An ophthalmic preparation for the treatment of macular edema, optic neuritis and non-infectious endophthalmitis.

Abstract

This invention provides an ophthalmic preparation for treating macular edema, optic neuritis, and non-infectious endophthalmitis. It is a preparation composed of an active ingredient for treating eye diseases and an ophthalmic preparation carrier or excipients. The active ingredient for treating eye diseases is a glucocorticoid and / or a non-steroidal anti-inflammatory drug (NSAID). The ophthalmic preparation carrier or excipients contain the following components: low-polymerization povidone, medium-polymerization povidone, and a solvent. This ophthalmic preparation of the present invention can be administered via eye drops, carrying (encapsulating) glucocorticoids and / or NSAIDs through the anterior segment of the eye and delivering them to the posterior segment to treat macular edema, optic neuritis, and non-infectious endophthalmitis. It has extremely excellent clinical value and very positive social significance.

Description

[0001] This application is a divisional application of the invention patent with application number 2021106720650 and application date of June 17, 2021. Technical Field

[0002] This invention belongs to the field of ophthalmic drugs, specifically relating to an ophthalmic preparation for the treatment of macular edema, optic neuritis and non-infectious endophthalmitis by eye drops. Background Technology

[0003] There are numerous patients with fundus diseases, with tens of millions in China alone. With an aging population and the widespread use of electronic products, the incidence rate is expected to rise annually. Common fundus diseases include diabetic macular edema, diabetic retinopathy, age-related macular degeneration, retinal vein occlusion, pathological myopia, geographic atrophy, ocular tumors, and endophthalmitis, which can lead to vision loss or even blindness, severely impacting people's quality of life. For example, approximately 6.8% of diabetic patients suffer from diabetic macular edema (DME), a leading cause of blindness in diabetic patients (Urias et al., Vision Research, V139:221-227, 2017; Mandal et al., Ocular delivery of proteins and peptides: Challenges and novel formulation approaches, Advanced Drug Delivery Reviews, 126:67-95, 2018).

[0004] Drug therapy is a major treatment method and research trend for fundus diseases. However, due to the complex physiological structures and barriers within the eye, it is difficult for drugs to enter the eyeball, especially the posterior segment, making it difficult to achieve an effective dose and thus hindering effective treatment. Finding an effective and safe agent or method for treating fundus diseases has been a persistent goal for researchers in this field.

[0005] There are generally three routes of drug administration in clinical ophthalmology: (1) Conjunctival sac administration (eye drops): After entering the aqueous humor through the cornea, the drug can diffuse to the iris and ciliary body, but the barrier effect of the lens and vitreous membrane makes it difficult for the drug to enter the lens and vitreous body; (2) Intraocular injection: This includes subconjunctival injection, anterior chamber injection, vitreous injection, retrobulbar injection, and orbital injection. Injection allows the drug to reach the treatment site directly, but injection is invasive and has potential risks, such as pain, photophobia, tearing, anterior chamber opacity, hemorrhage, corneal endothelial cell damage, traumatic cataract, etc., caused by anterior chamber injection; and lens opacity, vitreous organization, retinal / optic nerve damage, etc., caused by vitreous injection; (3) Systemic administration, including oral administration and intravenous administration: The drug generally accumulates in the liver, kidneys, or lungs in the body, and is hindered by the blood-retinal barrier (BRB), resulting in a low concentration reaching the ocular tissues. At the same time, the whole body, especially major organs, suffer unnecessary toxic side effects.

[0006] Currently, in clinical practice, techniques such as intravitreal injection or intravitreal implantation (internal insertion) are commonly used to deliver drugs to the vitreous body of patients in order to cross the ocular barrier and treat diseases of the posterior segment of the eye (Ling Peixue, ed., Ophthalmic Drugs and Preparations, China Light Industry Press, 2010, p. 3; Wang et al., Mediators of Inflammation, Vol. 2013, Article ID 780634; Luaces-Rodríguez et al., Pharmaceutics, 2018, 10, 66). Intravitreal injection or implantation of drugs is an invasive procedure that requires specially trained ophthalmologists to perform in a sterile environment such as an operating room. Due to the invasiveness of the procedure, complications may occur, such as high intraocular pressure, cataracts, iatrogenic endophthalmitis, vitreous hemorrhage, and retinal damage. The procedure requires high standards of conditions and environment and must be performed in hospitals with the necessary facilities. The production and usage costs of biopharmaceutical ophthalmic injections are high. Furthermore, there is a risk of delays in treatment due to limitations in medical resources, and the flexibility in adjusting the dosing regimen is poor (M. HATA et al., RETINA, 37:1320-1328, 2017).

[0007] In fact, existing ophthalmic injections for treating wet age-related macular degeneration (wAMD) and diabetic macular edema (DME) are administered via intravitreal injection, requiring injection every 1-3 months according to the drug instructions, and necessitating long-term administration. Furthermore, the glucocorticoid dexamethasone has also been developed as a vitreous implant (intraocular insert, trade name: [omitted]). Ozurdex is used to treat posterior segment eye diseases such as macular edema caused by retinal vein occlusion, and its efficacy can last for up to 6 months after implantation. However, some patients experience increased intraocular pressure after implantation, reaching its peak at 2 months. Patient Management Manual (Allergan, CN / 011 / 2018), there is a risk of side effects.

[0008] Conjunctival sac administration is the most convenient and safest method of ocular drug delivery. However, the cornea has a multi-layered structure, roughly divided from the outside in: a lipid-rich epithelial layer, a water-rich stroma layer, and a lipid-rich endothelial layer. After instillation, eye drops first come into contact with the tear film on the ocular surface, and then need to cross the epithelial, stroma, and endothelial layers to reach the posterior segment of the eye. During this process, due to tear dilution, the ocular surface barriers of the cornea and conjunctiva, and the anatomical positions of the lens and vitreous body, the eye drops often have a high concentration in the anterior segment of the eye, making it difficult to reach the posterior segment and achieve an effective therapeutic concentration. Therefore, although conjunctival sac administration is safe, its poor drug delivery makes it difficult to effectively treat fundus diseases.

[0009] In summary, current methods of administering medication for treating macular edema and other fundus diseases are insufficient to balance safety and effectiveness. Summary of the Invention

[0010] Compared with intravenous injection and intravitreal injection, eye drops have significant advantages in terms of safety and convenience. Inventing an ophthalmic preparation that can deliver drugs to the posterior segment of the eye to treat macular edema is a technical problem that urgently needs to be solved in clinical practice, and it has great clinical treatment value and social significance.

[0011] The purpose of this invention is to provide an ophthalmic drug for eye drops that can deliver active ingredients for treating eye diseases to the posterior segment of the eye, treating macular edema, optic neuritis, and non-infectious endophthalmitis.

[0012] This invention provides an ophthalmic preparation for eye drop administration, which is a preparation composed of an active ingredient for treating eye diseases and an ophthalmic preparation carrier or excipients;

[0013] The active ingredient for treating eye diseases is a glucocorticoid and / or a nonsteroidal anti-inflammatory drug;

[0014] The ophthalmic formulation carrier or excipient contains the following components: surfactant, ionic polymer and solvent;

[0015] Alternatively, the ophthalmic formulation carrier or excipient may contain the following components: low-polymerization degree povidone, medium-polymerization degree povidone, and solvent.

[0016] Furthermore, the mass ratio of surfactant to ionic polymer in the carrier or excipient of the above-mentioned ophthalmic preparation is (1-100):(0.1-50); the ratio of surfactant to solvent is 5-3000 mg surfactant per 100 mL of solvent.

[0017] Furthermore, the mass ratio of surfactant to ionic polymer in the carrier or excipient of the above-mentioned ophthalmic preparation is (12-31):(2-7.5); the ratio of surfactant to solvent is 880-1240 mg surfactant per 100 mL of solvent.

[0018] Furthermore, the surfactant is a nonionic surfactant; preferably, the nonionic surfactant is a Span, polysorbate, poloxamer, alkyl glucoside, vitamin E polysuccinate, sucrose stearate, or azone; preferably a Span or polysorbate.

[0019] Furthermore, the aforementioned ionic polymer is selected from at least one of carboxymethyl cellulose and its salts, sodium glycolate starch, hyaluronic acid and its salts, xanthan gum, alginate and its salts, and polyethylene glycol diacetate PEG-(COOH)2; preferably, the ionic polymer is selected from at least one of carboxymethyl cellulose and its salts and hyaluronic acid and its salts.

[0020] Furthermore, the mass ratio of low-polymerization degree povidone to medium-polymerization degree povidone in the carrier or excipient of the above-mentioned ophthalmic preparation is (0.1-10):1, and the ratio of low-polymerization degree povidone to solvent is 5-3000 mg of low-polymerization degree povidone per 100 mL of solvent.

[0021] Preferably, the mass ratio of the low-polymerization degree polyvinylpyrrolidone to the medium-polymerization degree polyvinylpyrrolidone is (0.24-0.8):1, and the ratio of the low-polymerization degree polyvinylpyrrolidone to the solvent is 240-840 mg of low-polymerization degree polyvinylpyrrolidone per 100 mL of solvent.

[0022] Furthermore, the aforementioned low-polymerization-degree polyvinyl chloride is a polyvinyl chloride with a weight-average molecular weight of 2000 to 5000, preferably a polyvinyl chloride PVP K12 with a weight-average molecular weight of 3500.

[0023] Furthermore, the aforementioned medium-degree-of-polymerization polyvinyl chloride is a polyvinyl chloride with a weight-average molecular weight of 20,000 to 60,000, preferably a polyvinyl chloride PVP K30 with a weight-average molecular weight of 35,000 to 50,000.

[0024] Furthermore, the solvent in the carrier or excipient of the above-mentioned ophthalmic preparation is a polar solvent, preferably water.

[0025] Furthermore, the carriers or excipients of the above-mentioned ophthalmic preparations also contain the following components: thickeners and / or solubilizers.

[0026] Furthermore, the tackifier is at least one of polyethylene glycol, carbomer, poloxamer, povidone, hydroxypropyl cellulose, methylcellulose, hydroxyethyl cellulose, polyvinyl alcohol, xanthan gum, polyoxyethylene fatty alcohols, hyaluronic acid and its salts, or hydroxypropyl methylcellulose; the cosolvent is propylene glycol, glycerol, liquid polyethylene glycol, or castor oil; the mass ratio of tackifier to surfactant, or tackifier to low-polymerization-degree povidone is 1:(0.1-100); the mass ratio of cosolvent to surfactant, or cosolvent to low-polymerization-degree povidone is (1-10):1.

[0027] Preferably, the mass ratio of the thickener to the surfactant is 1:(1.2-30), and the mass ratio of the cosolvent to the surfactant is (2.56-9):1;

[0028] Furthermore, the mass ratio of the above-mentioned surfactant or low-polymerization degree povidone to the active ingredient for treating eye diseases is (12-31):1.

[0029] Furthermore, the aforementioned glucocorticoid drugs are at least one of dexamethasone, hydrocortisone, prednisolone, and betamethasone; the aforementioned nonsteroidal drugs are at least one of diclofenac, pranoprofen, indomethacin, and bromfenac sodium.

[0030] Furthermore, the carrier or excipient of the above-mentioned ophthalmic preparation contains nanobody, which is formed by the self-assembly of the components of the carrier or excipient of the ophthalmic preparation; the nanobody encapsulates an active ingredient for treating eye diseases.

[0031] Furthermore, the aforementioned nanoparticles are spherical with a particle size of 1–100 nm; preferably, the particle size of the nanoparticles is 5–30 nm.

[0032] Furthermore, the above formulation also contains nanospheres, which are spherical with a particle size of 10–2000 nm. The nanospheres are formed by the self-assembly of nanobody particles; preferably, the particle size of the nanospheres is 100–2000 nm.

[0033] The present invention also provides a method for preparing the above-mentioned formulation, comprising the following steps:

[0034] (1) Prepare a solution by adding surfactants and / or thickeners to a solvent;

[0035] (2) Disperse the active ingredients for treating eye diseases and / or solubilizers in the solution obtained in step (1), and then add ionic polymers or their solutions to disperse and mix them to obtain a preliminary suspension;

[0036] (3) Stir or homogenize the initial suspension obtained in step (2) to obtain the final product;

[0037] Or may include the following steps:

[0038] (a) Prepare a solution by adding low-polymerization-degree polyvinyl ketone and / or a thickener to a solvent;

[0039] (b) Disperse the active ingredient for treating eye diseases and / or the solubilizer in the solution obtained in step (a), and then add medium-polymerized povidone or its solution, and disperse and mix to obtain a preliminary suspension;

[0040] (c) Grind or homogenize the mixture obtained in step (b) to obtain the final product.

[0041] Further, the dispersion described in step (2) or step (b) is selected from at least one of mechanical stirring dispersion, magnetic stirring dispersion, vortex shaking dispersion, shearing dispersion, homogenization dispersion, grinding dispersion, and ultrasonic dispersion.

[0042] The present invention also provides the use of the above-described formulation in the preparation of a medicament for treating fundus diseases; preferably, the medicament for treating fundus diseases is a medicament for treating macular edema, and / or optic neuritis, and / or non-infectious endophthalmitis.

[0043] Furthermore, the aforementioned medications for treating macular edema are medications for treating macular edema caused by fundus vascular diseases, central retinal vein occlusion macular edema, retinal branch vein occlusion macular edema, diabetic macular edema, pathological myopia macular edema, and / or macular edema caused by wet age-related macular degeneration.

[0044] Experimental results show that the ophthalmic preparation prepared by this invention is stable, easy to store, and can effectively deliver the active ingredients for treating eye diseases to the posterior segment of the eye, achieving an effective (expected) concentration in the fundus, thus treating fundus diseases such as macular edema. It overcomes the problems of existing intravitreal injections, intravitreal implants, oral administration, and systemic injections, and solves serious complications such as intraocular hemorrhage and pain. It greatly reduces the suffering of patients with fundus diseases, increases medical compliance, improves the quality of life of patients and their families, or avoids the systemic toxic side effects of systemic administration.

[0045] This invention can avoid complications caused by local injection or implantation in the eye.

[0046] The formulation developed in this invention has a small dosage and few toxic side effects, and can be used not only as a therapeutic drug, but also as a means of preventing and controlling ophthalmic diseases.

[0047] The formulation of this invention can meet the clinical need for long-term administration.

[0048] The eye drop drug delivery system of the present invention uses small molecule drugs that are already used in clinical practice and have a clear mechanism of action. The quality is controllable, the product is easy to use, the patient compliance is good, and the physician can flexibly adjust the dosing regimen according to the patient's condition.

[0049] The nanobody referred to in this invention is a nanoscale spherical aggregate formed by the self-assembly of components of ophthalmic formulation carriers or excipients in a solvent.

[0050] The nanospheres referred to in this invention are spherical self-assembled structures formed by the self-assembly of nanoparticles in a solvent.

[0051] The solvent referred to in this invention is a liquid capable of dissolving components of ophthalmic formulation carriers or excipients.

[0052] The surfactant referred to in this invention is a substance that can significantly reduce the surface tension of a liquid; the nonionic surfactant referred to in this invention is a surfactant that does not dissociate in water.

[0053] The ionic polymers referred to in this invention are polymers containing cations or anions.

[0054] The low degree of polymerization polyvinyl chloride referred to in this invention is polyvinyl chloride with a molecular weight of less than 10,000 Dalton, while medium degree of polymerization polyvinyl chloride refers to polyvinyl chloride with a molecular weight of more than 10,000 Dalton and less than 100,000 Dalton.

[0055] The "active ingredient for treating eye diseases" referred to in this invention is: an active substance that can be used to treat eye diseases, that is, an active substance that is currently used as an ophthalmic drug, and whose mechanism of action and target indicate that it can treat eye diseases, but is not currently used as an ophthalmic drug (Active Pharmaceutical Ingredient, API).

[0056] The ophthalmic drug delivery method described in this invention is a method of drug delivery by instilling the drug solution into the eye, which belongs to the corneal drug delivery route.

[0057] Obviously, based on the above description of the present invention, and according to common technical knowledge and conventional methods in the field, various other modifications, substitutions or alterations can be made without departing from the basic technical concept of the present invention.

[0058] The following detailed embodiments further illustrate the above-described content of the present invention. However, this should not be construed as limiting the scope of the present invention to the following examples. All technologies implemented based on the above-described content of the present invention fall within the scope of the present invention. Attached Figure Description

[0059] Figure 1 (A) Transmission electron microscopy (TEM) image (scale bar: 200 nm) of the sample prepared in Example 1; (B) TEM image after staining with dye (scale bar: 1 μm).

[0060] Figure 2 The full-thickness retina was measured at approximately 750 μm and 1500 μm to the left and right of the occluded vein at different examination time points in each group of animals. The horizontal axis represents the vertical distance (μm) between the measurement point and the center of the optic disc in the OCT image; the vertical axis represents the full-thickness retina (μm). Data are expressed as mean ± standard deviation (where 2A represents the full-thickness retina of each group before modeling, N=8; 2B represents the full-thickness retina of each group at D2, N=6 for the solvent control group and low-dose group, N=8 for the medium and high-dose groups and aflibercept group, and N=7 for the dexamethasone group; 2C represents the full-thickness retina of each group at D4, N=6 for the solvent control group, N=7 for the test sample dose group, and N=8 for the medium and high-dose groups of the test sample, aflibercept group, and dexamethasone group; compared with the solvent control group at the same time point, *p≤0.05).

[0061] Figure 3 Examples of fundus color photography and angiography.

[0062] Figure 4 Example of optical coherence tomography (retinal full-thickness measurement). Detailed Implementation

[0063] The reagents or instruments used in this invention can be purchased commercially. Unless otherwise specified, they should be used under conventional conditions or conditions recommended by the manufacturer.

[0064] Some of the instruments and equipment are as follows:

[0065] ES225SM-DR(E) electronic analytical balance, Precisa (Switzerland);

[0066] DF-101S Thermal Collector Type Constant Temperature Heating Magnetic Stirrer, Gongyi Yingyu High-Tech Instrument Factory (Henan, China);

[0067] WH-2 Miniature Vortex Mixer, Shanghai Huxi Analytical Instruments Co., Ltd. (Shanghai, China);

[0068] Disperser: T25 easy clean digital, IKA (Germany);

[0069] KQ-500 Ultrasonic Cleaner, Kunshan Ultrasonic Instrument Co., Ltd. (Kunshan, China);

[0070] JP-010T Ultrasonic Cleaner, Shenzhen Jiemeng Cleaning Equipment Co., Ltd.;

[0071] AH-NANO Plus High Pressure Homogenizer, Antos Nanotechnology (Suzhou) Co., Ltd. (China);

[0072] PM-DK2 planetary ball mill, ZhuoDe Instruments (Shanghai) Co., Ltd. (Shanghai, China);

[0073] Mettler Toledo FE20 pH meter, Mettler Toledo (Switzerland);

[0074] NS-90 Nanoparticle Size Analyzer, Zhuhai Omec Instruments Co., Ltd. (Zhuhai, China);

[0075] Agilent 1100HPLC high performance liquid chromatograph, Agilent Technologies (USA);

[0076] API4000 triple quadrupole mass spectrometer (Applied Biosystems, USA);

[0077] STY-1A Osmometer, Tianjin Tianda Tianfa Technology Co., Ltd. (Tianjin, China).

[0078] The method for detecting the properties of the formulation of the present invention is as follows:

[0079] Particle size detection methods:

[0080] Transfer 1 mL of the sample prepared in the examples or comparative examples into the sample cell. Set the detection temperature to 40°C and place the sample cell into the NS-90 nanoparticle size analyzer to start the detection. Each sample is tested three times, and the average of the three test results is taken as the sample's detection result, expressed as particle size (as light intensity distribution and percentage) and polydispersity index (PdI).

[0081] Osmotic pressure testing methods:

[0082] The osmolality of a solution is determined by measuring its freezing point depression. Procedure: Cleaning the STY-1A osmolarity meter probe: Take three 100μL portions of distilled water into three sample tubes. After the instrument has preheated, screw the sample tube containing 100μL of distilled water onto the instrument probe, select "Clean 3 times," and click "Clean." Repeat three times. Testing: Enter the sample information in the instrument information table and click "Test." Use a pipette to transfer 100μL of sample into the sample tube, gently screw it onto the instrument, and click "Start" to begin testing. Repeat the test three times, and take the average of the three results as the final result.

[0083] pH value testing methods:

[0084] The FE20 pH meter was calibrated using pH buffer solutions (pH values ​​of 4.00, 6.86, and 9.18). After rinsing the electrode with pure water and absorbing excess water with lint-free paper, the electrode was immersed in the liquid sample to be tested. The measurement was started by pressing the reading button. The data obtained after the reading stabilized was the pH value of the sample.

[0085] If the pH of the solution obtained from the test is <5 or >9, it needs to be adjusted to pH 6-8 with acid or alkali. Commonly used pH adjusters are NaOH and HCl, phosphoric acid and phosphates (such as sodium dihydrogen phosphate and disodium hydrogen phosphate), citric acid and citrates (such as sodium citrate), boric acid and borax. If the osmotic pressure of the liquid obtained from the test is not isotonic, an appropriate amount of sodium chloride is added to make it reach or approach isotonicity.

[0086] Methods for verifying the effectiveness of drug delivery to the posterior segment of the eye:

[0087] Experimental instruments and equipment: High performance liquid chromatograph, model: LC-20AD (Shimadzu, Japan); Mass spectrometer: model: API4000 triple quadrupole mass spectrometer (Applied Biosystems, USA); Chromatographic column: Fortis Pace C18 5μM, 2.1X30mm (Fortis, UK).

[0088] Healthy adult Sprague Dawley (SD) rats were selected and divided into a treatment group and a control group, with 6 eyes in each group. The treatment group was given the ophthalmic preparation prepared according to the present invention, while the control group was given a suspension of 2 mg drug / 5 mL physiological saline (vortexed and shaken well before use), 20 μL per eye. Animals were euthanized 0.5 hours or 1 hour after administration, and vitreous humor was rapidly collected. The vitreous humor samples were homogenized and stored at -80℃. 10 μL of the vitreous humor homogenate was taken, 90 μL of 95% ethanol was added, and the mixture was sonicated for 2 minutes and vortexed for 1 minute to obtain a vitreous humor homogenate. 50 μL of the homogenate was taken, 175 μL of methanol was added, and the mixture was vortexed for 3 minutes. The mixture was centrifuged at 12000 rpm for 10 minutes at 4℃. The supernatant was filtered through a 0.45 μm syringe filter, and the filtrate was used for LC / MS / MS (positive ion mode, MRM SCAN) analysis.

[0089] Example 1: Preparation of the ophthalmic formulation of the present invention

[0090] According to Table 1, weigh 0.24g of CMC-Na (sodium carboxymethyl cellulose, an ionic polymer) and add it to a glass Erlenmeyer flask containing 40mL of purified water. Stir magnetically for 2 hours to obtain solution 1. Weigh 1.0g of polysorbate 80 (surfactant) and 0.24g of... HPMC (hydroxypropyl methylcellulose, a thickener) was added to a glass Erlenmeyer flask containing 60 mL of purified water. A magnetic stirrer was turned on, and the mixture was heated in a water bath at approximately 40°C for 1.5 hours to obtain solution 2. 40 mg of dexamethasone and 4 mL of PEG400 (4.3 times the amount of surfactant, w / w) were weighed and added to solution 2. Heating and stirring continued for 30 minutes. Solution 1 was then added, and the mixture was stirred for another 30 minutes to obtain a mixed solution. The mixed solution was dispersed in a disperser at 9500 rpm for 5 minutes. After the foam disappeared, the mixture was filtered under reduced pressure using a Buchner funnel to obtain a dispersion. The dispersion was transferred to a high-pressure homogenizer, and homogenized at approximately 400 Bar for 3 minutes while maintaining a temperature of 15±5°C. The pressure was then increased to >800 Bar and homogenized for 25 minutes. The pressure was then reduced to 300 Bar and homogenized for 2 minutes before being discharged to obtain a colorless and clear solution. Further sterilization and removal of mechanical impurities by vacuum filtration yielded a purified colorless and clear solution.

[0091] pH and osmotic pressure adjustment: Add 700 mg NaH2PO4 and 400 mg Na2HPO4 to adjust the pH to pH=6.3; add sodium chloride to adjust the osmotic pressure to 282 mOsmol / kg.

[0092] HPLC detection: Detection instrument: Agilent 1100 high performance liquid chromatograph, operating software: OpenLab CDSc.01.10(201)Chemstation Edition;

[0093] Chromatographic conditions: Waters XBridge C18 5μm, 4.6x250mm column; column temperature 35℃; flow rate 1.0mL / min; detection wavelength 240nm; mobile phase: 0.1% phosphoric acid aqueous solution (72.0%) - acetonitrile (28.0%) isocratic elution. The sample was diluted 5-fold with the mobile phase, and 10μL was injected into the HPLC system. Detection result: 96.2%.

[0094] The particle size was 20.6 nm (85.6%), and the PdI was 0.266. After being stored at room temperature in the dark for one month, the appearance and content of the sample remained unchanged.

[0095] Example 2: Preparation of the ophthalmic formulation of the present invention

[0096] The preparation method is the same as in Example 1, and the raw materials and amounts are shown in Table 1. A colorless and clear solution after impurity removal was obtained.

[0097] pH adjustment: Adjust to pH 6.5 with 0.2N NaOH and / or 0.1N HCl.

[0098] The HPLC detection method was the same as in Example 1. The HPLC content detection results were: 95.1%, particle size 12.9 nm (92.1%), PdI: 0.509. The stability was good. There were no obvious changes in appearance and content after one month of storage at room temperature in the dark. A small amount of precipitation appeared after two months.

[0099] One hour after eye drops, the concentration of API in the vitreous humor of rats was 13.9 ng / g, with an RSD of 17.2%.

[0100] Example 3: Preparation of the ophthalmic formulation of the present invention

[0101] The preparation method and pH and osmotic pressure adjustment method are the same as in Example 1. The raw materials and dosages are shown in Table 1. A clear, slightly yellow solution after impurity removal is obtained.

[0102] HPLC analysis: Column: ZORBAX Eclipse Plus C18, 4.6 x 100 mm, 3.5 μm; Mobile phase A: 0.1% phosphoric acid, Mobile phase B: methanol (80:20) isocratic elution; Temp: 35℃; Detection wavelength: 280 nm; Flowrate: 0.8 ml / min; Result: 98.4%. Particle size 39.7 nm (95.5%), PdI: 0.318; No change in appearance or content after 1 month of storage at room temperature in the dark.

[0103] The concentration of API in the vitreous humor of rats was 78.3 ng / g 0.5 hours after eye drops.

[0104] Example 4: Preparation of the ophthalmic formulation of the present invention

[0105] The preparation method and pH and osmotic pressure adjustment were as described in Example 3, and the raw materials and amounts are shown in Table 1. A clear, slightly yellow solution after impurity removal was obtained.

[0106] The HPLC detection method was the same as in Example 3. The detection results were: 97.8%; particle size: 46.2 nm (95.5%), PdI: 0.343; there was no significant change in appearance and content after being stored at room temperature in the dark for 1 month.

[0107] Example 5: Preparation of the ophthalmic formulation of the present invention

[0108] The preparation method is the same as in Example 1. The raw materials and amounts are shown in Table 1. The amount of co-solvent PEG400 is 5 times (w / w) of the surfactant, and a colorless and clear solution after impurity removal is obtained.

[0109] pH and osmotic pressure adjustment: Adjust the pH to 6.2 with 1M Na2HPO4 solution, and adjust the osmotic pressure to 295 mOsmol / kg with sodium chloride;

[0110] The HPLC detection method was the same as in Example 1. The detection results were: 97.3%; particle size 22.4 nm (91.4%), PdI: 0.293; after being stored at room temperature in the dark for 1 month, there were no significant changes in appearance and content.

[0111] The concentration of API in the vitreous humor of rats was 42.7 ng / g 0.5 hours after eye drops.

[0112] Example 6: Preparation of the ophthalmic formulation of the present invention

[0113] The preparation method is the same as in Example 1. The raw materials and amounts are shown in Table 1. The amount of co-solvent PEG400 is 5 times (w / w) of the surfactant, and a colorless and clear solution after impurity removal is obtained.

[0114] pH 6.5, no adjustment required; osmotic pressure adjustment is as described in Example 1.

[0115] The HPLC detection wavelength was 245 nm, and the method was the same as in Example 1. The detection result was 98.1%.

[0116] Particle size 18.6 nm (96.9%), PdI: 0.257; after being stored at room temperature in the dark for 1 month, there was no significant change in appearance and content.

[0117] The concentration of API in the vitreous humor of rats was 43.8 ng / g 0.5 hours after eye drops.

[0118] Example 7: Preparation of the ophthalmic formulation of the present invention

[0119] The preparation method and pH and osmotic pressure adjustment were as described in Example 1, and the raw materials and amounts are shown in Table 1. A colorless and clear solution after impurity removal was obtained.

[0120] The HPLC detection method was the same as in Example 6, and the detection result was 98.3%.

[0121] Particle size 18.5 nm (97.6%), PdI: 0.208; after being stored at room temperature in the dark for 1 month, there was no significant change in appearance and content.

[0122] Example 8: Preparation of the ophthalmic formulation of the present invention

[0123] The preparation method and pH and osmotic pressure adjustment were as described in Example 1, and the raw materials and amounts are shown in Table 1. A colorless and clear solution after impurity removal was obtained.

[0124] The HPLC detection method was the same as in Example 1, and the detection result was 96.8%.

[0125] Particle size 18.7 nm (89.2%), PdI: 0.255; after being stored at room temperature in the dark for 1 month, there was no significant change in appearance and content.

[0126] Example 9: Preparation of the ophthalmic formulation of the present invention

[0127] The preparation method and pH and osmotic pressure adjustment were as described in Example 1. The raw materials and amounts are shown in Table 1. The amount of co-solvent PEG400 was 1 mL, which is 8 times (w / w) of the surfactant. A colorless and clear solution after impurity removal was obtained.

[0128] The HPLC detection method was the same as in Example 1, and the detection result was 97.2%.

[0129] Particle size 19.6 nm (97.0%), PdI: 0.289; after being stored at room temperature in the dark for 1 month, there was no significant change in appearance and content.

[0130] Comparative Example 1

[0131] The raw materials and dosages are shown in Table 1. Low-polymerization degree polyvinyl ketone (PVP12) was used to replace the surfactant, and hydroxypropyl cellulose (HPC) was used to replace the ionic polymer. The preparation method is the same as in Example 1. 1 mL of PEG400 was used as a co-solvent to obtain a milky white liquid.

[0132] Test results: Particle size 899nm (92.9%), PdI: 0.188, white precipitate formed after standing overnight.

[0133] Comparative Example 2

[0134] The raw materials and dosages are shown in Table 1. Medium-polymerization degree polyvinyl ketone (PVP30) was used to replace the surfactant. The preparation method was the same as in Example 1, and a colorless and clear liquid was obtained.

[0135] Test results: Particle size 241nm (53.1%) and 89.4nm (32.6%), PdI: 1.000, precipitation occurred after 1 week of storage.

[0136] Comparative Example 3

[0137] The raw materials and dosages are shown in Table 1. Medium-polymerization degree polyvinyl ketone (PVP30) was used to replace the surfactant. The preparation method was the same as in Example 1, and a colorless and clear liquid was obtained.

[0138] Test results: Particle size 483nm (47.6%) and 117nm (33.9%), PdI: 1.000, precipitation occurred after 1 week of storage.

[0139] Comparative Example 4

[0140] The raw materials and dosages are shown in Table 1. By changing the dosages of HPMC and CMC-Na, and following the preparation method in Example 1, a white emulsion was obtained.

[0141] Test results: Particle size 26.9 nm (40.5%) and 2043 nm (31.8%), PdI: 1.000, white precipitate formed after standing overnight.

[0142] Comparative Example 5

[0143] The raw materials and dosages are shown in Table 1. The dosages of HPMC and CMC-Na were varied, and the preparation method was the same as in Example 1. 3 ml of PEG400 and 1 ml of ethanol were added as co-solvents, and water was added to 100 mL to obtain a colorless solution. Sodium chloride was added to adjust the osmotic pressure to 262 mOsmol / kg.

[0144] Test results: Particle size 416nm (80%) and 18.1nm (20%), PdI: 0.462, after being left at room temperature for 2 weeks, it turned into a white emulsion.

[0145] Example 10: Preparation of the ophthalmic formulation of the present invention

[0146] The raw materials and dosages are shown in Table 2. 0.52 g of povidone (PVP) K30 was weighed and added to a 100 mL glass Erlenmeyer flask containing 25 mL of purified water. The mixture was magnetically stirred for 2 hours to obtain solution 1. 260 mg of HPMC and 420 mg of povidone (PVP) K12 were weighed and added to a 100 mL glass Erlenmeyer flask containing 25 mL of purified water. The mixture was magnetically stirred and heated in a water bath at 40 °C for 2 hours to obtain solution 2. 1 mL of PEG400 (co-solvent, the amount of which is equivalent to 2.56 times that of PVP K12) and 20 mg of dexamethasone were weighed and added to solution 2. The mixture was heated and stirred for 30 minutes. Solution 1 was then added and stirred for 30 minutes to obtain a mixed solution. A colorless and clear solution after impurity removal was obtained by high-speed dispersion, high-pressure homogenization, and membrane filtration similar to those in Example 1.

[0147] pH and osmotic pressure adjustment: pH 6.5 does not require adjustment. Adjust the osmotic pressure to 293 mmol / kg using sodium chloride.

[0148] The HPLC detection method was the same as in Example 1. The detection results were: 99.2%; particle size 575nm (92.6%), PdI: 0.211. After being stored at room temperature in the dark for 1 month, there were no obvious changes in appearance and content; after 2 months, a precipitate was formed.

[0149] The concentration of API in the vitreous humor of rats was 5.1 ng / g 0.5 hours after eye drops.

[0150] Example 11: Preparation of the ophthalmic formulation of the present invention

[0151] The raw materials and dosages are shown in Table 2. The amount of co-solvent PEG400 is 8.9 times (w / w) of PVP K12. The preparation method and pH and osmotic pressure adjustment methods are the same as in Example 10. A colorless and clear solution after impurity removal was obtained.

[0152] The HPLC detection method was the same as in Example 1, and the detection result was 98.5%.

[0153] Particle sizes of 125.6 nm (63.5%) and 13.6 nm (33.1%), PdI: 0.255; after being stored at room temperature in the dark for one month, there were no significant changes in appearance and content.

[0154] Comparative Example 6

[0155] The raw materials and dosages are shown in Table 2. The preparation method is the same as in Example 10. Ionic polymers are added to obtain a light white emulsion.

[0156] Test results: Particle size 1299nm (92.4%), PdI: 0.175, white precipitate formed after standing overnight.

[0157] Comparative Example 7

[0158] The raw materials and dosages are shown in Table 2. The preparation method is the same as in Example 10. 1 mL of PEG400 was added as a co-solvent (the amount of which is 20 times (w / w) of the surfactant) to obtain a colorless solution.

[0159] Test results: Particle size 637nm (85.9%), PdI: 0.258, white precipitate formed after standing overnight.

[0160] Comparative Example 8

[0161] The raw materials and dosages are shown in Table 2. The preparation method is the same as in Example 10, without povidone, with the addition of ionic polymers to obtain a colorless solution.

[0162] Test results: pH 5.2, particle size 46.2nm (96.9%), PdI: 0.343, HPLC content test result: 98.9%, fine crystals precipitated after refrigeration for 2 weeks.

[0163] The results of measuring the concentration of API in the vitreous body of rats after eye drop administration demonstrate that the ophthalmic formulation of the present invention can carry the active ingredient for treating eye diseases across the barrier of the eye structure and deliver an effective dose of drug to the vitreous body through conjunctival sac administration (eye drop administration), avoiding invasive administration methods such as intravitreal injection, and also significantly reducing the total amount of drug, reducing the absorption of the drug in the system, and avoiding toxic side effects.

[0164] Table 1

[0165]

[0166]

[0167] Table 2

[0168]

[0169]

[0170] The following experimental examples demonstrate the beneficial effects of the ophthalmic drug formulation of the present invention.

[0171] Experimental Example 1: Transmission Electron Microscopy Observation Results of the Carrier of the Invention

[0172] Transmission electron microscope (JEM-2100Plus, JEOL Ltd., Japan)

[0173] One drop of the liquid sample prepared in Example 1 was placed on a copper sample grid. After standing for 5 minutes, excess liquid sample was aspirated, and the sample was allowed to air dry. The grid was then placed in the electron microscope sample chamber for analysis. For sample staining: One drop of the liquid sample was placed on a copper sample grid. After removing excess sample from the grid, one drop of 2% phosphomolybdic acid was added. After standing for 5 minutes, excess liquid was aspirated, and the sample was allowed to air dry. The grid was then placed in the electron microscope for analysis. Results are shown below. Figure 1 It can be seen that the drug-loaded carrier prepared by this invention forms spherical structures (nanosomes) with a particle size of 1-100 nm in the solvent. Figure 1 A) Nanoparticles can further self-assemble into spheres with a particle size of 10–2000 nm (nanospheres). Figure 1 B).

[0174] Experimental Example 2: Particle Size, Content, and Stability Detection

[0175] 1. Experimental Methods

[0176] Transfer 1 mL of the samples prepared in the examples and comparative examples into the sample cell. Set the detection temperature to 40°C and place the sample cell into the NS-90 nanoparticle size analyzer to begin detection. Each sample was tested three times, and the average of the three results was taken as the sample's particle size (expressed as light intensity distribution, percentage) and polydispersity index (PdI). After testing, the samples were stored in the dark, and changes in appearance were observed, followed by re-detection of particle size.

[0177] The HPLC content of the ophthalmic formulation samples prepared in this invention was determined using an Agilent 1100 high-performance liquid chromatograph.

[0178] 2. Experimental Results

[0179] See Table 3:

[0180] Table 3

[0181]

[0182]

[0183]

[0184] The above results show that the pharmaceutical preparation prepared by this invention has a small particle size, high content of active ingredients as detected by HPLC, and stable morphology and content over a long period of time; indicating that the preparation of this invention has a high encapsulation efficiency and good stability. In contrast, the preparations prepared using comparative proportions with different excipients than those of this invention exhibit very poor stability, showing precipitation or deterioration within a short period.

[0185] Experimental Example 3: Verification of the efficacy of the ophthalmic formulation of the present invention in treating macular edema via eye drops.

[0186] 1. Experimental Methods

[0187] Forty-eight SD rats with no obvious abnormalities in both eyes were randomly divided into 6 groups (n=8 per group, half male and half female). Immediately after a tail vein injection of the photosensitizer Bengal Red (40 mg / mL, 40 mg / kg) on ​​day 1, laser photocoagulation of a single retinal vein in the right eye was performed. Approximately 15 minutes after photocoagulation, the occlusion of the target retinal vein was confirmed. After successful occlusion confirmation, drug treatment was administered according to Table 4. The test sample was used in the high-dose group (prepared in Example 1), the medium-dose group (prepared in Example 1 diluted 3 times with medical saline), and the low-dose group (prepared in Example 1 diluted 9 times with medical saline). Aflibercept intraocular injection solution (Bayer, Germany) and dexamethasone sodium phosphate injection (Sinopharm Rong Sheng Pharmaceutical Co., Ltd., China) were used as control drugs. Fundus photography (FP), fundus fluorescein angiography (FFA), and optical coherence tomography (OCT) were performed on the right eye of all animals on D-1 (before modeling), D1 (immediately after modeling), D2 (day 2 after modeling), and D4 (day 4 after modeling). FP and FFA images were analyzed, and the full-thickness retinal thickness was measured at approximately 750 and 1500 μm to the left and right of the obstructed vein on the OCT scan images. The left eye of all animals was not subjected to modeling, drug administration, or corresponding ophthalmic examinations.

[0188] Table 4 Animal grouping and drug administration table

[0189]

[0190]

[0191] 2. Experimental Results

[0192] like Figure 2 As shown, the full-thickness retinal layer increased significantly after modeling, while a significant decrease in full-thickness retinal layer was observed on days 2 and 4 after administering the formulation of the present invention via eye drops. Moreover, its effect was comparable to that of commercially available drugs administered via intravitreal injection, indicating that the formulation of the present invention can achieve a very good effect in improving retinal edema through eye drop administration.

[0193] like Figure 3 As shown, after modeling (D1), compared with before modeling, obvious venous thrombosis and blood stasis were observed at the laser photocoagulation site; while after administering the formulation of this invention by eye drops, although there was still some retinal vein occlusion, the venous thrombosis was significantly improved.

[0194] like Figure 4 As shown, after administering the formulation of this invention via eye drops, acute retinal edema and thickening occurred on day 2, but the edema began to gradually recover on day 4, approaching the normal state before modeling.

[0195] The above results indicate that repeated ocular instillation of the test substance (20 μL / eye, for 4 consecutive days, 4 times / day for the first 3 days, and a single instillation on the 4th day) at medium dose (0.133 mg / mL) and high dose (0.4 mg / mL) significantly inhibited / treated acute retinal edema in rats with retinal vein occlusion induced by laser photocoagulation combined with photosensitizer, and the effect was dose-dependent. The efficacy of the high-dose group was close to that of the commercially available control product, aflibercept intravitreal injection. In this study, a single intravitreal injection of the commercially available control product, dexamethasone sodium phosphate injection, did not show a clear inhibitory / treatment effect on acute retinal edema.

[0196] In summary, this invention provides an ophthalmic drug delivery formulation that, through ophthalmic administration, carries (encapsulates) glucocorticoids and / or nonsteroidal anti-inflammatory drugs through the anterior segment of the eye and delivers them to the posterior segment to exert a therapeutic effect. This achieves the goal of treating fundus diseases such as macular edema through ophthalmic drug delivery, solves a long-standing technical problem in the field, and has extremely high clinical value and significant social implications.

Claims

1. An ophthalmic preparation for eye drop administration, characterized in that, It is a formulation composed of active ingredients for treating eye diseases and ophthalmic formulation carriers or excipients; The active ingredient used to treat eye diseases is dexamethasone; The ophthalmic formulation carrier or excipient is composed of the following components: low degree of polymerization povidone, medium degree of polymerization povidone, thickener, co-solvent and solvent; The mass ratio of the low-polymerization degree povidone to the active ingredient for treating eye diseases is (12~31):1; The mass ratio of low-polymerization degree povidone to medium-polymerization degree povidone in the carrier or excipient of the ophthalmic preparation is (0.24~0.8):1, and the ratio of low-polymerization degree povidone to solvent is 240~840mg of low-polymerization degree povidone per 100mL of solvent. The mass ratio of the tackifier to the low-polymerization-degree polyvinyl ketone is 1:(1.2~30), and the mass ratio of the cosolvent to the low-polymerization-degree polyvinyl ketone is (2.56~9):1; The thickener is hydroxypropyl methylcellulose, and the cosolvent is liquid polyethylene glycol; The low-polymerization-degree polyvinyl ketone is polyvinyl ketone PVP K12; the medium-polymerization-degree polyvinyl ketone is polyvinyl ketone PVP K30; The solvent in the carrier or excipient of the ophthalmic preparation is water.

2. An ophthalmic preparation for eye drop administration, characterized in that, It is a formulation composed of active ingredients for treating eye diseases and ophthalmic formulation carriers or excipients; The active ingredient used to treat eye diseases is prednisolone; The ophthalmic formulation carrier or excipient is composed of the following components: low degree of polymerization povidone, medium degree of polymerization povidone, thickener, co-solvent and solvent; The mass ratio of the low-polymerization degree povidone to the active ingredient for treating eye diseases is 6:1; The mass ratio of low-polymerization degree povidone to medium-polymerization degree povidone in the carrier or excipient of the ophthalmic formulation is 6:25, and the ratio of low-polymerization degree povidone to solvent is 240 mg of low-polymerization degree povidone per 100 mL of solvent. The mass ratio of the tackifier to the low-polymerization-degree polyvinyl ketone is 1:1.2, and the mass ratio of the cosolvent to the low-polymerization-degree polyvinyl ketone is 8.9:

1. The thickener is hydroxypropyl methylcellulose, and the cosolvent is liquid polyethylene glycol; The low-polymerization-degree polyvinyl ketone is polyvinyl ketone PVP K12; the medium-polymerization-degree polyvinyl ketone is polyvinyl ketone PVP K30; The solvent in the carrier or excipient of the ophthalmic preparation is water.

3. The formulation according to claim 1 or 2, characterized in that, The carrier or excipient of the ophthalmic preparation contains nanobody, which is formed by the self-assembly of the components of the carrier or excipient of the ophthalmic preparation; the nanobody encapsulates an active ingredient for treating eye diseases.

4. The formulation according to claim 3, characterized in that: The nanoparticles are spherical with a particle size of 1~100nm.

5. The formulation according to claim 4, characterized in that: The nanoparticles have a particle size of 5~30nm.

6. The formulation according to claim 3, characterized in that: The formulation contains nanospheres, which are spherical with a particle size of 10-2000 nm; the nanospheres are formed by the self-assembly of nanobody.

7. The formulation according to claim 6, characterized in that: The nanospheres have a particle size of 100~2000nm.

8. A method for preparing the formulation according to any one of claims 1 to 7, characterized in that, Includes the following steps: (a) Prepare a solution by adding low-polymerization-degree polyvinyl ketone and a thickener to a solvent; (b) Disperse the active ingredient for treating eye diseases and the solubilizer in the solution obtained in step (a), and then add medium-polymerized povidone or its solution to disperse and mix to obtain a preliminary suspension; (c) Grind or homogenize the mixture obtained in step (b) to obtain the final product.

9. The method according to claim 8, characterized in that: The dispersion described in step (b) is selected from at least one of mechanical stirring dispersion, magnetic stirring dispersion, vortex shaking dispersion, shearing dispersion, homogenization dispersion, grinding dispersion, and ultrasonic dispersion.

10. The use of the formulation according to any one of claims 1 to 7 in the preparation of a medicament for treating fundus diseases, characterized in that, The treatment of fundus diseases includes macular edema, and / or optic neuritis, and / or non-infectious endophthalmitis.

11. The use according to claim 10, characterized in that, The macular edema referred to includes macular edema caused by fundus vascular diseases, central retinal vein occlusion macular edema, retinal branch vein occlusion macular edema, diabetic macular edema, pathological myopia macular edema, and / or macular edema caused by wet age-related macular degeneration.

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