Ophthalmic pharmaceutical composition for treating demodex blepharitis and method of preparing the same

Abstract

The present application relates to a kind of for treating demodex blepharitis ophthalmic pharmaceutical composition and its preparation method, wherein, ophthalmic pharmaceutical composition, comprising ivermectin, immunomodulator and ophthalmically acceptable carrier;The content of the ivermectin is 0.2% to 0.5% of the total weight of the pharmaceutical composition;The content of the immunomodulator is 0.001% to 0.4% of the total weight of the pharmaceutical composition, and the immunomodulator is selected from one of cyclosporine A and tacrolimus.The beneficial effect is that ivermectin is used in combination with immunomodulator, realizes the synergistic treatment strategy of insecticidal and anti-inflammatory.Both of them can be combined to simultaneously aim at the etiology and pathological consequences of demodex infection, exert synergistic effect, provide new drug selection for the clinical treatment of demodex blepharitis.

Description

Technical Field

[0001] This invention relates to the field of ophthalmic pharmaceutical formulation technology, and in particular to an ophthalmic pharmaceutical composition for treating Demodex blepharitis and its preparation method. Background Technology

[0002] Blepharitis is one of the most common chronic ocular surface diseases in clinical practice, often manifesting as eyelid congestion, itching, scaling, foreign body sensation, dryness, and burning sensation, severely impacting patients' quality of life. Recent studies have confirmed that *Demodex folliculorum* and *Demodex brevis* are important pathogens causing blepharitis, ocular erythema, meibomian gland dysfunction, dry eye, and eyelash loss. Demodex mites cause disease through multiple pathways, including mechanical damage, immune responses, and acting as carriers of pathogenic microorganisms; their infection is closely related to the occurrence and development of eyelid inflammation.

[0003] Currently, clinical treatments for Demodex blepharitis mainly include physical therapy, chemotherapy, and drug intervention, but all have limitations to varying degrees. Physical therapies such as warm compresses and eyelid cleaning can only play an auxiliary role in removal and are unlikely to completely eradicate the mites residing deep within the hair follicles and glands. Tea tree oil is a commonly used topical anti-Demodex mite preparation, but high concentrations of tea tree oil are often accompanied by adverse reactions such as eye irritation, burning sensation, and allergic reactions, and its efficacy is unstable. Furthermore, tea tree oil products have not yet been approved by regulatory agencies for the treatment of Demodex blepharitis.

[0004] Lotilaner ophthalmic solution, containing 0.25% loratadine, has been approved by the FDA, becoming the first drug specifically for the treatment of Demodex blepharitis. However, this drug also has adverse reactions such as stinging and burning at the instillation site, and it has not yet been approved for marketing in China, limiting its clinical accessibility.

[0005] Ivermectin, a macrolide derivative of the avermectin class, exhibits highly selective neurotoxicity in arthropods. It effectively kills parasites such as Demodex mites and scabies mites by acting on γ-aminobutyric acid (GABA)-gated chloride ion channels, causing neuromuscular hyperpolarization. In vitro and clinical studies have confirmed the good killing effect of ivermectin on Demodex folliculorum. However, current topical ivermectin preparations are mainly in cream or ointment form for dermatological use, and these preparations are not specifically designed for the eyelid margin, a unique anatomical location. Existing studies have shown that topical application of 1% ivermectin cream or ointment is effective in treating Demodex blepharitis, but the adhesion and retention of cream / ointment formulations on the eyelid margin are limited, and direct application to the eyes can easily cause irritation.

[0006] In conclusion, although ivermectin has a clear effectiveness against Demodex mites, there is currently no safe, stable, non-irritating ophthalmic formulation of ivermectin suitable for administration to the ocular surface and eyelid margin. Summary of the Invention

[0007] (a) Technical problems to be solved In view of the above-mentioned shortcomings and deficiencies of the prior art, the present invention provides an ophthalmic pharmaceutical composition for treating Demodex blepharitis and its preparation method, which solves the technical problem that there is currently no safe, stable, non-irritating ophthalmic preparation of ivermectin that is suitable for administration to the ocular surface and eyelid margin.

[0008] (II) Technical Solution To achieve the above objectives, the main technical solutions adopted by the present invention include: In a first aspect, embodiments of the present invention provide an ophthalmic pharmaceutical composition for treating Demodex blepharitis, comprising ivermectin, an immunomodulator, and an ophthalmologically acceptable carrier; The content of ivermectin is 0.2%-0.5% of the total weight of the pharmaceutical composition; The immunomodulator is selected from cyclosporine A and tacrolimus, and the content of the immunomodulator is 0.001%-0.4% of the total weight of the pharmaceutical composition.

[0009] The ivermectin is a mixture of ivermectin B1a and ivermectin B1b, wherein the content of ivermectin B1a is not less than 80% and the content of ivermectin B1b is not more than 20%.

[0010] It should be noted that, unless otherwise stated, the contents involved in this invention are all considered to be mass ratios.

[0011] In a preferred embodiment of the present invention, the immunomodulator is cyclosporine A, and its content is 0.01%-0.4%.

[0012] Cyclosporine A, as a calcineurin inhibitor, inhibits calcineurin activity by binding to cyclophilin, a cytoplasmic protein of T lymphocytes, thereby blocking the nuclear translocation of NFAT family members and inhibiting T cell activation and the release of inflammatory cytokines.

[0013] In a preferred embodiment of the present invention, the immunomodulator is tacrolimus, and its content is 0.001%-0.1%.

[0014] In inhibiting calcineurin activity, the half-maximal inhibitory concentration (IC50) of tacrolimus is approximately 30 times lower than that of cyclosporine A. In this invention, a significantly lower concentration of tacrolimus than cyclosporine A is used to achieve a comparable or stronger immunosuppressive effect.

[0015] As a preferred embodiment of the present invention, the ophthalmologically acceptable carrier comprises one or more of the following: solubilizer, thickener, tensioning agent, buffer, preservative, chelating agent, and pH adjuster.

[0016] In a preferred embodiment of the present invention, the content of the solubilizer is 0.5%-5.0% of the total weight of the pharmaceutical composition; The solubilizer is selected from one or more of polysorbate 80, polysorbate 65, polysorbate 60, polysorbate 40 or polysorbate 20, polyoxyethylene hydrogenated castor oil, poloxamer 188, and poloxamer 407.

[0017] In a preferred embodiment of the present invention, the content of the thickener is 0.1%-4% of the total weight of the pharmaceutical composition; The thickener is selected from one or more of hydroxypropyl methylcellulose, sodium carboxymethyl cellulose, hydroxypropyl cellulose, and carbomer.

[0018] In a preferred embodiment of the present invention, the content of the tensioning agent is 0.5%-5% of the total weight of the pharmaceutical composition; the content of the buffer is 0.1%-2.0% of the total weight of the pharmaceutical composition; the content of the preservative is 0.001%-0.1% of the total weight of the pharmaceutical composition; and the content of the chelating agent is 0.005%-0.1% of the total weight of the pharmaceutical composition. In a preferred embodiment of the present invention, the tensioning agent is selected from one or more of glycerol, dextran, mannitol, potassium chloride, and sodium chloride.

[0019] In a preferred embodiment of the present invention, the buffer is selected from one or more of acetate buffers, citrate buffers, phosphate buffers and borate buffers.

[0020] In a preferred embodiment of the present invention, the preservative is selected from one or more of thimerosal, benzalkonium chloride, benzalkonium bromide, potassium sorbate, phenoxyethanol, chlorobutanol, chlorhexidine, ethylparaben, boric acid, chlorobutanol, and polyhexamethylene biguanide hydrochloride.

[0021] In a preferred embodiment of the present invention, the chelating agent is disodium ethylenediaminetetraacetate and / or disodium edetate.

[0022] The pH of the pharmaceutical composition is adjusted to 6.5-8.0 using the pH adjuster. Preferably, the pH is 6.5-7.5.

[0023] In a preferred embodiment of the present invention, the dosage form of the pharmaceutical composition is an eye drop or an ophthalmic gel.

[0024] For eye drops, the dosage is 1-2 drops (approximately 25-50 μL) per eye, 1-2 times daily, with a total daily dose of ivermectin of 0.05 mg-0.5 mg. Administration is performed using a drop-and-apply technique: first, instill the drops into the conjunctival sac; then, close the eye to allow the preparation to contact the meibomian gland openings; finally, use a finger or applicator to spread the preparation accumulated outside the eyelid margin to the root of the eyelashes, achieving double coverage of both the eyelash follicle and the meibomian gland. The eye drops have a suitable viscosity to cover the meibomian gland openings when the patient blinks or closes their eyes, and to slow the expulsion of the preparation through the lacrimal puncta, thus prolonging the residence time of the preparation at the eyelid margin by at least 5, 10, 20, 30 seconds, or longer.

[0025] For the ophthalmic gel, the dosage is approximately 200 mg per administration, applied directly to the upper and lower eyelid margins, covering the root of the eyelashes to the meibomian gland opening area. The viscosity of the ophthalmic gel at 25°C is 500-50000 cP, preferably 2000-10000 cP, more preferably 4000-6000 cP, to further enhance the retention time on the eyelid margin and the contact time with the meibomian gland opening.

[0026] In a preferred embodiment of the present invention, the dosage form of the pharmaceutical composition is an ophthalmic gel, and the amount of the thickener is 0.5%-3% of the total weight of the pharmaceutical composition.

[0027] Secondly, embodiments of the present invention provide a method for preparing an ophthalmic pharmaceutical composition, wherein the dosage form of the pharmaceutical composition is an eye drop, and the preparation method includes the following steps: S1. Under aseptic conditions, heat the water for injection, add the solubilizer, tensioning agent and buffer, stir to dissolve, and obtain the first solution; S2. Cool the first solution to room temperature, add a thickener, stir to disperse and allow the thickener to swell, add a chelating agent, stir to dissolve, and obtain the second solution; S3. Take the prescribed amount of ivermectin and immunomodulator, slowly add them to the second solution, and stir until completely dissolved under light-protected conditions to obtain the third solution; S4. Adjust the pH of the third solution to 6.5-8.0, add preservative, add water for injection to the total volume, mix well, and obtain the fourth solution; S5. Under aseptic conditions, the fourth solution is filtered to remove bacteria, dispensed into sterile containers, and sealed to obtain the final product.

[0028] In a preferred embodiment of the present invention, the filtration and sterilization in step S5 uses a 0.22μm microporous filter membrane with a volume not exceeding 10ml / bottle, preferably 5ml / bottle.

[0029] Thirdly, embodiments of the present invention provide a method for preparing an ophthalmic pharmaceutical composition, wherein the dosage form of the pharmaceutical composition is an ophthalmic gel, and the preparation method includes the following steps: Step 1: Under aseptic conditions, take water for injection, add solubilizer and tensioning agent, and stir to dissolve; Step 2: Add the prescribed amount of ivermectin and immunomodulator, and stir until completely dissolved; Step 3: Add preservatives and chelating agents, and stir until homogeneous; Step 4: Adjust the pH to 6.5-8.0, filter to sterilize, and obtain the drug solution; Step 5: Disperse the thickener in an appropriate amount of water for injection, stir to swell, sterilize, and cool to obtain a gel matrix; Step 6: Under aseptic conditions, mix the drug solution with the gel matrix, stir evenly, add water for injection to the full volume, dispense into sterile containers, seal, and the product is obtained.

[0030] (III) Beneficial Effects The beneficial effects of this invention are as follows: This invention provides an ophthalmic pharmaceutical composition for treating Demodex mite-induced blepharitis and its preparation method, which combines ivermectin with an immunomodulator to achieve a synergistic therapeutic strategy of killing mites and reducing inflammation. On the one hand, ivermectin, as a macrolide antiparasitic drug, has a highly effective killing effect on Demodex mites; on the other hand, ivermectin itself also has clear anti-inflammatory properties, capable of dose-dependently inhibiting the expression of pro-inflammatory cytokines such as tumor necrosis factor-α, interleukin-1β, and IL-6, and exerting its anti-inflammatory effect by blocking the nuclear translocation of nuclear factor κB. The immunomodulator further reduces ocular surface inflammation by inhibiting T cell activation and the release of inflammatory mediators. The combination of these two agents can simultaneously target both the etiology and pathological consequences of Demodex mite infection, exerting a synergistic effect and providing a new drug option for the clinical treatment of Demodex mite-induced blepharitis.

[0031] The composition of this invention, administered topically to the eyelid margin, directly targets the affected area, effectively relieving clinical symptoms such as eyelid congestion, itching, scaling, foreign body sensation, dryness, and burning. It also improves meibomian gland function and promotes the recovery of tear film stability. The topical ophthalmic formulation allows the drug to act directly on the affected area, avoiding systemic side effects common with oral ivermectin, such as fever, headache, elevated liver enzymes, and drug interactions. Topical administration ensures an effective drug concentration at the site of action while reducing the amount of drug entering the systemic circulation, resulting in good local tolerability and safety.

[0032] The compositions of this invention, through reasonable excipient ratios and preparation processes, are formulated into clear and stable eye drops or ophthalmic gels with good physicochemical stability. The eye drops and ophthalmic gels are convenient to use, allowing patients to self-administer the medication, thus improving treatment adherence. Detailed Implementation

[0033] To better explain and facilitate understanding of the invention, specific embodiments are described in detail below.

[0034] While exemplary embodiments of the invention have been shown, it should be understood that the invention may be implemented in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided so that the invention may be understood more clearly and thoroughly, and so that the scope of the invention may be fully conveyed to those skilled in the art.

[0035] Example 1: This embodiment provides an eye drop for treating Demodex blepharitis and its preparation method. An exemplary prescription for the eye drop is shown in Table 1.

[0036] Table 1. Example prescription for eye drops (ivermectin content 0.2% w / w)

[0037] The method for preparing eye drops includes the following steps: (1) Under sterile conditions, take the prescribed amount of water for injection and heat it to 50-60°C. Add polysorbate 80 and glycerin, stir to dissolve, add sodium dihydrogen phosphate and disodium hydrogen phosphate, and stir until completely dissolved to obtain the first solution.

[0038] (2) Cool the first solution to room temperature, add hydroxypropyl methylcellulose, stir and disperse to allow it to swell fully and form a transparent solution, add disodium ethylenediaminetetraacetate, stir and dissolve to obtain the second solution.

[0039] (3) Take the prescribed amount of ivermectin and cyclosporine A, slowly add them to the second solution, and stir thoroughly under light-protected conditions until completely dissolved to obtain the third solution.

[0040] (4) Adjust the pH to 7.0-7.5 with sodium hydroxide and hydrochloric acid, add benzalkonium chloride, add water for injection to the total volume, mix thoroughly, and obtain the fourth solution.

[0041] (5) Under aseptic conditions, the fourth solution is filtered through a 0.22μm microporous membrane for sterilization, dispensed into sterile eye drop bottles, and sealed to obtain the solution.

[0042] Example 2: This embodiment provides an eye drop for treating Demodex blepharitis and its preparation method. An exemplary prescription for the eye drop is shown in Table 2.

[0043] Table 2. Example prescription for eye drops (ivermectin content 0.5% w / w)

[0044] The preparation method of the eye drops in this embodiment is the same as that in Example 1.

[0045] Example 3: This embodiment provides an eye drop for treating Demodex blepharitis and its preparation method. An exemplary prescription for the eye drop is shown in Table 3.

[0046] Table 3. Exemplary prescriptions for eye drops (ivermectin content 0.2% w / w)

[0047] The difference between the preparation method of the eye drops in this embodiment and that in Example 1 is that in step (3), the prescribed amount of ivermectin and tacrolimus are slowly added to the second solution and stirred thoroughly under light-protected conditions until completely dissolved to obtain the third solution; the remaining steps are the same as in Example 1.

[0048] Example 4: This embodiment provides an ophthalmic gel for treating Demodex blepharitis and a method for preparing the same. An exemplary formulation of the ophthalmic gel is shown in Table 4.

[0049] Table 4. Exemplary formulation of ophthalmic gel (ivermectin content 0.2% w / w)

[0050] The preparation method of ophthalmic gel includes the following steps: (1) Under sterile conditions, take the prescribed amount of water for injection, add propylene glycol and glycerin, stir to dissolve, and obtain the first solution.

[0051] (2) Take the prescribed amount of ivermectin and cyclosporine A, slowly add them to the first solution, and stir thoroughly under light-protected conditions until completely dissolved to obtain the second solution.

[0052] (3) Add benzalkonium chloride to the second solution, stir until homogeneous, add disodium edetate, and stir to dissolve.

[0053] (4) Adjust the pH to 7.0-7.5 with sodium hydroxide and hydrochloric acid, and filter the resulting solution through a 0.22μm microporous membrane to remove bacteria, and obtain the drug solution.

[0054] (5) Take carbomer and hydroxypropyl methylcellulose and disperse them in an appropriate amount of water for injection. Stir to fully swell and form a transparent gel matrix. Sterilize at 121°C for 20 min and cool to room temperature to obtain the gel matrix.

[0055] (6) Under aseptic conditions, mix the drug solution with the gel matrix, stir thoroughly to make it uniform, and add water for injection to the full volume to obtain ophthalmic gel; under aseptic conditions, dispense the obtained ophthalmic gel into sterile ophthalmic gel tubes, seal them, and the product is obtained. The filling amount of each container is 5g / tube.

[0056] Example 5: This embodiment provides an ophthalmic gel for treating Demodex blepharitis and a method for preparing the same. An exemplary formulation of the ophthalmic gel is shown in Table 5.

[0057] Table 5. Exemplary formulation of ophthalmic gel (ivermectin content 0.5% w / w)

[0058] The preparation method of the ophthalmic gel in this embodiment is the same as that in Example 4.

[0059] Example 6: This embodiment provides an ophthalmic gel for treating Demodex blepharitis and a method for preparing the same. An exemplary formulation of the ophthalmic gel is shown in Table 6.

[0060] Table 6. Exemplary formulation of ophthalmic gel (ivermectin content 0.2% w / w)

[0061] The difference between the ophthalmic gel preparation method in this embodiment and that in embodiment 4 is that in step (2), the prescribed amount of ivermectin and tacrolimus are slowly added to the first solution and stirred thoroughly under light-protected conditions until completely dissolved to obtain the second solution; the remaining steps are the same as in embodiment 4.

[0062] Comparative Example 1: This comparative example provides 0.25% loteranal eye drops as a control.

[0063] 0.25% Loteranal Eye Drops are a sterile, preservative-containing, multi-dose, pale yellow to slightly milky white ophthalmic solution. Its active ingredient is loteranal at a concentration of 0.25% (2.5 mg / mL). Excipients include: potassium sorbate (preservative), disodium edetate (chelating agent), hydroxypropyl methylcellulose (thickener), polyoxyethylene 35 castor oil (solvent / emulsifier), glycerin (strengthening agent / solvent), disodium hydrogen phosphate (pH adjuster / buffer), sodium dihydrogen phosphate (pH adjuster / buffer), and water for injection (solvent). The dosage regimen is one drop twice daily for six weeks.

[0064] Comparative Example 2: This comparative example provides an ivermectin monotherapy eye drop as a control. The difference between this ivermectin monotherapy eye drop and Example 1 is that it contains only ivermectin as the single active ingredient, does not contain immunomodulators (cyclosporine A or tacrolimus), and the ivermectin content is 0.25%. Its excipient composition is the same as in Example 1.

[0065] The preparation method is the same as in Example 1, except that the step of adding the immunomodulator is omitted and the amount of ivermectin is adjusted to 0.25%.

[0066] The dosing regimen was the same as in Example 1, i.e., 1 drop twice daily for 6 weeks.

[0067] 1. Quality Inspection Quality testing was conducted on Examples 1-6 above, and the testing was performed in accordance with the relevant provisions of General Chapter 0105, Ophthalmic Preparations, Part IV of the Pharmacopoeia of the People's Republic of China (2025 Edition). The test results are shown in Tables 7 and 8.

[0068] Table 7. Eye Drop Test Results

[0069] Table 8. Results of ophthalmic gel testing

[0070] As can be seen from the test results in Tables 7 and 8, the ophthalmic drug compositions prepared in Examples 1-6 of the present invention meet the relevant regulations for ophthalmic preparations in terms of appearance, pH value, osmotic pressure / viscosity, active ingredient content, sterility test and visible foreign matter.

[0071] 2. Stability Test The stability of the ophthalmic drug compositions prepared in Examples 1 and 4 was investigated under accelerated testing conditions (40℃±2℃, RH 75%±5%) for 6 months. Samples were taken at 0, 3, and 6 months, and the test indicators included appearance, pH value, active ingredient content, and sterility. The test results are shown in Table 9.

[0072] Table 9 Stability test results

[0073] As shown in Table 9, the ophthalmic drug composition prepared by this invention, after being placed under accelerated testing conditions for 6 months, met all the quality requirements for ophthalmic preparations, exhibiting good stability and controllable quality, and meeting the shelf-life requirements for clinical use. All other examples were also subjected to stability studies under the same conditions, and their properties, pH value, active ingredient content, and sterility tests all met the requirements, with trends basically consistent with those of Examples 1 and 4.

[0074] 3. Eye irritation test Eye irritation tests were conducted using New Zealand white rabbits (6 rabbits per group, half male and half female) for single and multiple administrations.

[0075] 1) Single administration: 50 μl of eye drops (0.5%, Example 2) or 200 mg of ophthalmic gel (0.5%, Example 5) was instilled into one eye of each rabbit, and an equal volume of blank carrier was given to the other eye as a control. The conjunctiva, cornea, and iris were scored according to the Draize scoring system at 1, 2, 4, 24, 48, and 72 hours after administration.

[0076] 2) Multiple administration: Administer twice daily for 28 consecutive days. Ophthalmological examinations should be performed on days 7, 14, 21, and 28, and the conjunctiva, cornea, and iris should be scored according to the Draize scoring system.

[0077] Draize Eye Irritation Score: The maximum total score is 110 points (cornea 80 points, iris 10 points, conjunctiva 20 points). Irritation level is determined based on the total score: 0-3 points indicate no irritation, 4-8 points indicate mild irritation, 9-12 points indicate moderate irritation, 13-16 points indicate moderate to severe irritation, and >16 points indicate severe irritation. Test results are shown in Tables 10 and 11.

[0078] Table 10 Results of single-dose eye irritation test (n=6, ±SD, total score)

[0079] Table 11 Results of eye irritation test after multiple administrations (28 days) (n=6, ±SD)

[0080] Results of the single-dose irritation test: As shown in Table 10, after a single dose, the Draize total score for both Example 2 (eye drops) and Example 5 (ocular gel) was less than 3 points. After reaching a peak at 1 hour, the score rapidly decreased, returning to near baseline levels (total score < 1) by 24 hours. At 48 and 72 hours, the score dropped to 0, with no corneal damage. These results indicate that the formulation of this invention is non-irritating to rabbit eyes, and the ocular irritation response completely recovers within 24 hours after administration, with all scores returning to baseline levels within 24 hours.

[0081] Results of repeated-dose irritation test: As shown in Table 11, after 28 days of continuous administration, there were no statistically significant differences in the corneal, iris, and conjunctival sub-scores and total scores at each time point between Example 2 (eye drops) and Example 5 (ophthalmic gel) and the blank control group. p >0.05). No obvious ocular congestion, edema, or increased secretions were observed in any of the formulation groups, and no corneal edema, neovascularization, or fluorescein-positive areas were observed.

[0082] Conclusion: The above eye irritation test results show that the eye drops and ophthalmic gel prepared in this invention have no obvious irritation or toxicity to rabbit eyes, exhibiting good ophthalmic safety and meeting the safety requirements for ophthalmic preparations, and can be safely used clinically. Other embodiments also demonstrate good ophthalmic safety.

[0083] 4. Evaluation of anti-Demodex mite efficacy A Demodex mite infection model was established using immunosuppressed New Zealand white rabbits. Specifically, systemic immunosuppression was induced by subcutaneous injection of cyclosporine A (15 mg / kg / day) and methylprednisolone (1 mg / kg / day) for 14 days, allowing Demodex mites to successfully colonize the rabbit eyelids. Subsequently, a suspension of human Demodex folliculorum was inoculated into the root of the eyelashes and the meibomian gland region of the rabbit eyelids. Successful model establishment was defined as an average of ≥5 mites per eyelash.

[0084] It should be noted that the cyclosporine A used in the modeling phase was administered systemically to induce systemic immunosuppression, and it was discontinued at the start of the trial; the test formulation containing cyclosporine A used in the treatment phase was administered topically (eye drops or ophthalmic gel), and the two have completely different routes of administration and pharmacological effects, and do not interfere with each other.

[0085] After successfully establishing the model, the animals (n=60) were randomly divided into 10 groups of 6 animals each: Group A (Example 1, n=6); Group B (Example 2, n=6); Group C (Example 3, n=6); Group D (Example 4, n=6); Group E (Example 5, n=6); Group F (Example 6, n=6); Group G (blank vector group, n=6): blank vector without active ingredients; Group H (positive control group, comparative example 1, n=6); Group I (comparative example 2, n=6); Group J (sham operation group, n=6): normal rabbits not inoculated with Demodex mites, given an equal volume of physiological saline.

[0086] Dosage regimen: Administer twice daily (approximately 12 hours apart) for 6 consecutive weeks. Every 2 weeks, sample eyelashes and count mites (using a direct microscope to count the number of mites in the shed eyelashes). Results are shown in Table 12.

[0087] Table 12. Changes in the density of eyelash mites in an in vivo rabbit model (mites / eyelash, mean ± standard deviation)

[0088] # indicates that compared with the blank vector group (G group), p < 0.05; # indicates that compared with the positive control group (H group), p < 0.05.

[0089] result: Group A: Mite density decreased from 6.8±1.2 at baseline to 1.5±0.9 at week 6, a reduction of 78% (p<0.001). By the end of week 6, 50% (3 / 6) of the animals had achieved the eradication criterion of ≤1 mite per eyelash.

[0090] Group B: Mite density decreased from 7.2±1.5 at baseline to 0.4±0.5 at week 6, a reduction of 94% (p<0.001). By the end of week 6, 83% (5 / 6) of the animals had achieved the eradication criterion of ≤1 mite per eyelash.

[0091] Group C: Mite density decreased from 7.0±1.4 at baseline to 1.0±0.6 at week 6, a reduction of 86% (p<0.001). By the end of week 6, 67% (4 / 6) of the animals had achieved the eradication criterion of ≤1 mite per eyelash.

[0092] Group D: Mite density decreased from 7.0±1.3 at baseline to 0.8±0.6 at week 6, a reduction of 89% (p<0.001). By the end of week 6, 67% (4 / 6) of the animals had achieved the eradication criterion of ≤1 mite per eyelash.

[0093] Group E: Mite density decreased from 7.1±1.4 at baseline to 0.5±0.5 at week 6, a reduction of 93% (p<0.001). By the end of week 6, 83% (5 / 6) of the animals had achieved the eradication criterion of ≤1 mite per eyelash.

[0094] Group F: Mite density decreased from 6.9±1.3 at baseline to 0.6±0.5 at week 6, a reduction of 91% (p<0.001). By the end of week 6, 83% (5 / 6) of the animals had achieved the eradication criterion of ≤1 mite per eyelash.

[0095] Group G: Mite density increased from 6.9±1.4 at baseline to 8.1±1.8 at week 6, with no improvement.

[0096] Group H: Mite density decreased from 7.1±1.4 at baseline to 1.1±0.8 at week 6, a reduction of 85% (p<0.001). By the end of week 6, 50% (3 / 6) of the animals had achieved the eradication criterion of ≤1 mite per eyelash.

[0097] Group I: Mite density decreased from 6.9±1.4 at baseline to 2.6±0.9 at week 6, a reduction of 62% (p<0.05). By the end of week 6, 17% (1 / 6) of the animals had achieved the eradication criterion of ≤1 mite per eyelash.

[0098] Group J: No Demodex mite colonization was observed throughout the entire process.

[0099] Results analysis: In both eye drops and gel formulations, the 0.5% ivermectin formulations (groups B and E) showed superior acaricidal efficacy (reduction rates of 94% and 93%, respectively) compared to the corresponding 0.2% formulations (groups A and D, reduction rates of 78% and 89%), demonstrating a concentration-dependent effect. The 0.2% ivermectin ophthalmic gel (group D, 89%) showed better acaricidal efficacy than the eye drops of the same concentration (group A, 78%), and was comparable to the 0.5% eye drops (group B, 94%). The gel formulation, by prolonging the residence time of the drug on the ocular surface and eyelid margin, achieves highly effective acaricidal activity at lower drug concentrations.

[0100] In 0.2% ivermectin eye drops, the addition of tacrolimus (Group C, 86%) resulted in a superior acaricidal effect compared to the addition of cyclosporine A (Group A, 78%), even though the concentration of tacrolimus was only 1 / 5 that of cyclosporine A. In the gel formulation, 0.2% ivermectin + tacrolimus (Group F, 91%) was comparable in efficacy to 0.2% ivermectin + cyclosporine A (Group D, 89%). These results indicate that tacrolimus, as an immunomodulator, can achieve synergistic anti-mite effects similar to or even better than cyclosporine A at lower concentrations. This may be related to the stronger immunosuppressive activity of tacrolimus (10-100 times more potent than cyclosporine A in vitro).

[0101] Comparing Group A (Example 1, 0.2% ivermectin + 0.05% cyclosporine A) with Group I (Comparative Example 2, 0.25% ivermectin eye drops), it was found that: the total concentration of active ingredients was the same in both groups (0.25%), but the mite density reduction rate in Group A was 78%, significantly higher than that in Group I (62%) (p < 0.05). These results indicate that, under the condition of consistent total drug dosage, the combined use of ivermectin and cyclosporine A is significantly more effective than ivermectin alone. If it were merely a simple additive effect, there should not be a significant difference in efficacy between the two; the observed difference in efficacy suggests a synergistic effect between ivermectin and cyclosporine A.

[0102] Groups B, E, and F showed significant mite density reductions by the end of week 2 (p < 0.05), while the positive control group (loteranar) only achieved the same reduction by the end of week 4, indicating that the formulation of this invention has a faster onset of action than the positive control drug. The 6-week mite-killing efficacy of groups B and E (94% and 93%, respectively) was superior to that of the positive control group (85%); the mite-killing efficacy of groups C and F was comparable to or better than that of the positive control group. Simultaneously, the eradication rate of groups B and E (83%) was also higher than that of the positive control group (50%).

[0103] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

[0104] The "range" disclosed in this application is defined by a lower limit and an upper limit. A given range is defined by selecting a lower limit and an upper limit, which define the boundaries of a particular range. Ranges defined in this way can include or exclude endpoints and can be arbitrarily combined; that is, any lower limit can be combined with any upper limit to form a range. For example, if ranges of 60-120 and 80-110 are listed for a specific parameter, it is expected that ranges of 60-110 and 80-120 are also included. Furthermore, if minimum range values ​​of 1 and 2 are listed, and if maximum range values ​​of 3, 4, and 5 are listed, then the following ranges are all expected: 1-3, 1-4, 1-5, 2-3, 2-4, and 2-5. In this application, unless otherwise stated, the numerical range "ab" represents a shortened representation of any combination of real numbers between a and b, where a and b are real numbers. For example, the numerical range "0-5" indicates that all real numbers between "0-5" have been listed in this article; "0-5" is simply a shortened representation of these numerical combinations. Furthermore, when a parameter is stated as an integer ≥2, it is equivalent to disclosing that the parameter is, for example, an integer such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, etc.

[0105] Unless otherwise specified, all embodiments and optional embodiments of this application can be combined to form new technical solutions.

[0106] Unless otherwise specified, all technical features and optional technical features of this application may be combined to form new technical solutions.

[0107] Unless otherwise specified, all steps in this application may be performed sequentially or randomly, for example, sequentially. For instance, the method includes steps (a) and (b), indicating that the method may include steps (a) and (b) performed sequentially, or it may include steps (b) and (a) performed sequentially. For instance, the method may also include step (c), indicating that step (c) may be added to the method in any order; for example, the method may include steps (a), (b), and (c), or it may include steps (a), (c), and (b), or it may include steps (c), (a), and (b), etc.

[0108] Unless otherwise specified, the terms "comprising" and "including" as used in this application can be open-ended or closed-ended. For example, "comprising" and "including" can mean that other components not listed may also be included, or that only the listed components may be included.

[0109] Unless otherwise specified, the term "or" is inclusive in this application. For example, the phrase "A or B" means "A, B, or both A and B". More specifically, the condition "A or B" is satisfied by any of the following conditions: A is true (or exists) and B is false (or does not exist); A is false (or does not exist) and B is true (or exists); or both A and B are true (or exist).

Claims

1. An ophthalmic pharmaceutical composition for treating Demodex blepharitis, characterized in that, It contains ivermectin, immunomodulators, and ophthalmologically acceptable carriers; The content of ivermectin is 0.2%-0.5% of the total weight of the pharmaceutical composition; The immunomodulatory agent is present in an amount of 0.001%-0.4% of the total weight of the pharmaceutical composition, and the immunomodulatory agent is selected from cyclosporine A and tacrolimus.

2. The ophthalmic pharmaceutical composition for treating Demodex blepharitis as described in claim 1, characterized in that, The immunomodulator is cyclosporine A, with a content of 0.01%-0.4%.

3. The ophthalmic pharmaceutical composition for treating Demodex blepharitis as described in claim 1, characterized in that, The immunomodulator is tacrolimus, with a content of 0.001%-0.1%.

4. The ophthalmic pharmaceutical composition for treating Demodex blepharitis as described in claim 1, characterized in that, The ophthalmologically acceptable carrier comprises one or more of the following: solubilizer, thickener, tensioning agent, buffer, preservative, chelating agent, and pH adjuster.

5. The ophthalmic pharmaceutical composition for treating Demodex blepharitis as described in claim 4, characterized in that, The content of the solubilizer is 0.5%-5.0% of the total weight of the pharmaceutical composition; The solubilizer is selected from one or more of polysorbate 80, polysorbate 65, polysorbate 60, polysorbate 40 or polysorbate 20, polyoxyethylene hydrogenated castor oil, poloxamer 188, and poloxamer 407.

6. The ophthalmic pharmaceutical composition for treating Demodex blepharitis as described in claim 4, characterized in that, The content of the thickener is 0.1%-4% of the total weight of the pharmaceutical composition; The thickener is selected from one or more of hydroxypropyl methylcellulose, sodium carboxymethyl cellulose, hydroxypropyl cellulose, and carbomer.

7. The ophthalmic pharmaceutical composition for treating Demodex blepharitis as described in claim 4, characterized in that, The content of the tensioning agent is 0.5%-5% of the total weight of the pharmaceutical composition; the content of the buffer is 0.1%-2.0% of the total weight of the pharmaceutical composition; the content of the preservative is 0.001%-0.1% of the total weight of the pharmaceutical composition; and the content of the chelating agent is 0.005%-0.1% of the total weight of the pharmaceutical composition.

8. The ophthalmic pharmaceutical composition for treating Demodex blepharitis as described in any one of claims 1-7, characterized in that, The dosage form of the pharmaceutical composition is eye drops or ophthalmic gel.

9. A method for preparing an ophthalmic pharmaceutical composition according to any one of claims 1-8, characterized in that, The dosage form of the pharmaceutical composition is eye drops, and the preparation method includes the following steps: S1. Under aseptic conditions, heat the water for injection, add the solubilizer, tensioning agent and buffer, stir to dissolve, and obtain the first solution; S2. Cool the first solution to room temperature, add a thickener, stir to disperse and allow the thickener to swell, add a chelating agent, stir to dissolve, and obtain the second solution; S3. Take the prescribed amount of ivermectin and immunomodulator, add them to the second solution, and stir under light-protected conditions until completely dissolved to obtain the third solution; S4. Adjust the pH of the third solution to 6.5-8.0, add preservative, add water for injection to the total volume, mix well, and obtain the fourth solution; S5. Under aseptic conditions, the fourth solution is filtered to remove bacteria, dispensed into sterile containers, and sealed to obtain the final product.

10. A method for preparing an ophthalmic pharmaceutical composition according to any one of claims 1-8, characterized in that, The dosage form of the pharmaceutical composition is an ophthalmic gel, and the preparation method includes the following steps: Step 1: Under aseptic conditions, take water for injection, add solubilizer and tensioning agent, and stir to dissolve; Step 2: Add the prescribed amount of ivermectin and immunomodulator, and stir until completely dissolved under light-protected conditions; Step 3: Add preservatives and chelating agents, and stir until homogeneous; Step 4: Adjust the pH to 6.5-8.0, filter to sterilize, and obtain the drug solution; Step 5: Disperse the thickener in an appropriate amount of water for injection, stir to swell, sterilize, and cool to obtain a gel matrix; Step 6: Under aseptic conditions, mix the drug solution with the gel matrix, stir evenly, add water for injection to the full volume, dispense into sterile containers, seal, and the product is obtained.

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