Eye drop containing syk and vegfr2 dual-target inhibitor and use thereof

Abstract

Disclosed are an eye drop containing a Syk and VEGFR2 dual-target inhibitor and use thereof, belonging to the field of pharmaceutical compositions. The eye drop contains an active ingredient, a buffer salt, an osmotic pressure regulator, and hydroxypropyl methylcellulose, and has a pH value of 5.9-7.6. The eye drop can not only effectively exert a drug effect, but also reduce irritation to the eyes, and it demonstrates good efficacy and administration compliance in the treatment of dry eyes. At the same time, the eye drop still has good stability under high temperature and light conditions, which is beneficial to improving the effectiveness and the safety of drugs during storage and use.

Description

An eye drop containing dual inhibitors of Syk and VEGFR2 and its application Technical Field

[0001] This invention belongs to the field of pharmaceutical compositions, specifically relating to an eye drop containing dual-target inhibitors of Syk and VEGFR2 and its application. Background Technology

[0002] Dry eye has a high global prevalence, and moderate to severe dry eye requires anti-inflammatory treatment. Current treatments include corticosteroids, nonsteroidal anti-inflammatory drugs (NSAIDs), and immunosuppressants. Currently, the main dry eye medications available domestically and internationally are artificial tears, corticosteroids, and cyclosporine. However, these medications only relieve dry eye symptoms and have a slow onset of action, and they also have certain side effects, failing to meet the huge demand in the dry eye medication market.

[0003] Therefore, given the large patient population suffering from dry eye disease, it is urgent to develop dry eye products with new medication mechanisms, faster onset of action, and better efficacy.

[0004] WO2021169958A1 discloses a dual-target inhibitor of Syk and VEGFR2, and discloses compounds relating to the following formula (III) or their medically acceptable salts. WO2022166548A1 further discloses the salt forms, crystal forms, and preparation methods of pyrazole-substituted imidazo[1,2-a]quinoxaline derivatives, and their applications in the preparation of drugs related to dual-target inhibitors of Syk and VEGFR2, and discloses crystal forms relating to compounds relating to the following formula (I).

[0005] The aforementioned molecules exhibit excellent target specificity and significant therapeutic and anti-inflammatory effects in animal dry eye models, with low biotoxicity, demonstrating promising potential for drug application. However, their high molecular structural stability leads to poor solubility in aqueous solutions, increasing the difficulty of forming effective drug formulations. Therefore, there is an urgent need to develop a formulation that can increase the solubility of these drugs or enable them to exert their therapeutic effects.

[0006] Summary of the Invention

[0007] The present invention first provides an eye drop containing: i) an active ingredient, said active ingredient being a compound of formula (I) or a pharmaceutically acceptable salt thereof; ii) a buffer salt; iii) an osmotic pressure regulator; and iv) hydroxypropyl methylcellulose (HPMC); and the pH value of said eye drop is 5.9 to 7.6.

[0008] The present invention further provides the use of the eye drops in the prevention, treatment or relief of dry eye syndrome.

[0009] The present invention further provides the use of the eye drops in the preparation of medicaments for the prevention, treatment or relief of dry eye syndrome.

[0010] The present invention also provides a medicine box containing a therapeutically effective amount of the eye drops, and illustrative material regarding the application of the eye drops to a patient suffering from dry eye syndrome.

[0011] This invention provides eye drops containing dual-target inhibitors of Syk and VEGFR2. These eye drops effectively exert their therapeutic effect while minimizing eye irritation, exhibiting good efficacy and administration compliance in the treatment of dry eye syndrome. Furthermore, these eye drops maintain good stability under high temperature and light conditions, which is beneficial for improving the efficacy and safety of the drug during storage and use. Attached Figure Description

[0012] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0013] Figure 1 shows the HE staining (100×) results of the right eye tissue of model group A♂01 in the experimental example of this invention.

[0014] Figure 2 shows the HE staining (100×) results of the right eye tissue of the eye drop group C♀03 in the experimental example of the present invention. Detailed Implementation

[0015] The following provides a detailed description of specific embodiments of the present invention. It should be understood that the specific embodiments described herein are for illustrative and explanatory purposes only and are not intended to limit the invention. Those skilled in the art can make various modifications and variations to the invention without departing from its scope or spirit. For example, features described or illustrated as part of one embodiment can be used in another embodiment to produce further embodiments.

[0016] Terminology Explanation

[0017] Unless otherwise stated, all terms used to disclose this invention (including technical and scientific terms) have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Further guidance is provided below for a better understanding of the teachings of this invention. The terminology used herein in the specification of this invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

[0018] The terms "and / or," "or / and," and "and / or" as used herein include any one of two or more of the related listed items, as well as any and all combinations of the related listed items. These arbitrary and all combinations include any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or," "or / and," and "and / or," it should be understood that in this application, the technical solution undoubtedly includes technical solutions connected by "logical AND," and also undoubtedly includes technical solutions connected by "logical OR." For example, "A and / or B" includes three parallel solutions: A, B, and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, a technical solution that is connected by "logical OR"), as well as any and all combinations of A, B, C, and D, that is, combinations of any two or three of A, B, C, and D, and also combinations of all four of A, B, C, and D (that is, a technical solution that is connected by "logical AND").

[0019] The terms “containing,” “comprising,” and “including” as used in this invention are synonyms and are inclusive or open-ended, not excluding additional, uncited members, elements, or method steps.

[0020] In this invention, the numerical range represented by endpoints includes all numerical values ​​and fractions contained within that range, as well as the endpoints mentioned.

[0021] This invention relates to concentration values, which include fluctuations within a certain range. For example, fluctuations are allowed within a corresponding precision range. For instance, 2% may fluctuate within ±0.1%. For larger values ​​or values ​​that do not require overly precise control, even greater fluctuations are permitted. For example, 100mM may fluctuate within ranges of ±1%, ±2%, ±5%, etc. Regarding molecular weight, fluctuations of ±10% are allowed.

[0022] In this invention, the terms "multiple" or "various" are used unless otherwise specified, referring to a quantity of 2 or more.

[0023] In this invention, the technical features described in an open-ended manner include both closed-ended technical solutions composed of the listed features and open-ended technical solutions that include the listed features.

[0024] In this invention, terms such as "preferred," "better," "more suitable," and "ideal" are merely descriptions of more effective implementation methods or embodiments, and should be understood not to limit the scope of protection of this invention.

[0025] In this invention, "optionally," "optionally," "optionally," "optionally," "optionally," and "optional" mean that they are optional, that is, they are selected from either "with" or "without." If multiple "optional" or "optional" terms appear in a technical solution, unless otherwise specified and there are no contradictions or mutual constraints, then each "optional" or "optional" term is independent.

[0026] In this invention, the term "dry eye syndrome," also known as keratoconjunctivitis sicca, is a chronic ocular surface disease caused by a variety of factors. It mainly involves abnormalities in the quality, quantity, and dynamics of tears. These abnormalities lead to tear film instability or imbalance of the ocular surface microenvironment, and may be accompanied by ocular surface inflammation, tissue damage, and nerve abnormalities, thereby causing a variety of ocular discomfort symptoms and / or visual dysfunction.

[0027] eye drops

[0028] The compound shown in Formula (I), or a pharmaceutically acceptable salt thereof, is a small molecule drug targeting tyrosine protein kinase. This compound, or its pharmaceutically acceptable salt, inhibits the 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, blocking the inflammatory cascade response caused by the immune response. The main preclinical characteristics of this compound, or its pharmaceutically acceptable salt, are its more significant target activity and better selectivity. At well-tolerated doses, both efficacy and inflammation improvement are significant. Eye drops containing this compound, or its pharmaceutically acceptable salt, as the active ingredient have a large safety window, good animal tolerability, and no significant local irritation, supporting their development for clinical treatment of dry eye.

[0029] However, this compound exhibits poor high-temperature stability and light stability in water. To address this, the present invention further investigates and first provides an eye drop containing: i) an active ingredient, which is a compound of formula (I) or a pharmaceutically acceptable salt thereof; ii) a phosphate buffer; iii) sodium chloride; and iv) hydroxypropyl methylcellulose (HPMC); and the pH value of the eye drop is 5.9 to 7.6.

[0030] The present invention has found that the above-mentioned eye drop formulation can effectively improve the solubility of the active ingredients, enhance their high-temperature stability and photostability, and reduce eye irritation. It has good efficacy and drug administration compliance in the treatment of dry eye syndrome.

[0031] In specific implementation, the active ingredient mentioned in this invention may be the compound shown in formula (I) or its pharmaceutically acceptable salt (such as hydrochloride, or the hydrate, maleate or gentianate disclosed in WO2022166548A1, etc.). At the same time, the compound or its pharmaceutically acceptable salt mentioned in this invention also includes its different crystal forms (such as crystal form A, crystal form B, crystal form C, crystal form D, crystal form E, etc. disclosed in WO2022166548A1).

[0032] In some embodiments, the active ingredient is a hydrochloride salt of the compound shown in formula (I). By preparing it as a hydrochloride salt, the solubility of the active ingredient can be further increased without affecting the efficacy, thereby improving the bioavailability of the drug. Moreover, when applied to the eye drops in the form of a hydrochloride salt, the stability and administration compliance of the drug can be further improved. In addition, the synthesis and preparation of this salt form are also simpler.

[0033] In some embodiments, the structure of the active ingredient is shown in formula (II):

[0034] In formula (II), p is selected from 0.7 to 2.3.

[0035] As an example, in some specific implementations, 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 in the range of 0.7 to 2.3.

[0036] In some specific implementations, p is 0.8, 1.0, or 1.7.

[0037] 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.

[0038] As an example, in some specific embodiments, the content of the active ingredient in the eye drops 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 in the range of 0.1 w / v% to 2.0 w / v%.

[0039] In some embodiments, the buffer salt is selected from one or more of phosphate buffer salts, citric acid buffer salts, acetate buffer salts, and borate buffer salts, more preferably phosphate buffer salts.

[0040] In some embodiments, the osmotic pressure regulator is selected from one or more of sodium chloride, mannitol, glucose, xylitol, phosphate, citrate, acetate, and borate, more preferably sodium chloride.

[0041] In some preferred embodiments, the buffer salt is a mixture of sodium dihydrogen phosphate and disodium hydrogen phosphate, wherein the content of sodium dihydrogen phosphate is 0.4 w / v% to 0.5 w / v, and the content of disodium hydrogen phosphate is 0.4 w / v% to 0.5 w / v%; the osmotic pressure regulator is sodium chloride, wherein the content of sodium chloride is 0.20 w / v% to 0.30 w / v. This results in better stability and more appropriate osmotic pressure in the eye drops, further preventing eye irritation.

[0042] As an example, in some specific embodiments, the sodium dihydrogen phosphate content is any value within the range of 0.40 w / v%, 0.41 w / v%, 0.42 w / v%, 0.43 w / v%, 0.44 w / v%, 0.45 w / v%, 0.46 w / v%, 0.47 w / v%, 0.48 w / v%, 0.49 w / v%, 0.50 w / v%, or 0.4 w / v% to 0.5 w / v%.

[0043] As an example, in some specific embodiments, the content of disodium hydrogen phosphate is any value within the range of 0.40 w / v%, 0.41 w / v%, 0.42 w / v%, 0.43 w / v%, 0.44 w / v%, 0.45 w / v%, 0.46 w / v%, 0.47 w / v%, 0.48 w / v%, 0.49 w / v%, 0.50 w / v%, or 0.4 w / v% to 0.5 w / v%.

[0044] As an example, in some specific embodiments, the sodium chloride content is any value within the range of 0.20 w / v%, 0.21 w / v%, 0.22 w / v%, 0.23 w / v%, 0.24 w / v%, 0.25 w / v%, 0.26 w / v%, 0.27 w / v%, 0.28 w / v%, 0.29 w / v%, 0.30 w / v%, or 0.20 w / v% to 0.30 w / v%.

[0045] In some embodiments, the amount of hydroxypropyl methylcellulose used is 0.8 w / v% to 1.2 w / v%. This results in an eye drop with an appropriate viscosity, which helps ensure an adequate residence time of the drug in the eye while reducing eye irritation.

[0046] As an example, in some specific embodiments, the amount of hydroxypropyl methylcellulose used is any value within the range of 0.8 w / v%, 0.9 w / v%, 1.0 w / v%, 1.1 w / v%, 1.2 w / v%, or 0.8 w / v% to 1.2 w / v%.

[0047] In some embodiments, the pH value of the eye drops is 6.5 to 7.0.

[0048] As an example, in some specific embodiments, the pH value of the eye drops is any value within the range of 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 5.9 to 7.6.

[0049] In some embodiments, the eye drops further contain a pH adjuster selected from one or more of hydrochloric acid, sulfuric acid, citric acid, sodium hydroxide, potassium hydroxide, sodium bicarbonate, disodium hydrogen phosphate, and sodium dihydrogen phosphate.

[0050] In some specific embodiments, the pH adjuster is selected from one or two of hydrochloric acid and sodium hydroxide.

[0051] 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, and reverse osmosis water.

[0052] In a preferred embodiment, the eye drops contain: i) 0.1 w / v% to 2.0 w / v% of an active ingredient, said active ingredient being the hydrochloride salt of the compound shown in 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 adjuster for adjusting the pH of the eye drops to 5.9 to 7.6; and vi) water.

[0053] In some embodiments, the osmotic pressure of the eye drops is 290 mmol·L⁻¹. -1 ~340mmol·L -1This helps to reduce eye irritation while maintaining the efficacy of the eye drops. In practice, the osmotic pressure of the eye drops can be adjusted by changing the amount of buffer salts (such as phosphate buffers) and osmotic pressure regulators (such as sodium chloride) in the eye drop formulation of this invention.

[0054] As an example, in some specific embodiments, the osmotic pressure of the eye drops is 290 mmol·L⁻¹. -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 ~340mmol·L -1 Any value within the range.

[0055] In some embodiments, the viscosity of the eye drops is 6 mPa·s to 10 mPa·s. Within this range, the active ingredient has an appropriate retention time in the eye and better bioavailability. In practice, the viscosity of the eye drops can be adjusted by changing the amount of hydroxypropyl methylcellulose in the eye drop formulation of this invention.

[0056] In some specific embodiments, the eye drops may also contain one or more of the following reagents: 1) chelating agents: such as disodium edetate, used to remove metal ions in the solution that may affect the stability of the drug; 2) antioxidants: such as ascorbic acid, isoascorbic acid, etc., used to prevent the preparation from deteriorating due to oxidation caused by the presence of oxygen free radicals or free metals in the composition.

[0057] Those skilled in the art can combine the above-mentioned embodiments with common sense to obtain more embodiments of the eye drops described in this invention.

[0058] Based on the above-described eye drop formulation and general knowledge in the field, those skilled in the art can confirm the preparation method of the eye drops.

[0059] In one specific embodiment, the preparation method of the eye drops includes: taking 70-85% of the formula amount of purified water, adding hydroxypropyl methylcellulose, phosphate and sodium chloride, stirring to dissolve, adding the active component and stirring to dissolve, then adjusting the pH to 5.9-7.6 with a pH adjuster, making up to volume with water, and measuring the pH to obtain the eye drops.

[0060] In some specific embodiments, the method for preparing the eye drops further includes the steps of sterilizing and / or filling the eye drops.

[0061] application

[0062] The present invention further provides the use of the aforementioned eye drops in the prevention, treatment, or relief of dry eye syndrome. In other words, the present invention also provides a method for the prevention, treatment, or relief of dry eye syndrome, the method comprising: administering a therapeutically effective amount of the aforementioned eye drops to a subject.

[0063] The present invention further provides the use of the eye drops in the preparation of medicaments for the prevention, treatment or relief of dry eye syndrome.

[0064] In some embodiments, the eye drops are used for at least one of the following purposes: 1) prolonging tear film breakup time; 2) increasing tear secretion; 3) reducing conjunctivitis cell infiltration; and 4) promoting goblet cell repair.

[0065] In some embodiments, the eye drops are suitable for topical application to the eye of a subject.

[0066] In this invention, "topical application" means applying something locally to the surface of a tissue, such as the eye, particularly any external aspect of the eye that is usually accessible between the eyelids. Topical application to the eye is typically done via eye drops, ointments, or sprays.

[0067] In some embodiments, the eye drops may be contained in rigid and / or squeeze-type bottles equipped with fitting caps configured to function as droppers. Human subjects may receive between 1 and 10 drops per day (e.g., 5 or 8 drops per day), and the dose may be repeated, for example, twice daily. The eye drops may be dispensed, for example, in 12 mL or 20 mL volumes per bottle.

[0068] In some embodiments, the eye drops may also be administered via a carrier medium, such as liquid drops, liquid lotion, gel, ointment, and spray, or combinations thereof.

[0069] In some embodiments, the local application may also occur by injecting the eye drops via a device (such as a pump-catheter system / continuous or selective release device, contact lens, or a combination thereof). The eye drops may also be administered in an injectable form, for example, such that injecting the eye drops retro-orbital and / or wherein the application involves intravitreal injection.

[0070] In this invention, the term "subject" refers to mammals such as humans, domestic animals (such as felines or canines), farm animals (e.g., cattle, horses, goats, sheep, and pigs), wild animals, or research animals (e.g., mice, rats, rabbits, goats, sheep, pigs, dogs, and cats, and bird species (such as chickens, turkeys, and songbirds)).

[0071] In some implementations, the subjects are humans.

[0072] The present invention also provides a medicine box containing a therapeutically effective amount of the eye drops, and illustrative material regarding the application of the eye drops to a patient suffering from dry eye syndrome.

[0073] In some embodiments, the illustrative material may include publications, charts, or any other medium of expression that can be used to convey the usefulness of the composition and its application. In some embodiments, the illustrative material may be attached to a container containing the eye drops of the present invention, or may otherwise be provided together with the container containing the composition. In some embodiments, the illustrative material may be provided separately, for example, by electronic transmission, such as by means of a computer (e.g., via email, or downloaded from a website).

[0074] In some embodiments, the kit also includes at least one adjuvant that can be used to prevent, treat or relieve dry eye syndrome.

[0075] Example

[0076] The embodiments of the present invention will be described in detail below with reference to examples. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. For experimental methods in the following embodiments where specific conditions are not specified, please refer to the guidelines given in this invention, or follow experimental manuals or conventional conditions in the art, or other experimental methods known in the art, or follow the conditions recommended by the manufacturer.

[0077] In the specific embodiments described below, the measurement parameters of the raw material components may have slight deviations within the weighing accuracy range unless otherwise specified. For temperature and time parameters, acceptable deviations due to instrument testing accuracy or operational precision are permissible. For pH values, slight deviations due to testing accuracy (e.g., within ±0.10) are permissible.

[0078] For ease of comparison, equal amounts of one active ingredient are used in the following examples and comparative examples, the structure of which is shown below, where p is 1.0. This active ingredient and its dosage do not constitute a limitation on the scope of protection of this invention. Active ingredients within the scope defined by this invention, when applied to the eye drops, all possess the technical effects described above.

[0079] Example 1

[0080] This embodiment first provides an eye drop containing 0.5 w / v% of the 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 adjusted to 5.95 by hydrochloric acid and sodium hydroxide, with the remainder being water for injection.

[0081] This embodiment further provides a method for preparing the above-mentioned eye drops, as follows: Take 80% of the prescribed amount of water for injection, add hydroxypropyl methylcellulose, sodium dihydrogen phosphate, disodium hydrogen phosphate and sodium chloride, stir to dissolve, add the active ingredient, stir to dissolve, adjust the pH to the specified value with 1 mol / L sodium hydroxide solution or hydrochloric acid solution, make up to volume, measure the pH, sterilize at 121°C for 15 minutes, and dispense into low-density polyethylene pharmaceutical eye drop bottles with stirring, 5 mL / bottle.

[0082] Example 2

[0083] This embodiment provides an eye drop containing 0.5 w / v% of the 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. The pH of the eye drop is adjusted to 6.77 by hydrochloric acid and sodium hydroxide, with the remainder being water for injection.

[0084] The preparation method is the same as in Example 1.

[0085] Example 3

[0086] This embodiment provides an eye drop containing 0.5 w / v% of the 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. The pH of the eye drop is adjusted to 7.59 by hydrochloric acid and sodium hydroxide, with the remainder being water for injection.

[0087] The preparation method is the same as in Example 1.

[0088] Example 4

[0089] This embodiment provides an eye drop containing 0.5 w / v% of the 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. The pH of the eye drop is adjusted to 6.77 by hydrochloric acid and sodium hydroxide, with the remainder being water for injection.

[0090] The preparation method is the same as in Example 1.

[0091] Example 5

[0092] This embodiment provides an eye drop containing 0.5 w / v% of the 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. The pH of the eye drop is adjusted to 6.77 by hydrochloric acid and sodium hydroxide, with the remainder being water for injection.

[0093] The preparation method is the same as in Example 1.

[0094] Example 6

[0095] This embodiment provides an eye drop containing 0.5 w / v% of the 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 adjusted to 6.77 by hydrochloric acid and sodium hydroxide, with the remainder being water for injection.

[0096] The preparation method is the same as in Example 1.

[0097] Example 7

[0098] This embodiment provides an eye drop containing 0.5 w / v% of the 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. The pH of the eye drop is adjusted to 6.77 by hydrochloric acid and sodium hydroxide, with the remainder being water for injection.

[0099] The preparation method is the same as in Example 1.

[0100] Comparative Example 1

[0101] This embodiment provides an eye drop solution, which differs from Embodiment 2 only in that sodium chloride is used instead of disodium hydrogen phosphate and sodium dihydrogen phosphate, and the osmotic pressure of the eye drop solution is adjusted to be close to that of Embodiment 2 by adjusting the amount of sodium chloride, while the pH value is adjusted to 5.00 with hydrochloric acid.

[0102] Experimental Example 1: Performance Testing of Eye Drops

[0103] 1. Stability

[0104] The eye drops from Examples 1-3 were respectively placed at high temperature (60°C) and exposed to light (5000Lx, UV 90uW / cm). 2 After 5 and 10 days of observation, the sample properties, pH value, osmotic pressure and related substances were checked. The test results are shown in Table 1.

[0105] After the configuration was completed, the relevant substances in Comparative Example 1 were tested, and the results are shown in Table 2 below.

[0106] Table 1

[0107] Table 2

[0108] Experimental results show that the samples prepared in this invention with a pH value of 5.95–7.59, after being subjected to high temperature (60℃) and light irradiation (5000 Lx, UV 90 uW / cm²), 2 After 5 and 10 days of observation, the physicochemical properties remained basically stable, but some subtle changes were noteworthy: ① Related substances: The increase in related substances in the light-exposed samples was slightly higher than that in the high-temperature-exposed samples (the increases were 0.05–0.06 and -0.01–0.03, respectively), indicating that the samples were less stable to light and should be stored away from light as much as possible; ② pH value: The pH value decreased slightly in the high-temperature-exposed samples (the decrease ranged from 0.05 to 0.13), while the pH value remained stable in the light-exposed samples; at the same time, the pH value decreased after sterilization at 121℃ during sample preparation, indicating that the pH of the samples tends to decrease at high temperatures, and the higher the initial pH value, the greater the decrease; ③ The osmolar concentration was slightly higher in the light-exposed samples than in the high-temperature-exposed samples, while the pH value remained stable in the light-exposed samples, which may be related to the water loss of the samples at high temperatures caused by the semi-permeable inner packaging material.

[0109] The content of related substances in Comparative Example 1 was significantly higher, and its stability was poor.

[0110] In general, the pH range of the eye drops of the present invention should be defined as 5.9 to 7.6 (intermediate limit is 6.5 to 7.0), within which the samples exhibit good stability. It can also be seen that in the eye drop system of the present invention, pH is the main factor affecting stability, and the pH values ​​of Examples 4 to 7 are the same as those of Example 2, with comparable stability performance data.

[0111] 2. Adjust the osmotic pressure after adjustment

[0112] The osmotic pressure of the eye drops in Examples 2, 4-5 was tested, and the results are shown in Table 3 below.

[0113] Table 3

[0114] The osmotic pressure of the above-mentioned eye drops is close to that of tears, ensuring that the osmotic pressure of the eye drops matches that of tears, thereby reducing discomfort and potential side effects. In other embodiments, the amounts of phosphate buffer and sodium chloride are the same as in Example 2, and the osmotic pressure is also close to that in Example 2.

[0115] 3. Viscosity

[0116] The viscosity of Examples 2, 6 and 7 was tested, and the results are shown in Table 4 below.

[0117] Table 4

[0118] Eye drops, as a special formulation, are easily diluted or washed away by tears after being instilled into the eye, which is the main reason for eye drop loss. The eye drops of this invention have an appropriate viscosity, which can reduce the loss of medication due to blinking, prolong the contact time between the medication and eye tissues, and also reduce the irritation of the eye drops. In Example 2, the viscosity of the eye drops is even better. In other examples, the content of hydroxypropyl methylcellulose is the same as in Example 2, and the viscosity values ​​of the eye drops are also close to those in Example 2.

[0119] Experimental Example 2: Effect of eye drops on ovalbumin-induced dry eye in guinea pigs

[0120] 1. Laboratory animals

[0121] See Table 5 below for details.

[0122] Table 5. Information on Laboratory Animals

[0123] 2. Experimental grouping and drug administration

[0124] Ten Hartley guinea pigs that passed the adaptation 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 albumin solution with a concentration of 10 mg / mL per animal. Sensitization was repeated every other day for three consecutive days. Fourteen days after the last sensitization, 20 μL of 20 mg / mL albumin solution was pipetted into the conjunctival sac of the guinea pigs using a pipette to challenge the model. Tear secretion (phenol red cotton thread method) (both eyes) and tear film breakup time (both eyes) were measured. Animals that were too light or too heavy for the model were excluded. The animals were divided into two groups according to the tear film breakup time (both eyes) after the first challenge. The model animals were divided into two groups: the model group and the eye drop group, with 4 animals (8 eyes) in each group, half male and half female.

[0125] The administration regimen consisted of 7 consecutive days of drug administration, with each dose administered via pipette at a rate of 20 μL / eye / time, 4 times / day (only one dose was administered on the day of grouping and the last treatment day), with an interval of approximately 2–3 hours between doses, as detailed in Table 6 below. During the experiment, challenge was performed on days 4 and 7 of the administration period, followed by drug administration approximately 5 minutes after challenge, and observation of ocular symptoms was conducted approximately 20 minutes later. Tear secretion (both eyes) and / or tear film breakup time (both eyes) were measured on days 4 and 7 of the administration period. After the last administration and all indicator observations and tests were completed, the animals were euthanized, and the palpebral conjunctiva and conjunctival fornix were harvested, fixed with Davison's solution, routinely stained with hematoxylin and eosin (HE), and the histological changes of the bulbar conjunctiva, palpebral conjunctiva, and conjunctival fornix were observed under a light microscope.

[0126] Table 6 Experimental Groups and Drug Administration

[0127] 3. Results Analysis

[0128] 3.1 Results of tear film breakup time in guinea pigs with dry eye syndrome

[0129] Table 7. Effects of the eye drops of this invention on tear film breakup time in an OVA-induced guinea pig dry eye model.

[0130] Note: Compared with the model group, * indicates P<0.05, and ** indicates P<0.01.

[0131] Tear film breakup time is a commonly used indicator for diagnosing dry eye syndrome. When the tear film is unstable, its breakup time is shortened, which can lead to a dry feeling in the eyes and increase the frequency of blinking. The experimental results in Table 7 show that: ① Before administration: Compared with the model group, there was no significant difference in tear film breakup time among the animals in the eye drop group. ② On day 4 after administration: Compared with the model group, there was no significant difference in tear film breakup time among the eye drop group (P>0.05); ③ On day 7 after administration: Compared with the model group, the tear film breakup time in the eye drop group was significantly prolonged, with statistically significant differences (P<0.05 or P<0.01). This indicates that the eye drop group can enhance tear film stability and prolong tear film breakup time.

[0132] 3.2 Results of tear secretion (length of phenol red cotton thread wetted) in guinea pigs with dry eye syndrome

[0133] Table 8. Effects of the eye drops of this invention on tear secretion in an OVA-induced guinea pig dry eye model.

[0134] Note: Compared with the model group, * indicates P<0.05, and ** indicates P<0.01.

[0135] Decreased tear secretion is also a significant cause of dry eye syndrome. Tear secretion was measured using the phenol red cotton thread test and the Schirmer I test. The results in Table 8 show that: ① Before administration: Compared with the model group, there was no significant difference in the wetted length of the phenol red cotton thread in the eye drop group. ② On day 4 after administration: Compared with the model group, the wetted length of the phenol red cotton thread in the eye drop group was significantly longer, with statistically significant differences (P<0.05 or P<0.01); ③ On day 7 after administration: Compared with the model group, the wetted length of the phenol red cotton thread in the eye drop group was significantly longer. This indicates that the eye drop group can increase tear secretion.

[0136] 3.3 Pathological examination results

[0137] Table 9. Statistical table of eyelid histological examination results for various animals.

[0138] Note: The degree of histological changes is indicated by -, ±, +, ++, and +++ to represent normal, very mild, mild, moderate, and severe, respectively.

[0139] 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) showed moderate to severe inflammatory cell infiltration in the conjunctival layer, with very mild to slight reduction in goblet cells; 1 eye (A♀04 left; 1 / 8) showed severe inflammatory cell infiltration in the conjunctival layer. The HE staining (100×) results of the right eye of A♂01 in the model group are shown in Figure 1. As can be seen from the figure, the right eye of A♂01 showed severe inflammatory cell infiltration in the conjunctival layer, with a slight reduction in goblet cells.

[0140] In the eye drop 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) showed very mild to moderate inflammatory cell infiltration in the conjunctival layer and very mild to mild reduction of goblet cells; 1 eye (C♂02 left; 1 / 8) showed very mild inflammatory cell infiltration in the conjunctival layer, with a normal degree of goblet cell reduction. Among them, the HE staining (100×) results of the right eye of C♀03 in the eye drop group are shown in Figure 2. As can be seen from the figure, the right eye of C♀03 showed mild inflammatory cell infiltration in the conjunctival layer and a mild reduction of goblet cells.

[0141] As shown in Table 9, after ovalbumin-induced dry eye in Hartley guinea pigs, the animals exhibited varying degrees of inflammatory cell infiltration and a reduction in goblet cells in the conjunctiva. The inflammatory cells were predominantly lymphocytes, indicating successful modeling. Compared to the model group, the eye drop group showed reduced inflammatory cell infiltration and a decreased reduction in conjunctival goblet cells, suggesting a role in goblet cell repair.

[0142] In summary, the eye drops of the present invention have a certain restorative effect on ovalbumin-induced dry eye syndrome in Hartley guinea pigs.

[0143] The embodiments described above are merely examples of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention.

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 5.9-7.

6.

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

3. The eye drop according to claim 2, wherein, The structure of the active ingredient is shown below as formula (II): In formula (II), p is selected from 0.7-2.

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%-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 phosphate buffer salt, citrate buffer salt, acetate buffer salt, and 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, phosphate, citrate, acetate, and borate.

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 sodium phosphate monobasic is 0.4 w / v%-0.5 w / v%, and the content of sodium phosphate dibasic is 0.4 w / v%-0.5 w / v%; the osmotic pressure adjusting agent is sodium chloride, and the content of sodium chloride is 0.20 w / v%-0.30 w / v%.

8. The eye drop according to any one of claims 1 to 7, wherein The amount of hydroxypropyl methylcellulose is 0.8 w / v%-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 6.5-7.

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%-2.0 w / v% of an active ingredient, which is hydrochloride of the compound of formula (I); ii) 0.4 w / v%-0.5 w / v% of sodium phosphate monobasic and 0.4 w / v%-0.5 w / v% of sodium phosphate dibasic; iii) 0.20 w / v%-0.30 w / v% of sodium chloride; iv) 0.8 w / v%-1.2 w / v% of hydroxypropyl methylcellulose; v) a pH adjusting agent for adjusting the pH of the eye drop to 5.9-7.6; 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-10 mPa-s.

14. Use of the eye drop of any one of claims 1-13 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 1-13, and instructional material for administering the eye drop to a patient suffering from dry eye syndrome.

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