CN101537010B - A kind of eye drop preparation and preparation method thereof - Google Patents

Abstract

The invention discloses a doxycycline eye drop preparation and a preparation method thereof. The doxycycline eye drop preparation contains 0.03-0.5 wt% doxycycline, distilled water, hydroxypropyl-β-cyclodextrin, The antioxidant has a pH of 5.0-6.0, wherein the weight ratio of doxycycline to hydroxypropyl-β-cyclodextrin is 1:1-50. The doxycycline eye drop preparation of the invention can improve the drug solubility and dissolution rate of doxycycline, and improve the stability of the drug.

Description

A kind of eye drop preparation and preparation method thereof

technical field

The invention belongs to the technical field of medicines, and relates to an eye drop preparation and a preparation method thereof.

Background technique

Dry eye is a general term for a class of diseases that cause ocular discomfort and ocular surface damage due to abnormal tear quality and (or) quantity and abnormal tear dynamics. It is one of the most common ocular surface diseases. The clinical manifestations are dryness, burning, physical sensation, itching, blurred vision, red eyes and visual fatigue. Severe dry eye can also cause severe damage to vision such as keratitis, corneal neovascularization, and corneal ulcer, and its impact on people's quality of life has attracted more and more attention. Its occurrence is closely related to blepharitis and meibomian gland dysfunction.

Current research suggests that dry eye may be a type of non-infectious inflammatory disease in nature, so anti-inflammation has become a new direction for dry eye treatment. The occurrence of dry eye is closely related to blepharitis and meibomian gland dysfunction. The study found that meibomian glands contain a large number of coryneform bacteria, coagulase-negative staphylococcus and propionibacterium. It is known that Propionibacterium can produce wax lipase and triglyceride, and coagulase-negative Staphylococcus can also produce cholesterol esterase. The presence of these enzymes can change the composition of the meibomian gland lipids. On the one hand, it can lead to an increase in the melting point of the meibomian gland secretions, which increases the viscosity of the meibomian gland lipids, causing narrowing or even blockage of the meibomian gland ducts, resulting in Plate gland dysfunction; on the other hand, can produce free fatty acids, destroy the stability of the tear film, causing dry eye. Doxycycline (doxycycline, also known as doxycycline, deoxyoxytetracycline) has a broad-spectrum antibacterial effect, oral administration of doxycycline can effectively inhibit the growth of bacteria that cause chronic blepharitis and meibomian gland dysfunction, and It can effectively inhibit the production and activity of coagulase-negative staphylococcus and propionibacterium lipase, so it has a good therapeutic effect on chronic blepharitis and meibomian gland dysfunction, thus playing the role of treating dry eye. At the same time, the long-term application of the commonly used anti-inflammatory drugs glucocorticoids has relatively large side effects, which can cause serious complications such as cataracts and glaucoma. Studies in recent years have shown that doxycycline, previously used as an antibiotic, can inhibit the expression of inflammation-related factors in human corneal epithelial cells and conjunctival epithelial cells cultured in vitro. Experiments have proved that doxycycline eye drops can reduce the inflammatory response of the dry eye rabbit ocular surface, and can effectively inhibit the corneal neovascularization induced by ocular chemical injury. For example, Chinese patent 200410027807.0 discloses eye drops with a content of 0.1wt% to 0.5wt% doxycycline hydrochloride and its preparation method. The preparation method is: dissolving doxycycline hydrochloride and an osmotic pressure regulator in distilled water; Distilled water is added to the required volume, and the pH value is adjusted to 5.5-6.0; membrane filtration is sterilized, and the eye drops are obtained by subpackaging.

Yet doxycycline is a kind of light yellow powder or yellow crystalline powder under dry condition, odorless and tasteless, bitter in the mouth, has slight hygroscopicity, stable at room temperature, meet light deterioration, but almost insoluble in chloroform; It is unstable in neutral and alkaline solutions, and its antibacterial activity decreases rapidly, but its activity increases in acidic solutions; when it is formulated into an aqueous solution, it is unstable and easily decomposes when exposed to light. The pH value of 1% aqueous solution is 2-3, and its irritation is relatively high. big.

The doxycycline molecule is a complex compound containing multifunctional groups and 5 asymmetric carbon atoms, including structures such as phenol-ketone, ketone-enol and phthaloamide. Its structure is composed of unstable carbon rings and diketones, so it is easily decomposed by factors such as light, heat, and water. The decomposition reactions of doxycycline hydrochloride mainly include base-catalyzed hydrolysis reactions, isomerization reactions at C1 and C6 positions, and oxidation reactions.

Among the currently marketed ophthalmic preparations, traditional ophthalmic preparations still dominate, of which eye drops account for about 70%, of which 62.4% are solution type, 8.7% are suspension type; there is also a small amount of ointment accounting for 17.4% %. However, eye drops generally only have a bioavailability of 1-10% due to factors such as tear drainage and short residence time, while eye ointments generally cause blurred vision, and patients have poor compliance.

Drainage of tears and blinking from eye irritation are the most common causes of periocular drug loss. The function of the gel system is to increase the viscosity of the preparation, reduce the fluidity, and prolong the contact time to achieve the improved effect.

In situ gel refers to a preparation that is in a liquid state before administration and forms a gel after being instilled into the fornix of the eye due to changes in the environment. Compared with bioadhesive gels, in situ gels can provide more accurate dosing, and the gel is spread more evenly, so there is better compliance and reproducibility. According to the different mechanism of gel formation, it is mainly divided into temperature sensitive type, ion sensitive type, pH sensitive type and mixed type.

Contents of the invention

One of the objects of the present invention is to provide a doxycycline eye drop preparation, which has stable properties and good absorption effect.

A doxycycline eye drop preparation, containing 0.03-0.5wt% doxycycline, distilled water, hydroxypropyl-β-cyclodextrin, antioxidant, pH 5.5-6.0, wherein doxycycline The weight ratio to hydroxypropyl-β-cyclodextrin is 1:1-50.

Preferably, the antioxidant is selected from one or a combination of sodium sulfite, sodium bisulfite, sodium metabisulfite, and sodium thiosulfate.

The eye drop preparation is preferably a temperature-sensitive in-situ gel eye drop, and the in-situ gel eye drop also contains poloxamer, and the content of the poloxamer in the preparation is 16-30wt%. More preferably, the poloxamer is poloxamer 407 and poloxamer 188, and the dosage ratio of poloxamer 407 and poloxamer 188 is 25:1-3:2.

Another object of the present invention is to provide a preparation method of the above-mentioned cyclocycline eye drop preparation.

A method for preparing the doxycycline eye drop preparation described in claim 1, comprising the following steps:

(1) Doxycycline, distilled water and hydroxypropyl-β-cyclodextrin are prepared into an aqueous solution of doxycycline-hydroxypropyl-β-cyclodextrin clathrate by ultrasonic method, stirring method or grinding method, Filter through a 0.45 μm microporous membrane to obtain solution A;

(2) Add an appropriate amount of antioxidant to adjust the pH value of the solution to 5.5-6.0, add distilled water to the required weight, and sterilize by membrane filtration.

Between steps (1) and (2), the following steps are also included:

Weigh the poloxamer in parts by weight, slowly disperse it into an appropriate amount of distilled water, store it in low-temperature refrigeration until a clear and uniform solution is formed, and obtain solution B, fully mix solution A and solution B; or follow the above-mentioned doxycycline eye drops The preparation weighs poloxamer in parts by weight, slowly disperses it into solution A, and stores it in low-temperature refrigeration until a clear and uniform solution is formed.

The pharmaceutically acceptable carrier suitable for making ophthalmic gel is a matrix that makes the ophthalmic gel formulation of doxycycline stable and less irritating, which can prolong the residence time of the drug in the eye and enhance drug absorption. Improve bioavailability, reduce the frequency of medication, enhance efficacy and drug safety. The present invention is optimized through a large number of experiments, and the preferred gel matrix of the present invention, poloxamer, is selected from many polymer materials.

The advantage of the present invention is that the clathrate of the medicine is prepared by using the hydroxypropyl-beta-cyclodextrin, so that the solubility and dissolution rate of the medicine are improved, and the stability of the medicine is improved. And using the temperature-sensitive properties of poloxamer aqueous solution and the combination of different types of poloxamers, an ophthalmic in-situ gel preparation with a suitable phase transition temperature was prepared, so that it can be administered in a liquid state at room temperature and Forming a gel on the surface of the cornea, compared with eye ointment, does not cause blurred vision, is convenient for patients to take medication, and improves medication compliance. Compared with eye drops, it reduces the loss of drugs, and at the same time delays drug elimination to improve local biological Utilization.

Description of drawings

Fig. 1 is the phase dissolution diagram of the hydroxypropyl-beta-cyclodextrin inclusion compound of doxycycline monohydrate;

Fig. 2 is the X-ray diffraction patterns of doxycycline monohydrate, hydroxypropyl-β-cyclodextrin, their physical mixture and clathrate.

Detailed ways

Cyclodextrin is a series of cyclic oligosaccharides formed by the action of cyclodextrin glucosyl translocase on starch. Because cyclodextrin and its derivatives are water-soluble, safe and low-toxic to human body, and will not cause immune reaction, they have attracted more and more attention and are widely used in inclusion technology.

The cyclodextrin inclusions of drugs are characterized by: increasing the solubility of drugs; increasing the stability of drugs; powdering volatile liquids, solids or oily liquids; reducing the irritation and side effects of drugs; bioavailability. In recent years, various inclusions made of drugs will often occur. In medicine, α and β cyclodextrins are more commonly used.

Cyclodextrins are cyclic compounds composed of 6, 7, 8 or more D-glucopyranose units linked by α-1,4 glycosidic bonds, respectively called α-CD, β-CD, γ-CD.

The hole size of β-CD is between the other two, the hole shape is prismatic, and the solubility in water is 18.5g/L. It is the easiest to precipitate crystals from water, and the solubility increases with the increase of water temperature. Therefore, the inclusion property is good, and the toxicity is small, which is good for absorption and decomposition. However, β-CD can only contain some groups of drugs, and its stability is not good. Solubilization ability is limited.

Semi-synthetic β-CD derivatives mainly carry out necessary modifications to their molecular structures. The water solubility of β-CD is small because there are 7 primary hydroxyl groups and 14 secondary hydroxyl groups at both ends of the hole, and the intermolecular or intramolecular hydrogen groups prevent the hydration of water molecules. Introducing methyl groups, hydroxypropyl groups, hydroxyethyl groups and other groups into β-CD molecules to carry out alkylation reactions with hydroxyl groups can destroy the formation of hydrogen bonds in β-CD molecules and change the water solubility.

Hydroxypropyl-β-cyclodextrin (HP-β-CD) is a hydrophilic β-CD derivative. Compared with β-CD, the physical and chemical properties have undergone the following changes: 1. From crystallinity to Amorphous. 2. Changes in solubility properties. The solubility of HP-β-CD in water is greater than 50%, and it is soluble in aqueous alcohol solution. 3. HP-β-CD is not hydrolyzed by gastric acid and α-amylase, and hardly participates in the metabolism of organisms and does not accumulate. After oral administration, almost all of them are excreted with the stool in a complete form. 4. The complex formed by β-CD and drug has a sustained release effect on the drug, while the complex formed by HP-β-CD and drug has a rapid release effect on the drug, so that the drug can be released rapidly in the organism. 5. β-CD has a hemolytic effect, and parenteral administration also has a certain degree of irritation. HP-β-CD has low surface activity, basically no hemolysis and irritation. 6. The selectivity of β-CD and HP-β-CD to the guest drug is somewhat different.

In many literature reports, HP-β-CD has the following effects: improving drug solubility, increasing dissolution rate, improving oral drug bioavailability, reducing drug irritation to the gastrointestinal tract, masking bad smell of drugs, and improving drug stability Some literatures mentioned the improvement of powder properties and targeting effect of HP-β-CD.

Hydroxypropyl-β-cyclodextrin is a derivative of β-cyclodextrin. It has a giant cylindrical molecular structure, is easily soluble in water, has a large inclusion capacity and low toxicity, and can be used for eye administration. According to tests, after doxycycline and hydroxypropyl-β-cyclodextrin are clathrated at a certain weight ratio, the solubility is improved and the stability is also greatly improved.

Example 1

The eye drop preparation described in this embodiment is an eye drop solution, based on 100 g, containing 0.10 g of doxycycline, 1.00 g of hydroxypropyl-β-cyclodextrin, 0.10 g of sodium bisulfite, and pH 5.5-6.0 .

Preparation method: Take doxycycline hydrochloride (containing 0.10g of doxycycline), 1.00g of hydroxypropyl-β-cyclodextrin, add a small amount of distilled water until moist, fully grind in a mortar for 3 hours, add an appropriate amount of distilled water to dilute, Obtain light yellow clear doxycycline clathrate solution, add sodium bisulfite 0.10g, add distilled water to 100g, adjust the pH value in the process to be 5.0-6.0 (pH regulator can be disodium hydrogen phosphate, dihydrogen phosphate sodium, sodium hydroxide, or sodium bicarbonate), membrane filter sterilization, doxycycline eye drops.

Example 2

The eye drop preparation described in this embodiment is an eye drop solution, based on 100 g, containing 0.10 g of doxycycline, 1.00 g of hydroxypropyl-β-cyclodextrin, 0.10 g of sodium bisulfite, and pH 5.5-6.0 .

Preparation method: Take 1.00 g of hydroxypropyl-β-cyclodextrin and dissolve it in 10 g of distilled water to obtain a solution of hydroxypropyl-β-cyclodextrin; take doxycycline (containing 0.10 g of doxycycline monohydrate ), dispersed in about 5.0 g of distilled water, slowly added dropwise to the hydroxypropyl-β-cyclodextrin solution while stirring, and ultrasonicated for 3 hours to obtain a pale yellow clear doxycycline inclusion compound solution. Analyzed by phase dissolution and X-ray diffraction, as shown in Figure 1 and Figure 2. Among them, Figure 1 (A) doxycycline monohydrate (B) hydroxypropyl-β-cyclodextrin (C) physical mixture of doxycycline monohydrate and hydroxypropyl-β-cyclodextrin (1 :24 w/w) (D) doxycycline monohydrate hydroxypropyl-β-cyclodextrin inclusion complex (1:24w/w).

As can be seen from Figure 1, hydroxypropyl-β-cyclodextrin can significantly improve the solubility of doxycycline monohydrate in water; as can be seen from the X-ray diffraction analysis results, doxycycline monohydrate and hydroxypropyl-β- - There are obvious drug characteristic peaks in the physical mixture of cyclodextrin, but not in the inclusion complex of the two, it can be inferred that doxycycline monohydrate and hydroxypropyl-β-cyclodextrin form an inclusion complex.

Add 0.10 g of sodium thiosulfate to the doxycycline clathrate solution, add distilled water to 100 g, adjust the pH value to 5.5-6.0 during the process, perform membrane filtration sterilization, and obtain doxycycline eye drops.

Example 3

The eye drop preparation described in this example is a temperature-sensitive in-situ gel eye drop, containing 0.03 g of doxycycline, 0.90 g of hydroxypropyl-β-cyclodextrin, and poloxamer 407 in 100 g. 16.00g, sodium bisulfite 0.10g, pH 5.5-6.0.

Preparation method: Take 0.90 g of hydroxypropyl-β-cyclodextrin, dissolve it in 10 g of distilled water to obtain a solution of hydroxypropyl-β-cyclodextrin; take doxycycline (containing 0.03 g of doxycycline monohydrate ), dispersed in about 1.5 g of distilled water, slowly added dropwise to the hydroxypropyl-β-cyclodextrin solution while stirring, and ultrasonicated for 3 hours to obtain a light yellow clear doxycycline inclusion complex solution, which was solution A.

Take 16.00g of Poloxamer 407, slowly disperse it into 50g of distilled water, store it in cold storage for 12-24h, until a clear and uniform solution is formed, which is solution B.

Fully mix the above solution A with solution B, add 0.10g of sodium bisulfite, add distilled water to 100g, adjust the pH value to 5.0-6.0 during the process, sterilize by membrane filtration, doxycycline ophthalmic temperature sensitive in situ Gel eye drops. The phase transition temperature is 29-30°C.

Example 4

The eye drop preparation of the present invention is a temperature-sensitive in-situ gel eye drop, containing 0.50 g of doxycycline, 25.00 g of hydroxypropyl-β-cyclodextrin, and 20.00 g of poloxamer 407 in 100 g. g, Poloxamer 188 3.00g, sodium bisulfite 0.10g, pH5.5-6.0.

Preparation method: Take 25.00 g of hydroxypropyl-β-cyclodextrin, dissolve it in 40 g of distilled water to obtain a solution of hydroxypropyl-β-cyclodextrin; take doxycycline (containing 0.50 g of doxycycline monohydrate ), dispersed in about 5 g of distilled water, slowly added dropwise into the hydroxypropyl-β-cyclodextrin solution while stirring, and magnetically stirred for 10 h to obtain a pale yellow clear doxycycline inclusion complex solution.

Take 20.00g of poloxamer 407 and 3.00g of poloxamer 188, and slowly disperse them into the above doxycycline inclusion complex solution, and store them in cold storage for 12-24h until a clear and uniform solution is formed. Add 0.10 g of sodium sulfite, add distilled water to 100 g, adjust the pH value to 5.0-6.0 during the process, sterilize by membrane filtration, and doxycycline ophthalmic temperature-sensitive in-situ gel eye drops. The phase transition temperature is 27-28°C, and the phase transition temperature is 36-37°C after dilution with simulated tear fluid (the gel-tear fluid volume ratio is about 40:7).

Example 5

The eye drop preparation of the present invention is a temperature-sensitive in-situ gel eye drop, containing 0.10 g of doxycycline, 3.00 g of hydroxypropyl-β-cyclodextrin, and 25.00 g of poloxamer 407 in 100 g. g, Poloxamer 188 10.00g, sodium bisulfite 0.10g, pH5.5-6.0.

Preparation method: Take 3.00 g of hydroxypropyl-β-cyclodextrin, dissolve it in 10 g of distilled water to obtain a solution of hydroxypropyl-β-cyclodextrin; take doxycycline (containing 0.10 g of doxycycline monohydrate ), dispersed in about 5 g of distilled water, and slowly added dropwise to the hydroxypropyl-β-cyclodextrin solution while stirring, and magnetically stirred for 10 hours to obtain a pale yellow clear doxycycline inclusion complex solution, which is solution A.

Take 25.00g of Poloxamer 407 and 10.00g of Poloxamer 188, slowly disperse them into 45g of distilled water, and store them in cold storage for 12-24h until a clear and uniform solution is formed, which is called solution B.

Fully mix the above solution A with solution B, add 0.10g of sodium bisulfite, add distilled water to 100g, adjust the pH value to 5.0-6.0 during the process, sterilize by membrane filtration, doxycycline ophthalmic temperature sensitive in situ Gel eye drops. The phase transition temperature is 29-30°C, and the phase transition temperature is 36-37°C after dilution with simulated tear fluid (the gel-tear fluid volume ratio is about 40:7).

Example 6

The eye drop preparation of the present invention is a temperature-sensitive in-situ gel eye drop, containing 0.1 g of doxycycline, 5.00 g of hydroxypropyl-β-cyclodextrin, and 25.00 g of poloxamer 407 in 100 g g, Poloxamer 188 10.00g, sodium bisulfite 0.10g, pH 5.0-6.0.

Preparation method: take 5.00 g of hydroxypropyl-β-cyclodextrin, dissolve it in 10 g of distilled water to obtain a solution of hydroxypropyl-β-cyclodextrin; take doxycycline (containing 0.10 g of doxycycline monohydrate ), dispersed in about 5 g of distilled water, and slowly added dropwise to the hydroxypropyl-β-cyclodextrin solution while stirring, and magnetically stirred for 10 hours to obtain a pale yellow clear doxycycline inclusion complex solution, which is solution A.

Take 25.00g of poloxamer 407 and 2.00g of poloxamer 188, slowly disperse them into 40g of distilled water, store them in cold storage for 12-24h, until a clear and uniform solution is formed, which is called solution B.

Fully mix the above solution A with solution B, add 0.10g of sodium bisulfite, add distilled water to 100g, adjust the pH value to 5.0-6.0 during the process, sterilize by membrane filtration, doxycycline ophthalmic temperature sensitive in situ Gel eye drops. The phase transition temperature is 25-26°C, and the phase transition temperature is 33-34°C after dilution with simulated tear fluid (gel-tear fluid volume ratio is about 40:7).

Example 7: Evaluation of curative effect of doxycycline gel on corneal neovascularization inhibition

Objective To study the therapeutic effect of topical doxycycline ophthalmic gel on corneal neovascularization in rats.

Methods Fourteen female SD rats were surgically implanted with bFGF slow-release membrane at the top of the corneal pouch. The surgical incision was closed without suturing. After the operation, 0.5% tobramycin ophthalmic ointment was added. Rats were randomly divided into treatment groups (0.1% doxycycline gel) and control groups (vehicle). The treatment group received 0.1% doxycycline drop gel, four times a day, 50ul each time, and continued to use the drug for 6 days after operation; the control group received excipients, four times a day, 50ul each time. The heart is perfused with ink, which completely turns the corneal rim, conjunctival vessels, and corneal neovascularization into black. Eyeballs were enucleated and fixed with 4% paraformaldehyde. After 24 hours, the fixed eyeballs were taken out. The cornea was removed along the cornea and sclera under the operating microscope, and the cornea was cut radially according to the growth zone of the CNV. The corneas were dynamically observed and photographed with a slit-lamp microscope 7 days after treatment. Image Pro-Plus 5.1 image processing software (MediaCybernetics, USA) was used to measure the length and area of CNV. The length of the blood vessel was based on the blood vessel growing vertically from the edge of the horn to the sustained-release tablet. The statistical analysis between each group was based on independent samples t test or Mann-Whitney U test. Statistical difference was defined as P<0.05. All statistical analyzes were performed using SPSS 13.0 software package (SPSS Inc., Chicago, IL, USA).

result

Table 1 The size of the sustained-release tablets of the two experimental groups and the data of their distance to the limbus ^*

^* Measured on the 7th day of the test ^** The results are described by arithmetic mean ± standard deviation

Table 2 The area of blood vessels and the average length of blood vessels in the two test groups

^* Mann-Whitney U test

There was no significant difference in the size of bFGF sustained-release tablets and the distance from the sustained-release tablets to the limbus between the treatment group and the control group (Table 1). Between the treatment group and the control group, there were statistically significant differences in vessel area and vessel length (Table 2).

Conclusion Topical application of doxycycline gel can inhibit corneal neovascularization.

Claims (3)

1. doxycycline eye drip preparation, contain the 0.03-0.5wt% doxycycline, distilled water, it is characterized in that: described eye drip preparation is the temperature-sensitive situ-gel eye drop, also contain HP-, antioxidant and poloxamer, pH is 5.0-6.0, wherein the weight ratio of doxycycline and HP-is 1: 1-50, the content of described poloxamer in preparation is 16-30wt%, and described poloxamer is that amount ratio is that 25: 1～3: 2 poloxamers 407 and poloxamer 188 are formed; Described doxycycline is to be present in the preparation with the doxycycline monohydrate.

2. doxycycline eye drip preparation according to claim 1 is characterized in that: described antioxidant is selected from more than one the combination in sodium sulfite, sodium sulfite, sodium pyrosulfite, the sodium thiosulfate.

3. method for preparing the described doxycycline eye drip of claim 1 preparation is characterized in that: mainly may further comprise the steps:

(1) with ultrasonic method, paddling process or polishing doxycycline, distilled water and HP-are mixed with doxycycline-hydroxypropyl-beta-cyclodextrin inclusion aqueous solution, use 0.45 μ m filtering with microporous membrane, get solution A;

(2) add an amount of antioxidant, the pH value of regulator solution is 5.0-6.0, adds distilled water to required weight, membrane filtration degerming;

Between step (1) and (2), also include following steps:

Take by weighing the poloxamer of weight portion by the described doxycycline eye drip of claim 1 preparation, slowly be distributed in an amount of distilled water, deepfreeze is preserved, until the solution that forms the clarification homogeneous, solution B; Solution A and solution B are fully mixed; Perhaps take by weighing the poloxamer of weight portion by the described doxycycline eye drip of claim 1 preparation, slowly be distributed in the solution A, deepfreeze is preserved, until the solution that forms the clarification homogeneous.

Images