / loxp mice with Rosa26CreER mice, GJB2loxp / loxp; Rosa26CreER mice and GJB2loxp / loxp mice were obtained respectively. On the day of birth and the next day of GJB2loxp / loxp; Rosa26CreER mice and GJB2loxp / loxp mice, tamoxifen solution was injected subcutaneously into the mice, and the injection dose was 1.5 μg / 10 g of mouse body weight. Mice with widespread knockout of the GJB2 gene were obtained ( Figure 1 )(GJB2loxp / loxp; Rosa26CreER) and control group mice (GJB2loxp / loxp).

[0046] (3) Genotyping:

[0047] On the fourth day after the mice were born, toe or tail tip tissues of the mice were cut, and genomic DNA was extracted using a one-step mouse genotype identification kit (purchased from Novoprotein Scientific Inc.) for genotyping. Identify the genotypes carried by the mice.

[0048] (4) Grouping and treatment methods for each group:

[0049] On the fifth day after the mice were born, thyroid hormone T3 solution (dose: 5 μl / 1 g of mouse body weight) or an equal volume of normal saline was injected subcutaneously into GJB2loxp / loxp; Rosa26CreER mice and GJB2loxp / loxp mice. Grouping was based on genotype and treatment method. Control group: GJB2loxp / loxp mice injected with normal saline. T3 treatment group: GJB2loxp / loxp mice injected with thyroid hormone T3 solution. Untreated GJB2 gene knockout group: GJB2loxp / loxp; Rosa26CreER mice injected with normal saline. T3-treated GJB2 gene knockout group: GJB2loxp / loxp; Rosa26CreER mice injected with thyroid hormone T3 solution.

[0050] (5) Hearing detection:

[0051] The hearing of mice was detected on the 18th day after birth. The mice were anesthetized with 1.25% tribromoethanol solution (Avertin), and the anesthetized mice were transferred to an acoustic and electrical shielding box. A hot water bag (37 °C) was used to maintain the body temperature of the anesthetized mice. During the hearing detection process, the status of the mice was observed. The skin at the root of the ear and the top of the skull of the mice was disinfected with 75% alcohol. The recording electrode disinfected with 75% alcohol was inserted subcutaneously under the skull between the two ears, the reference electrode was inserted subcutaneously behind the ear to be tested, and the ground electrode was inserted subcutaneously behind the ear of the contralateral ear. The MF-1 speaker was located 10 cm away from the external auditory canal of the ear to be tested to test the open-field hearing of the mice. The mice were stimulated with ToneBurst sounds of different frequencies (4 kHz, 8 kHz, 12 kHz, 16 kHz, 20 kHz, 24 kHz, 28 kHz, 32 kHz, 40 kHz). The ABR signals of the mice were amplified using an amplifier, and the hearing threshold of the mice was detected using a TDT RZ6 auditory physiology workstation. The latency of the ToneBurst scanning electrical signal was 10 ms, and the stimulation interval was 21 times / s. The initial intensity of the Click and ToneBurst stimulation sounds was 90 dB, which decreased by 5 dB, and the minimum stimulation intensity detected was 15 dB. The ABR waveforms of the mice detected under sound stimulation of different frequencies were recorded, and the minimum stimulation sound intensity that could induce similar ABR waveforms was the hearing threshold.

[0052] (6) Observation of cochlear tissue structure:

[0053] The mice whose hearing had been detected were sacrificed, and the cochleas of the mice were dissected. The cochlear tissues were fixed with freshly prepared 4% paraformaldehyde solution at room temperature for 2 - 3 hours. The cochleas of the mice fixed in the previous step were transferred into a decalcifying solution containing 10% EDTA-2Na for decalcification for 48 hours. After gradient dehydration of the cochlear samples with sucrose solution, the cochlear tissues were embedded with OCT frozen section embedding agent. The embedded cochlear tissues were cut into 10-μm-thick slices using a cryostat. Immunofluorescence staining was performed and observed and photographed under a laser confocal microscope. The decalcified cochlear tissues were directly dissected for immunofluorescence staining, and the cochlear structure was observed and photographed under a confocal microscope.

[0054] (7) Experimental results:

[0055] Result 1: Thyroid hormone T3 successfully treated congenital profound hearing loss caused by GJB2 gene mutation ( Figure 3)。The ABR hearing threshold waveforms of the control group mice showed a "V" shape. Their hearing was relatively poor at low frequencies (4 kHz, 8 kHz) and high frequencies (32 kHz, 40 kHz), and relatively good at medium frequencies (16 kHz, 20 kHz, 24 kHz, 28 kHz), with the lowest audible sound level being 25 dB SPL. The ABR hearing threshold waveforms of the T3 treatment group mice also showed a "V" shape, and the hearing thresholds at each test frequency were the same as those of the control group mice, indicating that the treatment with thyroid hormone T3 does not damage the hearing of normal mice and has good safety. The hearing threshold waveforms of the untreated GJB2 gene knockout group mice showed a "-" shape, and no hearing threshold waveforms could be evoked in the mice with sound stimulation at 90 dB SPL at each test frequency, indicating that the untreated GJB2 gene knockout group mice could not hear sounds and suffered from profound congenital hearing loss across all frequencies. The medium-frequency (16 kHz, 20 kHz, 24 kHz, 28 kHz) and high-frequency (32 kHz, 40 kHz) hearing thresholds of the GJB2 gene knockout group treated with T3 were significantly lower than those of the untreated GJB2 gene knockout group mice. The medium-frequency hearing could be improved by 50 - 65 dB SPL, and the high-frequency hearing could be improved by 30 - 45 dB SPL, indicating that T3 treatment can effectively rescue the profound congenital hearing loss caused by GJB2 gene mutation.

[0056] Result 2: Thyroid hormone T3 successfully treated the cochlear structural development disorder caused by GJB2 gene mutation ( Figure 4 )。In the cochlear specimens of the control group mice, an open Corti tunnel structure (marked with a yellow curve) could be observed. No Corti tunnel structure was observed in the cochlear specimens of the untreated GJB2 gene knockout group mice, indicating that GJB2 gene knockout led to cochlear structural development disorder. An open Corti tunnel structure (marked with a yellow curve) could be observed in the cochlear specimens of the GJB2 gene knockout group treated with T3.

[0057] Result 3: Thyroid hormone T3 successfully treated the loss of cochlear hair cells caused by GJB2 gene mutation ( Figure 5 )。Hair cells are the cells in the cochlea responsible for sound perception. Once hair cells are lost, they cannot be regenerated throughout life, which will lead to cochlear sound perception dysfunction and hearing loss. Intact inner hair cells and three rows of outer hair cells could be observed in the cochlear specimens of the control group mice. Different degrees of loss of inner and outer hair cells were observed in the cochlear specimens of the untreated GJB2 gene knockout group mice in the apical, middle, and basal turns of the cochlea, and the most obvious loss of inner and outer hair cells in the middle turn was the most significant (marked with a green square), indicating that GJB2 gene knockout led to the damage and loss of inner and outer hair cells. Intact inner hair cells and three rows of outer hair cells could be observed in the cochlear specimens of the GJB2 gene knockout group treated with T3, indicating that thyroid hormone T3 successfully treated the loss of cochlear hair cells caused by GJB2 gene mutation.

[0058] Example 2: Verification of the efficacy of T4 in treating congenital deafness caused by GJB2 gene mutation

[0059] 1. Experimental Materials

[0060] (1) Experimental drugs: Tetraiodothyronine /

[0061] L-Thyroxine), abbreviated as T4. This product is white powder. Purchased from MedChemExpress, USA, item number: HY-18341R.

[0062] (2) Other reagents:

[0063] Tamoxifen. Analytically pure, white fine particles. Purchased from Sigma, USA, item number: T5648.

[0064] Sodium hydroxide (NaOH). Analytical grade, white flakes. Purchased from Sinopharm Chemical Reagent Co., Ltd., catalog number: 10019718.

[0065] Hydrochloric acid (HCL) was purchased from Sinopharm Chemical Reagent Co., Ltd., catalog number: 10011028.

[0066] 0.9% physiological saline, colorless and transparent liquid, purchased from Sichuan Kelun Pharmaceutical Co., Ltd.

[0067] Corn oil. Yellow transparent liquid. Purchased from Dalian Meilun Biotechnology Co., Ltd., item number: 8001-30-7.

[0068] Experimental animals:

[0069] The strains of GJB2loxp / loxp mice and Rosa26CreER mice are both CBA strains, SPF grade. Animal source: donated by Professor Xi Lin of Emory University, USA.

[0070] Mouse feeding conditions: Mice were raised in the SPF animal room of the Animal Experiment Center of Huazhong University of Science and Technology. All mice were caged, 3-5 per cage. The breeding environment was kept at a constant temperature and humidity of 22-26°C and a relative humidity of 40% to 50%. The light and dark durations in the breeding environment were both 12 hours. The mouse feed was cobalt-60 irradiated breeding feed for rats and mice, provided by Sibeifu (Beijing) Biotechnology Co., Ltd. The mice were fed with autoclaved purified water.

[0071] This example verifies the efficacy of thyroid hormone (T4) in treating congenital deafness caused by GJB2 gene mutation. The experimental method is as follows:

[0072] (1) Drug configuration:

[0073] Preparation of sodium hydroxide solution: Dissolve sodium hydroxide in physiological saline to prepare a 40 mM / L sodium hydroxide solution.

[0074] Preparation of thyroid hormone T4 solution: Dissolve T4 powder in 0.9% sodium chloride solution with a pH of 7.4 - 7.7 to prepare a solution with a concentration of 1 μg / μL.

[0075] Preparation of tamoxifen solution: Dissolve tamoxifen powder in corn oil to prepare a 7.5 μg / ml solution. It can be heated in a 55°C metal bath to accelerate dissolution, and can be stored at -20°C after preparation.

[0076] (2) Construction of GJB2 gene - widely knocked - out mice:

[0077] Mate GJB2loxp / loxp mice with Rosa26CreER mice to obtain GJB2loxp / loxp; Rosa26CreER mice and GJB2loxp / loxp mice respectively. On the day of birth and the next day of GJB2loxp / loxp; Rosa26CreER mice and GJB2loxp / loxp mice, inject tamoxifen solution subcutaneously into the mice, and the injection dose is 1.5 μg / 10 g of mouse body weight. Obtain mice with widely knocked - out GJB2 gene ( Figure 1 )(GJB2loxp / loxp; Rosa26CreER) and control group mice (GJB2loxp / loxp).

[0078] (3) Genotyping:

[0079] On the fourth day after the mice are born, cut off the mouse toe or tail tip tissue, and use a one - step mouse genotype identification kit (purchased from Novoprotein Scientific Inc.) to extract genomic DNA for genotyping. Identify the genotype carried by the mice.

[0080] (4) Grouping and treatment methods for each group:

[0081] On the fifth day after the mice are born, microinject thyroid hormone T4 solution (dose: 2 μl / mouse weight) into the round window of the cochlea of GJB2loxp / loxp; Rosa26CreER mice.

[0082] (5) Cochlear pathological observation:

[0083] On the 18th day after the mice were born, the mice were sacrificed and the cochleas were dissected. The cochlear tissues were fixed with freshly prepared 4% paraformaldehyde solution at room temperature for 2 - 3 hours. The fixed cochleas of the mice in the previous step were transferred into a decalcifying solution containing 10% EDTA-2Na for 48 hours of decalcification. After gradient dehydration of the cochlear samples with sucrose solution, the cochlear tissues were embedded with OCT cryostat embedding medium. The embedded cochlear tissues were cut into 10-μm thin sections using a cryostat. Immunofluorescence staining was performed and observations were made and photographs were taken under a laser confocal microscope. The decalcified cochlear tissues were directly dissected for immunofluorescence staining, and the cochlear structure was observed and photographed under a confocal microscope.

[0084] (6) Experimental results:

[0085] Thyroid hormone T4 successfully treated the cochlear structural development disorder caused by GJB2 gene mutation ( Figure 6 ). An open Corti tunnel structure (marked with a yellow curve) could be observed in the bilateral cochlear specimens of the T4-treated GJB2 gene knockout group mice.

[0086] In summary, the present invention provides the use of thyroid hormones in the preparation of drugs for treating congenital deafness related to GJB2 gene mutation. Through thyroid hormone T3 or T4, congenital deafness and cochlear structural development disorder caused by GJB2 gene mutation can be effectively treated, expanding the new indications of thyroid hormones beyond hypothyroidism and goiter. While improving the patient's hearing, it can treat the cochlear development disorder of the patient, providing a new treatment approach for congenital deafness caused by GJB2 gene mutation.

[0087] Finally, it should be noted that the above embodiments are only used to illustrate the present invention and do not limit the protection scope of the present invention. Additionally, after reading the technical content of the present invention, those skilled in the art can make various changes, modifications or variations to the present invention, and all these equivalent forms also fall within the protection scope defined by this application.

Claims

1. Application of thyroid hormone in the preparation of drugs for the treatment of congenital deafness associated with GJB2 gene mutation.

2. The use of the thyroid hormone according to claim 1 in the preparation of a drug for treating congenital deafness associated with GJB2 gene mutation, characterized in that: The thyroid hormone is T3 or T4, wherein the T3 is triiodothyronine≡O-(4-hydroxy-3-iodophenyl)-3,5-diiodo-L-tyrosine, and the T4 is levothyroxine≡O-(4-hydroxy-3,5-diiodophenyl)-3,5-diiodo-L-tyrosine.

3. The use of the thyroid hormone according to claim 1 in the preparation of a drug for treating congenital deafness associated with GJB2 gene mutation, characterized in that: The drug is an aqueous dispersion of thyroid hormone T3 or T4. The preparation method of the aqueous dispersion of T3 is: dissolving T3 powder in 40mM sodium hydroxide to prepare a liquid with a concentration of 1μg / μL, and adjusting the solution to pH=7.4-7.7; the preparation method of the aqueous dispersion of T4 is: dissolving T4 powder in physiological saline to prepare a liquid with a concentration of 1μg / μL, and adjusting the solution to pH=7.4-7.

7.

4. The use of thyroid hormone according to claim 1 in the preparation of a drug for treating congenital deafness associated with GJB2 gene mutation, characterized in that: The drug is prepared into an oral dosage form or an injection by adding conventional excipients, or by combining with other drugs for treating deafness. The excipients include disintegrants, lubricants, and adhesives. The other drugs for treating deafness include: dexamethasone, PARP-1 inhibitors, acetylcysteine, and gene therapy drugs.

5. The use of the thyroid hormone according to claim 1 in the preparation of a drug for treating congenital deafness associated with GJB2 gene mutation, characterized in that: The drug is used to treat congenital deafness or cochlear structure development disorder caused by GJB2 gene mutation.

6. The use of thyroid hormone according to claim 1 in the preparation of a drug for treating congenital deafness associated with GJB2 gene mutation, characterized in that: The administration method of the drug includes subcutaneous administration, digestive tract administration or local cochlear administration.

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