Baicalin eye drops as well as preparation method and application thereof

Abstract

The invention belongs to the field of biological medicine preparations, and particularly relates to baicalin eye drops as well as a preparation method and application thereof. The formula of the baicalin eye drops comprises the following raw material components: baicalin, mannitol and ultrapure water. The baicalin eye drops provided by the invention are reasonable in formula, mild and non-irritant in use, and free of pricking and discomfort, and a convenient and effective non-surgical treatment means is provided for patients with diabetic cataract.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to eye drops, a preparation method and application thereof. Background Art

[0002] Since the beginning of the 21st century, people's living standards have continued to improve. Changes in lifestyle and eating habits, environmental pollution, mental stress and other factors have led to an upward trend in the prevalence of diabetes. Diabetes seriously endangers human health and has become a global public health problem. Diabetic cataract (DC) is one of the most common and earliest complications of diabetes. Clinically, cataracts caused by abnormal glucose metabolism such as diabetes, galactosemia, and xylosemia are collectively referred to as diabetic cataracts, or sugar cataracts for short.

[0003] DC is not only an important factor in blindness in diabetic patients, but also seriously affects their daily lives. The formation of DC leads to turbidity of the refractive medium, which often makes it impossible for patients to further identify fundus lesions such as diabetic retinopathy, ultimately leading to irreversible visual impairment or even blindness, and increasing medical costs. Although there are many studies on the mechanism and drug intervention treatment of cataracts at home and abroad, and various different pharmacological effects and even traditional Chinese medicine have confirmed the potential protective effect of traditional Chinese medicine on the lens, there is still no effective drug method other than surgery. The cost of surgery is a heavy burden on patients, and the risk of delayed wound healing, corneal and macular edema, and infection after surgery is increased. Therefore, although surgery is the fastest and most effective treatment, it is also necessary to find non-surgical methods to delay the development of DC. So far, the exact pathogenesis of diabetic cataracts is still unclear. Preliminary results show that metabolic abnormalities such as increased serum tryptophan, lens uric acid, and polyols are related to lens opacities in diabetes. At present, the clinical treatment of diabetic cataracts mostly adopts surgical methods such as phacoemulsification and femtosecond laser surgery. There are also oral preparations such as capsules and tablets, but there are few topical ophthalmic preparations for the treatment of diabetic cataracts. According to previous reports and related studies, the Chinese medicine ingredient baicalin can delay the development of diabetic cataracts by inhibiting aldose reductase, but it is limited to mechanism research, and the clinically available drug dosage form is not disclosed. Therefore, based on the above problems, the present invention uses the unique natural active ingredients of traditional Chinese medicine to develop a baicalin eye drop that can effectively delay the development of diabetic cataracts. Summary of the invention

[0004] In order to solve the problem of single treatment method and difficult prognosis for treating cataract, especially diabetic cataract, the purpose of the present invention is to provide a non-surgical method for delaying the development of cataract, especially diabetic cataract, using baicalin eye drops.

[0005] The technical purpose of the present invention is achieved through the following technical solutions:

[0006] The baicalin eye drops of the present invention include the following raw material components in parts by weight:

[0007] 0.5-5 parts of baicalin; 9-10 parts of mannitol, 200 parts of ultrapure water.

[0008] Preferably, the formula of the baicalin eye drops of the present invention comprises the following raw material components in parts by weight: 1-2 parts of baicalin, 9.5-9.7 parts of mannitol, and 200 parts of ultrapure water.

[0009] Another technical object of the present invention is to provide a method for preparing baicalin eye drops, comprising the following steps:

[0010] 1) Prepare the raw material components according to the above weight proportions;

[0011] 2) Add mannitol to some ultrapure water and stir until completely dissolved;

[0012] 3) adding baicalin to the solution obtained in step 2) and stirring until baicalin is dissolved;

[0013] 4) Add ultrapure water to make up to the full volume.

[0014] Wherein, the stirring time of step 2) is 2-10 minutes, preferably 4-6 minutes, most preferably 5 minutes.

[0015] The stirring time in step 3) is 20-100 minutes, preferably 40-80 minutes, and most preferably 60 minutes.

[0016] Preferably, after step 4), the method further comprises sterilizing and sealing the prepared baicalin eye drops, preferably by filtering with a 0.22 μm filter membrane.

[0017] The present invention also aims to provide the above-mentioned eye drops for preparing a drug for delaying the development of cataracts, especially diabetic cataracts, by non-surgical methods.

[0018] The beneficial effects that can be achieved by the present invention using the above technical solution are:

[0019] The formula of the baicalin eye drops of the present invention is simple and reasonable. After auxiliary material screening research, the mannitol in the prescription can better regulate the osmotic pressure. As a non-electrolyte, mannitol generates osmotic pressure in the solution through molecular weight, so the osmotic pressure of the solution can be effectively regulated at a low concentration. At the same time, mannitol has non-ionicity in the solution, good chemical stability, is not easy to react with other components, and helps to maintain the activity of baicalin, and can maintain the long-term stability of the eye drops. The eye drops are very gentle when used, without stinging and discomfort.

[0020] Furthermore, animal experiments have shown that, compared with the diabetes model group and the Shapuaisi eye drops group, the rats treated with the baicalin eye drops of the present invention for 30 days had the lowest incidence of cataract and the lowest degree of lens opacity. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 These are the effects of normal rats, diabetic model rats and different drug groups on diabetic model rats after treatment for 1, 2, 3 and 4 weeks.

[0022] Figure 2 Cataract scores of rats in different groups after 4 weeks of treatment DETAILED DESCRIPTION

[0023] The technical solution of the present invention will be described clearly and completely below in conjunction with the accompanying drawings and specific embodiments. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them; based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention. It should be noted that, unless otherwise specified, the raw materials involved in the present invention are purchased through conventional commercial channels.

[0024] Example 1 Baicalin Eye Drops

[0025] 1. Formula of baicalin eye drops: baicalin 2g, mannitol 9g, ultrapure water 200ml.

[0026] 2. Preparation process of baicalin eye drops:

[0027] 1) Prepare the raw material components according to the formula;

[0028] 2) Take a beaker, add the prescribed amount of mannitol to 180 ml of ultrapure water and stir for 5 minutes until it is completely dissolved;

[0029] 3) Weigh baicalin, add it to the solution obtained in step 2), and stir and disperse for 60 minutes until the baicalin is dissolved.

[0030] 4) Add ultrapure water to make up to the full volume, filter with a 0.22μm filter membrane, and seal.

[0031] [Dosage and Usage of Baicalin Eye Drops] Apply to eyes, 1 drop at a time, 4 times a day.

[0032] Example 2 Baicalin Eye Drops

[0033] 1. Formula of baicalin eye drops: baicalin 2g, mannitol 9.57g, ultrapure water 200ml.

[0034] 2. Processing of baicalin eye drops:

[0035] 1) Prepare the raw material components according to the formula;

[0036] 2) Take a beaker, add the prescribed amount of mannitol to 180 ml of ultrapure water and stir for 5 minutes until it is completely dissolved;

[0037] 3) Weigh baicalin, add it to the solution obtained in step 2), and stir and disperse for 60 minutes until the baicalin is dissolved.

[0038] 4) Add ultrapure water to make up to the full volume, filter with a 0.22μm filter membrane, and seal.

[0039] [Dosage and Usage of Baicalin Eye Drops] Apply to eyes, 1 drop at a time, 4 times a day.

[0040] Example 3 Baicalin Eye Drops

[0041] 1. Formula of baicalin eye drops: baicalin 2g, mannitol 10g, ultrapure water 200ml.

[0042] 2. Processing of baicalin eye drops:

[0043] 1) Prepare the raw material components according to the formula;

[0044] 2) Take a beaker, add the prescribed amount of mannitol to 180 ml of ultrapure water and stir for 5 minutes until it is completely dissolved;

[0045] 3) Weigh baicalin, add it to the solution obtained in step 2), and stir and disperse for 60 minutes until the baicalin is dissolved.

[0046] 4) Add ultrapure water to make up to the full volume, filter with a 0.22μm filter membrane, and seal.

[0047] [Dosage and Usage of Baicalin Eye Drops] Apply to eyes, 1 drop at a time, 4 times a day.

[0048] Example 4 Baicalin Eye Drops

[0049] 1. Formula of baicalin eye drops: baicalin 0.5g, mannitol 9.57g, ultrapure water 200ml.

[0050] 2. Processing of baicalin eye drops:

[0051] 1) Prepare the raw material components according to the formula;

[0052] 2) Take a beaker, add the prescribed amount of mannitol to 180 ml of ultrapure water and stir for 5 minutes until it is completely dissolved;

[0053] 3) Weigh baicalin, add it to the solution obtained in step 2), and stir and disperse for 60 minutes until the baicalin is dissolved.

[0054] 4) Add ultrapure water to make up to the full volume, filter with a 0.22μm filter membrane, and seal.

[0055] [Dosage and Usage of Baicalin Eye Drops] Apply to eyes, 1 drop at a time, 4 times a day.

[0056] Example 5 Baicalin Eye Drops

[0057] 1. Formula of baicalin eye drops: baicalin 5g, mannitol 9.57g, ultrapure water 200ml.

[0058] 2. Process of baicalin eye drops:

[0059] 1) Prepare the raw material components according to the formula;

[0060] 2) Take a beaker, add the prescribed amount of mannitol to 180 ml of ultrapure water and stir for 5 minutes until it is completely dissolved;

[0061] 3) Weigh baicalin, add it to the solution obtained in step 2), and stir and disperse for 60 minutes until the baicalin is dissolved.

[0062] 4) Add ultrapure water to make up to the full volume, filter with a 0.22μm filter membrane, and seal.

[0063] [Dosage and Usage of Baicalin Eye Drops] Apply to eyes, 1 drop at a time, 4 times a day.

[0064] Experimental study on the treatment of diabetic cataract with baicalin eye drops

[0065] Evaluation of the therapeutic effect of baicalin eye drops prepared in Example 2 on diabetic cataract in rats:

[0066] a. Construction of diabetic cataract rat model

[0067] (1) After SD rats were adaptively fed for 7 days, they were weighed and recorded as initial body weight. The rats were divided into a normal group (n=6) and a diabetic group (n=40). The rats in the diabetic and normal groups were fasted for 12-16 hours, but not water. The diabetic group received a single intraperitoneal injection of STZ solution at a dose of 70 mg / kg. Note: The STZ solution was injected within 30 minutes.

[0068] (2) After the injection of STZ solution, the rats were kept normally for three days and observed for mortality. Then the diabetic rats were divided into groups.

[0069] (3) After all rats were injected and continued to be fed for 5-7 days, random blood glucose values ​​were measured (without fasting). If ≥16.7mmol / L, the diabetic rat model was successfully established. Since fasting blood glucose testing is the most accurate, subsequent tests were still based on fasting blood glucose values ​​(without fasting for 8-12 hours), that is, ≥11.1mmol / L was used to determine that the diabetic rat model was successfully established.

[0070] (4) Starting on the third day after injection, the patient should be examined with a slit lamp after dilating the pupil with tropicamide eye drops (1 drop / time). If the animal's image is blurred and initial lens opacity is observed using a handheld slit lamp, it indicates that the animal has developed early cataracts after the formation of diabetic lesions.

[0071] b. Processing group settings

[0072] Normal group: The mice were fed normally and no medication was given daily.

[0073] Model group: After the diabetic model was successfully established in rats, no medication was given daily.

[0074] Shapuaisi eye drops group: After the rats successfully established the diabetic model, the drug was administered to the rats' eyes four times a day, one drop each time. Low-dose baicalin eye drops group: After the rats successfully established the diabetic model, the drug was administered to the rats' eyes twice a day, one drop each time.

[0075] High-dose baicalin eye drops group: After the diabetic model was successfully established in rats, the drug was administered to the rats' eyes four times a day, one drop each time.

[0076] c. After successful establishment of the diabetes model: measure the fasting blood glucose of rats every week, and use a handheld slit lamp to examine the lens opacity of rats.

[0077] d. Animal experiment results:

[0078] The effect of normal rats, diabetic model rats and different drug groups on diabetic model rats after 1, 2, 3 and 4 weeks of treatment (see Figure 1 )

[0079] The number and score of cataractous eyes of the surviving rats in each group at the fourth week are shown in Tables 1 and Figure 2 The results showed that 4 weeks after administration, the model group had the highest number of eye disease incidences, with an incidence rate of 100%, while the high-dose group had the lowest number of eye disease incidences, with an incidence rate of 70%, followed by the low-dose group with an incidence rate of 80%, and the positive drug group had an incidence rate of 93.75%.

[0080] Table 1 Number and score of cataractous eyes in surviving rats in each group at the fourth week

[0081]

[0082] Note: Compared with the normal group, * indicates P < 0.05, compared with the model group.

[0083] In terms of morbidity, the morbidity in both the high-dose group and the low-dose group was significantly higher than that in the model group (P 高 =0.012, P 低=0.048), and the positive drug group was not statistically significant (P>0.05), indicating that baicalin eye drops can significantly delay the onset of diabetic cataract, and its effect is better than that of Shapuaisi eye drops, but it is not statistically significant (P>0.05).

[0084] In terms of the cumulative number of eyeballs with disease and their scores, compared with the normal group, the cataract score in the model group was significantly higher, with a significant difference (P<0.05), indicating that the diabetic cataract model was successfully established; the scores of the drug-treated groups also showed significant differences (P<0.05), but the statistical chart showed a clear trend, indicating that baicalin eye drops and Shapuaisi eye drops have a certain delaying effect on diabetic cataract in rats. Compared with the model group, the scores of the high-dose group and the low-dose group were both significant (P<0.05), and the high-dose group was more significant, indicating that baicalin eye drops had a good effect; compared with the positive drug group, the scores of the high-dose group and the low-dose group were significantly lower (P<0.05), indicating that baicalin eye drops were better than Shapuaisi eye drops in delaying the onset of diabetic cataract, and the high-dose effect was better than the low-dose.

[0085] In summary, the baicalin eye drops prepared in Example 2 can effectively delay the onset and progression of cataract in diabetic rats.

[0086] a. Preparation of baicalin eye drops:

[0087] Sample 1: Baicalin eye drops prepared according to Example 2 above

[0088] Sample 2: Baicalin eye drops prepared as in Comparative Example 1

[0089] 1. Formula of baicalin eye drops: baicalin 2g, sodium chloride 4.5g, ultrapure water 200ml.

[0090] 2. Processing of baicalin eye drops:

[0091] 1) Prepare the raw material components according to the formula;

[0092] 2) Take a beaker, add the prescribed amount of sodium chloride to 180 ml of ultrapure water and stir for 5 minutes until it is completely dissolved;

[0093] 3) Weigh baicalin, add it to the solution obtained in step 2), and stir and disperse for 60 minutes until the baicalin is dissolved.

[0094] 4) Add ultrapure water to make up to the full volume, filter with 0.22μm filter membrane, and seal.

[0095] b. Content determination

[0096] 1) Chromatographic conditions

[0097] A C18 column (4.6 mm × 250 mm, 5 μm) was used as the test chromatographic column, the mobile phase was methanol-0.1% phosphoric acid solution (47:53), the detection wavelength was 280 nm, the column temperature was 30°C, the flow rate was 1.0 mL / min, and the injection volume was 5 μL. The theoretical plate number calculated according to the baicalin peak area should be no less than 2500, and the resolution should be greater than 1.5.

[0098] 2) Treatment of reference solution

[0099] Accurately weigh 0.5 g of baicalin reference substance, place it in a 50 mL brown volumetric flask, add methanol to dissolve, take 5 mL of the above solution, place it in a 25 mL brown volumetric flask, add methanol to the scale, shake well, filter, and obtain

[0100] 3) Treatment of test solution

[0101] Take 5 mL of sample 1 and sample 2 solutions respectively, place them in two 25 mL brown volumetric flasks, add methanol to the mark, shake well, and filter to obtain the solution.

[0102] 4) Samples 1 and 2 were subjected to a long-term stability study (25°C ± 2°C, RH 60% ± 5%), stored away from light, and sampled and tested at 0, 6, 12, and 16 months. The results are shown in Table 2.

[0103] Table 2 Results of long-term stability test of baicalin eye drops

[0104]

[0105] C. Results

[0106] As can be seen from the results in Table 2, using mannitol as an osmotic pressure regulator for baicalin eye drops can significantly improve the stability of eye drops. Compared with sodium chloride as an osmotic pressure regulator, mannitol, as a non-electrolyte, produces osmotic pressure in the solution through molecular weight, so it can effectively regulate the osmotic pressure of the solution at low concentrations. At the same time, mannitol has non-ionicity in the solution, good chemical stability, is not easy to react with other ingredients, and helps to maintain the activity of baicalin, and can maintain the long-term stability of eye drops. These properties all help to reduce the irritation of eye drops to the ocular surface and improve the stability of eye drops, so this patent uses mannitol as an osmotic pressure regulator.

Claims

1. A baicalin eye drop, characterized in that: In parts by weight, the raw material components include: 0.5-5 parts of baicalin; 9-10 parts of mannitol, 200 parts of ultrapure water.

2. The baicalin eye drops according to claim 1, characterized in that The preparation comprises the following raw material components in parts by weight: 1-2 parts of baicalin, 9.5-9.7 parts of mannitol, and 200 parts of ultrapure water.

3. The baicalin eye drops according to claim 1, characterized in that The preparation method comprises the following raw material components in parts by weight: 0.5 parts of baicalin, 9.57 parts of mannitol and 200 parts of ultrapure water.

4. The baicalin eye drops according to claim 1, characterized in that The preparation comprises the following raw material components in parts by weight: 2 parts of baicalin, 10 parts of mannitol and 200 parts of ultrapure water.

5. The baicalin eye drops according to claim 1, characterized in that The preparation method comprises the following raw material components in parts by weight: 2 parts of baicalin, 9 parts of mannitol and 200 parts of ultrapure water.

6. The baicalin eye drops according to claim 1, characterized in that The preparation method comprises the following raw material components in parts by weight: 2 parts of baicalin, 9.57 parts of mannitol and 200 parts of ultrapure water.

7. The baicalin eye drops according to claim 1, characterized in that The preparation method comprises the following raw material components in parts by weight: 5 parts of baicalin, 9.57 parts of mannitol and 200 parts of ultrapure water.

8. A method for preparing the baicalin eye drops according to any one of claims 1 to 7, characterized in that: The following steps are involved: 1) Prepare each raw material component by weight; 2) Add mannitol to some ultrapure water and stir until completely dissolved; 3) adding baicalin to the solution obtained in step 2) and stirring until baicalin is dissolved; 4) Add ultrapure water to make up to the full volume.

9. The method for preparing baicalin eye drops according to claim 8, characterized in that: in, The stirring time of step 2) is 2-10 minutes, and the stirring time of step 3) is 20-100 minutes.

10. The method for preparing baicalin eye drops according to claim 9, characterized in that: in, The stirring time of step 2) is 4-6 minutes, and the stirring time of step 3) is 40-80 minutes.

11. The method for preparing baicalin eye drops according to claim 10, characterized in that: in, The stirring time of step 2) is 5 minutes, and the stirring time of step 3) is 60 minutes.

12. The method for preparing baicalin eye drops according to any one of claims 8 to 11, characterized in that: After step 4), the method further includes the steps of sterilizing and sealing the prepared baicalin eye drops.

13. The method for preparing baicalin eye drops according to claim 12, characterized in that: Sterilize by filtering with a 0.22 μm membrane.

14. A baicalin eye drop for preventing or treating diabetic cataract, characterized in that: Made from the following raw materials: 2g baicalin, 9.57g mannitol, 200ml ultrapure water; the preparation process is as follows: 1) Prepare various raw material components; 2) Add the prescribed amount of mannitol to 180 ml of ultrapure water and stir for 5 minutes until completely dissolved; 3) Add baicalin to the solution obtained in step 2) and stir and disperse for 60 minutes until the baicalin is dissolved; 4) Add ultrapure water to make up to the full volume, filter with a 0.22μm filter membrane, and seal.

15. Use of the baicalin eye drops according to any one of claims 1 to 7, the baicalin eye drops prepared by the preparation method according to any one of claims 8 to 13, or the baicalin eye drops according to claim 14 in preparing a medicament for preventing or treating cataract.

16. The use according to claim 15, characterized in that The cataract is diabetic cataract.

17. The use according to claim 15 or 16, characterized in that The baicalin eye drops are administered by eye drops, 1 drop at a time, 4 times a day.

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