CN116172885A

CN116172885A - Pharmaceutical preparation for resisting after-cataract and preparation method thereof

Info

Publication number: CN116172885A
Application number: CN202111549091.0A
Authority: CN
Inventors: 张颖; 李景果; 栗占荣; 刘瑞星
Original assignee: Henan Provincial Peoples Hospital
Current assignee: Henan Provincial Peoples Hospital
Priority date: 2021-12-17
Filing date: 2021-12-17
Publication date: 2023-05-30

Abstract

The invention belongs to the crossing field of biomedical materials and medicine, and discloses a pharmaceutical preparation for resisting after-cataract and a preparation method thereof. The preparation method comprises the following steps: (1) Weighing polyethylene glycol 15-hydroxystearate and tripterine, and dissolving the polyethylene glycol 15-hydroxystearate and the tripterine in ethanol; wherein the dosage ratio of polyethylene glycol 15-hydroxystearate to tripterine to ethanol is 10: 10 g:0.1-1.5 g:2-20 mL; (2) Dropwise adding the mixture obtained in the step (1) into water for injection under ultrasonic stirring, and removing ethanol by rotary evaporation to obtain a drug-loaded polymer micelle; the dosage of the water for injection in the step is 5-15 times of the dosage of the ethanol in the step (1); (3) And uniformly mixing 0.05-5 g of drug-loaded polymer micelle and 1-50 g of thermosensitive polymer hydrogel, and adding water for injection to a constant volume of 1000 mL to obtain the drug preparation for resisting the secondary cataract. The pharmaceutical preparation obtained by the invention can effectively inhibit the development of PCO after cataract operation.

Description

Pharmaceutical preparation for resisting after-cataract and preparation method thereof

Technical Field

The invention belongs to the crossing field of biomedical materials and medicine, and particularly relates to a drug preparation for resisting after-cataract and a preparation method thereof.

Background

Cataract is the first blinding eye disease in the world, accounting for more than 50% of all blinding eye diseases. According to the estimated world health organization, the number of people blinded by cataract is about 2000 ten thousand in the world at present, and the people continue to rise along with the aging process of society, and the number of people can reach 5000 ten thousand in 2050. Surgery is the only effective means to treat cataracts. The secondary cataract (posterior capsular opacification, PCO) is the most common complication after cataract surgery, the postoperative incidence rate is 12% -67% for adults and is as high as 100% in infants. PCO obviously reduces the eyesight and contrast sensitivity of the patient, causes inconvenient life, and simultaneously affects the eyesight development of the infant patient, thereby causing amblyopia. PCO can be treated by YAG laser, but laser treatment risks causing glaucoma, macular cystoid edema, retinal detachment, intraocular lens shift, while increasing the economic burden on the patient. For infant patients who cannot be treated with YAG laser, surgical excision is again required. People also pass through a variety of other methods including: methods such as intraocular lens surface modification, postoperative topical drug application, etc. have attempted to reduce or even eliminate PCO, but due to aqueous circulation, about 2 hours of anterior chamber drug will be diluted and expelled from the trabecular meshwork; the existence of the local biological barrier of the eye ensures that the bioavailability of the medicine locally spotted on the eye after operation is only 5 percent, and the methods can not effectively prevent the occurrence and development of PCO. How to effectively inhibit the formation of PCO and search for new therapeutic targets and specific medicaments, thereby developing novel anti-PCO medicaments with low side effects and strong specificity and having important clinical significance.

Celastrol (CEL) is bioactive monomer extracted from herba Polygoni Perfoliati, and has molecular formula of C ₂₉ H ₃₈ O ₄ The molecular weight is 450.61.Celastrol has antiinflammatory, antitumor and antitumor effectsThe composition has various biological activities such as neovascularization, obesity and the like, and has strong therapeutic effects on autoimmune diseases, tumors, obesity and neurodegenerative diseases. Recent studies suggest that Celastrol can inhibit various tissue fibrosis such as lung. Although Celastrol has strong biological activity, its further application is limited by the extremely poor water solubility. How to enhance the water solubility and the drug concentration and the bioavailability of the eye target tissue are key problems to be solved urgently. There is no report of the application of its pharmaceutical formulation to the surgery for the treatment of PCO.

Disclosure of Invention

Aiming at the defects and shortcomings of the prior art, the invention aims to provide an anti-secondary cataract pharmaceutical preparation and a preparation method thereof.

In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:

a preparation method of a pharmaceutical preparation for treating after-cataract comprises the following preparation steps:

(1) Weighing polyethylene glycol 15-hydroxystearate and tripterine, and dissolving the polyethylene glycol 15-hydroxystearate and the tripterine in ethanol; wherein the dosage ratio of polyethylene glycol 15-hydroxystearate to tripterine to ethanol is 10: 10 g:0.1-1.5 g:2-20 mL;

(2) Dropwise adding the mixture obtained in the step (1) into water for injection under ultrasonic stirring, and removing ethanol by rotary evaporation to obtain a drug-loaded polymer micelle; the dosage of the water for injection in the step is 5-15 times of the dosage of the ethanol in the step (1);

(3) And uniformly mixing 0.05-5 g of drug-loaded polymer micelle and 1-50 g of thermosensitive polymer hydrogel, and adding water for injection to a constant volume of 1000 mL to obtain the drug preparation for resisting the secondary cataract.

Preferably, the temperature-sensitive polymer hydrogel is one or a combination of a plurality of poloxamer hydrogel, poly isopropyl acrylamide hydrogel, poly [ 2- (N, N-dimethylamino) ethyl methacrylate hydrogel, polyethylene glycol hydrogel and chitosan-based polymer hydrogel. In the present invention, chitosan-based polymer hydrogels can be prepared according to the prior art.

Preferably, the ethanol is 95 v% ethanol or absolute ethanol.

A pharmaceutical preparation for treating after-cataract prepared by the above preparation method is provided.

The beneficial effects are that: the tripterine has extremely poor water solubility, the solubility in water at 37 ℃ is only 3.8 micrograms per milliliter, and the tripterine is not prepared into an injectable preparation for treating the secondary cataract in the operation in the prior art; according to the invention, a nano technology is adopted, firstly, a medicine-carrying polymer micelle carrying the tripterine is prepared to enhance the apparent solubility of the medicine-carrying polymer micelle, then, the medicine-carrying polymer micelle and the temperature sensitive polymer hydrogel are self-assembled to form a medicine-carrying injectable injection preparation for treating the post-cataract carrying the tripterine, and the medicine-carrying injection preparation is injected into a bag in cataract extraction, so that the slow release of the tripterine in a target tissue is realized, and the development of PCO after cataract operation is effectively inhibited.

Drawings

Fig. 1: day 7 post-surgery (first row), day 28 (first row), control group, TSG group, and TSG/CEL group anterior ocular segment photographs.

Fig. 2: on day 28 post-surgery, control, TSG, and TSG/CEL cornea, retina HE staining results.

Detailed Description

The present invention will be described in further detail below for the purpose of making the present invention clearer and more specific. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.

Chitosan-based polymer hydrogel references ACS Applied Materials & Interfaces 2021, 13, 49369-49379 in the examples below were prepared.

Example 1

Weighing polyethylene glycol 15-hydroxystearate (Kolliphor HS 15) 10 g and tripterine 0.75 g, and dissolving in 10 mL absolute ethanol; then, dripping the mixture into 100 mL injection water under ultrasonic stirring, and removing ethanol by rotary evaporation to obtain a drug-loaded polymer micelle; and (3) uniformly mixing the drug-carrying polymer micelle 1 g and the 15 g chitosan-based polymer hydrogel, and adding water for injection to 100 mL to obtain the drug preparation for resisting the secondary cataract.

Example 2

The difference from example 1 is that: the amount of tripterine was changed to 1.5. 1.5 g, otherwise the same as in example 1.

Example 3

The difference from example 1 is that: the amount of tripterine was changed to 0.3. 0.3 g, otherwise the same as in example 1.

Example 4

The difference from example 1 is that: the amount of tripterine was changed to 0.1. 0.1 g, otherwise the same as in example 1.

Animal test

The anti-secondary cataract effect of the pharmaceutical preparation is evaluated by taking a New Zealand white rabbit secondary cataract model as an animal model. For specific methods, refer to published article (ACS Applied Bio Materials 2021 (4), 3579-3586), the detailed steps are as follows:

3% (g/mL) of pentobarbital sodium salt solution is injected into the ear margin for inducing the general anesthesia of New Zealand white rabbits, the dosage is calculated according to 3 mL/kg, and no obvious reflex of the jaw lip is taken as the anesthesia effect; 0.5% (g/mL) of compound topicarb-amine eye drops are used for mydriasis, sterile normal saline 5 mL is used for flushing conjunctival sac of the eye, iodophor is used for disinfecting the eyes, and 0.5% (g/mL) of procaine hydrochloride eye drops are used for carrying out local anesthesia on the eye surface; making a 3 mm transparent cornea incision under a microscope, injecting 100 mu L of viscoelastic medical sodium hyaluronate gel into the anterior chamber, continuously annularly tearing a capsule, and ensuring that the diameter of a capsule tearing opening is 5 mm, sucking crystal nuclei and cortex by an injection needle head, injecting medicine into a crystal sac to form the anterior chamber, and suturing 1 needle by 10-0 lines of the transparent cornea incision to close the incision; the eye gel of the Bi Jiati of the art is coated in conjunctival sac to prevent infection; surgery was performed on the right eye of the animal; the eye drops of tobramycin and dexamethasone are locally spotted on conjunctival sac of the experimental animal after operation three times a day, one drop at a time for seven days.

Experimental animals were divided into three groups: the first group (labeled TSG/CEL group), after intraoperative cortical aspiration, was injected with 5. Mu.L of the anti-posterior cataract drug formulation of example 1; a second group (labeled TSG group), in which 5 μl of chitosan-based polymer hydrogel without any drug is injected into the capsular bag after the cortex is aspirated; a third group (labeled as Control group), in which 5. Mu.L of physiological saline was injected into the capsular bag after the cortex was aspirated; the animal's eyes were observed under the slit lamp every day after the operation, and cornea, anterior chamber and posterior obstacle were recorded. The observation period is 28 days; three animals per group were subjected to statistics of PCO scores, which were based on EPCO standards, as shown in Table 1. Clinical assessment and PCO score calculation were performed on days 7, 28, respectively, by area, type and density analysis. Rabbits were sacrificed by air embolism at the end of the experiment by intravenous injection of 20-40 mL air from the ear margin. And (5) fixing the animal eyeball specimen, and carrying out HE staining.

Fig. 1 is a photograph of the anterior ocular segment of the Control, TSG, and TSG/CEL groups at post-operative day 7 (first row), day 28 (second row). As can be seen from fig. 1; anterior ocular segment photographs showed no significant difference in capsular opacification after day 7 in the three groups; on day 28, white turbidity appeared around the posterior capsule of Control and TSG groups, suggesting lens epithelial cell fibrosis, capsule wrinkling, where posterior cataract occurred in both groups, the posterior capsule of TSG/CEL group remained almost transparent and smooth; according to analysis and evaluation of posterior capsule turbidity images, the turbidity density is from 0 to 4, namely no (0), no (1), no (2), no (3) and no (4) turbidity, the turbidity of partial areas of a Control group and a TSG group reaches 4 levels or no (dark gray area) turbidity after 28 days after operation, PCO scores are respectively 1.5 and 1.7, and the difference between the two groups has no statistical significance; the posterior capsule of the TSG/CEL group had only marginal posterior capsule turbidity (grade 1) in a few light grey areas, with a PCO score of 0.07. The PCO score difference of the TSG/CEL group compared with the Control group of physiological saline and the TSG group without drug has statistical significance. The above shows that: the development of PCO in the TSG/CEL group is obviously lower than that in the Control group and the TSG group, namely the pharmaceutical preparation obtained by the invention can effectively inhibit the development of PCO after cataract operation.

Furthermore, we further evaluated the drug safety of TSG/CEL and TSG, and the results showed that no significant abnormal ocular inflammatory response (such as corneal thickening, corneal stromal fibrosis, hemorrhage or edema, etc.) occurred in either TSG/CEL or TSG groups during the 28 day post-operative observation period. The cornea tissue and the retina tissue are separated and observed 28 days after the transparent crystal extirpation, the cornea and retina HE staining results of the Control group, the TSG group and the TSG/CEL group are shown in figure 2, the cell morphology among the groups is not obviously different, and the cornea endothelial layer is complete, so that the pharmaceutical preparation obtained by the invention has good pharmaceutical safety.

In conclusion, animal experiments show that the injection of the pharmaceutical preparation can obviously reduce the degree of capsular opacification after cataract surgery, has no toxicity to tissues such as cornea and retina, and has safe effect of resisting the posterior cataract.

Claims

1. The preparation method of the anti-after cataract pharmaceutical preparation is characterized by comprising the following preparation steps:

2. The method for preparing the anti-secondary cataract pharmaceutical formulation according to claim 1, wherein: the temperature-sensitive polymer hydrogel is one or a combination of a plurality of poloxamer hydrogel, poly isopropyl acrylamide hydrogel, poly [ 2- (N, N-dimethylamino) ethyl methacrylate hydrogel, polyethylene glycol hydrogel and chitosan-based polymer hydrogel.

3. The method for preparing the anti-secondary cataract pharmaceutical formulation according to claim 1, wherein: the ethanol is 95 v% ethanol or absolute ethanol.

4. An anti-posterior cataract pharmaceutical formulation prepared by the method of any one of claims 1-3.