A pharmaceutical preparation for preventing after-cataract and a method for preparing the same

Abstract

This invention belongs to the interdisciplinary field of biomedical materials and medicine, and discloses an anti-secondary cataract drug preparation and its preparation method. The preparation steps are as follows: (1) Weigh polyethylene glycol 15-hydroxystearate and triptolide, and dissolve them together in ethanol; wherein, the ratio of polyethylene glycol 15-hydroxystearate, triptolide and ethanol is 10 g: (0.1-1.5) g: (2-20) mL; (2) Under ultrasonic stirring, add the mixture obtained in step (1) dropwise to water for injection, remove the ethanol by rotary evaporation, and obtain drug-loaded polymer micelles; the amount of water for injection in this step is 5-15 times the volume of ethanol in step (1); (3) Mix 0.05-5 g of drug-loaded polymer micelles and 1-50 g of thermosensitive polymer hydrogel evenly, and add water for injection to make up to 1000 mL, thus obtaining the anti-secondary cataract drug preparation. The drug preparation obtained by this invention can effectively inhibit the development of post-cataract surgery PCO.

Description

Technical Field

[0001] This invention belongs to the interdisciplinary field of biomedical materials and medicine, specifically relating to a drug formulation for treating secondary cataracts and its preparation method. Background Technology

[0002] Cataracts are the leading cause of blindness worldwide, accounting for more than 50% of all blindness cases. The World Health Organization estimates that approximately 20 million people worldwide are currently blind due to cataracts, and this number is expected to rise to 50 million by 2050 due to an aging population. Surgery is the only effective treatment for cataracts. Posterior capsular opacification (PCO) is the most common complication after cataract surgery, with an incidence rate of 12% to 67% in adults and as high as 100% in infants and young children. PCO significantly reduces patients' vision and contrast sensitivity, causing inconvenience in daily life and affecting the visual development of infants and young children, leading to amblyopia. PCO can be treated with YAG laser therapy, but laser treatment carries risks such as glaucoma, macular edema, retinal detachment, and intraocular lens displacement, while also increasing the financial burden on patients. For infants and young children who cannot cooperate with YAG laser treatment, further surgical removal is necessary. Various other methods have been used to attempt to reduce or even eliminate PCO, including intraocular lens surface modification and intraoperative and postoperative topical drug application. However, due to the presence of aqueous humor circulation, the medication in the anterior chamber is diluted and excreted through the trabecular meshwork within approximately 2 hours. Furthermore, the presence of local ocular biological barriers results in a bioavailability of only 5% for postoperative topical medication. Therefore, none of these methods can effectively prevent the occurrence and development of PCO. Finding effective ways to inhibit PCO formation, exploring new therapeutic targets and specific drugs, and developing novel anti-PCO drugs with low side effects and high specificity are of significant clinical importance.

[0003] Celastrol (CEL) is a bioactive monomer extracted from the traditional Chinese medicine Tripterygium wilfordii, with the molecular formula C2. 29 H 38 O4, with a molecular weight of 450.61, is a bioactive compound called Celastrol. Celastrol possesses various biological activities, including anti-inflammatory, anti-tumor, angiogenesis inhibition, and obesity-related activity, demonstrating potent therapeutic effects on autoimmune diseases, tumors, obesity, and neurodegenerative diseases. Recent studies suggest that Celastrol can inhibit fibrosis in various tissues, including the lungs. Despite its strong biological activity, Celastrol's extremely poor water solubility limits its further application. Enhancing its water solubility and improving drug concentration and bioavailability in ocular target tissues are key challenges that urgently need to be addressed. Currently, there are no reports of its formulations being used intraoperatively to treat polycystic ovary syndrome (PCO). Summary of the Invention

[0004] In view of the defects and deficiencies of the prior art, the purpose of this invention is to provide a drug preparation for treating secondary cataracts and its preparation method.

[0005] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0006] A method for preparing a drug formulation for treating secondary cataracts, comprising the following steps:

[0007] (1) Weigh out polyethylene glycol 15-hydroxystearate and triptolide, and dissolve them together in ethanol; wherein the ratio of polyethylene glycol 15-hydroxystearate, triptolide and ethanol is 10 g: (0.1-1.5) g: (2-20) mL.

[0008] (2) Add the mixture obtained in step (1) dropwise to water for injection under ultrasonic stirring, remove ethanol by rotary evaporation, and obtain drug-loaded polymer micelles; the amount of water for injection in this step is 5-15 times the volume of ethanol in step (1);

[0009] (3) Mix 0.05~5 g of drug-loaded polymer micelles and 1~50 g of thermosensitive polymer hydrogel evenly, then add water for injection to make up to 1000 mL to obtain the drug preparation for anti-secondary cataract.

[0010] Preferably, the temperature-sensitive polymer hydrogel is one or a combination of several selected from poloxamer hydrogel, polyisopropylacrylamide hydrogel, poly(2-(N,N-dimethylamino)ethyl methacrylate) hydrogel, polyethylene glycol hydrogel, and chitosan-based polymer hydrogel. In this invention, the chitosan-based polymer hydrogel can be prepared according to existing techniques.

[0011] Preferably, the ethanol is 95% ethanol or anhydrous ethanol.

[0012] A drug formulation for treating secondary cataracts prepared by the method described above.

[0013] Beneficial effects: Tripterygium wilfordii has extremely poor water solubility, with a solubility of only 3.8 micrograms per milliliter in water at 37°C. Currently, there is no existing technology that can formulate it into an injectable preparation for intraoperative treatment of secondary cataracts. This invention utilizes nanotechnology to first prepare drug-loaded polymer micelles containing triptolide to enhance its apparent solubility. Then, the drug-loaded polymer micelles are self-assembled with a thermosensitive polymer hydrogel to form a drug-loaded, injectable anti-secondary cataract drug formulation containing triptolide. During cataract extraction, this formulation is injected into the capsular bag, achieving slow release of triptolide into the target tissue and effectively inhibiting the development of postoperative cataract occlusion (PCO). Attached Figure Description

[0014] Figure 1 Anterior segment images of the Control group, TSG group, and TSG / CEL group on postoperative day 7 (first row) and day 28 (first row).

[0015] Figure 2 HE staining results of cornea and retina in the Control group, TSG group and TSG / CEL group on the 28th day after surgery. Detailed Implementation

[0016] To make the present invention clearer and more explicit, the present invention will be further described in detail below. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

[0017] The chitosan-based polymer hydrogels in the following examples were prepared in accordance with ACS Applied Materials & Interfaces 2021, 13, 49369-49379.

[0018] Example 1

[0019] Weigh 10 g of polyethylene glycol 15-hydroxystearate (Kolliphor HS 15) and 0.75 g of triptolide, and dissolve them together in 10 mL of anhydrous ethanol. Then, under ultrasonic stirring, add the mixture dropwise to 100 mL of water for injection, and remove the ethanol by rotary evaporation to obtain drug-loaded polymer micelles. Mix 1 g of drug-loaded polymer micelles and 15 g of chitosan-based polymer hydrogel evenly, and add water for injection to 100 mL to obtain the anti-secondary cataract drug formulation.

[0020] Example 2

[0021] The difference from Example 1 is that the amount of triptolide is changed to 1.5 g, while the rest is the same as in Example 1.

[0022] Example 3

[0023] The difference from Example 1 is that the amount of triptolide is changed to 0.3 g, and the rest is the same as in Example 1.

[0024] Example 4

[0025] The difference from Example 1 is that the amount of triptolide is changed to 0.1 g, while the rest is the same as in Example 1.

[0026] animal testing

[0027] Using the New Zealand white rabbit model of secondary cataract as an animal model, the efficacy of the drug formulation in preventing secondary cataract was evaluated. The specific methods were described in the published article (ACS Applied Bio Materials 2021 4 (4), 3579-3586), and the detailed steps are as follows:

[0028] General anesthesia was induced in New Zealand white rabbits by intravenous injection of 3% (g / mL) sodium pentobarbital solution at a dose of 3 ml / kg. Anesthesia was considered effective when there was no obvious reflex when the lips were clamped. Mydriasis was achieved with 0.5% (g / mL) compound tropicamide eye drops, followed by irrigation of the conjunctival sac with 5 mL of sterile saline. The periorbital area was disinfected with povidone-iodine, and local anesthesia of the ocular surface was administered with 0.5% (g / mL) promecaine hydrochloride eye drops. A 3 mm clear corneal incision was made under a microscope, and 100 μL of viscoelastic medical-grade sodium hyaluronate gel was injected into the anterior chamber. Continuous circular capsulorhexis was performed, with a capsulorhexis opening diameter of 5 mm. mm, the lens nucleus and cortex were aspirated with an aspiration needle, and the drug was injected into the lens capsule to form the anterior chamber. The transparent corneal incision was closed with one 10-0 suture. After the operation, gatifloxacin eye gel was applied to the conjunctival sac to prevent infection. All surgeries were performed on the right eye of the animals. After the operation, the experimental animals were given tobramycin dexamethasone eye drops three times a day, one drop each time, for a total of seven days.

[0029] The experimental animals were divided into three groups: Group 1 (labeled TSG / CEL group), after intraoperative cortical aspiration, 5 μL of the anti-posterior cataract drug preparation obtained in Example 1 was injected into the lens capsule; Group 2 (labeled TSG group), after intraoperative cortical aspiration, 5 μL of chitosan-based polymer hydrogel without any drug loading was injected into the lens capsule; Group 3 (labeled Control group), after intraoperative cortical aspiration, 5 μL of physiological saline was injected into the lens capsule. Postoperatively, the operated eyes of the animals were observed daily under a slit lamp, and the cornea, anterior chamber, and posterior cataract were recorded. The observation period was 28 days; three animals were used in each group, and the PCO score was calculated using the EPCO standard (see Table 1). Clinical assessment and PCO score calculation were performed on days 7 and 28 by area, type, and density analysis. After the experiment, rabbits were euthanized using the air embolization method, which involved injecting 20-40 mL of air into the marginal ear vein. Animal eyeball specimens were collected, fixed, and stained with hematoxylin and eosin (HE).

[0030]

[0031] Figure 1 Anterior segment photographs of the Control group, TSG group, and TSG / CEL group taken on postoperative day 7 (first row) and day 28 (second row). Figure 1As can be seen, the anterior segment images showed no significant difference in posterior capsule opacity among the three groups on day 7. On day 28, white opacity appeared around the posterior capsule in the Control and TSG groups, indicating fibrosis of the lens epithelial cells and capsular wrinkling. Secondary cataracts occurred in these two groups. The posterior capsule in the TSG / CEL group remained almost transparent and smooth. According to the image analysis of posterior capsule opacity, the opacity density was scored from 0 to 4, which were none (0), very (1), mild (2), moderate (3), and severe (4), respectively. On day 28 postoperatively, some areas of the Control and TSG groups had opacity of grade 4 or severe (dark gray area), with PCO scores of 1.5 and 1.7, respectively. There was no statistically significant difference between the two groups. In the TSG / CEL group, only a few light gray areas of the posterior capsule showed marginal posterior capsule opacity (grade 1), with a PCO score of 0.07. The PCO scores of the TSG / CEL group were significantly different from those of the Control group with saline and the TSG group without drug loading. The above indicates that the development of PCO in the TSG / CEL group was significantly lower than that in the Control group and the TSG group, meaning that the drug formulation obtained in this invention can effectively inhibit the development of PCO after cataract surgery.

[0032] Furthermore, we evaluated the drug safety of TSG / CEL and TSG. The results showed that during the 28-day postoperative observation period, no significant abnormal ocular inflammatory reactions (such as corneal thickening, corneal stromal fibrosis, hemorrhage, or edema) occurred in either the TSG / CEL group or the TSG group. Twenty-eight days after clear lens extraction, corneal and retinal tissues were separated and observed. HE staining results of the cornea and retina in the Control group, TSG group, and TSG / CEL group are as follows: Figure 2 As shown, there were no significant differences in cell morphology among the groups, and the corneal endothelium remained intact, indicating that the drug formulation obtained in this invention has good drug safety.

[0033] In summary, animal experiments have shown that injection of the drug formulation of this invention can significantly reduce the degree of posterior capsule opacification after cataract surgery, and it is non-toxic to tissues such as the cornea and retina, and has a safe anti-secondary cataract effect.

Claims

1. A method for preparing a drug formulation for treating secondary cataracts, characterized in that, The preparation steps are as follows: (1) Weigh out polyethylene glycol 15-hydroxystearate and triptolide, and dissolve them together in ethanol; wherein the ratio of polyethylene glycol 15-hydroxystearate, triptolide and ethanol is 10 g: (0.1-1.5) g: (2-20) mL; (2) Add the mixture obtained in step (1) dropwise to water for injection under ultrasonic stirring, remove ethanol by rotary evaporation, and obtain drug-loaded polymer micelles; the amount of water for injection in this step is 5-15 times the volume of ethanol in step (1); (3) After mixing 0.05~5g of drug-loaded polymer micelles and 1~50g of chitosan-based polymer hydrogel evenly, add the remaining amount for injection. The volume was adjusted to 1000 mL with water to obtain the anti-secondary cataract drug preparation.

2. The method for preparing the anti-secondary cataract drug formulation as described in claim 1, characterized in that: The ethanol is 95% ethanol or anhydrous ethanol.

3. A drug formulation for treating secondary cataracts prepared by the method described in claim 1 or 2.

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