Pharmaceutical composition and application thereof in preparation of medicine for treating sudden deafness

Abstract

The invention discloses a pharmaceutical composition and application thereof in preparation of a medicine for treating sudden deafness, pharmacodynamic data shows that all components of the composition have a synergistic effect, the solubility and bioavailability of the composition in water can be remarkably improved, and the pharmaceutical composition can be used for treating sudden deafness. Through an oral administration mode, the hearing threshold of cochlear ischemia reperfusion model animals can be remarkably reduced, release of inflammatory factors such as serum IL-1beta and TNF-alpha is inhibited, oxidative stress is relieved (the malondialdehyde level is reduced), so that multi-target synergistic treatment of sudden deafness is realized, the effect is remarkable, the defects that an existing drug is single in target and inconvenient to administrate are overcome, and the application prospect is wide. The important clinical application value is realized.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a pharmaceutical composition and its application in the preparation of a drug for treating sudden deafness. Background Technology

[0002] Sudden Sensorineural Hearing Loss (SSNHL), also known as sudden deafness, refers to the sudden onset of sensorineural hearing loss within 72 hours of unknown cause, typically with a hearing loss greater than 30 decibels at at least three consecutive frequencies. Patients are often admitted to the hospital with unilateral hearing loss, which may be accompanied by symptoms such as tinnitus, dizziness, and nausea. The medical community generally believes that it is not caused by a single factor, but rather by the combined effect of multiple factors. In summary, the formation mechanism of sudden deafness is complex, and it is likely that the patient has pre-existing vascular or immune predispositions. Under the influence of triggers such as fatigue, stress, or infection, this ultimately leads to impaired blood circulation or inflammatory damage to the inner ear.

[0003] Currently, treatment for sudden deafness primarily relies on medication. The American Clinical Practice Guidelines for Sudden Deafness (2019) only support glucocorticoids as a treatment, limiting their use to within 6 weeks of onset. Systemic or intratympanic injections of hormones are recommended within 2 weeks of onset. For cases lasting more than 2 weeks, intratympanic injections of hormones are recommended as supplementary therapy, and the use of other medications is strongly discouraged. my country's Guidelines for the Diagnosis and Treatment of Sudden Deafness (2015) list glucocorticoids as the first-line treatment, recommending systemic oral or intravenous administration within 2 weeks of onset. Hormones are often used in combination with drugs that improve inner ear microcirculation. Based on the type of hearing loss, batroxobin is recommended for flat and total deafness. The German guidelines prioritize drugs that improve inner ear hemorheology as the primary treatment, with hormone therapy as the second choice. Therefore, in summary, the main treatments for sudden deafness currently include glucocorticoids, drugs that improve inner ear circulation, and fibrinolytic drugs. However, these single-target drugs are insufficient to cover multiple causes, and clinical feedback on treatment effectiveness is not optimistic.

[0004] Some studies have shown that puerarin injection can treat sudden deafness, but existing injections require intravenous administration, which leads to poor patient compliance and can easily cause side effects such as phlebitis with long-term use.

[0005] Therefore, developing a drug composition that is orally effective and can synergistically improve multiple pathological mechanisms has important clinical value. Summary of the Invention

[0006] To address the shortcomings of the prior art, the present invention aims to provide an application of a pharmaceutical composition in the preparation of a drug for treating sudden deafness. The components of this composition have synergistic effects, which can significantly improve the solubility and bioavailability in water. It can improve cochlear ischemia-reperfusion injury and inhibit inflammation and oxidative stress through oral administration, thereby achieving the goal of multi-target synergistic treatment of sudden deafness.

[0007] This invention is achieved through the following technical solution: The use of a pharmaceutical composition in the preparation of a drug for treating sudden deafness, said pharmaceutical composition being made from the following components in parts by weight: Mangiferin-7-O-glucoside 4-6 parts, daidzein 6-9 parts, 3'-methoxypuerarin 4-6 parts, puerarin 44-54 parts, puerarin B 1-3 parts, daidzein-8-C-apigenyl-(1-6)-glucoside 17-21 parts, puerarin xyloside 2-4 parts, daidzein-7,4'-diglucoside 0.5-1.5 parts, 3'-hydroxypuerarin 4-6 parts, puerarin-4'-O-glucoside 3-5 parts, mefosine-4'-glucoside 0.1-0.5 parts, 4'-methoxygenin 0.5-1.5 parts, puerarin C 0.5-1.5 parts.

[0008] As a preferred technical solution, the weight parts of each component in the pharmaceutical composition are as follows: 5 parts of gentianin-7-O-glucoside, 7 parts of daidzein, 5 parts of 3'-methoxypuerarin, 49 parts of puerarin, and puerarin B 2 parts, daidzein-8-C-apigenyl-(1-6)-glucoside 19 parts, puerarin xyloside 3 parts, daidzein-7,4'-diglucoside 1 part, 3'-hydroxypuerarin 5 parts, puerarin-4'-O-glucoside 3.5 parts, mefosine-4'-glucoside 0.4 parts, 4'-methoxygenyl genistein 1 part, puerarin C 1 part.

[0009] In the pharmaceutical composition of this invention, puerarin primarily improves inner ear microcirculation. 3'-methoxypuerarin and 3'-hydroxypuerarin are derivatives of puerarin, which can enhance the vasodilatory effect of puerarin and synergistically improve inner ear microcirculation with puerarin. Puerarin xylose and puerarin-4'-O-glucoside optimize water solubility through glycan structure optimization, promote intestinal absorption, and rapidly reach the inner ear target tissue, enhancing the bioavailability of puerarin-like components and solving the problem of poor oral absorption of puerarin alone. Daidzin, daidzin-8-C-apigenyl-(1-6)-glucoside, and 4'-methoxygenyl genistein synergistically inhibit IL-1β and TNF-α. Inflammatory factors; puerarin B, puerarin C, gentianin-7-O-glucoside and daidzein-7,4'-diglucoside synergistically reduce oxidative stress; the synergistic effect between the components in this drug composition can significantly improve the solubility and bioavailability in water, and treat sudden deafness through multi-target synergistic therapy.

[0010] The sudden hearing loss described in this invention includes low-frequency hearing loss, mid-frequency hearing loss, high-frequency hearing loss, flat hearing loss, and total hearing loss. Its clinical manifestations include sudden hearing loss in one or both ears, auditory nerve inflammation or sensory nerve damage, or tinnitus, dizziness, ear fullness, ear discomfort, or balance disorder.

[0011] The pharmaceutical composition of the present invention can exert a therapeutic effect by synergistically improving cochlear ischemia-reperfusion injury, inhibiting inflammatory response and reducing oxidative stress.

[0012] In a preferred embodiment of the present invention, the medicament comprises an effective amount of a pharmaceutical composition and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers of the present invention include, but are not limited to, lubricants, pH adjusters, penetration enhancers, plasticizers, antioxidants, preservatives, etc., in amounts conventionally used in the art.

[0013] The drug of the present invention can be prepared in any pharmaceutically applicable dosage form according to conventional methods in the art.

[0014] As a preferred embodiment of the present invention, the dosage form of the drug includes tablets, capsules, granules, oral liquids, pills, tinctures, or ointments.

[0015] As a preferred embodiment of the present invention, the drug of the present invention can be administered orally. The safe and effective dose of the drug composition of the present invention is 250-1000 mg / time, twice a day.

[0016] Compared with the prior art, the present invention has the following advantages: This invention provides a novel use of a pharmaceutical composition in the preparation of drugs for treating sudden deafness. Pharmacodynamic data show that the components of this composition have a synergistic effect, significantly improving solubility and bioavailability in water. Oral administration can significantly reduce the hearing threshold in cochlear ischemia-reperfusion model animals, inhibit the release of inflammatory factors such as serum IL-1β and TNF-α, and alleviate oxidative stress (by reducing malondialdehyde levels). This achieves multi-target synergistic treatment of sudden deafness with significant efficacy, overcoming the shortcomings of existing drugs with single targets and inconvenient administration, and has important clinical application value. Attached Figure Description

[0017] Figure 1 The effect of each group on the auditory brainstem response threshold of guinea pigs with cochlear ischemia-reperfusion injury; Figure 2 The effects of each group on serum IL-1β in guinea pigs with cochlear ischemia-reperfusion injury; Figure 3 The effects of each group on serum TNF-α in guinea pigs with cochlear ischemia-reperfusion injury; Figure 4 The effect of each group on serum malondialdehyde in guinea pigs with cochlear ischemia-reperfusion injury; Figure 5 The pathological scores of the cochlea of ​​guinea pigs with cochlear ischemia-reperfusion injury in each group are shown. Detailed Implementation

[0018] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. The described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0019] The raw materials used in the embodiments of the present invention can be obtained by separation and purification using conventional methods in the art.

[0020] Example: Pharmacodynamic study of a pharmaceutical composition in a guinea pig model of cochlear ischemia-reperfusion. 1. Preparation of pharmaceutical compositions Weigh the following components in the indicated weight proportions: 5 parts of daidzein-7-O-glucoside, 7 parts of genistein, 5 parts of 3'-methoxypuerarin, 49 parts of puerarin, 2 parts of puerarin B, 19 parts of daidzein-8-C-apigenyl-(1-6)-glucoside, 3 parts of puerarin xyloside, 1 part of daidzein-7,4'-diglucoside, 5 parts of 3'-hydroxypuerarin, 3.4 parts of puerarin-4'-O-glucoside, 0.4 parts of mefosine-4'-glucoside, 1 part of 4'-methoxygenyl genistein, and 1 part of puerarin C. Mix the above components thoroughly to obtain the pharmaceutical composition.

[0021] 2. Experimental Methods: Hartley guinea pigs were acclimatized for 7 days after purchase. Before the formal experiment, all animals were anesthetized with salbutamol / xilazine and their hearing was confirmed to be normal by auditory brainstem response testing. Subsequently, 60 guinea pigs were randomly selected to establish a cochlear ischemia-reperfusion model by severing both vertebral arteries with a monopolar electrocoagulator and clamping the right common carotid artery with an arterial clamp for 30 minutes before opening it. The hearing of the right cochlea of ​​the animals was tested using a small animal auditory brainstem testing instrument after surgery. After confirming the successful establishment of the model, the model animals were randomly divided into 5 groups: model control group, positive drug group (prednisone, 5.42 mg / kg), puerarin monomer group, low-dose group of the formulation (composition, 40 mg / kg), medium-dose group of the formulation (composition, 80 mg / kg), and high-dose group of the formulation (composition, 160 mg / kg). Ten normal guinea pigs were used as a normal control group. All experimental animals were administered the drugs orally for 7 consecutive days.

[0022] 3. Evaluation indicators: Physiological indicator: Auditory brainstem response threshold.

[0023] Biochemical indicators: serum IL-1β, TNF-α, and malondialdehyde levels.

[0024] Pathological indicators: HE staining was used to observe the morphology and structure of the cochlea.

[0025] 4. Statistical methods: Experimental results are expressed as mean ± standard deviation. The results indicate that the data between the two groups were analyzed using SPSS 25.0 software with a two-way ANOVA and the Sidak test to determine that there was a highly significant difference between the two groups, with P < 0.05 defined as a statistically significant difference.

[0026] 5. Experimental Results 5.1 Auditory brainstem response threshold After 7 days of continuous administration, the results were as follows: Figure 1As shown, compared with the normal group, the hearing threshold of the model group was significantly increased, with statistically significant differences (P<0.01 vs. model group). Compared with the model group, the hearing threshold of the positive control drug prednisone was significantly decreased, with statistically significant differences (P<0.01 vs. model group). Compared with the model group, the low-dose group showed a significant decrease in hearing threshold, with statistically significant differences (P<0.01 vs. model group). Compared with the model group, the medium-dose group showed a significant decrease in hearing threshold, with statistically significant differences (P<0.01 vs. model group). Compared with the model group, the high-dose group showed a significant decrease in hearing threshold, with statistically significant differences (P<0.01 vs. model group). Furthermore, the effect of the composition on reducing hearing threshold increased with increasing dosage, exhibiting a dose-dependent trend.

[0027] Compared with the model group, the hearing threshold of the puerarin control group did not show a downward trend. Statistical results showed no significant difference (P>0.05, vs. model group), indicating that oral administration of puerarin to the control group had no significant effect on improving hearing in guinea pigs with cochlear ischemia-reperfusion.

[0028] 5.2 Biochemical index detection After 7 days of continuous administration, the results were as follows: Figure 2 As shown, compared with the normal group, the serum inflammatory marker IL-1β was increased in the model group, and the statistical results showed a significant difference (P<0.05 vs. model group). Compared with the model group, the serum inflammatory marker IL-1β was decreased in the positive control group (prednisone), and the statistical results showed a significant difference (P<0.05 vs. model group). Compared with the model group, the low, medium, and high dose groups of the formulation showed a decreasing trend in serum IL-1β. The statistical results showed no significant difference in low dose, a significant difference in medium dose (P<0.05 vs. model group), and a significant difference in high dose (P<0.01 vs. model group). Compared with the model group, the puerarin control group showed no decreasing trend in IL-1β, and the statistical results showed no significant difference (P>0.05 vs. model group).

[0029] like Figure 3As shown, compared with the normal group, the serum inflammatory marker TNF-α in the model group was significantly increased, with statistical results showing a highly significant difference (P<0.01 vs. model group). Compared with the model group, the serum inflammatory marker TNF-α in the positive control group (prednisone) was decreased, with statistical results showing a significant difference (P<0.05 vs. model group). Compared with the model group, the serum inflammatory marker TNF-α in the low, medium, and high dose groups of the formulation was significantly decreased in the low, medium, and high dose groups of the formulation, with statistical results showing a highly significant difference (P<0.01 vs. model group). Compared with the model group, the puerarin control group showed no decreasing trend in TNF-α, with statistical results showing no significant difference (P>0.05 vs. model group). This indicates that oral administration of puerarin to the control group could not reduce the inflammatory response in guinea pigs with cochlear ischemia-reperfusion.

[0030] like Figure 4 As shown, compared with the normal group, the serum oxidative stress index malondialdehyde (MDA) was significantly increased in the model group, with statistically significant differences (P < 0.01 vs. model group). Compared with the model group, the serum MDA in the positive control group (prednisone) was decreased, with statistically significant differences (P > 0.05 vs. model group). Compared with the model group, the serum MDA in the low-dose group was decreased, with statistically significant differences (P > 0.05 vs. model group). Compared with the model group, the serum MDA in the medium- and high-dose groups was significantly decreased, with statistically significant differences (P < 0.05 vs. model group).

[0031] Compared with the model group, the MDA of the puerarin control group did not show a decreasing trend. Statistical results showed no significant difference (P>0.05, vs. model group), indicating that oral administration of puerarin to the control group could not alleviate the oxidative stress response in guinea pigs with cochlear ischemia-reperfusion.

[0032] 5.3 Pathological Results After seven consecutive days of drug administration, cochlear tissue samples were collected from different groups of guinea pigs, fixed with paraformaldehyde, and stained with hematoxylin and eosin (HE). The staining results were scored according to the following criteria: Table 1 below. Table 1 Scoring Criteria

[0033] The results are as follows Figure 5As shown, compared with the normal group, the pathological score of the model group was significantly higher, and the statistical results showed a highly significant difference (P<0.01 vs. model group). Compared with the model group, the pathological score of the positive control drug prednisone was significantly lower, and the statistical results showed a significant difference (P<0.01 vs. model group). Compared with the model group, the pathological scores of the low, medium, and high dose groups of the formulation were significantly lower, and the statistical results showed a highly significant difference (P<0.01 vs. model group). With the increase of dose, the pathological damage was significantly reduced, showing a good dose-dependent effect. Compared with the model group, the puerarin control group showed no decreasing trend, and the statistical results showed no significant difference (P>0.05 vs. model group), indicating that oral puerarin control group could not reduce the pathological damage of cochlear ischemia-reperfusion guinea pigs.

[0034] In summary, the pharmaceutical composition of this invention, when administered orally, significantly reduces the hearing threshold in cochlear ischemia-reperfusion model animals, inhibits inflammatory factors such as serum IL-1β and TNF-α, and alleviates oxidative stress (by reducing malondialdehyde levels), exhibiting a dose-dependent synergistic effect, thus demonstrating its therapeutic efficacy for sudden deafness. Furthermore, compared with oral administration of puerarin alone, the efficacy of the pharmaceutical composition of this invention is significantly superior to that of puerarin alone.

Claims

1. The use of a pharmaceutical composition in the preparation of a drug for treating sudden deafness, characterized in that, The pharmaceutical composition is made of the following components by weight fraction: Formononetin-7-O-glucoside 4-6 parts, daidzin 6-9 parts, 3'-methoxy puerarin 4-6 parts, puerarin 44-54 parts, puerarin glycoside B 1-3 parts, daidzein-8-C-apioyl-(1-6)-glucoside 17-21 parts, puerarin xyloside 2-4 parts, daidzein-7, 4'-diglucoside 0.5-1.5 parts, 3'-hydroxypuerarin 4-6 parts, puerarin-4'-O-glucoside 3-5 parts, methoxy genistin 0.1-0.5 parts, 4'-methoxy genistin 0.5-1.5 parts, puerarin glycoside C 0.5-1.5 parts.

2. Use according to claim 1, characterized in that, The weight fraction of each component is: Formononetin-7-O-glucoside 5 parts, daidzin 7 parts, 3'-methoxy puerarin 5 parts, puerarin 49 parts, puerarin glycoside B 2 parts, daidzein-8-C-apioyl-(1-6)-glucoside 19 parts, puerarin xyloside 3 parts, daidzein-7, 4'-diglucoside 1 part, 3'-hydroxypuerarin 5 parts, puerarin-4'-O-glucoside 3.5 parts, methoxy genistin 0.4 parts, 4'-methoxy genistin 1 part, puerarin glycoside C 1 part. The sudden deafness includes low frequency drop type, medium frequency drop type, high frequency drop type, flat drop type and total deafness type, and the clinical manifestations include unilateral or bilateral sudden drop of ear hearing, inflammation of auditory nerve or damage of sensory nerve, or accompanied by tinnitus, dizziness, ear congestion, ear discomfort or balance disorder.

3. Use according to claim 1 or 2, characterized in that, The pharmaceutical composition plays a therapeutic role by synergistically improving cochlear ischemia-reperfusion injury, inhibiting inflammatory response and reducing oxidative stress.

4. Use according to claim 1 or 2, characterized in that, The medicine includes an effective amount of the pharmaceutical composition and a pharmaceutically acceptable carrier.

5. Use according to claim 1 or 2, characterized in that, The medicine is administered by oral administration.

6. Use according to claim 5, characterized in that, The dosage form of the medicine includes tablets, capsules, granules, oral liquids, dripping pills, tinctures or ointments.

7. Use according to claim 6, characterized in that, ​

Images