Eye drops containing Syk and VEGFR2 double-target inhibitor and application thereof

Abstract

The invention belongs to the field of pharmaceutical compositions, and particularly relates to eye drops containing Syk and VEGFR2 double-target inhibitors and application of the eye drops. The eye drops contain active ingredients, buffer salt, an osmotic pressure regulator and hydroxypropyl methyl cellulose, and the pH value of the eye drops is 5.9-7.6. The eye drops not only can effectively play a drug role, but also can reduce irritation to eyes, and have good drug effect and administration compliance in the aspect of treating xerophthalmia. Meanwhile, the eye drops still have good stability under the conditions of high temperature and illumination, and the effectiveness and safety of the medicine in the storage and use process are improved.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of pharmaceutical compositions, and particularly relates to an eye drop containing a Syk and VEGFR2 dual-target inhibitor and application thereof. BACKGROUND

[0002] The global prevalence of dry eye is high, and moderate to severe dry eye requires anti-inflammatory treatment. Current treatment drugs include glucocorticoids, non-steroidal anti-inflammatory drugs and immunosuppressants. Current dry eye drugs at home and abroad mainly include artificial tears, glucocorticoid drugs and cyclosporine. The above drugs can only relieve dry eye symptoms and have a slow onset, and have certain side effects, which cannot meet the huge demand for dry eye drugs.

[0003] Therefore, in the face of a large patient population with dry eye disease, it is urgent to develop new drug mechanisms, faster-acting and more effective dry eye products.

[0004] In WO2021169958A1, a Syk and VEGFR2 dual-target inhibitor is disclosed, and a compound having the following formula (III) or a pharmaceutically acceptable salt thereof is disclosed. In WO2022166548A1, the salt type, crystal form and preparation method of the pyrazole-substituted imidazo[1,2-a]quinoxaline derivative are further disclosed, and its application in the preparation of Syk and VEGFR2 dual inhibitors related drugs is disclosed, and a crystal form having the compound of formula (I) is disclosed.

[0005]

[0006]

[0007] The above-mentioned molecule exhibits excellent target specificity, and exhibits significant therapeutic and anti-inflammatory effects in animal dry eye models, and has less biological toxicity, and exhibits excellent drug application prospects. However, due to the strong stability of the molecular structure, the solubility of the drug in aqueous solution is poor, which increases the difficulty of forming an effective drug preparation. Therefore, it is urgent to develop a preparation formula that can increase the solubility of the above-mentioned drug or can make it play a therapeutic effect of the drug. SUMMARY

[0008] The present application first provides an eye drop containing: i) an active ingredient, the active ingredient being a compound represented by formula (I) or a pharmaceutically acceptable salt thereof; ii) a buffer salt; iii) an osmotic pressure regulator; and iv) hydroxypropyl methyl cellulose (HPMC); and the pH value of the eye drop is 5.9-7.6.

[0009]

[0010] The application further provides the eye drops in the prevention, treatment or alleviation of dry eye.

[0011] The application further provides the use of the eye drops in the preparation of a medicine for the prevention, treatment or alleviation of dry eye.

[0012] The application further provides a kit comprising a therapeutically effective amount of the eye drops, and instructional material relating to the administration of the eye drops to a patient suffering from dry eye.

[0013] The application provides eye drops containing a Syk and VEGFR2 dual-target inhibitor, which can effectively exert a drug effect and reduce irritation to the eye, and has good drug efficacy and drug compliance in the treatment of dry eye. Meanwhile, the eye drops have good stability under high temperature and light conditions, which is conducive to improving the effectiveness and safety of the drug during storage and use. BRIEF DESCRIPTION OF DRAWINGS

[0014] In order to more clearly illustrate the specific embodiments of the application or the technical solutions in the prior art, the drawings needed in the specific embodiments or prior art description will be briefly introduced below. Obviously, the drawings in the following description are some embodiments of the application, and those skilled in the art can obtain other drawings according to these drawings without creative labor.

[0015] Figure 1 The results of HE staining of the right eye tissue of model group A♂01 in the test example of the application (100x).

[0016] Figure 2 The results of HE staining of the right eye tissue of eye drop group C♀03 in the test example of the application (100x). DETAILED DESCRIPTION

[0017] The specific embodiments of the application will be described in detail below. It should be understood that the specific embodiments described herein are only used to illustrate and explain the application, and are not used to limit the application, and those skilled in the art can make various modifications and changes to the application without departing from the scope or spirit of the application. For example, features described or illustrated as part of one embodiment can be used in another embodiment to produce further embodiments.

[0018] Terminology

[0019] Unless otherwise defined, all terms (including technical and scientific terms) used in the disclosure of the present application have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. By further guidance, the following definitions are used to better define the present teachings. The terms used in the specification of the present application herein are only for the purpose of describing specific embodiments and are not intended to limit the present application.

[0020] The selection scope of the terms "and / or", "or / and", "and / or" used herein includes any one of two or more related listed items, and also includes any and all combinations of the related listed items, including any two related listed items, any more related listed items, or all related listed items. It should be noted that when at least two conjunctions selected from "and / or", "or / and", "and / or" are combined to connect at least three items, it should be understood that in the present application, the technical solution undoubtedly includes the technical solution connected by "logical and", and also undoubtedly includes the technical solution connected by "logical or". For example, "A and / or B" includes three parallel solutions of A, B and A+B. For another example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C and D (i.e. the technical solution connected by "logical or"), and also includes any and all combinations of A, B, C and D, i.e. includes the combination of any two or any three of A, B, C and D, and also includes the four-item combination of A, B, C and D (i.e. the technical solution connected by "logical and").

[0021] The terms "containing", "including" and "comprising" used in the present application are synonymous and are inclusive or open-ended and do not exclude additional, unrecited members, elements or method steps.

[0022] The numerical ranges used in the present application expressed in endpoints include all values and fractions within that range as well as the recited endpoints.

[0023] In the present application, the concentration values involved include fluctuations within a certain range. For example, it can fluctuate within a corresponding accuracy range. For example, 2% can be allowed to fluctuate within ±0.1%. For larger values or values that do not need to be controlled too finely, it is also allowed that the meaning includes larger fluctuations. For example, 100mM can be allowed to fluctuate within ±1%, ±2%, ±5%, etc. For molecular weight, it is allowed that the meaning includes a fluctuation of ±10%.

[0024] In the present application, the descriptions such as "a plurality of", "a plurality of" and the like are used without special limitation, which means greater than or equal to 2 in quantity.

[0025] In the present application, the technical features described in an open way include both the closed technical solution consisting of the listed features and the open technical solution containing the listed features.

[0026] In the present application, "preferably", "more preferably", "more preferably", "preferably" only describes the better effect of the implementation or embodiment, and it should be understood that it does not constitute a limitation on the protection scope of the present application.

[0027] In the present application, "optionally", "optional", "optional", "optionally", "optional", "optional" means optional, that is, selected from any one of the two parallel schemes of "yes" or "no". If there are multiple "options" or "options" in a technical solution, unless otherwise specified, and there is no contradiction or mutual restriction, each "option" or "option" is independent.

[0028] In the present application, the term "dry eye" is also known as keratoconjunctival xerosis, which is a chronic ocular surface disease caused by multiple factors, mainly involving the quality, quantity and dynamics of tear fluid. These abnormalities lead to unstable tear film or imbalance of ocular surface microenvironment, and may be accompanied by ocular surface inflammatory response, tissue damage and nerve abnormalities, thereby causing various uncomfortable symptoms and / or visual dysfunction.

[0029] Eye drops

[0030] The compound represented by formula (I) or its pharmaceutically acceptable salt is a small molecule drug targeting tyrosine protein kinase. The compound or its pharmaceutically acceptable salt can inhibit the protein phosphorylation of spleen tyrosine kinase (Syk) / vascular endothelial growth factor receptor 2 (VEGFR-2), thereby inhibiting the release of various inflammatory cytokines and immune cell-mediated signal transduction, and blocking the inflammatory cascade reaction caused by immune response. The main preclinical characteristics of the compound or its pharmaceutically acceptable salt are more significant target activity and better selectivity. At a well-tolerated dose, the drug efficacy and inflammation improvement are both obvious. The eye drop containing the compound or its pharmaceutically acceptable salt as an active ingredient has a large safety window, good animal tolerance, and no obvious local irritation, which supports its development for clinical treatment of dry eye.

[0031] However, the high-temperature stability and light stability of the compound in water are not good. In this regard, the present application further explores and first provides an eye drop containing: i) an active ingredient, the active ingredient being a compound represented by formula (I) or a pharmaceutically acceptable salt thereof; ii) a phosphate buffer salt; iii) sodium chloride; and iv) hydroxypropyl methyl cellulose (HPMC); and the pH value of the eye drop is 5.9-7.6.

[0032]

[0033] The present application finds that the above eye drop formula can better improve the solubility of the active ingredient, and improve its high temperature stability and light stability, and is beneficial to reducing the irritation to the eye, and has good drug efficacy and drug compliance in the treatment of dry eye.

[0034] In specific implementation, the active ingredient mentioned in the present application can be a compound represented by formula (I), or a pharmaceutically acceptable salt thereof (such as a hydrochloride, or a hydrate, a maleate or a gentisate disclosed in WO2022166548A1, etc.), and the compound mentioned in the present application or the pharmaceutically acceptable salt thereof also includes different crystal forms thereof (such as A crystal form, B crystal form, C crystal form, D crystal form, E crystal form disclosed in WO2022166548A1, etc.).

[0035] In some embodiments, the active ingredient is a hydrochloride of the compound represented by formula (I). By preparing it as a hydrochloride, the solubility of the active ingredient can be further increased without affecting the drug efficacy, thereby improving the bioavailability of the drug. Moreover, when applied in the eye drop in the form of a hydrochloride, the stability and drug compliance of the drug can be further improved. In addition, the synthesis and preparation of this salt type are also more simple.

[0036] In some embodiments, the active ingredient has a structure represented by formula (II) as follows:

[0037]

[0038] In formula (II), p is selected from 0.7 to 2.3.

[0039] As an example, in some specific embodiments, p is selected from 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3 or any value within the range of 0.7 to 2.3.

[0040] In some specific embodiments, p is 0.8, 1.0 or 1.7.

[0041] In some embodiments, the content of the active ingredient is 0.1 w / v% to 2.0 w / v%, more preferably 0.3 w / v% to 1.0 w / v%.

[0042] As an example, in some specific embodiments, the content of the active ingredient in the eye drop can be 0.1 w / v%, 0.3 w / v%, 0.5 w / v%, 0.7 w / v%, 1.0 w / v%, 1.5 w / v%, 2.0 w / v%, or any value within the range of 0.1 w / v% to 2.0 w / v%.

[0043] In some embodiments, the buffer salt is selected from one or more of phosphate buffer salt, citrate buffer salt, acetate buffer salt, borate buffer salt, more preferably phosphate buffer salt.

[0044] In some embodiments, the osmotic pressure regulator is selected from one or more of sodium chloride, mannitol, glucose, xylitol, phosphate, citrate, acetate, borate, more preferably sodium chloride.

[0045] In some preferred embodiments, the buffer salt is a mixture of sodium phosphate monobasic and sodium phosphate dibasic, the content of sodium phosphate monobasic is 0.4w / v% to 0.5w / v%, the content of sodium phosphate dibasic is 0.4w / v% to 0.5w / v%; the osmotic pressure regulator is sodium chloride, the content of sodium chloride is 0.20w / v% to 0.30w / v%. In this way, the stability of the eye drops is better, and the osmotic pressure is more appropriate, which can further avoid the irritation to the eye.

[0046] For example, in some specific embodiments, the content of sodium phosphate monobasic is 0.40w / v%, 0.41w / v%, 0.42w / v%, 0.43w / v%, 0.44w / v%, 0.45w / v%, 0.46w / v%, 0.47w / v%, 0.48w / v%, 0.49w / v%, 0.50w / v% or any value within the range of 0.4w / v% to 0.5w / v%.

[0047] For example, in some specific embodiments, the content of sodium phosphate dibasic is 0.40w / v%, 0.41w / v%, 0.42w / v%, 0.43w / v%, 0.44w / v%, 0.45w / v%, 0.46w / v%, 0.47w / v%, 0.48w / v%, 0.49w / v%, 0.50w / v% or any value within the range of 0.4w / v% to 0.5w / v%.

[0048] For example, in some specific embodiments, the content of sodium chloride is 0.20w / v%, 0.21w / v%, 0.22w / v%, 0.23w / v%, 0.24w / v%, 0.25w / v%, 0.26w / v%, 0.27w / v%, 0.28w / v%, 0.29w / v%, 0.30w / v% or any value within the range of 0.20w / v% to 0.30w / v%.

[0049] In some embodiments, the amount of hydroxypropyl methyl cellulose is 0.8w / v% to 1.2w / v%. In this way, the viscosity of the obtained eye drops is appropriate, which helps to ensure the appropriate residence time of the drug in the eye while reducing the irritation to the eye.

[0050] As an example, in some specific embodiments, the amount of hydroxypropyl methylcellulose is 0.8 w / v %, 0.9 w / v %, 1.0 w / v %, 1.1 w / v %, 1.2 w / v % or any value within the range of 0.8 w / v % to 1.2 w / v %.

[0051] In some embodiments, the pH of the eye drops is 6.5 to 7.0.

[0052] As an example, in some specific embodiments, the pH of the eye drops is 5.9, 6.0, 6.3, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.2, 7.5, 7.6 or any value within the range of 5.9 to 7.6.

[0053] In some embodiments, the eye drops further comprise a pH adjusting agent selected from one or more of hydrochloric acid, sulfuric acid, citric acid, sodium hydroxide, potassium hydroxide, sodium bicarbonate, disodium hydrogen phosphate, sodium dihydrogen phosphate.

[0054] In some specific embodiments, the pH adjusting agent is selected from one or both of hydrochloric acid and sodium hydroxide.

[0055] In some embodiments, the water in the eye drops is selected from one or more of Purified Water, Water for Injection, Deionized Water, Sterile Water, Distilled Water, Reverse Osmosis Water.

[0056] In a preferred embodiment, the eye drops comprise: i) 0.1 w / v % to 2.0 w / v % of an active ingredient, which is hydrochloride salt of the compound of formula (I); ii) 0.4 w / v % to 0.5 w / v % of sodium dihydrogen phosphate and 0.4 w / v % to 0.5 w / v % of disodium hydrogen phosphate; iii) 0.20 w / v % to 0.30 w / v % of sodium chloride; iv) 0.8 w / v % to 1.2 w / v % of hydroxypropyl methylcellulose; v) a pH adjusting agent for adjusting the pH of the eye drops to 5.9 to 7.6; and vi) water.

[0057] In some embodiments, the osmolality of the eye drops is 290 mmol·L -1 to 340 mmol·L -1Thus, the eye drops can reduce irritation to the eye while maintaining the drug efficacy. In specific embodiments, the osmotic pressure of the eye drops can be adjusted by adjusting the amount of buffer salt (e.g., phosphate buffer salt) and osmotic pressure regulator (e.g., sodium chloride) in the eye drop formulation.

[0058] For example, in some embodiments, the osmotic pressure of the eye drops is in the range of any value between 290 mmol·L -1 , 295 mmol·L -1 , 300 mmol·L -1 , 305 mmol·L -1 , 310 mmol·L -1 , 315 mmol·L -1 , 320 mmol·L -1 , 325 mmol·L -1 , 330 mmol·L -1 , 335 mmol·L -1 , 340 mmol·L -1 , or 290 mmol·L -1 to 340 mmol·L -1 .

[0059] In some embodiments, the viscosity of the eye drops is in the range of 6 mPa·s to 10 mPa·s. The retention time of the active ingredient in the eye is appropriate in this range, and the bioavailability is better. In specific embodiments, the viscosity of the eye drops can be adjusted by adjusting the amount of hydroxypropyl methyl cellulose in the eye drop formulation.

[0060] In some embodiments, the eye drops can further contain one or more of the following agents: 1) chelating agent: such as disodium edetate, which is used to remove metal ions in the solution that can affect the stability of the drug; 2) antioxidant: such as ascorbic acid, erythorbic acid, etc., which is used to prevent the deterioration of the preparation due to oxidation caused by the presence of oxygen free radicals or free metals in the composition.

[0061] Those skilled in the art can combine the above-mentioned embodiments according to common sense to obtain more examples of the eye drops according to the present application.

[0062] Based on the above-mentioned eye drop formulation and general common sense in the art, those skilled in the art can confirm the preparation method of the eye drops.

[0063] In one embodiment, the method of preparing the eye drops comprises: taking purified water in an amount of 70-85% of the formulation, adding hydroxypropyl methylcellulose, phosphate and sodium chloride, stirring and dissolving, then adding the active ingredient and stirring and dissolving, and then adjusting the pH to 5.9-7.6 with a pH adjuster, making up the volume with water, and measuring the pH to obtain the eye drops.

[0064] In some embodiments, the method of preparing the eye drops further comprises the steps of sterilizing and / or filling the eye drops.

[0065] Applications

[0066] The present application further provides the use of the eye drops in preventing, treating or alleviating dry eye. In other words, the present application also provides a method for preventing, treating or alleviating dry eye, which comprises administering to a subject a therapeutically effective amount of the eye drops.

[0067] The present application further provides the use of the eye drops in the preparation of a medicament for preventing, treating or alleviating dry eye.

[0068] In some embodiments, the eye drops are used for at least one of the following: 1) prolonging tear film break-up time; 2) increasing tear secretion; 3) reducing the degree of conjunctival cell infiltration; and 4) promoting the repair of goblet cells.

[0069] In some embodiments, the eye drops are suitable for topical administration to the eye of a subject.

[0070] In the present application, "topical administration" means administration to the surface of a tissue such as the eye, in particular any external aspect of the eye that is generally accessible between the eyelids. Topical administration to the eye can generally be by means of eye drops, ointments or sprays.

[0071] In some embodiments, the eye drops can be present in a rigid and / or squeeze-type bottle equipped with a fitting cap configured to serve as a dropper. A human subject can receive between 1 and 10 drops per day (for example, 5 or 8 drops per day), and the dose can be repeated, for example, twice a day. The eye drops can be dispensed, for example, in a volume of 12 mL per bottle or in a volume of 20 mL per bottle.

[0072] In some embodiments, the eye drops can also be administered by a carrier vehicle such as liquid drops, liquid lotions, gels, ointments and sprays, or combinations thereof.

[0073] In some embodiments, the topical administration can also occur by injection of the eye drop via a device such as a pump-catheter system / continuous or selective release device, a contact lens, or a combination thereof. The eye drop can also be administered in an injectable form, e.g., such that the eye drop is injected into the posterior of the eye and / or wherein administration involves intravitreal injection.

[0074] In the present application, the term "subject" refers to a mammal, such as a human, a domestic animal (such as a feline or canine), a farm animal (e.g., bovine, equine, caprine, ovine, and porcine subjects), a wild animal, or a research animal (e.g., mice, rats, rabbits, goats, sheep, pigs, dogs and cats, avian species such as chickens, turkeys, and songbirds).

[0075] In some embodiments, the subject is a human.

[0076] The present application also provides a kit comprising a therapeutically effective amount of the eye drop described, and instructional material relating to the administration of the eye drop to a patient suffering from dry eye.

[0077] In some embodiments, the instructional material can include a publication, a chart, or any other medium that can be used to convey the usefulness of the composition and its administration. In some embodiments, the instructional material can be attached to the container that houses the eye drop of the present application or can be otherwise provided with the container that houses the composition. In some embodiments, the instructional material can provide the instructional material separately, e.g., by electronic transmission, e.g., by means of a computer (such as by email, or download from a website).

[0078] In some embodiments, the kit further comprises at least one additional agent useful for preventing, treating, or ameliorating dry eye.

[0079] Examples

[0080] Embodiments of the present application will be described in detail with reference to the following examples. It is understood that these examples are intended to be illustrative only and are not intended to limit the scope of the present application. Unless otherwise indicated, the experimental methods in the following examples follow the guidelines set forth in the present application, and can also follow the protocols or routine conditions in the art, or other experimental methods known in the art, or as suggested by the manufacturer.

[0081] In the following specific examples, the amount of the raw material components is described in terms of measurement parameters. Unless otherwise specified, minor variations within the range of weighing accuracy are possible. In terms of temperature and time parameters, acceptable variations are allowed due to instrument testing accuracy or operational accuracy. In terms of pH, minor variations due to testing accuracy (e.g. within ±0.10) are allowed.

[0082] For the purpose of comparison, the following examples and comparative examples use the same amount of one active ingredient, which has the following structure, wherein p is 1.0. The active ingredient and its amount do not limit the scope of protection of the present application, and active ingredients within the scope of the present application have the technical effects described above when applied to the eye drops.

[0083]

[0084] Example 1

[0085] This example first provides an eye drop containing 0.5 w / v% of an active ingredient, 0.43 w / v% of sodium dihydrogen phosphate, 0.46 w / v% of disodium hydrogen phosphate, 0.25 w / v% of sodium chloride, and 1.0 w / v% of hydroxypropyl methyl cellulose, and the pH of the eye drop is 5.95 by means of hydrochloric acid and sodium hydroxide, and the balance is water for injection.

[0086] This example further provides a preparation method of the above eye drop, which is specifically as follows: take 80% of the prescription amount of water for injection, add hydroxypropyl methyl cellulose, sodium dihydrogen phosphate, disodium hydrogen phosphate, and sodium chloride, stir and dissolve, add the active ingredient, stir and dissolve, adjust the pH to the specified value with 1 mol / L sodium hydroxide solution or hydrochloric acid solution, dilute to volume, measure the pH, sterilize at 121°C for 15 minutes, and under stirring, dispense into low-density polyethylene pharmaceutical eye drop bottles, 5 mL per bottle.

[0087] Example 2

[0088] This example provides an eye drop containing 0.5 w / v% of an active ingredient, 0.43 w / v% of sodium dihydrogen phosphate, 0.46 w / v% of disodium hydrogen phosphate, 0.25 w / v% of sodium chloride, and 1.0 w / v% of hydroxypropyl methyl cellulose, and the pH of the eye drop is 6.77 by means of hydrochloric acid and sodium hydroxide, and the balance is water for injection.

[0089] The preparation method is the same as that of Example 1.

[0090] Example 3

[0091] The present example provides an eye drop containing 0.5 w / v% of an active ingredient, 0.43 w / v% of sodium dihydrogen phosphate, 0.46 w / v% of disodium hydrogen phosphate, 0.25 w / v% of sodium chloride, and 1.0 w / v% of hydroxypropyl methylcellulose, and the pH of the eye drop is 7.59 by hydrochloric acid and sodium hydroxide, with the balance being water for injection.

[0092] The preparation method is the same as that of Example 1.

[0093] Example 4

[0094] The present example provides an eye drop containing 0.5 w / v% of an active ingredient, 0.43 w / v% of sodium dihydrogen phosphate, 0.46 w / v% of disodium hydrogen phosphate, 0.20 w / v% of sodium chloride, and 1.0 w / v% of hydroxypropyl methylcellulose, and the pH of the eye drop is 6.77 by hydrochloric acid and sodium hydroxide, with the balance being water for injection.

[0095] The preparation method is the same as that of Example 1.

[0096] Example 5

[0097] The present example provides an eye drop containing 0.5 w / v% of an active ingredient, 0.43 w / v% of sodium dihydrogen phosphate, 0.46 w / v% of disodium hydrogen phosphate, 0.30 w / v% of sodium chloride, and 1.0 w / v% of hydroxypropyl methylcellulose, and the pH of the eye drop is 6.77 by hydrochloric acid and sodium hydroxide, with the balance being water for injection.

[0098] The preparation method is the same as that of Example 1.

[0099] Example 6

[0100] The present example provides an eye drop containing 0.5 w / v% of an active ingredient, 0.43 w / v% of sodium dihydrogen phosphate, 0.46 w / v% of disodium hydrogen phosphate, 0.25 w / v% of sodium chloride, and 0.5 w / v% of hydroxypropyl methylcellulose, and the pH of the eye drop is 6.77 by hydrochloric acid and sodium hydroxide, with the balance being water for injection.

[0101] The preparation method is the same as that of Example 1.

[0102] Example 7

[0103] The present example provides an eye drop containing 0.5 w / v% of an active ingredient, 0.43 w / v% of sodium dihydrogen phosphate, 0.46 w / v% of disodium hydrogen phosphate, 0.25 w / v% of sodium chloride, and 1.5 w / v% of hydroxypropyl methylcellulose, and the pH of the eye drop is 6.77 by hydrochloric acid and sodium hydroxide, with the balance being water for injection.

[0104] The preparation method is the same as that of Example 1.

[0105] Comparative Example 1

[0106] The eye drops of the present example are different from those of Example 2 only in that sodium chloride is used instead of disodium hydrogen phosphate and sodium dihydrogen phosphate, and the osmotic pressure of the eye drops is adjusted to be close to that of Example 2 by adjusting the amount of sodium chloride, and the pH value is adjusted to 5.00 by using hydrochloric acid.

[0107] Test Example 1: Performance detection of eye drops

[0108] 1. Stability

[0109] The eye drops of Examples 1-3 were respectively placed under high temperature (60℃) and light (5000Lx, ultraviolet 90uW / cm 2 ) for 5 days and 10 days, and the sample properties, pH value, osmotic pressure and related substances were checked after 5 days and 10 days, and the test results are shown in Table 1.

[0110] After the preparation was completed, the related substances of Comparative Example 1 were detected, and the results are shown in Table 2 below.

[0111] Table 1

[0112]

[0113] Table 2

[0114] Samples Maximum single impurities (%) Total impurities (%) Comparative Example 1 0.32 0.58

[0115] The test results show that: the samples prepared in the present application have a pH value of 5.95-7.59, and after 5 days and 10 days of high temperature (60℃) and light (5000Lx, ultraviolet 90uW / cm 2 ) investigation, the physicochemical properties are basically stable, but some subtle changes can be concerned: ① related substances: the light investigation sample is slightly higher than the high temperature investigation sample (the growth rate is 0.05-0.06 and -0.01-0.03 respectively), which indicates that the sample is more unstable to light and should be stored in the dark as much as possible; ② pH value: the high temperature investigation sample decreases slightly (the decrease rate is 0.05-0.13), and the light investigation sample remains stable; at the same time, the pH value decreases after 121℃ sterilization during sample preparation, indicating that the pH value of the sample tends to decrease at high temperature, and the higher the initial pH value, the greater the decrease rate; ③ osmotic pressure molarity: the high temperature investigation sample increases slightly, and the light investigation sample remains stable, which may be related to the water loss of the sample at high temperature caused by the semi-permeable inner packaging material.

[0116] The related substance content of Comparative Example 1 is obviously higher, and the stability is poor.

[0117] In general, the pH value of the eye drops of the present application should be in the range of 5.9-7.6 (with a middle limit of 6.5-7.0), in which pH value range the sample stability is better. At the same time, it can also be seen that, in the eye drop system of the present application, the pH value is the main factor affecting the stability, and the pH value of Examples 4-7 is the same as that of Example 2, and the effect data in terms of stability is equivalent to that of Example 2.

[0118] 2. Adjusted osmotic pressure

[0119] The osmotic pressure of the eye drops of Examples 2, 4-5 was detected, and the results are shown in Table 3 below.

[0120] Table 3

[0121] Samples Osmotic pressure (mmol L -1 ) Example 2 294 Example 4 318 Example 5 337

[0122] The osmotic pressure of the above eye drops is close to the osmotic pressure of tears, which can ensure that the osmotic pressure of the eye drops matches that of tears, thereby reducing discomfort and potential side effects. The amount of phosphate buffered salt and sodium chloride in other examples is the same as that of Example 2, and the osmotic pressure is also close to that of Example 2.

[0123] 3. Viscosity

[0124] The viscosity of Examples 2, 6-7 was detected, and the results are shown in Table 4 below.

[0125] Table 4

[0126]

[0127]

[0128] As a special preparation, eye drops are easily diluted or washed away by tears after being dropped into the eye, which is also the main reason for the loss of eye drops. The viscosity of the eye drops of the present application is appropriate, which can reduce the loss of drug solution due to blinking, prolong the contact time of the drug with the eye tissue, and also reduce the irritation of the eye drops to the eye. Among them, the viscosity of the eye drops in Example 2 is better. The content of hydroxypropyl methylcellulose in other examples is the same as that of Example 2, and the viscosity value of the eye drops is also close to that of Example 2.

[0129] Test Example 2: Study on the effect of eye drops on guinea pig dry eye syndrome caused by ovalbumin

[0130] 1. Experimental animals

[0131] The specific information is shown in Table 5 below.

[0132] Table 5 Information Table of Experimental Animals

[0133]

[0134] 2. Experimental grouping and administration

[0135] Ten Hartley guinea pigs that passed the adaptive observation were selected for the experiment. Four animals were randomly selected as the negative control group, and the remaining animals were sensitized by intraperitoneal injection of 0.5 mL of an albumin solution with a concentration of 10 mg / mL, once every other day for 3 times. Fourteen days after the last sensitization, 20 mg / mL of albumin solution was dropped into the conjunctival sac of the guinea pigs using a pipette, 20 μL / eye, for challenge, and the tear secretion (phenol red cotton thread method) and tear film break-up time (both eyes) were measured. Animals with mild or severe modeling were excluded, and the animals were grouped according to the tear film break-up time (both eyes) after the first challenge. The modeling animals were divided into two groups: a model group and a drop group, each with 4 animals (8 eyes), half male and half female.

[0136] The administration scheme was continuous administration for 7 days, all using a pipette for administration, 20 μL / eye / time, 4 times / day (only 1 time on the grouping day and the last treatment day), with an interval of about 2-3 hours. The specific administration scheme is shown in Table 6 below. During the experiment, the animals were challenged on the 4th and 7th days of the administration period, and then administered about 5 minutes after the challenge, and the ocular symptoms were observed about 20 minutes later. The tear secretion (both eyes) and / or tear film break-up time (both eyes) were measured on the 4th and 7th days of the administration period. After the last administration, the animals were sacrificed after the observation and detection of each index were completed, and the palpebral conjunctiva, conjunctival fornix, and conjunctiva were cut and fixed with Davison's solution, and the histological changes of the bulbar conjunctiva, palpebral conjunctiva, and fornix conjunctiva were observed under a light microscope.

[0137] Table 6 Experimental grouping and administration

[0138]

[0139] 3. Results analysis

[0140] 3.1 Tear film break-up time of guinea pig dry eye model

[0141] Table 7 Effect of the eye drops of the application on the tear film break-up time of the OVA-induced guinea pig dry eye model

[0142]

[0143] Note: Compared with the model group, * indicates P<0.05, and ** indicates P<0.01.

[0144] The tear film break-up time is one of the indicators commonly used to diagnose dry eye. When the tear film is unstable, its break-up time is shortened, and when the tear film break-up time is shortened, the eye has a dry and itchy feeling, thereby increasing the frequency of blinking. The test results in Table 7 show that: ① Before administration: compared with the model group, the tear film break-up time of the eye drop group animals had no significant difference. ② On the 4th day of administration: compared with the model group, the tear film break-up time of the eye drop group had no significant difference (P>0.05); ③ On the 7th day of administration: compared with the model group, the tear film break-up time of the eye drop group was significantly prolonged, and there was a statistically significant difference (P<0.05 or P<0.01). This indicates that the eye drop group can enhance the stability of the tear film and prolong the tear film break-up time.

[0145] 3.2 Tear secretion (phenol red cotton thread wet length) results of guinea pig dry eye model

[0146] Table 8 Effect of the eye drops of the application on tear secretion of OVA-induced guinea pig dry eye model

[0147]

[0148] Note: compared with the model group, * indicates P<0.05, and ** indicates P<0.01.

[0149] The decrease in tear secretion is also one of the important causes of dry eye. The tear secretion was determined by the phenol red cotton thread test and Schirmer I test. The test results in Table 8 show that: ① Before administration: compared with the model group, the phenol red cotton thread wet length of the eye drop group had no significant difference. ② On the 4th day of administration: compared with the model group, the phenol red cotton thread wet length of the eye drop group animals was significantly longer, and there was a statistically significant difference (P<0.05 or P<0.01); ③ On the 7th day of administration: compared with the model group, the phenol red cotton thread wet length of the eye drop group animals was significantly longer. This indicates that the eye drop group can increase the tear secretion.

[0150] 3.3 Pathological examination results

[0151] Table 9 Statistical table of histological examination results of eyelid tissues of each animal

[0152]

[0153] Note: The degree of histological change is represented by normal, very mild, mild, moderate and severe, indicated by -, +, +, ++ and +++.

[0154] The model group: 7 eyes (A♂01 left, A♂01 right, A♂02 left, A♂02 right, A♀03 left, A♀03 right, A♀04 right; 7 / 8) had moderate to severe inflammatory cell infiltration in the conjunctival layer, and very mild to mild reduction of goblet cells; 1 eye (A♀04 left; 1 / 8) had severe inflammatory cell infiltration in the conjunctival layer. Among them, the HE staining (100x) results of the right eye of the model group A♂01 are shown inFigure 1 As shown in the figure, the conjunctival layer of A♂01 right eye has severe inflammatory cell infiltration, and the goblet cells are slightly reduced.

[0155] The eye drops group: 7 eyes (C♂01 left, C♂01 right, C♂02 right, C♀03 left, C♀03 right, C♀04 left, C♀04 right; 7 / 8) have extremely mild to moderate inflammatory cell infiltration in the conjunctival layer, and the goblet cells are extremely mild to mild reduced; 1 eye (C♂02 left; 1 / 8) has extremely mild inflammatory cell infiltration in the conjunctival layer, and the reduction degree of goblet cells is normal. Among them, the HE staining (100x) result of C♀03 right eye in the eye drop group is shown in Figure 2 As shown in the figure, the conjunctival layer of C♀03 right eye has mild inflammatory cell infiltration, and the goblet cells are slightly reduced.

[0156] As shown in Table 9, after the Hartley guinea pig dry eye model is established by using ovalbumin, the conjunctival layer of the animal eye has different degrees of inflammatory cell infiltration, and the goblet cells are reduced, wherein the inflammatory cells are mainly lymphocytes, indicating that the modeling is successful. Compared with the model group, the inflammatory cell infiltration degree of the eye drop group is reduced, and the reduction degree of the conjunctival goblet cells is also reduced, which has a repairing effect on the goblet cells.

[0157] As can be known from the above, the eye drop of the present application has a certain recovery effect on the ovalbumin-induced Hartley guinea pig dry eye.

[0158] The above-described embodiments only express several embodiments of the present application, and the description is more specific and detailed, but it cannot be understood as limiting the scope of the patent. It should be pointed out that for ordinary skilled in the art, without departing from the concept of the present application, a number of modifications and improvements can be made, which belong to the protection scope of the present application.

Claims

1. An eye drop comprising: i) an active ingredient, which is a compound represented by formula (I) or a pharmaceutically acceptable salt thereof; ii) a buffer salt; iii) an osmotic pressure adjusting agent; and iv) hydroxypropyl methylcellulose; and the pH of the eye drop is 5.9 to 7.

6.

2. The eye drop according to claim 1, wherein, The active ingredient is a hydrochloride salt of a compound represented by formula (I).

3. The eye drop according to claim 2, wherein, The active ingredient has a structure represented by formula (II): In formula (II), p is selected from 0.7 to 2.

3.

4. The eye drop according to any one of claims 1 to 3, wherein, The content of the active ingredient is 0.1 w / v% to 2.0 w / v%.

5. The eye drop according to any one of claims 1 to 4, wherein The buffer salt is selected from one or more of a phosphate buffer salt, a citrate buffer salt, an acetate buffer salt, and a borate buffer salt.

6. The eye drop according to any one of claims 1 to 5, wherein The osmotic pressure adjusting agent is selected from one or more of sodium chloride, mannitol, glucose, xylitol, a phosphate salt, a citrate salt, an acetate salt, and a borate salt.

7. The eye drop according to any one of claims 1 to 4, wherein The buffer salt is a mixture of sodium phosphate monobasic and sodium phosphate dibasic, the content of the sodium phosphate monobasic is 0.4 w / v% to 0.5 w / v%, and the content of the sodium phosphate dibasic is 0.4 w / v% to 0.5 w / v%; and the osmotic pressure adjusting agent is sodium chloride, the content of which is 0.20 w / v% to 0.30 w / v%.

8. The eye drop according to any one of claims 1 to 7, wherein The amount of the hydroxypropyl methylcellulose is 0.8 w / v% to 1.2 w / v%.

9. The eye drop according to any one of claims 1 to 8, wherein, The pH of the eye drop is 6.5 to 7.

0.

10. The eye drop according to any one of claims 1 to 9, wherein The eye drop further comprises a pH adjusting agent selected from one or more of hydrochloric acid, sulfuric acid, citric acid, sodium hydroxide, potassium hydroxide, sodium bicarbonate, sodium phosphate dibasic, and sodium phosphate monobasic.

11. The eye drop according to any one of claims 1 to 4, 9 to 10, wherein, The eye drop comprises: i) 0.1 w / v% to 2.0 w / v% of an active ingredient, which is a hydrochloride salt of a compound represented by formula (I); ii) 0.4 w / v% to 0.5 w / v% of sodium phosphate monobasic and 0.4 w / v% to 0.5 w / v% of sodium phosphate dibasic; iii) 0.20 w / v% to 0.30 w / v% of sodium chloride; iv) 0.8 w / v% to 1.2 w / v% of hydroxypropyl methylcellulose; v) a pH adjusting agent for adjusting the pH of the eye drop to 5.9 to 7.6; and vi) water.

12. The eye drop according to any one of claims 1 to 11, wherein, The osmotic pressure of the eye drop is 290mmol·L -1 ~ 340mmol·L -1 .

13. The eye drop according to any one of claims 1 to 12, wherein, The viscosity of the eye drop is 6 mPa-s to 10 mPa-s.

14. Use of the eye drop of any one of claims 1 to 13 in any one of: I) preventing, treating, or alleviating dry eye syndrome; II) preparing a medicament for preventing, treating, or alleviating dry eye syndrome.

15. Use according to claim 14, wherein, The eye drop is used in at least one of: 1) prolonging tear film break-up time; 2) increasing tear secretion; 3) reducing conjunctival cell infiltration; 4) promoting goblet cell repair.

16. Use according to claim 14 or 15, wherein, The eye drop is suitable for topical administration to the eye of a subject.

17. A kit comprising a therapeutically effective amount of the eye drop of any one of claims 1 to 13, and instructional material for administering the eye drop to a patient suffering from dry eye syndrome.

Images