Protein tyrosine kinase inhibitor pharmaceutical composition, and preparation method therefor and use thereof

Abstract

A protein tyrosine kinase inhibitor pharmaceutical composition, and a preparation method therefor and the use thereof. The pharmaceutical composition is a liquid composition, containing compound TKI or a pharmaceutically acceptable salt thereof as an active ingredient, a surfactant, a solubilizer, and a viscosity modifier, wherein the compound TKI is a protein tyrosine kinase inhibitor. The pharmaceutical composition can be used as eye drops for treating related ocular diseases.

Description

A protein tyrosine kinase inhibitor pharmaceutical composition, its preparation method and application Technical Field

[0001] This invention relates to the field of pharmaceutical formulation technology, specifically to a protein tyrosine kinase inhibitor pharmaceutical composition, its preparation method, and its application. Background Technology

[0002] Age-related macular degeneration (AMD) and diabetic retinopathy (DR) are the most common eye diseases and leading causes of blindness in the elderly and working-age populations. These diseases are associated with neovascularization and extravasation in the posterior segment of the eye. In particular, wet AMD is characterized by the formation of new blood vessels from the choroidal microvascular bed and their invasion into the subretinal space, while DR preferentially involves extravasation and neovascularization at the retinal level. Both AMD and DR are characterized by endothelial cell (EC) proliferation and migration, increased vascular permeability, and inflammation, in which vascular endothelial growth factor-A (VEGF-A) and its corresponding receptor (VEGFR) play crucial roles. In addition, tumor growth and metastasis depend on tumor vascular networks to provide adequate oxygen and nutrients. Tumor angiogenesis relies on a highly complex process involving growth factor signaling, endothelial cell proliferation, extracellular matrix (ECM) remodeling, and stromal cell interactions. Among these, vascular endothelial growth factor (VEGF) is the most important driver of angiogenesis. It has been shown that many proliferative disorders, such as those in ophthalmic diseases, tumors, and cancers, involve the overexpression or upregulation of receptor tyrosine kinase (RTK) activity. Receptor tyrosine kinases are a class of kinases that modify proteins by chemically adding phosphate groups (phosphorylation). Phosphorylation typically leads to functional changes in target proteins by altering enzyme activity, cellular localization, or binding to other proteins. Kinases are known to regulate most cellular pathways, especially those involved in signal transduction. To date, one approach to inhibiting the VEGF pathway is to inhibit receptor tyrosine kinase (RTK) activity. In the treatment of age-related macular degeneration (AMD) and diabetic retinopathy (DR), protein tyrosine kinase inhibitor therapy aims to counteract pathological neovascularization and disease progression, preventing visual impairment. Simultaneously, the importance of VEGFR as a pro-angiogenic inducer in tumor growth, invasion, and extravasation makes it an excellent therapeutic target for various cancers.

[0003] PCT application WO2024032584 discloses a series of protein tyrosine kinase inhibitor compounds, which hold promise for the treatment of related eye diseases. The eye has a complex physiological and anatomical structure, with numerous physiological barriers such as tears, cornea, conjunctiva, blood-aqueous humor, and blood-retina. Clinically, eye diseases can be treated with oral, topical, and injectable medications. However, oral or injectable administration requires crossing the blood-eye barrier, resulting in large doses. While achieving effective therapeutic concentrations, these doses can easily cause toxic side effects. Therefore, topical ocular administration is a more ideal method. However, due to the eye's unique structure, ocular drug delivery often has several drawbacks. For example, administration is inconvenient; the drug is easily washed away from the ocular surface due to tear flushing and blinking reflexes; and the presence of numerous physiological barriers such as the cornea makes it difficult for the drug to penetrate ocular tissues, resulting in low exposure to retinal tissues and low bioavailability. Currently, there are no reports of marketed ocular drug delivery formulations of the aforementioned protein tyrosine kinase inhibitor compounds. Summary of the Invention

[0004] To overcome the shortcomings of the prior art, the present invention provides a protein tyrosine kinase inhibitor pharmaceutical composition, its preparation method and application.

[0005] In a first aspect of the invention, a pharmaceutical composition is provided, which is a liquid composition comprising: a compound TKI or a pharmaceutically acceptable salt thereof as an active ingredient, a surfactant, a solubilizer, and a viscosity modifier.

[0006] Specifically, the surfactant is selected from one or more of polyoxyethylene castor oil, poloxamer, tyloxapine, Tween, and polyoxyethylene 40 hydrogenated castor oil, particularly tyloxapine; its concentration in the pharmaceutical composition is 0.1%-5% (w / v, g / 100ml) (e.g., 0.1%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 4%, 5%), particularly 0.5%-2%, for example 1%.

[0007] Specifically, the solubilizer is a cyclodextrin, such as a β-cyclodextrin derivative, a γ-cyclodextrin derivative, or a combination thereof. A preferred β-cyclodextrin derivative is a hydroxyalkyl-β-cyclodextrin, such as hydroxypropyl-β-cyclodextrin (HP-β-CD), and another preferred β-cyclodextrin derivative is a sulfonyl ether-β-cyclodextrin, such as sulfobutyl ether-β-cyclodextrin (SBE-β-CD). The concentration of the solubilizer in the pharmaceutical composition is 0.5%-30% (w / v, g / 100 ml) (e.g., 0.5%, 1%, 5%, 10%, 12%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 22%, 25%, 30%), particularly 10%-25%, 15%-25%, for example, 20%.

[0008] Specifically, the viscosity modifier is selected from: cellulose derivatives (e.g., methylcellulose, hydroxypropylcellulose, hydroxypropyl methylcellulose), biocompatible polysaccharides (e.g., gellan gum, xanthan gum, chitosan and its derivatives), alginate or its salts (e.g., sodium alginate), hyaluronic acid or its salts (e.g., sodium hyaluronate)), carbomer, polyvinylpyrrolidone, polyvinyl alcohol, or combinations thereof, especially biocompatible polysaccharides. Depending on the type, gellan gum, sodium hyaluronate, and hydroxypropyl methylcellulose are preferred from the above viscosity modifiers. Gellan gum has excellent ion-sensitive gel properties; in the presence of monovalent or divalent cations (e.g., sodium ions, potassium ions, calcium ions), it rapidly gels and covers the surface to form a film structure, which is beneficial for the long-term retention and release of drugs. Hyaluronic acid or its salts (such as sodium hyaluronate) possess excellent hydrophilicity and water-retention properties, forming a hydrated gel network to maintain ocular surface moisture. Combined with pseudoplasticity, they maintain high viscosity during their stay on the ocular surface, prolonging drug residence time and improving bioavailability. Furthermore, ocular tissues contain varying concentrations of hyaluronic acid, thus exhibiting good biocompatibility and membrane affinity, which facilitates compound penetration. Hydroxypropyl methylcellulose, a commonly used viscosity modifier, possesses good biocompatibility and stability. It can also swell with water or body fluids to form a gel layer, helping to prolong drug residence time in the eye and potentially improving bioavailability.

[0009] In some embodiments of the invention, the viscosity modifier is gellan gum, and its concentration in the pharmaceutical composition is 0.01%-1% (w / v, g / 100ml) (e.g., 0.01%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.4%, 0.5%, 0.6%, 0.8%, 1%), particularly 0.1%-0.2%, for example, 0.15%.

[0010] In other embodiments of the invention, the viscosity modifier is hyaluronic acid or a salt thereof (such as sodium hyaluronate) at a concentration of 0.05%-1% (w / v, g / 100 ml) in the pharmaceutical composition (e.g., 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.4%, 0.5%, 0.6%, 0.8%, 1%), particularly 0.1%-0.4%, for example 0.2%.

[0011] Furthermore, the molecular weight of the hyaluronic acid is 1,000 to 2,000,000 Daltons (e.g., 1,000, 5,000, 10,000, 50,000, 100,000, 200,000, 400,000, 500,000, 600,000, 1,000,000, 1,200,000, 1,400,000, 1,500,000, 1,600,000, 1,800,000, 2,000,000 Daltons), for example, 100,000 to 1,000,000, or 100,000 to 500,000 Daltons.

[0012] In other embodiments of the invention, the viscosity modifier is hydroxypropyl methylcellulose, and its concentration in the pharmaceutical composition is 0.1%-5% (w / v, g / 100 ml) (e.g., 0.1%, 0.3%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 5%), particularly 0.4%-0.6%, for example, 0.5%.

[0013] Further, the viscosity of the hydroxypropyl methylcellulose is 3-5000 mPa·s (e.g., 5, 10, 20, 50, 100, 150, 200, 400, 500, 600, 800, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000 mPa·s), for example 100-1000 mPa·s, 200-600 mPa·s.

[0014] Specifically, the compound TKI is a protein tyrosine kinase inhibitor, such as that described in PCT application WO2024032584, for example, compounds shown in formulas II, III, and IV, and compounds shown in Table 1 (excluding compounds I-12) (the preparation of the compound TKI can be referred to WO2024032584).

[0015] Equation IV, as described in WO2024032584, is as follows:

[0016] in,

[0017] R3 is selected from: H, C1-C6 alkyl groups;

[0018] R5 is selected from: H, halogen, cyano, amino, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl;

[0019] L1 is a C2-C6 alkenyl or C2-C6 ynyl;

[0020] The E ring is a benzene ring or a five- or six-membered nitrogen-containing heteroaromatic ring. Some have the following structure: R6 is selected from: H, halogen, cyano, amino, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl;

[0021] L2 is a single bond or a C1-C6 alkylene group;

[0022] Y is selected from: H, -NR7R8, or optionally substituted nitrogen-containing heterocyclic groups, wherein the nitrogen-containing heterocyclic group may optionally be substituted by one or more R9 groups;

[0023] R7 and R8 are independently selected from: H, C1-C6 alkyl, -(C0-C6 alkylene)-(C3-C6 cycloalkyl);

[0024] R9 is selected from: H, C1-C6 alkyl groups (e.g., -CH3, ...). ), C1-C6 hydroxysubstituted alkyl groups (e.g. ), C1-C6 amino-substituted alkyl groups (e.g. ), C1-C6 alkoxy-substituted alkyl groups (e.g. ), C1-C6 alkylamine substituted alkyl groups (e.g. -(C0-C6 alkylene)-(C3-C6 cycloalkyl) (e.g.) );

[0025] Part of in,

[0026] R 4a Selected from: -C(O)NR 401 R 402 -(C0-C6 alkylene)-NR 401 R 402 -C(O)R 401 -C(O)OR 401 R 401 and R 402 Independently selected from: H, C1-C6 alkyl, -(C0-C6 alkylene)-(C3-C6 cycloalkyl); wherein the C0-C6 alkylene, C1-C6 alkyl, and C3-C6 cycloalkyl may optionally be substituted by one or more groups selected from: halogen, cyano, amino, hydroxyl, C1-C6 alkyl, and C1-C6 haloalkyl;

[0027] R 4b It is one or more independent substituents on the benzene ring, selected from: H, halogen, cyano, amino, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl.

[0028] In some embodiments of the present invention, R3 is H.

[0029] In some embodiments of the present invention, R5 is H.

[0030] In some embodiments of the present invention, R6 is H.

[0031] In some embodiments of the present invention, L1 is... In some embodiments of the present invention, L1 is...

[0032] In some embodiments of the present invention, the compound TKI has the following structure:

[0033] in,

[0034] R1 is selected from: -C(O)NR 101 R 102 -(C0-C6 alkylene)-NR 101 R 102 -C(O)R 101 -C(O)OR 101 R 101 and R 102 Independently selected from: H, C1-C6 alkyl, -(C0-C6 alkylene)-(C3-C6 cycloalkyl); wherein the C0-C6 alkylene, C1-C6 alkyl, and C3-C6 cycloalkyl may optionally be substituted by one or more groups selected from: halogen, cyano, amino, hydroxyl, C1-C6 alkyl, and C1-C6 haloalkyl;

[0035] R2 is one or more independent substituents on the benzene ring, selected from: H, halogen, cyano, amino, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl;

[0036] L is a single bond or a C1-C6 alkylene group;

[0037] Y is selected from: H, -NR7R8, or optionally substituted nitrogen-containing heterocyclic groups, wherein the nitrogen-containing heterocyclic group may optionally be substituted by one or more R9 groups;

[0038] R7 and R8 are independently selected from: H, C1-C6 alkyl, -(C0-C6 alkylene)-(C3-C6 cycloalkyl);

[0039] R9 is selected from: H, C1-C6 alkyl, C1-C6 hydroxy-substituted alkyl, C1-C6 amino-substituted alkyl, C1-C6 alkoxy-substituted alkyl, C1-C6 alkylamino-substituted alkyl, -(C0-C6 alkylene)-(C3-C6 cycloalkyl).

[0040] In some embodiments of the present invention, L represents a single bond.

[0041] In some embodiments of the present invention, L is a C1-C6 alkylene group, such as...

[0042] In some embodiments of the present invention, Y is -NR7R8, and R7 and R8 are independently selected from: H, C1-C6 alkyl groups, for example,

[0043] In other embodiments of the invention, Y is an optionally substituted saturated nitrogen-containing heterocyclic group, for example, (where R9 can be substituted at any suitable carbon or nitrogen atom); more specifically, Y is selected from:

[0044] In some embodiments of the present invention, R9 is selected from: H, C1-C6 alkyl groups (e.g., -CH3, ...). ).

[0045] In some embodiments of the present invention, Y is selected from:

[0046] In some embodiments of the present invention, R1 is selected from:

[0047] In some embodiments of the present invention, R2 is H or a halogen (such as F).

[0048] More specifically, the compound TKI is compounds I-9, I-11, I-13, I-25, I-32, I-43, I-44, I-46, I-60, I-61, I-69, I-76, I-101, I-102, I-103, I-104, and I-106 as described in WO2024032584. In some embodiments of the present invention, the compound TKI has the following structure:

[0049] Specifically, the concentration of the compound TKI or a pharmaceutically acceptable salt thereof in the pharmaceutical composition is 0.01-50 mg / ml (e.g., 0.5, 5, 10, 12, 14, 15, 16, 18, 20, 25, 30, 35, 40, 45, 50 mg / ml), particularly 10-30 mg / ml, 10-20 mg / ml.

[0050] Preferably, the active ingredient is a pharmaceutically acceptable salt of the compound TKI, such as hydrochloride or phosphate.

[0051] Specifically, the pharmaceutical composition also contains a solvent, particularly water, such as water for injection.

[0052] Specifically, the pharmaceutical composition is a homogeneous formulation, such as a solution or emulsion, preferably eye drops. Further, the viscosity (at 25°C) of the solution or emulsion is 1-120 cp (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120 cp), for example, viscosities at 25°C of 1-100 cp, 1-80 cp, 1-60 cp, 1-50 cp, 1-40 cp, 5-50 cp, 5-40 cp; and viscosities at 37°C of 1-100 cp, 1-80 cp, 1-60 cp, 1-50 cp, 1-40 cp, 1-30 cp, 1-20 cp.

[0053] In some embodiments of the present invention, the pharmaceutical composition further includes an osmotic pressure regulator, such as glycerol, mannitol, 1,2-propanediol, glucose, sodium chloride, sorbitol, sodium citrate, etc.; the osmotic pressure regulator is added as needed, and further, the amount of the osmotic pressure regulator is such that the final osmotic pressure of the pharmaceutical composition is in the range of 250-450 mOsmol / kg (e.g., 260, 270, 280, 285, 290, 295, 300, 305, 310, 315, 320, 330, 340, 350, 360, 370, 400, 420, 440, 450 mOsmol / kg). In some embodiments of the invention, the osmotic pressure regulator is glycerol, which is present in the pharmaceutical composition at a concentration of 0.1%-0.5% (w / v, g / 100 ml) (e.g., 0.1%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.5%), particularly 0.2%-0.4%, e.g., 0.3%.

[0054] Specifically, the pH of the pharmaceutical composition is in the range of 5.0 to 9.0 (e.g., 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0), and more specifically in the range of 6.0 to 8.0, for example, around 6.5.

[0055] In some embodiments of the present invention, the pharmaceutical composition further includes a pH adjuster, such as borate, phosphate, bicarbonate, carbonate, citrate, tetraborate, hydrogen phosphate, tromethamine, hydroxyethylmorpholine, L-carnosine, etc.; in some embodiments of the present invention, the pH adjuster is tromethamine; in some embodiments of the present invention, the pH adjuster is L-carnosine. Further, the amount of the pH adjuster is such that the final pH of the pharmaceutical composition is 6.0-8.0, for example, around 6.5.

[0056] In some embodiments of the present invention, the pharmaceutical composition may further include preservatives such as sorbic acid, chlorobutanol, sodium chlorite, sodium perborate, quaternary ammonium salts, hydroxyphenyl esters, phenylmercuric nitrate, etc., to inhibit the growth and reproduction of microorganisms and ensure the stability and safety of the drug.

[0057] In some embodiments of the present invention, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of the compound TKI as the active ingredient, tylosap, hydroxypropyl-β-cyclodextrin, and a viscosity modifier.

[0058] In some embodiments of the present invention, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of a TKI compound as the active ingredient, teroxaprol, hydroxypropyl-β-cyclodextrin, and gellan gum; further, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of a TKI compound as the active ingredient, 0.5%-2% teroxaprol, 10%-25% hydroxypropyl-β-cyclodextrin, and 0.1%-0.2% gellan gum; in one embodiment of the present invention, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of a TKI compound as the active ingredient, 1% teroxaprol, 20% hydroxypropyl-β-cyclodextrin, and 0.15% gellan gum; even further, the pharmaceutical composition may further comprise L-carnosine as a pH adjuster; even further, the pharmaceutical composition may further comprise glycerol as an osmotic pressure adjuster.

[0059] In some embodiments of the present invention, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of compound TKI as the active ingredient, teroxaprol, hydroxypropyl-β-cyclodextrin, and sodium hyaluronate; further, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of compound TKI as the active ingredient, 0.5%-2% teroxaprol, 10%-25% hydroxypropyl-β-cyclodextrin, and 0.1%-0.4% sodium hyaluronate; in one embodiment of the present invention, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of compound TKI as the active ingredient, 1% teroxaprol, 20% hydroxypropyl-β-cyclodextrin, and 0.2% sodium hyaluronate; in one embodiment of the present invention, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of compound TKI as the active ingredient, 1% teroxaprol, 20% hydroxypropyl-β-cyclodextrin, and 0.2% sodium hyaluronate; in another embodiment of the present invention, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of compound TKI as the active ingredient, 0.5%-2% teroxaprol, 10%-25% hydroxypropyl-β-cyclodextrin, and 0.2% sodium hyaluronate. The pharmaceutical composition comprises: a pharmaceutically acceptable salt of a TKI as the active ingredient, 1% teroxaprol, 10% hydroxypropyl-β-cyclodextrin, and 0.2% sodium hyaluronate; in one embodiment of the invention, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of a TKI as the active ingredient, 1% teroxaprol, 20% hydroxypropyl-β-cyclodextrin, and 0.1% sodium hyaluronate; in another embodiment of the invention, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of a TKI as the active ingredient, 1% teroxaprol, 20% hydroxypropyl-β-cyclodextrin, and 0.3% sodium hyaluronate; further, the pharmaceutical composition may also include tromethamine as a pH adjuster; further still, the pharmaceutical composition may also include glycerol as an osmotic pressure adjuster.

[0060] In some embodiments of the present invention, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of compound TKI as the active ingredient, teroxaprol, hydroxypropyl-β-cyclodextrin, hydroxypropyl methylcellulose, and glycerin; further, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of compound TKI as the active ingredient, 0.5%-2% teroxaprol, 10%-25% hydroxypropyl-β-cyclodextrin, and 0.4%-0.6% hydroxypropyl methylcellulose; in one embodiment of the present invention, the pharmaceutical composition comprises: compound TKI The pharmaceutical composition comprises: a pharmaceutically acceptable salt of compound TKI as the active ingredient, 1% teroxaprol, 20% hydroxypropyl-β-cyclodextrin, and 0.5% hydroxypropyl methylcellulose; further, the pharmaceutical composition may further comprise tromethamine as a pH adjuster; further still, the pharmaceutical composition may further comprise glycerol as an osmotic pressure adjuster.

[0061] In some embodiments of the present invention, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of a compound TKI as an active ingredient, tylosap, hydroxypropyl-β-cyclodextrin, a viscosity modifier, and an osmotic pressure modifier.

[0062] In some preferred embodiments of the present invention, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of a TKI compound as the active ingredient, teroxaprol, hydroxypropyl-β-cyclodextrin, gellan gum, and glycerol; further, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of a TKI compound as the active ingredient, 0.5%-2% teroxaprol, 15%-25% hydroxypropyl-β-cyclodextrin, 0.1%-0.2% gellan gum, and 0.2%-0.4% glycerol; in one embodiment of the present invention, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of a TKI compound as the active ingredient, 1% teroxaprol, 20% hydroxypropyl-β-cyclodextrin, 0.15% gellan gum, and 0.3% glycerol; even further, the pharmaceutical composition may further comprise L-carnosine as a pH adjuster.

[0063] In some preferred embodiments of the present invention, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of a TKI compound as the active ingredient, teroxaprol, hydroxypropyl-β-cyclodextrin, sodium hyaluronate, and glycerin; further, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of a TKI compound as the active ingredient, 0.5%-2% teroxaprol, 15%-25% hydroxypropyl-β-cyclodextrin, 0.1%-0.4% sodium hyaluronate, and 0.2%-0.4% glycerin; in one embodiment of the present invention, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of a TKI compound as the active ingredient, 1% teroxaprol, 20% hydroxypropyl-β-cyclodextrin, 0.2% sodium hyaluronate, and 0.3% glycerin; even further, the pharmaceutical composition may further comprise tromethamine as a pH adjuster.

[0064] In some preferred embodiments of the present invention, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of a TKI compound as the active ingredient, teroxaprol, hydroxypropyl-β-cyclodextrin, hydroxypropyl methylcellulose, and glycerol; further, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of a TKI compound as the active ingredient, 0.5%-2% teroxaprol, 15%-25% hydroxypropyl-β-cyclodextrin, 0.4%-0.6% hydroxypropyl methylcellulose, and 0.2%-0.4% glycerol; in one embodiment of the present invention, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of a TKI compound as the active ingredient, 1% teroxaprol, 20% hydroxypropyl-β-cyclodextrin, 0.5% hydroxypropyl methylcellulose, and 0.3% glycerol; even further, the pharmaceutical composition may further comprise tromethamine as a pH adjuster.

[0065] In a second aspect of the invention, a method for preparing the pharmaceutical composition described in the first aspect is provided, comprising the step of mixing the components.

[0066] Specifically, the preparation method includes the following steps:

[0067] (1) Add solubilizer, surfactant and viscosity modifier to water to obtain a solution;

[0068] (2) Add the compound TKI or its pharmaceutically acceptable salt to the solution obtained in step (1).

[0069] Specifically, step (1) further includes heating and / or stirring to promote dissolution; more specifically, the heating temperature is 60-80°C, such as 70°C.

[0070] In some embodiments of the present invention, step (1) further includes adding an isotonic conditioner to the water.

[0071] Specifically, step (2) also includes the step of measuring the pH value of the solution, and if necessary, step (2) also includes adding a pH adjuster to adjust the pH value of the solution to a suitable value.

[0072] Specifically, the preparation method further includes step (3): adding water to the designed amount.

[0073] In a third aspect of the invention, the use of the pharmaceutical composition described in the first aspect in the preparation of a medicament for the prevention and / or treatment of diseases is provided.

[0074] Specifically, the disease is a proliferative disorder mediated by protein tyrosine kinases, particularly an eye disease.

[0075] In some embodiments of the present invention, the disease is an ocular disease, including, but not limited to, diseases accompanied by pathological neovascularization, retinal ischemia, retinal edema, and diabetic retinopathy.

[0076] In some embodiments of the present invention, the disease is an ocular disease, including, but not limited to, diabetic retinopathy (including simple (background) diabetic retinopathy, proliferative diabetic retinopathy, and diabetic macular edema); age-related macular degeneration (AMD) (including neovascular (wet / exudative) AMD, dry AMD, and geographic atrophy); pathological choroidal neovascularization (CNV) originating from any pathological mechanism (i.e., high myopia, trauma, sickle cell (anemia) disease; ocular histoplasmosis, angiostriate striae, traumatic choroidal rupture, optic nerve drusen, and certain retinal dystrophys); Pathological retinal neovascularization of any pathological mechanism (i.e., sickle cell retinopathy, Eales disease (retinal periphlebitis), ocular ischemia syndrome, carotid-cavernous fistula, familial exudative vitreoretinopathy, hyperviscosity syndrome, idiopathic arteritis obliterans, birdshot retinochoroidopathy, retinal vasculitis, sarcoidosis, or toxoplasmosis); uveitis; retinal vein occlusion (central or branch); cystoid macular edema; ocular ischemia; retinopathy of prematurity; Coat's disease; sickle cell retinopathy and / or neovascularizing glaucoma.

[0077] In some embodiments of the present invention, the eye disease is a posterior segment disease, that is, a disease occurring in the vitreous body, retina, choroid, sclera, or optic nerve.

[0078] In some embodiments of the present invention, the disease is diabetic retinopathy, including simple (background) diabetic retinopathy (non-proliferative diabetic retinopathy), proliferative diabetic retinopathy, and diabetic macular edema, particularly non-proliferative diabetic retinopathy.

[0079] In other embodiments of the invention, the disease is age-related macular degeneration (AMD), including neovascular (wet / exudative) AMD, dry AMD, and geographic atrophy.

[0080] In a fourth aspect of the invention, a pharmaceutical composition described in the first aspect is provided for the prevention and / or treatment of disease.

[0081] Specifically, the disease is as described in the third aspect of the present invention.

[0082] In a fifth aspect of the invention, a method of treating a disease is provided, comprising the step of administering the pharmaceutical composition of the first aspect of the invention to a subject in need of such treatment.

[0083] Specifically, the disease is as described in the third aspect of the present invention.

[0084] Specifically, the subjects are mammals, such as humans.

[0085] Specifically, the method of administration can be eye drops.

[0086] This invention provides a protein tyrosine kinase inhibitor pharmaceutical composition, its preparation method, and its application. The pharmaceutical composition is a liquid composition that can be used as eye drops for the treatment of related eye diseases. It exhibits good formulation properties, moderate viscosity, high stability, and high exposure to retinal tissue, which is beneficial for the active ingredient to exert its efficacy, demonstrating excellent application prospects. Detailed Implementation

[0087] Unless otherwise defined, all scientific and technical terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art.

[0088] The term "alkyl" refers to a straight-chain or branched hydrocarbon radical that does not contain unsaturated bonds and is connected to the rest of the molecule by single bonds. Typical alkyl groups contain 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) carbon atoms, preferably 1 to 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, n-hexyl, isohexyl, etc. In this invention, CO alkyl refers to H, i.e., C 0-10 Alkyl (or C0-C) 10 Alkyl groups include H and C. 1-10 Alkyl (or C1-C) 10 alkyl).

[0089] The term "alkylene" refers to a hydrocarbon group (divalent alkyl) formed by the loss of two hydrogen atoms from an alkane molecule. It can be straight-chain or branched and is connected to the rest of the molecule by a single bond. Typical alkylene groups described herein have 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) carbon atoms, preferably 1 to 6 carbon atoms, such as methylene (-CH2-), ethylene, propylene, butylene, etc. In this invention, CO alkylene refers to a single bond, i.e., C... 0-10 Alkylene (or C0-C) 10 Alkylenes include single bonds and C bonds. 1-10 Alkylene (or C1-C) 10 (alkylene).

[0090] The term "cycloalkyl" refers to alicyclic hydrocarbons, such as those containing 1 to 4 monocyclic and / or fused rings, containing 3 to 18 carbon atoms, preferably 3 to 10 (e.g., 3, 4, 5, 6, 7, 8, 9, 10) carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or adamantyl.

[0091] The term "alkoxy" refers to a substituent formed when the hydrogen in a hydroxyl group is replaced by an alkyl group, such as alkoxy groups containing 1-10 carbon atoms, such as methoxy, ethoxy, propoxy, butoxy, etc.

[0092] The term "alkylamine" refers to a substituent formed when one or two hydrogen atoms of an amino group (-NH2) are replaced by an alkyl group, such as an alkylamine group containing 1-10 carbon atoms, for example...

[0093] The term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0094] The term "haloalkyl" refers to a group formed by replacing one or more hydrogen atoms in an alkyl group with a halogen atom (such as fluorine, chlorine, bromine or iodine), such as -CHF2, -CH2F, -CF3, -CH2-CF3, -CH2CH2-CF3, -CH2CH2CH2-CF3.

[0095] The term "heterocyclic group" refers to a 3- to 18-membered non-aromatic ring group containing 2 to 17 carbon atoms and 1 to 10 heteroatoms. Heterocyclic groups can be monocyclic, bicyclic, tricyclic, or tetracyclic ring systems, and can include fused, spirocyclic, or bridged ring systems. Heterocyclic groups can be partially saturated (heteroaryl) or fully saturated (heterocyclic alkyl). Suitable heteroaryl groups in the compounds of the present invention contain one, two, or three heteroatoms selected from N, O, S, and P atoms. These heteroaryl groups include, for example, coumarin (including 8-coumarin), quinolinyl (including 8-quinolinyl, isoquinolinyl, pyridinyl, pyrazinyl, pyrazolyl, pyrimidinyl, furanyl, pyrroloyl, thiopheneyl, thiazolyl, isothiazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, imidazoleyl, indolyl, isoyndolyl, indazoleyl, inazinyl, phthalazinyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furazolidyl, pyridazinyl, triazinyl, cenolinyl, benzimidazolyl, benzofuranyl, benzofuranyl, benzothiopheneyl, benzothiazolyl, benzooxazolyl, quinazolinyl, quinoxolinyl, naphridinyl, and furanopyridinyl. Suitable heterocyclic alkyl groups in the compounds of the present invention contain one, two, or three heteroatoms selected from N, O, or S atoms. These heterocyclic alkyl groups include, for example, pyrrolidinyl, tetrahydrofuranyl, dihydrofuran, tetrahydrothiophenyl, tetrahydrothiophenyl, piperidinyl, morpholinyl, thiomorpholinyl, oxothiohexacyclohexyl, piperazine, aziridine, oxothiobutyl, thiohexacyclobutyl, homopiperidinyl, oxopropane, thiopropane, acrylonitrile, oxazinyl, diaziridine, etc. Heptyl, triacetyl, 1,2,3,6-tetrahydropyridyl, 2-pyrrolinyl, 3-pyrrolinyl, dihydroindolyl, 2H-pyranyl, 4H-pyranyl, dioxacyclohexyl, 1,3-dioxapentyl, pyrazolinyl, dithiaalkyl, dithiopentyl, dihydropyranyl, dihydrothiophenyl, pyrazolinyl, imidazolinyl, imidazolinyl, 3-azabicyclo[3.1.0]hexyl, 3-azabicyclo[4.1.0]heptyl, 3H-indolyl, and quinazinyl. In this invention, for optionally substituted heterocyclic groups, the substituted position can be any suitable carbon atom or heteroatom.

[0096] The term "pharmaceutically acceptable salt" generally refers to a salt that is safe and / or effective when used in mammals and has the intended biological activity. The compounds described in this invention can form salts with acids. Non-limiting examples of pharmaceutically acceptable salts include: hydrochloride, hydrobromide, hydroiodide, sulfate, hydrogen sulfate, citrate, succinate, ascorbate, oxalate, nitrate, phosphate, hydrogen phosphate, dihydrogen phosphate, salicylate, tartrate, maleate, fumarate, formate, acetate, propionate, benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, benzenesulfonate, etc.

[0097] The terms “patient” or “subject”, etc., may be used interchangeably herein to refer to any animal or its cells, whether in vitro or in situ, treated according to the methods described herein. Specifically, the aforementioned animals include mammals, such as rats, mice, guinea pigs, rabbits, dogs, monkeys, or humans, especially humans.

[0098] The term "treatment" refers to the prevention, cure, reversal, reduction, mitigation, minimization, suppression, cessation, and / or cessation of one or more clinical symptoms of a disease after its onset.

[0099] The term "prevention" refers to the treatment taken before a disease develops to avoid, minimize, or prevent the disease from developing or progressing.

[0100] All publications, patents, and published patent specifications cited in this article are incorporated herein in their entirety through citation.

[0101] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0102] Compound I described in the following examples has the following structure, and its preparation is based on Example 1 of PCT application WO2024032584:

[0103] Some of the instruments and equipment used in the following embodiments:

[0104] SevenExcellence pH meter (Mettler-Toledo)

[0105] DVNXLVCJG Viscometer (BROOKFIELD)

[0106] OM819.C Osmometer

[0107] The formulation property detection methods used in the following examples are as follows:

[0108] (1) pH value detection method:

[0109] Before using the SevienExcellence pH meter, it needs to be calibrated with a pH buffer solution. After the measurement begins, rinse the electrode probe with pure water and remove the water. Insert it below the surface of the sample liquid and press the reading key. After the reading stabilizes, the pH value of the sample is obtained.

[0110] (2) Viscosity testing method:

[0111] Turn on the DVNXLVCJG viscometer, set the water bath temperature to maintain constant experimental conditions, transfer 0.5 mL of liquid sample into the stirring cup, set the rotation speed to start the measurement, repeat the measurement 3 times, and take the average of the 3 test data as the viscosity test result.

[0112] (3) Osmotic pressure testing methods:

[0113] The OM819.C osmoremeter works by measuring the freezing point depression of a solution to determine its osmolar concentration. Before use, zero-point calibration and standard calibration must be performed using pure water and standard solution respectively. Before each use, the electrode probe must be cleaned with pure water and the water absorbed. Use a pipette to transfer 100 μL of sample into the sample tube, close the instrument, start the detection, and repeat the measurement three times. The average of the three measurements is taken as the osmolarity result.

[0114] Some of the reagents used in the following examples:

[0115] Sodium hyaluronate, unless otherwise specified, was purchased from Shandong Zhongshan Biotechnology Co., Ltd., product number SH06211201_04.

[0116] Hydroxypropyl methylcellulose (HPMC), unless otherwise specified, was purchased from Shanghai Carexon Coating Technology Co., Ltd., under the product name Methodox E5 PREMIUM LV (METHOCEL). TM E5 Premium LV).

[0117] Gellan gum, purchased from Luofu Pharmaceutical Technology, manufactured by CP Kelco, item number: 20052448.

[0118] Hydroxypropyl β-cyclodextrin, purchased from Luofu Pharmaceutical Technology, manufactured by Gangwal Healthcare, product number: KD-689.

[0119] Polyoxyethylene castor oil (ELP), purchased from Sigma, item number: 30906.

[0120] Unless otherwise stated, the percentage content of each component in the formulation composition tables of the following examples are all mass-volume ratios.

[0121] Example 1

[0122] The formulation of compound I was prepared according to the composition shown in the table below, and the preparation method is as follows:

[0123] Weigh 15g of hydroxypropyl-β-cyclodextrin, 1g of polyoxyethylene castor oil, and 1g of poloxamer 188 and add them to 50mL of water for injection. Stir magnetically until dissolved to obtain solution 1. Weigh 0.6g of tromethamine (Tris) and add it to 100mL of water for injection. Stir until dissolved and adjust the pH to 6.8 with HCl solution to obtain 50mM tromethamine-hydrochloric acid buffer. Weigh 0.3g of gellan gum and add it to the buffer (pre-heated to 70℃ in a water bath). Stir magnetically until dissolved to obtain solution 2. Transfer 2mL of 98% methanesulfonic acid to 28.6mL of water for injection and vortex to mix to obtain 1M methanesulfonic acid solution. Weigh 3.03g of tromethamine and add it to approximately 45mL of water for injection. Stir until dissolved and bring the volume to 50mL to obtain 0.5M tromethamine solution. Add the free compound I powder of the prescribed amount to solution 1 at 50% of the total volume (v / v), stir and mix well, and then sonicate to disperse. Next, add an appropriate volume of 1M methanesulfonic acid solution, stir and mix well, and sonicate until clear. Then, add solution 2 to dilute and immediately vortex to mix well. Finally, add an appropriate volume of 0.5M tromethamine solution to adjust the pH of the solution to about 7, and prepare a clear solution formulation with a concentration of 20 mg / mL.

[0124] Table 1. Formulation composition and state

[0125] The compound used in Example 1 is a free compound. Key control points in the formulation preparation process are: multiple pre-solutions need to be prepared for use; the compound cannot be dissolved by conventional stirring and multiple ultrasonic-assisted operations are required; the pH adjustment step requires first controlling the pH to the point where the compound dissolves, and then adjusting the pH to the target range. The preparation process is relatively complex and requires precise control of pH to avoid gelation during the process.

[0126] Examples 2-7

[0127] The formulation was prepared using compound I salt form according to the composition shown in the table below, and the preparation method is as follows:

[0128] The preparation methods for Examples 3, 4, 6, and 7 are the same, as detailed below:

[0129] Weigh 24.22 g of tromethorphan and add it to water for injection. Stir to dissolve and bring the volume to 100 mL to obtain a 2 M tromethorphan solution. Weigh 0.22 g of tylosap, 0.075 g of glycerol, 4 g of hydroxypropyl-β-cyclodextrin and viscosity modifier (Examples 3 and 6: weigh 20 mg of sodium hyaluronate; Examples 4 and 7: weigh 100 mg of hydroxypropyl methylcellulose) and add them to water for injection preheated to 70 °C. Vortex to dissolve and bring the volume to 20 mL to obtain solution 1. Weigh the amount of compound I salt into a 5 mL volumetric flask, add solution 1, and stir in a 60 °C water bath to dissolve. Measure the pH of the solution. If the pH is within the range of 6.5 ± 0.2, no adjustment is needed. If the pH is not within the range of 6.5 ± 0.2, it needs to be adjusted with 2 M tromethamine solution to 6.5 ± 0.2. Make up to 5 mL, vortex to mix, and prepare a clear solution with a concentration of 15 mg / mL.

[0130] The preparation methods for Example 2 and Example 5 are the same and similar to those described above, as detailed below:

[0131] Weigh 0.22 g tylosap, 0.075 g glycerol, 4 g hydroxypropyl-β-cyclodextrin, 30 mg gellan gum, and 12.71 mg carnosine, and add them to preheated water for injection at 70°C. Dissolve by vortexing and bring the volume to 20 mL to obtain solution 1. Weigh the amount of compound I salt (acid-base molar ratio 1:1) into a 5 mL volumetric flask, add solution 1, and dissolve by stirring in a 60°C water bath. Measure the pH of the solution; if it is within the range of 6.5 ± 0.2, bring the volume to 5 mL, vortex to mix, and prepare a clear solution with a concentration of 15 mg / mL.

[0132] Table 2. Formulation Composition and State

[0133] Viscosity measurements were performed on formulation samples at 25℃ and 37℃ and on samples after the addition of artificial tears (STF). The results are shown in Table 2.

[0134] In the above examples, all compounds were salt-type compounds. Examples 2, 3, and 4 were hydrochloride salts, while Examples 5, 6, and 7 were phosphate salts. The examples show that both salt-type compounds can be dissolved by stirring, easily prepared into clear solutions, and the process is simple and easy to implement.

[0135] Compared with the free compound in Example 1, the preparation process is significantly simplified, the salt-type compound is easy to dissolve, and shows higher solubility than in-situ salt formation.

[0136] In the above examples, different types of viscosity modifiers were used: gellan gum, sodium hyaluronate, and hydroxypropyl methylcellulose. Comparative results from Examples 3 and 4 show that sodium hyaluronate exhibits better viscosity than hydroxypropyl methylcellulose in hydrochloride-type compound formulations, which is related to its excellent water retention and pseudoplasticity. In Examples 2 and 5, the viscosity increased rapidly after the addition of artificial tears at 37°C, indicating that gellan gum gels quickly upon contact with metal cations, meeting the expected design.

[0137] Examples 8-11

[0138] The preparation methods for Examples 8 and 10 are the same, as detailed below:

[0139] Weigh 24.22 g of tromethamine and add it to water for injection. Stir to dissolve and bring the volume to 100 mL to obtain a 2M tromethamine solution. Weigh 0.075 g of glycerol, 30 mg of gellan gum, 0.22 g of tylosap, and 4 g of hydroxypropyl-β-cyclodextrin and add them to preheated water for injection at 70 °C. Vortex to dissolve and bring the volume to 20 mL to obtain solution 1. Weigh the required amount of compound I salt and add it to a 5 mL volumetric flask. Add solution 1 and stir to dissolve in a 60 °C water bath. Measure the pH of the solution. If the pH is within the range of 6.5 ± 0.2, no adjustment is needed. If the pH is not within the range of 6.5 ± 0.2, it needs to be adjusted with 2M tromethamine solution to 6.5 ± 0.2. Bring the volume to 5 mL, vortex to mix, and prepare a clear solution with a concentration of 15 mg / mL.

[0140] The preparation methods for Examples 9 and 11 are the same, as detailed below:

[0141] Weigh 24.22 g of tromethamine and add it to water for injection. Stir to dissolve and bring the volume to 100 mL to obtain a 2M tromethamine solution. Weigh 0.075 g of glycerol, 30 mg of gellan gum, 0.22 g of polyoxyethylene castor oil (ELP), and 2 g of sulfobutyl-β-cyclodextrin. Add them to preheated water for injection at 70 °C. Vortex to dissolve and bring the volume to 20 mL to obtain solution 1. Weigh the required amount of compound I salt and add it to a 5 mL volumetric flask. Add solution 1 and stir to dissolve in a 60 °C water bath. Measure the pH of the solution. If the pH is within the range of 6.5 ± 0.2, no adjustment is needed. If the pH is not within the range of 6.5 ± 0.2, it needs to be adjusted with 2M tromethamine solution to 6.5 ± 0.2. Bring the volume to 5 mL, vortex to mix, and prepare a clear solution with a concentration of 15 mg / mL.

[0142] Table 3. Formulation Composition and State

[0143] Viscosity measurements were performed on formulation samples at 25℃ and 37℃ and on samples after the addition of artificial tears (STF). The results are shown in Table 3.

[0144] Based on the results of Examples 10 and 11, when gellan gum is used as a viscosity modifier and tromethamine is used as a pH modifier, phosphate compounds are more prone to gelling. In addition, no gelling occurred in Example 5 when gellan gum and carnosine were used together, no gelling occurred in Example 6 when hydroxypropyl methylcellulose and tromethamine were used together, and no gelling occurred in Example 7 when sodium hyaluronate and tromethamine were used together. This may indicate that the use of phosphate compounds is limited when gellan gum and tromethamine are used together, and they are more sensitive to pH. Carnosine, as a pH modifier, can reduce the risk of gelling.

[0145] In Examples 8 and 9, the compounds used were hydrochloride salts, and the formulations were in solution form. After standing for 7 days, the solutions remained in solution form without gelation. Compared to phosphate salts, the formulations exhibited greater stability, wider applicability to excipients and pH levels, and a wider range of applicable formulations.

[0146] The above examples used hydroxypropyl-β-cyclodextrin and sulfobutyl-β-cyclodextrin respectively. Comparing the results of Examples 8 and 9, both were in solution state and showed a solubilizing effect.

[0147] In Example 9, when polyoxyethylene castor oil ELP was used in combination with sulfobutyl-β-cyclodextrin, its viscosity was close to that of artificial tears at 37°C, and it did not exhibit the ionic gel characteristics of gellan gum. In addition, according to literature reports, the solubilizing effect of tyloxapine is comparable to that of polyoxyethylene castor oil ELP, and long-term experiments show that it is safer.

[0148] Based on the results of the above embodiments, the surfactant is preferably tylosap, and the solubilizer is preferably hydroxypropyl-β-cyclodextrin.

[0149] Examples 12-17

[0150] The preparation method of Example 12 is as follows:

[0151] Weigh 24.22 g of tromethamine and add it to water for injection. Stir to dissolve and bring the volume to 100 mL to obtain a 2M tromethamine solution. Weigh 100 mg of hydroxypropyl methylcellulose and add it to preheated water for injection at 70°C. Vortex to dissolve and bring the volume to 20 mL to obtain solution 1. Weigh the prescribed amount of compound I hydrochloride and add it to a 5 mL volumetric flask. Add solution 1 and stir to dissolve in a 60°C water bath. Measure the pH value of the solution. If the pH value is within the range of 6.5 ± 0.2, no adjustment is needed. If the pH value is not within the range of 6.5 ± 0.2, it needs to be adjusted with 2M tromethamine solution to 6.5 ± 0.2. Bring the volume to 5 mL, vortex to mix, and prepare a clear solution with a concentration of 15 mg / mL.

[0152] The preparation methods of Examples 13, 14, 15, 16, and 17 are the same as those of Examples 2, 3, and 4.

[0153] Table 4. Formulation Composition and State

[0154] Viscosities of formulation samples at 25℃ and 37℃, as well as samples with added artificial tears (STF), were measured, and the results are shown in Table 4. The osmotic pressure of some formulation samples was also measured, and the results are shown in Table 4.

[0155] Example 12 shows that the hydrochloride-type compound was prepared into a basically clear, slightly turbid solution of 15 mg / mL without surfactants and solubilizers, confirming that the salt type has high solubility, which is in line with the design expectation.

[0156] Example 13 shows that by adding the surfactant tyloxapine and the solubilizer hydroxypropyl-β-cyclodextrin, a clear solution of 15 mg / mL can be prepared, confirming that the selected surfactant and solubilizer are preferred and meet expectations.

[0157] Example 14 yielded a clear solution at 20 mg / mL, while Examples 15 and 16 showed gelation at a concentration of 25 mg / mL, demonstrating the upper limit of the concentration of hydrochloride-type compounds.

[0158] In both Examples 15 and 16, gelling occurred when gellan gum was used. In Example 15, the amount of gellan gum used was 0.4% and the pH adjuster was tromethamine. In Example 16, the amount of gellan gum used was 0.15% and the pH adjuster was carnosine (as confirmed in Example 5, carnosine can reduce the risk of gelling by adjusting the pH). Gelling still occurred. Therefore, it can be determined that when gellan gum is used as a viscosity modifier, setting the upper limit of the concentration of hydrochloride compounds at 20 mg / mL is more reasonable.

[0159] In Example 17, sodium hyaluronate was used as the viscosity modifier, which could prepare a clear solution of 25 mg / mL. No gelation occurred after 7 days of standing, indicating good stability. This suggests that sodium hyaluronate is another preferred viscosity modifier. Viscosity testing showed that its viscosity was moderate, but the osmotic pressure was higher than the upper limit of 450 mOsmol / kg, indicating room for further optimization. This could be improved by adjusting the amount of glycerol used as the osmotic pressure modifier.

[0160] Examples 18-25

[0161] The formulation was prepared using compound I hydrochloride according to the composition shown in the table below, and the preparation method was carried out in accordance with Examples 3 and 4.

[0162] Table 5. Formulation Composition and State

[0163] Sodium hyaluronate 0.2M, purchased from Aladdin, item number: H293507; Sodium hyaluronate 1M, purchased from Shandong Zhongshan Biotechnology Co., Ltd., item number: SH06211201_04; HPMC 400cp, purchased from Aladdin, item number: H108819.

[0164] Examples 18 and 19 were prepared using different viscosity modifiers, resulting in suitable final viscosity and osmotic pressure. A clear solution of 30 mg / mL compound I hydrochloride was obtained and maintained for at least one month. Example 20, based on Example 19, attempted to reduce the hydroxypropyl β-cyclodextrin to 10%, also achieving good solubility and suitable viscosity. The osmotic pressure could be adjusted to near physiological conditions by adding an appropriate osmotic pressure modifier (such as glycerol). Examples 21 and 22, based on Example 18, reduced or increased the sodium hyaluronate content, resulting in a linear change in viscosity and minimal change in osmotic pressure. Examples 23 to 25 adjusted the proportion of hydroxypropyl β-cyclodextrin. The results showed that 5% hydroxypropyl β-cyclodextrin was insufficient to achieve a clear formulation, indicating that at least 10% hydroxypropyl β-cyclodextrin was suitable.

[0165] Example 26: Animal Experiment

[0166] Experimental plan:

[0167] Male Dutch Black-banded rabbits were purchased from Pizhou Oriental Breeding Co., Ltd., and the animals weighed 1.5-2.5 kg. All procedures performed on the experimental animals in this experiment complied with the relevant Standard Operating Procedures (SOPs) of Suzhou Shengsu New Drug Development Co., Ltd., and were approved by the Laboratory Animal Use Management Committee (IACUC) of Suzhou Shengsu New Drug Development Co., Ltd.

[0168] Solution A is the formulation prepared in Example 2 above. Solution B is the formulation obtained by slight adjustment based on Example 3 (the sodium hyaluronate content was adjusted to 0.2%, while everything else remained the same as in Example 3). Solution C is the formulation prepared in Example 12 above.

[0169] On the day of the experiment, animals were administered the drug at a dose of 15 mg / mL (0.75 mg / eye, 50 μL / eye). Two animals were in each group. At 0.25, 0.5, 1, 2, 4, 12, and 24 hours after drug administration, the animals were sacrificed, and blood and ocular tissues (cornea, conjunctiva, retina, choroid, sclera, and iris) were collected. One mL of collected whole blood was placed in an EDTA-K2 anticoagulant tube, thoroughly mixed by inverting several times, and centrifuged (1500–1600 g) for 10 minutes within 30 minutes to separate the plasma. The resulting plasma sample was stored at -80°C for biological sample analysis. Ocular tissues were cleaned with pre-cooled physiological saline, dried, and stored at -80°C or under equivalent conditions for subsequent tissue homogenization and sample analysis. For ocular tissue requiring homogenization, all or part of the tissue was selected, accurately weighed, and homogenized with 20% methanol-water at a 1:10 ratio. The tissue homogenate was stored at -80°C for sample analysis. The concentrations of analytes in rabbit ocular tissue and plasma obtained in this experiment were determined using LC-MS / MS. The matrix consisted of Dutch rabbit plasma (EDTA-K2 anticoagulant) and rabbit ocular tissue (1:10, 20% methanol-water). Samples from the left and right eyes were tested separately. The BLOQ (lower limit of quantitation) is the lowest analyte concentration quantified with acceptable precision and accuracy. Compounds were diluted 1:10 with the homogenate before analysis; if the concentration was less than 0.2 ng / ml, it was labeled with the BLOQ.

[0170] The results of the exposure of solutions A, B, and C to the eye tissues of experimental animals are shown in the table below.

[0171] Table 6 Comparison of rabbit eye tissue exposure levels of different formulations Solution A: 20% hydroxypropyl β-cyclodextrin, 1% teroxaprol, 0.3% glycerol, 0.15% gellan gum, 7.5 mM L-carnosine; Solution B: 20% hydroxypropyl β-cyclodextrin, 1% teroxaprol, 0.3% glycerol, 0.2% sodium hyaluronate; Solution C: 20% hydroxypropyl β-cyclodextrin, 1% teroxaprol, 0.3% glycerol, 0.5% hydroxypropyl methylcellulose.

[0172] The results show that solutions using gellan gum, sodium hyaluronate, or hydroxypropyl methylcellulose can achieve good exposure of fundus tissues. Solutions A and B are more effective for the cornea, conjunctiva, and iris, while solution C is more effective for the retina and choroid.

[0173] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

[0174] The foregoing embodiments and methods described in this invention may vary based on the capabilities, experience, and preferences of those skilled in the art.

[0175] Listing the steps of the method in a certain order in this invention does not constitute any restriction on the order of the method steps.

Claims

1. A pharmaceutical composition, which is a liquid composition, comprising: a compound TKI or a pharmaceutically acceptable salt thereof as an active ingredient, a surfactant, a solubilizer, and a viscosity modifier; wherein the compound TKI is a protein tyrosine kinase inhibitor; Preferably, the surfactant is selected from one or more of the following: polyoxyethylene castor oil, poloxamer, tyloxaparin, Tween, and polyoxyethylene 40 hydrogenated castor oil, especially tyloxaparin; Preferably, the solubilizer is a cyclodextrin, more preferably a hydroxyalkyl-β-cyclodextrin, especially a hydroxypropyl-β-cyclodextrin.

2. The pharmaceutical composition of claim 1, wherein, The surfactant has a mass-volume concentration of 0.1%-5% in the pharmaceutical composition, preferably 0.5%-2%, and more preferably 1%.

3. The pharmaceutical composition of claim 1, wherein The solubilizer has a mass-volume concentration of 0.5%-30% in the pharmaceutical composition, preferably 10%-25% or 15%-25%, and more preferably 20%.

4. The pharmaceutical composition according to any one of claims 1 to 3, wherein The viscosity modifier is selected from: gellan gum, hyaluronic acid or its salt, and hydroxypropyl methylcellulose; Preferably, the viscosity modifier is gellan gum, and its concentration in the pharmaceutical composition is 0.01%-1%, more preferably 0.1%-0.2%, and more preferably 0.15%; or, Preferably, the viscosity modifier is hyaluronic acid or a salt thereof, and its concentration in the pharmaceutical composition is 0.05%-1%, preferably 0.1%-0.4%, more preferably 0.2%; or, Preferably, the viscosity modifier is hydroxypropyl methylcellulose, and its concentration in the pharmaceutical composition is 0.1%-5%, more preferably 0.4%-0.6%, and more preferably 0.5%.

5. The pharmaceutical composition of claim 1, wherein The compound TKI has the following structure: in, R1 is selected from: -C(O)NR 101 R 102 -(C0-C6 alkylene)-NR 101 R 102 -C(O)R 101 -C(O)OR 101 R 101 and R 102 Independently selected from: H, C1-C6 alkyl, -(C0-C6 alkylene)-(C3-C6 cycloalkyl); wherein the C0-C6 alkylene, C1-C6 alkyl, and C3-C6 cycloalkyl may optionally be substituted by one or more groups selected from: halogen, cyano, amino, hydroxyl, C1-C6 alkyl, and C1-C6 haloalkyl; R2 is one or more independent substituents on the benzene ring, selected from: H, halogen, cyano, amino, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl; L is a single bond or a C1-C6 alkylene group; Y is selected from: H, -NR7R8, or optionally substituted nitrogen-containing heterocyclic groups, wherein the nitrogen-containing heterocyclic group may optionally be substituted by one or more R9 groups; R7 and R8 are independently selected from: H, C1-C6 alkyl, -(C0-C6 alkylene)-(C3-C6 cycloalkyl); R9 is selected from: H, C1-C6 alkyl, C1-C6 hydroxy-substituted alkyl, C1-C6 amino-substituted alkyl, C1-C6 alkoxy-substituted alkyl, C1-C6 alkylamino-substituted alkyl, -(C0-C6 alkylene)-(C3-C6 cycloalkyl); Preferably, the compound TKI is selected from the following structures: Preferably, the concentration of the compound TKI or its pharmaceutically acceptable salt in the pharmaceutical composition is 0.01-50 mg / ml, more preferably 10-30 mg / ml or 10-20 mg / ml.

6. The pharmaceutical composition of claim 5, wherein The active ingredient is a pharmaceutically acceptable salt of the compound TKI, such as a hydrochloride or phosphate.

7. The pharmaceutical composition according to any one of claims 1 to 6, wherein The pharmaceutical composition further includes an osmotic pressure regulator and / or a pH regulator; Preferably, the osmotic pressure regulator is selected from: glycerol, mannitol, 1,2-propanediol, glucose, sodium chloride, sorbitol, and sodium citrate; Preferably, the pH adjuster is selected from: borate, phosphate, bicarbonate, carbonate, citrate, tetraborate, hydrogen phosphate, tromethamine, hydroxyethylmorpholine, and L-carnosine.

8. The pharmaceutical composition of claim 1, wherein The pharmaceutical composition comprises: a pharmaceutically acceptable salt of a compound TKI as the active ingredient, 0.5%-2% tylosap, 10%-25% hydroxypropyl-β-cyclodextrin, and 0.1%-0.2% gellan gum; or, The pharmaceutical composition comprises: a pharmaceutically acceptable salt of a compound TKI as the active ingredient, 0.5%-2% tylosap, 10%-25% hydroxypropyl-β-cyclodextrin, and 0.1%-0.4% sodium hyaluronate; or, The pharmaceutical composition comprises: a pharmaceutically acceptable salt of compound TKI as the active ingredient, 0.5%-2% tylosap, 10%-25% hydroxypropyl-β-cyclodextrin, and 0.4%-0.6% hydroxypropyl methylcellulose; Preferably, the pharmaceutical composition comprises: a pharmaceutically acceptable salt of a compound TKI as the active ingredient, 1% tylosap, 20% hydroxypropyl-β-cyclodextrin, 0.15% gellan gum, and 0.3% glycerol; or, The pharmaceutical composition comprises: a pharmaceutically acceptable salt of a compound TKI as the active ingredient, 1% tylosap, 20% hydroxypropyl-β-cyclodextrin, 0.2% sodium hyaluronate, and 0.3% glycerin; or, The pharmaceutical composition comprises: a pharmaceutically acceptable salt of compound TKI as the active ingredient, 1% tylosap, 20% hydroxypropyl-β-cyclodextrin, 0.5% hydroxypropyl methylcellulose, and 0.3% glycerol.

9. A method for preparing the pharmaceutical composition according to any one of claims 1-8, comprising the following steps: (1) Add solubilizer, surfactant and viscosity modifier to water to obtain a solution; (2) Add the compound TKI or its pharmaceutically acceptable salt to the solution obtained in step (1).

10. Use of the pharmaceutical composition according to any one of claims 1-8 in the preparation of a medicament for the prevention and / or treatment of diseases, wherein, The disease is a proliferative disorder mediated by protein tyrosine kinase, particularly an ocular disease; Preferably, the disease is selected from: diabetic retinopathy, age-related macular degeneration (AMD), pathological choroidal neovascularization (CNV), pathological retinal neovascularization, uveitis, retinal vein occlusion, cystoid macular edema, ocular ischemia, retinopathy of prematurity, Coxs disease, sickle cell retinopathy, and neovascular glaucoma. Preferably, the eye disease is a posterior segment eye disease.

Images