Salt form of protein tyrosine kinase inhibitor

Abstract

Provided are a salt form of a protein tyrosine kinase inhibitor, a preparation method therefor, and use thereof. The protein tyrosine kinase inhibitor has the ability to inhibit an anti-angiogenesis tyrosine kinase. In addition to effectively antagonizing VEGFR1, VEGFR2, and VEGFR3 tyrosine kinase activities, the protein tyrosine kinase inhibitor has high selectivity for inhibiting EGFR tyrosine kinase activity, and can effectively reduce and avoid side effects, especially the side effects on the eyes, making it safer to use. Compared with a free base of the protein tyrosine kinase inhibitor, the aforementioned salt form of the protein tyrosine kinase inhibitor has significantly improved solubility, relatively high solid stability, and relatively low hygroscopicity, and has very good application value in drug development and industrialization.

Description

Salt form of a protein tyrosine kinase inhibitor TECHNICAL FIELD

[0001] The present application relates to the field of biochemical medicine, in particular to a salt form of a protein tyrosine kinase inhibitor, its preparation method and application. BACKGROUND

[0002] Receptor tyrosine kinases play an important role in developmental biology, tissue homeostasis and cancer biology. Receptor tyrosine kinases consist of an extracellular ligand-binding domain, a transmembrane domain and an intracellular catalytic domain. Dimerization of two receptor tyrosine kinases after ligand binding leads to autophosphorylation of tyrosine residues in the intracellular catalytic domain, which leads to the active conformation and subsequent activation of the intracellular signal transduction cascade. Due to its important influence on cells, tyrosine kinases are highly regulated. When these kinases are constitutively activated by mutation or overexpression and are independent of ligand, many diseases (such as cancer or diabetic retinopathy and other ophthalmic diseases) will develop through unregulated cell proliferation and other mechanisms. For this reason, tyrosine kinase inhibitors can be used to treat some diseases by interfering with this unregulated process. The development of inhibitors targeting pro-angiogenic receptor tyrosine kinases (mainly the vascular endothelial growth factor receptor (VEGFR) family) (RTKi) has significantly improved the prognosis of some types of cancer (such as renal cell carcinoma, hepatocellular carcinoma and colorectal cancer), becoming an effective treatment for tumor-related angiogenesis.

[0003] Diabetic retinopathy (DR) and age-related macular degeneration (AMD) are the leading causes of blindness worldwide. These pathologies are associated with neovascularization in the posterior segment of the eye, with DR preferentially presenting changes in neovascularization at the retinal level, while the hallmark of wet AMD is the formation of new blood vessels from the choroidal microvascular bed and invasion into the subretinal space. Both DR and AMD are characterized by endothelial cell (EC) proliferation and migration, increased vascular permeability, and inflammation. Vascular endothelial growth factor-A (VEGF-A) and its corresponding receptors (VEGFRs) play a key role in these processes. Many proliferative disorders, such as ophthalmic diseases, tumors, and cancers, involve overexpression or upregulation of receptor tyrosine kinase (RTK) activity. Receptor tyrosine kinases are a class of kinase enzymes that modify proteins by chemically adding a phosphate group (phosphorylation). Phosphorylation often leads to functional changes in the target protein 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 of the ways to inhibit the VEGF pathway is to inhibit receptor tyrosine kinase (RTK) activity. In the treatment of ocular diseases such as diabetic retinopathy (DR) and age-related macular degeneration (AMD), the goal of protein tyrosine kinase inhibitor therapy is to counteract pathological neovascularization and disease progression, preventing visual impairment. At the same time, the importance of VEGFR as a pro-angiogenic inducer in tumor growth, invasion, and extravasation makes it an excellent therapeutic target for a variety of cancers. However, existing VEGFR receptor tyrosine kinase inhibitors (RTKi) often also inhibit EGFR activity. This cross-reactivity can lead to adverse side effects by inhibiting biological functions associated with one or more of these off-target receptors. This problem greatly affects the application of VEGFR receptor tyrosine kinase inhibitors in the treatment of ophthalmic diseases. SUMMARY

[0004] To overcome the deficiencies of the prior art, the inventors prepared a protein tyrosine kinase inhibitor compound (as described in patent application PCT / CN2023 / 142499, which is incorporated by reference in its entirety), and on this basis, further studied the salt type of the compound, obtaining the technical solution of the present application.

[0005] Specifically, the technical solution of the present application is as follows:

[0006] In the first aspect of the present application, a salt type of a compound having the following structure is provided, the salt type being selected from the group consisting of hemi-sulfate, hemi-citrate, hemi-fumarate, hemi-succinate, hemi-L-tartrate, hemi-adipate,

[0007] wherein,

[0008] R 1b selected from: C1-C6haloalkyl, cyano, nitro, azido, -OR 101 , -C(O)R 101 , -C(O)OR 101 , -NHC(O)OR 101 , -OC(O)R 101 , -NHSO2R 101 , -SO2NR 101 R 102 , -NHC(O)R 101 , -C(O)NR 101 R 102 , -NR 101 R 102 , -SR 101 , -S(O)2R 101 , -SO3H, -(Co-C6alkylene)-(phenyl), -(Co-C6alkylene)-(4-10 membered heterocyclyl), -(Co-C6alkylene)-(C3-C 10 cycloalkyl);

[0009] R 1c selected from: H, F, C1-C6alkyl, cyano, nitro, azido, -OR 101 , -C(O)R 101 , -C(O)OR 101 , -NHC(O)OR 101 , -OC(O)R 101 , -NHSO2R 101 , -SO2NR 101 R 102 , -NHC(O)R 101 , -C(O)NR 101 R 102 , -NR 101 R 102 , -SR 101 , -S(O)2R 101 , -SO3H, -(Co-C6alkylene)-(phenyl), -(Co-C6alkylene)-(4-10 membered heterocyclyl), -(Co-C6alkylene)-(C3-C 10 cycloalkyl);

[0010] R 1a , R 1d , R 1e are independently selected from: H, C1-C6alkyl, halogen, C1-C6haloalkyl, cyano, nitro, azido, -OR 101 , -C(O)R 101 , -C(O)OR101 -NHC(O)OR 101 -OC(O)R 101 -NHSO2R 101 -SO2NR 101 R 102 -NHC(O)R 101 -C(O)NR 101 R 102 -NR 101 R 102 -SR 101 -S(O)2R 101 -SO3H, -(Co-C6 alkylene)-(phenyl), -(Co-C6 alkylene)-(4-10 membered heterocyclyl), -(Co-C6 alkylene)-(C3-C 10 cycloalkyl);

[0011] each R 101 and R 102 is independently selected from the group consisting of H, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxysubstituted alkyl, C1-C6 alkoxy- substituted alkyl, C1-C6 aminosubstituted alkyl, C1-C6 alkylamino- substituted alkyl, C3-C 10 cycloalkyl, C4-C 10 cycloalkylalkyl, substituted or unsubstituted 4-10 membered heterocyclyl, phenyl, benzyl, C1-C 10 silyl-substituted alkyl, C1-C 10 silyl;

[0012] L2 is Co-C 10 alkylene, wherein one or more H atoms in the alkylene are optionally and independently substituted with C1-C6 alkyl, halogen, cyano, nitro, trifluoromethyl, trifluoromethoxy, azido, hydroxy, C1-C6 alkoxy, C3-C6 cycloalkyl, C4-C 10 cycloalkylalkyl;

[0013] R0 is selected from the group consisting of H, C1-C 10 alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl, halogen, cyano, nitro, trifluoromethyl, trifluoromethoxy, azido, -OR 001 -C(O)R 001 -C(O)OR 001 -NR 002 C(O)OR 001 -OC(O)R 001 -NR 002 SO2R 001 -SO2NR 001R 002 -NR 002 C(O)R 001 -C(O)NR 001 R 002 -NR 001 R 002 -S(O) i R 002 Where i is an integer from 0 to 2, -SO3H, -NR 002 (CR 003 R 004 ) t OR 001 , t is an integer from 1 to 5;

[0014] Among them, ring E is C6-C 10 Aromatic rings or 4-10 quintone heterocyclic rings;

[0015] R2 is selected from: H, =O, Cl-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne, halogen, cyano, nitro, trifluoromethyl, trifluoromethoxy, azide, -OR 201 -C(O)R 201 -C(O)OR 201 -NR 202 C(O)OR 201 -OC(O)R 201 -NR 202 SO2R 201 -SO2NR 201 R 202 -NR 202 C(O)R 201 -C(O)NR 201 R 202 -NR 201 R 202 -S(O) i R 201 Where i is an integer from 0 to 2, -SO3H, -(C0-C6 alkylene)-(C6-C 10 aryl), -SO2-(C0-C6 alkylene)-(C6-C 10 aryl), -S-(C0-C6 alkylene)-(C6-C 10 aryl), -O-(C0-C6 alkylene)-(C6-C 10aryl), -(C0-C6alkylene)-(4-10 membered heterocyclyl), -SO2-(C0-C6alkylene)-(4-10 membered heterocyclyl), -S-(C0-C6alkylene)-(4-10 membered heterocyclyl), -O-(C0-C6alkylene)-(4-10 membered heterocyclyl), -(C0-C6alkylene)-(C3-C 10 cycloalkyl), -SO2-(C0-C6alkylene)-(C3-C 10 cycloalkyl), -S-(C0-C6alkylene)-(C3-C 10 cycloalkyl), -O-(C0-C6alkylene)-(C3-C 10 cycloalkyl); wherein said C1-C 10 alkyl, C6-C 10 aryl, 4-10 membered heterocyclyl are optionally substituted with a member selected from halogen, cyano, nitro, azido, -OR', -C(O)R', -C(O)OR', -OC(O)R', -NR'C(O)OR", -NR'SO2R", -SO2NR'R", -NR'C(O)R", -C(O)NR'R", -NR'R", -SR', -SOR', -SO2R', -SO3H, C1-C 10 cycloalkyl, phenyl, 4-10 membered heterocyclyl;

[0016] n is an integer from 1-5 (e.g., 1, 2, 3, 4, 5, if valence allows);

[0017] each R 001 to R 004 are independently selected from the group consisting of: H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkylalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted heterocyclylalkyl, C1-C 10 silyl, wherein said alkyl, cycloalkyl, heterocyclyl are optionally substituted with a member selected from halogen, cyano, nitro, azido, -OR', -C(O)R', -C(O)OR', -OC(O)R', -NR'C(O)OR", -NR'SO2R", -SO2NR'R", -NR'C(O)R", -C(O)NR'R", -NR'R", -SR', -SOR', -SO2R', -SO3H, C1-C 10 alkyl, C1-C 10 haloalkyl, C1-C 10 silyl, C3-C 10 cycloalkyl, phenyl, 4-10 membered heterocyclyl;

[0018] each R 201 to R 204independently selected from the group consisting of: H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkylalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted heterocyclylalkyl, C1-C 10 silyl, wherein said alkyl, cycloalkyl, heterocyclyl are optionally substituted with a group selected from halogen, cyano, nitro, azido, -OR', -C(O)R', -C(O)OR', -OC(O)R', -NR'C(O)OR", -NR'SO2R", -SO2NR'R", -NR'C(O)R", -C(O)NR'R", -NR'R", -SR', -SOR', -SO2R', -SO3H, C1-C 10 alkyl, C1-C 10 haloalkyl, C1-C 10 silyl, C3-C 10 cycloalkyl, phenyl, 4-10 membered heterocyclyl;

[0019] R' and R" are independently selected from the group consisting of: H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkylalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted heterocyclylalkyl.

[0020] In particular, the molar ratio of the compound (free base) to the acid in the salt form is 1 : 0.4-0.6, in particular 1 : 0.45-0.55, 1 : 0.5.

[0021] In particular, the salt form can be amorphous or crystalline.

[0022] In particular, each R 101 and R 102 may be independently selected from the group consisting of: H, methyl, ethyl, n-propyl, i-propyl, -CF3, -CHF2, -CH2F,

[0023] More particularly, each R 101 and R 102 are independently selected from the group consisting of: H, methyl, ethyl, n-propyl, i-propyl, -CF3, -CHF2, -CH2F,

[0024] In some embodiments of the application, R 102 is H.

[0025] In particular, R 1b may be selected from the group consisting of: -CF3, -CHF2, -CH2F, cyano, nitro, azido, -OH,

[0026] In some embodiments of the application, R 1b is -C(O)NR 101 R 102 , wherein R 101 and R 102 are independently selected from the group consisting of H, C1-C6 alkyl, C3-C6 cycloalkyl, C4-C 10 cycloalkylalkyl.

[0027] In particular, R 1a , R 1e may be independently selected from the group consisting of H, halogen (such as F, Cl), C1-C3 alkyl (e.g. methyl).

[0028] In particular, R 1c may be selected from the group consisting of H, F.

[0029] In particular, R 1d may be selected from the group consisting of H, halogen (such as F, Cl), C1-C3 alkyl (e.g. methyl).

[0030] In some embodiments of the application, the compound has the following structure:

[0031] In particular, in formula V, R 101 is selected from the group consisting of H, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxy-substituted alkyl, C3-C6 cycloalkyl, C4-C 10 cycloalkylalkyl; in some embodiments of the application, R 101 is selected from the group consisting of H, C1-C3 alkyl (such as methyl, ethyl, n-propyl, i-propyl), C3-C6 cycloalkyl (such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl).

[0032] In particular, L2 is C0-C6 alkylene, such as C2-C6 straight chain alkylene.

[0033] In some embodiments of the application, R0 is wherein E ring is a 4-10 membered (e.g. 4, 5, 6, 7, 8, 9, 10 membered) heterocyclic ring, in particular a 4-8 membered saturated nitrogen-containing heterocyclic ring (including monocyclic, polycyclic, such as fused, spiro or bridged polycyclic). In some embodiments of the application, moiety is

[0034] In particular, n is 1 or 2.

[0035] ​In particular, R2is selected from: H, halogen, cyano, nitro, azido, hydroxyl, C1-C6alkyl, C1-C6haloalkyl, C1-C6hydroxysubstituted alkyl, C1-C6alkoxysubstituted alkyl, C3-C6cycloalkyl, C4-C 10 cycloalkylalkyl, -C(O)(C1-C6alkyl), -C(O)O(C1-C6alkyl).

[0036] More particularly, R2is selected from: H, methyl, ethyl, n-propyl, i-propyl, -CF3, -CHF2, -CH2F, F, Cl, Br, I, cyano, nitro, azido, hydroxyl, methoxy, ethoxy,

[0037] In some embodiments of the application, R0is selected from:

[0038] In some embodiments of the application, R0is -NR 001 R 002 wherein R 001 and R 002 are independently selected from: H, C1-C6alkyl, C1-C6haloalkyl, C1-C6hydroxysubstituted alkyl, C1-C6alkoxysubstituted alkyl, C1-C6aminosubstituted alkyl, C1-C6alkaminosubstituted alkyl, C3-C6cycloalkyl, C4-C 10 cycloalkylalkyl; in particular, R 001 and R 002 are independently selected from: H, methyl, ethyl, n-propyl, i-propyl, -CF3, -CHF2, -CH2F, In some embodiments of the application, R0is selected from:

[0039] In other embodiments of the application, R0is selected from: H, cyano, -O(C0-C 10 alkyl), -O(C1-C 10 silyl), -S(C0-C 10 alkyl), -C(O)(C0-C 10 alkyl), -C(O)O(C0-C 10 alkyl), -OC(O)(C0-C 10 alkyl), -N(C0-C 10 alkyl)SO2(C0-C 10 alkyl), -SO2N(C0-C 10 alkyl)(C0-C 10alkyl), -N(Co-C 10 alkyl), -N(Co-C 10 alkyl), -N(Co-C 10 alkyl), -N(Co-C 10 alkyl), -N(Co-C 10 alkyl), -N(Co-C H, cyano, -OH,

[0040] In some embodiments of the application, the compound has the following structure:

[0041] In some embodiments of the application, the compound has the following structure:

[0042] (the preparation and activity test of the compound, except T078, T116, are as described in patent application PCT / CN2023 / 142499, which is incorporated by reference in its entirety).

[0043] In the second aspect of the application, a preparation method of the salt form of the first aspect is provided, which comprises the following steps: mixing the compound with an acid, adding an organic solvent.

[0044] Specifically, the molar ratio of the compound to the acid is about 1:0.4-0.9, for example, 1:0.5, 1:0.55, 1:0.6, 1:0.65, 1:0.7, 1:0.8.

[0045] Specifically, the organic solvent is selected from acetone, methanol, ACN, and particularly methanol.

[0046] Specifically, the preparation method further comprises: suspending the obtained mixture, and then separating the obtained suspension to obtain a solid, and drying the obtained solid.

[0047] Specifically, the suspension step comprises: suspending the obtained mixture at 40-60°C (for example, 45, 48, 50, 52, 55°C) (1-4 hours, for example, 2 hours), and then cooling to 20-30°C (for example, 25°C) and suspending (for example, at least 48 hours).

[0048] Specifically, the solid separation can employ filtration, centrifugation, or a combination of both.

[0049] In some embodiments of the present application, the filtration step comprises passing the resulting suspension through a 0.45 μιη nylon filter membrane.

[0050] In some embodiments of the present application, the centrifugation step comprises centrifuging the resulting filtrate at a speed of 14000 rpm.

[0051] Specifically, the drying is vacuum drying, for example, vacuum drying at 45-60 °C (e.g., 45, 48, 50, 52, 55 °C) (1-4 hours, e.g., 2 hours).

[0052] In a third aspect of the present application, there is provided a pharmaceutical composition comprising the salt form of the first aspect, and one or more pharmaceutically acceptable excipients.

[0053] Specifically, the pharmaceutically acceptable excipients can be selected from one or more of disintegrants, binders, lubricants, suspending agents, stabilizers, fillers, absorption promoters, surfactants, flavoring agents, antioxidants, preservatives, pH adjusting agents, osmotic pressure adjusting agents, buffers, solubilizers, and the like.

[0054] Specifically, in the pharmaceutical composition, the salt form of the first aspect can be used alone, or in combination with other kinds of active ingredients.

[0055] Specifically, the pharmaceutical composition can employ any suitable administration route, for example, gastrointestinal administration (e.g., oral, rectal administration) or non-gastrointestinal administration (e.g., intravenous, intramuscular, intranasal, intraocular, intracerebral, intravaginal, intraperitoneal, transdermal, subcutaneous, intradermal, respiratory tract administration, etc.). In some embodiments of the present application, the pharmaceutical composition employs an intraocular administration route (e.g., intraocular administration, ocular surface administration, periocular administration, specifically, intravitreal injection administration, anterior chamber injection, intravitreal implant administration; eye drop administration, eye ointment administration, eye gel administration; subconjunctival injection administration, retrobulbar injection, periocular administration, subtenon administration, etc.).

[0056] Specifically, the pharmaceutical composition can be in any suitable dosage form, for example, a gastrointestinal administration dosage form, for example, including, but not limited to, a tablet, a pill, a powder, a granule, a capsule, a lozenge, a syrup, a liquid, an emulsion, a suspension, etc.; a non-gastrointestinal administration dosage form, for example, an injection administration dosage form: such as an injection (for example, for subcutaneous injection, intravenous injection, intramuscular injection, intraperitoneal injection), a respiratory administration dosage form: such as a spray, an aerosol, a powder spray, etc., a skin administration dosage form: such as an external solution, a lotion, an ointment, a plaster, a paste, a patch, etc., a mucous membrane administration dosage form: such as an eye drop, an eye ointment, a nose drop, a gargle, a sublingual tablet, etc., a cavity administration dosage form: such as a suppository, an aerosol, an effervescent tablet, a drop, a drop pill, etc., for rectum, vagina, urethra, nasal cavity, ear canal, etc.

[0057] In some embodiments of the present application, the pharmaceutical composition is an ophthalmic preparation, for example, an eye drop (solution, emulsion, suspension), an eye ointment, an eye gel.

[0058] Specifically, the various dosage forms of the pharmaceutical composition can be prepared according to the conventional production method in the pharmaceutical field. For example, the active ingredient is mixed with one or more pharmaceutically acceptable excipients, and then it is made into the desired dosage form.

[0059] Specifically, in the pharmaceutical composition, the weight percentage of the salt form of the first aspect (in terms of the compound) can be 0.1-99.5%, for example, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 6%, 8%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, in particular, 0.1-30%.

[0060] In the fourth aspect of the present application, the use of the salt form of the first aspect in the preparation of a medicament for preventing and / or treating a disease related to protein tyrosine kinase is provided.

[0061] Specifically, the tyrosine kinase is VEGFR, for example, one or more of VEGFR1, VEGFR2, VEGFR3.

[0062] Specifically, the disease is a disease that can be beneficial to its prevention and / or treatment by inhibiting protein tyrosine kinase, for example, a tumor, an autoimmune disease, an inflammatory disease, a disease associated with pathogen infection, a neurodegenerative disease, a cardiovascular disease, a metabolic disease, a fibrotic disease, an ocular disease, in particular, a proliferative disease mediated by protein tyrosine kinase.

[0063] In some embodiments of the application, the disease is a tumor, in particular a malignant tumor (cancer), including, but not limited to, breast cancer, lung cancer (in particular non-small cell lung cancer), colorectal cancer, renal cancer, liver cancer, pancreatic cancer, ovarian cancer, prostate cancer, glioblastoma, glioma, myeloid dysplasia, mesothelioma, sarcoma, myelodysplastic syndrome, hematological malignancies.

[0064] In particular, the hematological malignancies include leukemia, lymphoma, multiple myeloma (MM).

[0065] In particular, the leukemia can be chronic lymphocytic leukemia (CLL), chronic myelocytic leukemia (CML), acute lymphocytic leukemia (ALL), acute myelocytic leukemia (AML), acute monocytic leukemia.

[0066] In particular, the lymphoma can be Hodgkin's lymphoma (HL) and non-Hodgkin's lymphoma (NHL) (e.g., diffuse large cell lymphoma (DLCL) (e.g., diffuse large B-cell lymphoma), follicular lymphoma, chronic lymphocytic leukemia / small lymphocytic lymphoma (CLL / SLL), mantle cell lymphoma (MCL), marginal zone B-cell lymphoma, primary mediastinal B-cell lymphoma, Burkitt's lymphoma, lymphoplasmacytic lymphoma, hairy cell leukemia (HCL), immunoblastic large cell lymphoma, precursor B-lymphoblastic lymphoma, and primary central nervous system (CNS) lymphoma and T-cell NHL, such as precursor T-lymphoblastic lymphoma / leukemia, peripheral T-cell lymphoma (PTCL), angioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma, enteropathy-type T-cell lymphoma, subcutaneous panniculitis-like T-cell lymphoma, and anaplastic large cell lymphoma, NK / T-cell lymphoma, in particular diffuse large B-cell lymphoma (DLBCL). In particular, in the application, the treatment of the tumor includes killing the tumor, preventing metastatic spread of the tumor and growth of micrometastases.

[0067] In some embodiments of the application, the tumor is selected from the group consisting of renal cell carcinoma, hepatocellular carcinoma, colorectal cancer, lung cancer, gastric cancer, leukemia.

[0068] In some embodiments of the application, the disease is an ocular disease, including, but not limited to, diabetic retinopathy (including nonproliferative (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); pathologic choroidal neovascularization (CNV) from any pathologic mechanism (i.e., high myopia, trauma, sickle cell (anemia) disease; ocular histoplasmosis, angioid streaks, traumatic choroidal rupture, optic nerve drusen, and certain retinal dystrophies); pathologic retinal neovascularization from any pathologic mechanism (i.e., sickle cell retinopathy, Eales disease, ocular ischemic syndrome, carotid-cavernous fistula, familial exudative vitreoretinopathy, hyperviscosity syndrome, idiopathic retinal vasculitis, birdshot retinochoroidopathy, retinal vasculitis, sarcoid, or toxoplasmosis); uveitis; retinal vein occlusion (central or branch); ocular trauma; surgical-induced edema; surgical-induced neovascularization; cystoid macular edema; ocular ischemia; retinopathy of prematurity; Coat's disease (Coats' disease, also known as exudative retinopathies, retinal telangiectasia); sickle cell retinopathy and / or neovascular glaucoma; retinoblastoma.

[0069] In one embodiment of the application, the disease is diabetic retinopathy, including nonproliferative (background) diabetic retinopathy, proliferative diabetic retinopathy, and diabetic macular edema.

[0070] In another embodiment of the application, the disease is age-related macular degeneration (AMD), including neovascular (wet / exudative) AMD, dry AMD, geographic atrophy.

[0071] In some embodiments of the application, the disease is an autoimmune disease, including, but not limited to: organ-specific autoimmune diseases, systemic lupus erythematosus, rheumatoid arthritis, systemic vasculitis, scleroderma, pemphigus, dermatomyositis, mixed connective tissue disease, autoimmune hemolytic anemia, autoimmune thyroid disease, ulcerative colitis, and the like.

[0072] In some embodiments of the present application, the disease is an inflammatory disease, including but not limited to: osteoarthritis, acute gout, multiple sclerosis, inflammatory bowel disease (such as Crohn's disease and ulcerative colitis), neuroinflammation, asthma, chronic obstructive airway disease, lung inflammation, myositis, eczema, dermatitis, acne, cellulitis, occlusive disease, thrombosis, alopecia, nephritis, vasculitis, retinitis, uveitis, scleritis, sclerosing cholangitis, hypophysitis, thyroiditis, septic shock, systemic inflammatory response syndrome (SIRS), toxic shock syndrome, acute lung injury, ARDS (adult respiratory distress syndrome), acute kidney failure, burn, pancreatitis (such as acute pancreatitis), postoperative syndrome, sarcoidosis, Herxheimer reaction, encephalitis, myelitis, meningitis, malaria, and the like.

[0073] In some embodiments of the present application, the disease is a neurodegenerative disease, including but not limited to: Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), different types of spinocerebellar ataxia (SCA), Pick's disease, and the like.

[0074] In some embodiments of the present application, the disease is a pathogen infection related disease, including but not limited to: influenza, SARS, COVID-19, viral hepatitis (such as hepatitis A, hepatitis B, hepatitis C, hepatitis D, and the like), AIDS, rabies, dengue fever, Ebola virus disease, and the like.

[0075] In some embodiments of the present application, the disease is a cardiovascular disease, including but not limited to: coronary heart disease, peripheral arterial disease, atherosclerosis, ischemic heart disease, ischemic cardiomyopathy, myocardial infarction, heart failure, angina pectoris, myocarditis, hypercholesterolemia, hypertension, ischemia-reperfusion injury, cerebrovascular ischemia (stroke), embolism (such as pulmonary embolism, renal embolism, hepatic embolism, gastrointestinal embolism, or peripheral limb embolism), or myocardial ischemia, and the like.

[0076] In some embodiments of the present application, the disease is a metabolic disease, including but not limited to: diabetes (such as type I diabetes, type II diabetes, or gestational diabetes), obesity, fatty liver (NASH or others), cachexia, hypercholesterolemia, gout, and the like.

[0077] In some embodiments of the present application, the disease is a fibrotic disease, including but not limited to: myocardial fibrosis, pulmonary fibrosis, renal fibrosis, post-surgical stenosis, keloid formation, cirrhosis, biliary cirrhosis, scleroderma, and the like.

[0078] In the fifth aspect of the present application, a method for preventing and / or treating a protein tyrosine kinase-mediated proliferative disease is provided, which comprises the step of administering to a subject in need thereof an effective amount of the salt form of the first aspect, or the pharmaceutical composition of the third aspect.

[0079] Specifically, the disease is as described in the fourth aspect of the present application.

[0080] Specifically, the subject is a mammal, for example, a human.

[0081] Specifically, the administration can employ any suitable administration route, for example, a gastrointestinal administration (e.g., oral administration) or a non-gastrointestinal administration (e.g., intravenous, intramuscular, intranasal, intraocular, intracerebral, intravaginal, intraperitoneal, transdermal, subcutaneous, intradermal, respiratory tract administration, etc.) route. In some embodiments of the present application, the administration route is an intraocular administration.

[0082] The present application provides a salt form of a protein tyrosine kinase inhibitor. In the development process, the inventors previously prepared a protein tyrosine kinase inhibitor compound, which has the ability to inhibit anti-angiogenic tyrosine kinases, in addition to effectively antagonizing VEGFR1, VEGFR2, VEGFR3 tyrosine kinase activity, has high selectivity for inhibiting EGFR tyrosine kinase activity, can effectively reduce and avoid side effects, especially ocular side effects (such as epithelial degeneration and defects, ulcers, corneal epithelial thinning, erosion, and / or corneal edema, keratitis), and has a higher maximum tolerated dose and safer use in subjects. On this basis, the inventors further studied the salt form (hemisulfate, hemicitrate, hemifumarate, hemisuccinate, hemi-L-tartrate, hemihexandioate) of the drug, which has significantly improved solubility, higher solid stability, and lower hygroscopicity compared to the free base, and has very good application value in drug development and industrialization. BRIEF DESCRIPTION OF DRAWINGS

[0083] Figure 1 shows the H-NMR spectrum of the hemifumarate salt of compound T116. 1 Figure 1 shows the H-NMR spectrum of the hemifumarate salt of compound T116.

[0084] Figure 2 shows the H-NMR spectrum of the hemisuccinate salt of compound T116. 1 Figure 2 shows the H-NMR spectrum of the hemisuccinate salt of compound T116.

[0085] Figure 3 shows the H-NMR spectrum of the hemi-L-tartrate salt of compound T116. 1 Figure 3 shows the H-NMR spectrum of the hemi-L-tartrate salt of compound T116.

[0086] Figure 4 shows the H-NMR spectrum of the hemicitrate salt of compound T078. 1 Figure 4 shows the H-NMR spectrum of the hemicitrate salt of compound T078.

[0087] Figure 5 shows the H-NMR spectrum of the hemifumarate salt of compound T078.1 H-NMR spectrum.

[0088] Figure 6 shows the H-NMR spectrum of compound T078 hemi-succinate salt. 1 H-NMR spectrum.

[0089] Figure 7 shows the H-NMR spectrum of compound T078 hemi-tartrate salt. 1 H-NMR spectrum.

[0090] Figure 8 shows the H-NMR spectrum of compound T078 hemi-adipate salt. 1 H-NMR spectrum. DETAILED DESCRIPTION

[0091] Unless defined otherwise, all scientific and technical terms used in the present application have the same meaning as is commonly understood by one of skill in the art to which the application pertains.

[0092] The term "alkyl" refers to a straight-chain or branched-chain hydrocarbon radical, which is not unsaturated and which is attached to the rest of the molecule by a single bond. Typical alkyl groups contain from 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, i-propyl, n-butyl, i-butyl, t-butyl, n-pentyl, i-pentyl, neopentyl, t-pentyl, n-hexyl, i-hexyl, and the like. If the alkyl group is substituted with a cycloalkyl group, it corresponds to a "cycloalkylalkyl", such as cyclopropylmethyl, cyclopropylethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, and the like. If the alkyl group is substituted with an aryl group, it corresponds to an "aralkyl", such as benzyl, benzhydryl, or phenethyl. If the alkyl group is substituted with a heterocyclyl group, it corresponds to a "heterocyclylalkyl". In the present application, a Co alkyl group refers to H, i.e. Co-C6alkyl includes H and C1-C6alkyl. 10 alkyl includes H and C1-C6alkyl. 10 alkyl.

[0093] The term "alkylene" refers to a hydrocarbon radical (divalent alkyl) resulting from the loss of two hydrogen atoms from an alkane molecule, which can be straight-chain or branched and which is attached to the rest of the molecule by a single bond. Typical alkylene groups herein contain from 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, and the like. In the present application, a Co alkylene group refers to a single bond, i.e. Co-C6alkylene includes a single bond and C1-C6alkylene.

[0094] The term "cycloalkyl" refers to an alicyclic hydrocarbon, such as a monocyclic and / or fused ring radical containing from 1 to 4 rings and / or containing from 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, and the like.

[0095] The term "alkoxy" refers to a substituent formed by the replacement of a hydrogen in a hydroxyl group with an alkyl group, such as an alkoxy group containing 1-10 carbon atoms, for example methoxy, ethoxy, propoxy, butoxy, and the like.

[0096] The term "alkylamino" refers to a substituent formed by the replacement of one or both hydrogens in an amino group (-NH2) with an alkyl group, such as an alkylamino group containing 1-10 carbon atoms, for example

[0097] The term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0098] The term "haloalkyl" refers to a group formed by the replacement of one or more hydrogens in an alkyl group with a halogen atom (e.g., fluorine, chlorine, bromine, or iodine), for example -CHF2, -CH2F, -CF3, -CH2-CH2F, -CH2-CHF2, -CH2-CF3, -CH2CH2-CF3, -CH2CH2CH2-CF3.

[0099] The term "aryl" refers to a monocyclic or polycyclic radical, including polycyclic radicals containing single aryl groups and / or fused aryl groups, such as C6-Ci8(e.g., C6, C8, C10, C12, C14, C16, C18) aryl groups containing 1-3 single or fused rings and 6-18 (e.g., 6, 8, 10, 12, 14, 16, 18) carbon ring atoms, as described herein. 12 The term "aryl" refers to a monocyclic or polycyclic radical, including polycyclic radicals containing single aryl groups and / or fused aryl groups, such as C6-Ci8(e.g., C6, C8, C10, C12, C14, C16, C18) aryl groups containing 1-3 single or fused rings and 6-18 (e.g., 6, 8, 10, 12, 14, 16, 18) carbon ring atoms, as described herein.

[0100] The term "heterocyclyl" refers to a 3- to 18-membered ring radical comprising 2 to 17 carbon atoms and 1 to 10 heteroatoms selected from N, O, or S atoms. Heterocyclyl groups can be monocyclic, bicyclic, tricyclic, or tetracyclic, etc. polycyclic ring systems, which can include fused (two rings share two ring atoms), spiro (two rings share one ring atom), or bridged (two rings share more than two ring atoms) ring systems (excluding annulated rings). Heterocyclyl groups can be partially saturated (heteroaryl) or fully saturated (heterocycloalkyl). Suitable heteroaryl groups in the compounds of the present application contain 1, 2, or 3 heteroatoms selected from N, O, or S atoms, and include, for example, coumarinyl, including 8-coumarinyl, quinolinyl, including 8-quinolinyl, isoquinolinyl, pyridyl, pyrazinyl, pyrazolyl, pyrimidinyl, furanyl, pyrrolyl, thienyl, thiazolyl, isothiazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, imidazolyl, indolyl, isoindolyl, indazolyl, indolizinyl, phtalazinyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furopyridinyl, pyridazinyl, triazinyl, cinnolinyl, benzimidazolyl, benzofuranyl, benzofuropyridinyl, benzothienyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, and furopyridinyl. Suitable heterocycloalkyl groups in the compounds of the present application contain 1, 2, or 3 heteroatoms selected from N, O, or S atoms, and include, for example, pyrrolidinyl, tetrahydrofuranyl, dihydrofuran, tetrahydrothienyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, oxathianyl, piperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxiranyl, thiiranyl, azepinyl, oxazepinyl, diazepinyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 3-azabicyclo[3.1.0]hexyl, 3-azabicyclo[4.1.0]heptyl, 3H-indolizinyl, and quinolizinyl.

[0101] The term "optionally substituted" group can be halogen, -CN, -NO2, -OR', -NR'R", -S(O)t-R', -S(O)t-NR'R", -COR', -C(O)OR', -C(O)NR'R", -C(O)N(R')OR", -OC(O)R', -OC(O)NR'R", -NR'C(O)R", -N(R')C(O)NR'R', -N(R')C(NR')NR'R', -NR'-S(O)t-R', -NR'-S(O)t-NR'R', -N=S(O)R'R", -S(NR')(O)R", -N(R')CN, -P(O)(R')NR'R", -P(O)(R')OR", or -P(O)R'R", C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkenyl, C 1-6 alkynyl, C 3-6 cycloalkyl, C 4-10 cycloalkylalkyl, C 6-10 aryl, C 6-10 arylalkyl, C 3-8 heterocyclyl, C 3-8 heterocyclylalkyl; t is 0, 1 or 2; each R' and R" is independently selected from the group consisting of H, halogen, -CN, -NO2, alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclyl, heterocyclylalkyl; or R' and R" attached to the same nitrogen, together with the nitrogen atom, form a heterocyclic ring.

[0102] The terms "patient" or "subject" and the like are used interchangeably herein and refer to any animal or cell thereof, whether in vitro or in situ, treated according to the methods described herein. In particular, the aforementioned animal includes a mammal, for example, a rat, a mouse, a guinea pig, a rabbit, a dog, a monkey, a human, and particularly a human.

[0103] The term "treatment" refers to preventing, curing, reversing, attenuating, alleviating, minimizing, inhibiting, arresting and / or stopping one or more clinical symptoms of a disease after the onset of the disease.

[0104] The term "prevention" refers to avoiding, minimizing or making it difficult for a disease to occur or develop by treatment before the onset of the disease.

[0105] The term "tumor" refers to an abnormal mass of tissue in which the growth of the mass exceeds and is not coordinated with the growth of normal tissue. Tumors can be "benign" or "malignant," depending on the following characteristics: degree of cellular differentiation (including morphology and function), rate of growth, local invasion, and metastasis. "Benign tumors" are generally well-differentiated, characterized by slower growth than malignant tumors, and remain confined to the site of origin. In addition, benign tumors do not have the ability to infiltrate, invade, or metastasize to distant sites. In some cases, certain "benign" tumors can later give rise to malignant tumors, possibly due to additional genetic changes in a subpopulation of neoplastic cells of the tumor, and these tumors are referred to as "pre-malignant tumors." "Malignant tumors" are generally poorly differentiated (anaplastic), and are characterized by rapid growth with concomitant progressive infiltration, invasion, and destruction of surrounding tissue. In addition, malignant tumors generally have the ability to metastasize to distant sites.

[0106] The term "cancer" refers to a malignant tumor (Stedman's Medical Dictionary, 25th ed.; Hensyl ed.; Williams & Wilkins: Philadelphia, 1990).

[0107] The term "protein tyrosine kinase inhibitor" refers to a molecule that decreases, inhibits, or otherwise reduces one or more biological activities of a protein tyrosine kinase. The inhibition using a protein tyrosine kinase inhibitor does not necessarily indicate a complete ablation of protein tyrosine kinase activity. Protein tyrosine kinase activity can be reduced by a significant amount, e.g., at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% compared to a control.

[0108] The disclosures of various publications, patents and published patent specifications referenced herein are hereby incorporated by reference in their entireties.

[0109] The technical solutions of the present application will be described clearly and completely below in combination with the embodiments of the present application. Obviously, the described embodiments are only some of the embodiments of the present application, but not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor are within the scope of protection of the present application.

[0110] Example 1: Synthesis of compound T078

[0111] The synthetic route is as follows:

[0112] First step

[0113] 2-fluoro-5-(hydroxymethyl)benzoic acid

[0114] 2-fluoro-5-(hydroxymethyl)benzoic acid

[0115] 2-fluoro-5-(hydroxymethyl)benzoic acid

[0116] The product was confirmed by LCMS and H-NMR.

[0117] 1 H NMR (400 MHz, MeOD) δ 7.80 (d, J = 6.4 Hz, 1H), 7.48 (s, 1H), 7.15 - 7.09 (m, 1H), 4.61 (s, 2H).

[0118] Second Step

[0119] 5-(chloromethyl)-2-fluoro-benzoyl chloride

[0120] 5-(chloromethyl)-2-fluoro-benzoyl chloride

[0121] The product was confirmed by LCMS and H-NMR.

[0122] The product was confirmed by LCMS and H-NMR.

[0123] Third Step

[0124] 5-(chloromethyl)-2-fluoro-benzoyl chloride

[0125] 5-(chloromethyl)-2-fluoro-benzoyl chloride

[0126] Dissolve 5-(chloromethyl)-2-fluoro-benzoyl chloride I337 (150 mg, 724.54 pmol) in DCM (4 mL), add cyclopropylamine (62 mg, 1.09 mmol) and triethylamine (220 mg, 2.17 mmol) at 0 °C, then stir the reaction for about 2 hours. After complete reaction, the mixture is directly concentrated under reduced pressure to obtain the crude product, which is purified by preparative thin layer chromatography (PE / EA = 1 / 1) to obtain the product 5-(chloromethyl)-N-cyclopropyl-2-fluoro-benzamide I338 (50 mg, 219.62 pmol, 30.31% yield) as a white solid.

[0127] The product is confirmed by LCMS and H-NMR.

[0128] 1 H NMR (400 MHz, CDC13) δ 8.13 (dd, J = 7.2, 2.4 Hz, 1H), 7.55 - 7.49 (m, 1H), 7.12 (dd, J = 11.6, 8.4 Hz, 1H), 6.82 (d, J = 10.4 Hz, 1H), 4.61 (s, 2H), 2.96 (td, J = 6.4, 3.6 Hz, 1H), 0.90 (t, J = 6.4 Hz, 2H), 0.68 - 0.60 (m, 2H).

[0129] Fourth step

[0130] methyl 3-[[3-(cyclopropylcarbamoyl)-4-fluoro-phenyl]methoxy]-5-methylsulfonyl-isothiazole-4-carboxylate

[0131] 3-[[3-(cyclopropylcarbamoyl)-4-fluoro-phenyl]methoxy]-5-methylsulfonyl-isothiazole-4-carboxylate

[0132] Methyl 3-hydroxy-5-(methylsulfonyl)isothiazole-4-carboxylate I009 (1.1 g, 4.64 mmol) and potassium carbonate (2.0 g, 14.33 mmol) were dissolved in DMF (20 mL), 5-(chloromethyl)-N-cyclopropyl-2-fluoro-benzamide I338 (1.4 g, 6.15 mmol) was added, the reaction system was stirred at room temperature for 2 hours. After complete reaction, water (100 mL) was added to the reaction solution, the mixture was extracted with ethyl acetate (50 mL x 3), the organic phase was washed with water, saturated brine successively, dried over anhydrous sodium sulfate, concentrated under reduced pressure to obtain the crude product, the crude product was stirred in MTBE (30 mL), filtered, and the filter cake was dried to obtain the yellow solid product methyl 3-[[3-(cyclopropylcarbamoyl)-4-fluoro-phenyl]methoxy]-5-methylsulfonylisothiazole-4-carboxylate I339 (1 g, 2.33 mmol, 50.34% yield).

[0133] The product was confirmed by LCMS and H-NMR.

[0134] 1 H NMR (400 MHz, CDCl3) δ 8.25 (dd, J = 7.6, 2.4 Hz, 1H), 7.60-7.56 (m, 1H), 7.15 (dd, J = 11.6, 8.4 Hz, 1H), 6.85 (d, J = 13.2 Hz, 1H), 5.50 (s, 2H), 4.01 (s, 3H), 3.51 (s, 3H), 2.99-2.94 (m, 1H), 0.93-0.90 (m, 2H), 0.67-0.64 (m, 2H).

[0135] Fifth step

[0136] methyl 3-[[3-(cyclopropylcarbamoyl)-4-fluoro-phenyl]methoxy]-5-[(2,4-dimethoxyphenyl)methylamino]isothiazole-4-carboxylate

[0137] methyl 3-[[3-(cyclopropylcarbamoyl)-4-fluoro-phenyl]methoxy]-5-[(2,4-dimethoxyphenyl)methylamino]isothiazole-4-carboxylate

[0138] Methyl 3-[[3-(cyclopropylcarbamoyl)-4-fluoro-phenyl]methoxy]-5-methylsulfonylisothiazolium-4-carboxylate I339 (700 mg, 1.63 mmol) was dissolved in THF (15 mL), and 2,4-dimethoxybenzylamine (2.7 g, 16.34 mmol) was added. The reaction mixture was stirred at 65 °C for 1 hour. After the reaction was completed, the reaction solution was added to water (30 mL) and stirred for about 20 minutes. Then, the mixture was filtered, and the filter cake was dried to give a yellow solid product, methyl 3-[[3-(cyclopropylcarbamoyl)-4-fluoro-phenyl]methoxy]-5-[(2,4-dimethoxyphenyl)methylamino]isothiazolium-4-carboxylate I340 (500 mg, 969.83 μmol, 59.36% yield).

[0139] The product was confirmed by LCMS and H-NMR.

[0140] 1 H NMR (400MHz, CDCl3) δ8.21 (dd, J=7.6, 2.4Hz, 1H), 8.13 (t, J=5.6Hz, 1H), 7.59–7.52 (m,1H),7.17(d,J=8.4Hz,1H),7.10(dd,J=11.6,8.4Hz,1H),6.86(d,J=12.4Hz,1H) ,6.49–6.44(m,2H),5.41(s,2H),4.28(d,J=6.0Hz,2H),3.87(s,3H),3.86(s,3H),3 .81(s,3H),2.95(dt,J=10.4,3.6Hz,1H),0.89(q,J=6.8Hz,2H),0.66–0.60(m,2H).

[0141] Step 6

[0142] methyl 5-amino-3-[[3-(cyclopropylcarbamoyl)-4-fluoro-phenyl]methoxy]isothiazole-4-carboxylate

[0143] 5-Amino-3-[[3-(cyclopropylcarbamoyl)-4-fluoro-phenyl]methoxy]isothiazol-4-carboxylic acid methyl ester

[0144] Methyl 3-[[3-(cyclopropylcarbamoyl)-4-fluoro-phenyl]methoxy]-5-[3-(4- methylpiperazin-1-yl)propylcarbamoylamino]isothiazole-4-carboxylate I340 (600 mg, 1.16 mmol) was dissolved in DCM / H20 (10 / 2 mL), DDQ (528 mg, 2.33 mmol) was added portionwise at 0 °C, the reaction was stirred at room temperature for 2 hours. After the reaction was completed, water (30 mL) was added to the reaction solution, which was extracted with ethyl acetate (20 mL x 3), the combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (neutral alumina, PE / EA = 10 / 1-5 / 1) to obtain white solid product methyl 5-amino-3-[[3-(cyclopropylcarbamoyl)-4-fluoro-phenyl]methoxy]isothiazole-4- carboxylate I341 (340 mg, 930.54 μmol, 79.96% yield).

[0145] The product was confirmed by LCMS and H-NMR.

[0146] 1 H NMR (400 MHz, CDC13) δ 8.25 (dd, J = 7.6, 2.0 Hz, 1H), 7.60 - 7.55 (m, 1H), 7.12 (dd, J = 11.8, 8.4 Hz, 1H), 6.84 (d, J = 12.0 Hz, 1H), 6.50 (s, 2H), 5.44 (s, 2H), 3.92 (s, 3H), 2.96 (dt, J = 10.4, 3.6 Hz, 1H), 0.91 (q, J = 6.8 Hz, 2H), 0.67 - 0.62 (m, 2H).

[0147] Seventh step

[0148] methyl

[0149] 3-[[3-(cyclopropylcarbamoyl)-4-fluoro-phenyl]methoxy]-5-[3-(4-methylpiperazin-1-yl)propylcarbamoylamino]isot hiazole-4-carboxylate

[0150] 3-[[3-(cyclopropylcarbamoyl)-4-fluoro-phenyl]methoxy]-5-[3-(4-methylpiperazin-1-yl)propylcarbamoylamino]isot hiazole-4-carboxylate

[0151] Dissolve 3-(4-methylpiperazin-1-yl)propan-1 -amine (116 mg, 738.96 μmol) in anhydrous THF (5 mL), under N2protection, reduce to 0 °C, add CDI (120 mg, 738.96 μmol), stir the reaction at room temperature for 1 hour, add DMSO (5 mL), remove most of the THF under reduced pressure, add 5-amino-3-[[3-(cyclopropylcarbamoyl)-4-fluoro-phenyl]methoxy]isothiazole-4- carboxylic acid methyl ester 1341 (180 mg, 492.64 μmol) and potassium carbonate (136 mg, 985.28 μmol), stir the reaction at room temperature for 1 hour. After complete reaction, add water (20 mL) to the reaction, extract the mixture with ethyl acetate (10 mL x 3), wash the organic phase with water, saturated brine successively, dry over anhydrous sodium sulfate, concentrate under reduced pressure to obtain the crude product, purify the crude product using preparative thin layer chromatography (DCM / MeOH = 10 / 1, 1% NH3.H2O) to obtain the product 3-[[3-(cyclopropylcarbamoyl)-4-fluoro-phenyl]methoxy]-5-[3-(4-methylpiperazin-1-yl)propylcarbamoylamino]isothiazole-4-carboxylic acid methyl ester 1342 (90 mg, 164.05 μmol, 33.30% yield) as a white solid.

[0152] The product was confirmed by LCMS and H-NMR.

[0153] 1 H NMR (400 MHz, CDC13) δ 10.39 (s, 1H), 8.29 (dd, J = 7.6, 2.0 Hz, 1H), 8.11 (s, 1H), 7.59 - 7.53 (m, 1H), 7.12 (dd, J = 11.6, 8.4 Hz, 1H), 6.84 (d, J = 12.8 Hz, 1H), 5.45 (s, 2H), 3.96 (d, J = 5.2 Hz, 3H), 3.47 (d, J = 5.2 Hz, 2H), 2.97 (dd, J = 7.2, 3.6 Hz, 1H), 2.61 (dd, J = 12.8, 7.2 Hz, 10H), 2.40 (s, 3H), 1.78 - 1.73 (m, 2H), 0.90 (t, J = 6.4 Hz, 2H), 0.65 (q, J = 6.8 Hz, 2H).

[0154] Eighth step

[0155] 3-[[3-(cyclopropylcarbamoyl)-4-fluoro-phenyl]methoxy]-5-[3-(4-methylpiperazin-1-yl)propylcarbamoylamino]isot hiazole-4-carboxamide

[0156] 3-[[3-(cyclopropylcarbamoyl)-4-fluoro-phenyl]methoxy]-5-[3-(4-methylpiperazin- 1 -yl)propylcarbamoylamino]isothiazole-4-carboxamide

[0157] Methyl 3-[[3-(cyclopropylcarbamoyl)-4-fluoro-phenyl]methoxy]-5-[3-(4- methylpiperazin-1-yl)propylcarbamoylamino]isothiazole-4-carboxylate I342 (100 mg, 182.27 pmol) was dissolved in NH3 / MeOH (6 mL, 9 mol / L) in a microwave tube, and the mixture was stirred at 60 °C for 16 h. The reaction solution was concentrated under reduced pressure to give a crude product, which was purified by high performance preparative chromatography to give 3-[[3-(cyclopropylcarbamoyl)-4-fluoro-phenyl]methoxy]-5-[3-(4- methylpiperazin-1-yl)propylcarbamoylamino]isothiazole-4-carboxamide T078 (17.8 mg, 33.36 pmol, 18.30% yield) as a white solid.

[0158] The product was confirmed by LCMS, H-NMR and C-NMR.

[0159] 1 H NMR (400 MHz, CD3OD) δ 7.80 (dd, J = 6.8, 2.0 Hz, 1H), 7.67 (d, J = 3.2 Hz, 1H), 7.23 (m, 1H), 5.50 (s, 2H), 3.28 (d, J = 6.8 Hz, 2H), 2.88 (dd, J = 7.2, 3.6 Hz, 1H), 2.51 (dd, J = 31.6, 24.4 Hz, 10H), 2.36 (s, 3H), 1.79 - 1.73 (m, 2H), 0.86 - 0.81 (m, 2H), 0.66 - 0.62 (m, 2H).

[0160] 13 C NMR (100 MHz, CDC13) 169.56, 165.85, 164.26, 164.23, 161.67, 161.52, 159.20, 154.04, 133.48, 133.38, 132.90, 132.87, 131.98, 131.96, 121.29, 121.16, 116.81, 116.36, 69.08, 56.32, 54.77, 52.69, 45.66, 29.69, 25.26, 23.19, 6.88.

[0161] Example 2: Synthesis of compound T116

[0162] The synthesis route is as follows:

[0163] First step

[0164] 3-[(3S)-3-fluoropyrrolidin-1-yl]propanenitrile

[0165] 3-[(3S)-3-fluoropyrrolidin-1-yl]propanenitrile

[0166] 3-[(3S)-3-fluoropyrrolidin-1-yl]propanenitrile

[0167] Product was confirmed by LCMS and H-NMR

[0168] 1 H NMR (400 MHz, CDC13): δ (ppm) 5.28 - 5.06 (m, 1H), 3.00 - 2.73 (m, 5H), 2.64 - 2.44 (m, 3H), 2.26 - 1.97 (m, 2H).

[0169] Second step

[0170] 3-[(3S)-3-fluoropyrrolidin-1-yl]propan-1-amine

[0171] 3-[(3S)-3-fluoropyrrolidin-1-yl]propan-1-amine

[0172] 3-[(3S)-3-fluoropyrrolidin-1-yl]propan-1-amine

[0173] Product was confirmed by LCMS and H-NMR.

[0174] 1 H NMR (400 MHz, CDC13): δ (ppm) 5.83-4.93 (m, 1H), 2.93-2.34 (m, 7H), 2.17-1.48 (m, 6H).

[0175] Third Step

[0176] methyl

[0177] 3-[[5-(cyclopropylcarbamoyl)-2-fluoro-phenyl]methoxy]-5-[3-[(3S)-3-fluoropyrrolidin-1-yl]propylcarbamoylamin o]isothiazole-4-carboxylate

[0178] 3-[[5-(cyclopropylcarbamoyl)-2-fluoro-phenyl]methoxy]-5-[3-[(3S)-3-fluoropyrrolidin-1-yl]propylcarbamoylamin o]isothiazole-4-carboxylate

[0179] Dissolve 3-[(3S)-3-fluoropyrrolidin-1-yl]propylamine I463 (196.1 mg, 1.3 mmol) in anhydrous THF (5 mL), under N2protection, add CDI (217.5 mg, 1.3 mmol), stir the reaction at 25 °C for 1.0 hour, add DMSO (5 mL), remove THF under reduced pressure, add 5-amino-3-[[5-(cyclopropylcarbamoyl)-2-fluoro-phenyl]methoxy]isothiazole-4-carboxylic acid methyl ester I376 (350 mg, 957.9 μmol) and potassium carbonate (397.2 mg, 2.9 mmol), stir the reaction at 25 °C for 1.0 hour. Pour the reaction into water (60 mL), extract the mixture with ethyl acetate (20 mL x 3), wash the combined organic phase with saturated brine (20 mL), dry over anhydrous sodium sulfate, concentrate under reduced pressure to give the crude product, purify the crude product using preparative thin layer chromatography (DCM / MeOH = 10 / 1) to give the product 3-[[5-(cyclopropylcarbamoyl)-2-fluoro-phenyl]methoxy]-5-[3-[(3S)-3-fluoropyrrolidin-1-yl]propylcarbamoylamino]isothiazole-4-carboxylic acid methyl ester T161 (175 mg, 325.5 μmol, 33.98% yield) as a yellow solid.

[0180] The product is confirmed by LCMS.

[0181] Fourth Step

[0182] 3-[[5-(cyclopropylcarbamoyl)-2-fluoro-phenyl]methoxy]-5-[3-[(3S)-3-fluoropyrrolidin-1-yl]propylcarbamoylamino]isothiazole-4-carboxamide

[0183] 3-[[5-(cyclopropylcarbamoyl)-2-fluoro-phenyl]methoxy]-5-[3-[(3S)-3-fluoropyrrolidin-1-yl]propylcarbamoylamino]isothiazole-4-carboxamide

[0184] Methyl 3-[[5-(cyclopropylcarbamoyl)-2-fluoro-phenyl]methoxy]-5-[3-[(3S)-3- fluoropyrrolidin-1-yl]propylcarbamoylamino]isothiazole-4-carboxylate I464 (175 mg, 325.5 μmol) was dissolved in NH3 / MeOH (10 mL, 10 mol / L) and DMSO (0.3 mL). The mixture was stirred at 40 °C for 16 hours. The reaction solution was concentrated under reduced pressure to give the crude product, which was purified using preparative thin layer chromatography (DCM / MeOH = 10 / 1) to give the product 3-[[5-(cyclopropylcarbamoyl)-2-fluoro-phenyl]methoxy]-5-[3-[(3S)-3- fluoropyrrolidin-1-yl]propylcarbamoylamino]isothiazole-4-carboxamide T116 (40 mg, 76.55 μmol, 23.51% yield) as a white solid.

[0185] The product was confirmed by LCMS, H-NMR, C-NMR and F-NMR.

[0186] 1 H NMR (400 MHz, CD3OD): δ (ppm) 8.00 (dd, J = 7.2, 2.4 Hz, 1H), 7.88-7.82 (m, 1H), 7.24 (t, J = 7.2 Hz, 1H), 5.56 (s, 2H), 5.26-5.06 (m, 1H), 3.27 (t, J = 6.8 Hz, 2H), 3.02-2.88 (m, 2H), 2.86-2.79 (m, 1H), 2.72-2.64 (m, 2H), 2.64-2.53 (m, 2H), 2.46-2.36 (m, 1H), 2.27-2.11 (m, 1H), 2.09-1.91 (m, 1H), 1.83-1.70 (m, 1H), 0.84-0.75 (m, 2H), 0.67-0.57 (m, 2H).

[0187] 13C NMR (101 MHz, CD3OD) δ 170.35, 167.16, 165.71, 163.19, 162.97, 156.10, 132.09, 131.57, 131.52, 131.18, 131.08, 125.08, 124.93, 116.86, 116.63, 98.73, 95.18, 93.44, 65.16, 61.76, 61.53, 54.71, 53.40, 40.43, 39.41, 33.57, 33.34, 29.60, 24.07, 6.55.

[0188] 19 F NMR (400 MHz, CD3OD) δ (ppm) -115.43, -168.98.

[0189] Example 3: Detection of the ability of compounds to inhibit tyrosine kinases

[0190] The assay uses Mobility Shift Assays / IMAP TM The ability of compounds to inhibit kinase phosphorylation is monitored directly by measuring unphosphorylated and phosphorylated substrates in the kinase reaction.

[0191] Experimental procedure:

[0192] The kinase and DTT were added to 1x kinase buffer to make a 2.5x kinase solution; 10 μΐ of the 2.5x kinase solution was transferred to the 384 well plate reaction plate, and 1x kinase buffer was added to the negative control wells. Incubate at room temperature for 10 minutes; the FAM labeled peptide, MgCl2, ATP, etc. were added to 1x kinase buffer to make a 2.5x substrate solution; 10 μΐ of the 2.5x substrate solution was transferred to the 384 well plate reaction plate; incubate at 28°C for different time, and 25 μΐ of stop solution was added to stop the reaction. Read the conversion rate data on the Caliper EZ Reader II. Convert the conversion rate to inhibition rate data.

[0193] "Min" is the reading of the control well with no enzyme; "Max" is the reading of the control well with DMSO. IC50values were fitted using XLFit excel add-in version 5.4.0.8. 50

[0194] X is the log value of the compound concentration, and Y is the inhibition rate (% inhibition).

[0195] IC50values of compounds against tyrosine kinases 50 ​The results are shown in the following table, with the compound PAN90806 (commercial product) as positive control.

[0196] Table 1 Experimental results

[0197] From the results in Table 1, it can be seen that compounds T078 and T116 have a significant improvement in selectivity for inhibiting TIE2 and EGFR, not only to improve the antagonistic activity of all VEGFR receptors (VEGFR1, VEGFR2, VEGFR3) tyrosine kinase, but also significantly improve the selectivity of inhibiting TIE2 and EGFR receptor activity, which is conducive to reducing and avoiding side effects.

[0198] Example 4: Detection of the ability of compounds to bind to melanin

[0199] Pigmented ocular tissues contain melanin in intracellular melanosomes. Drugs bind to melanin to varying degrees, from no binding to extensive binding. Therefore, melanin binding is an important feature that affects ocular drug delivery and biodistribution. The ability of compounds to bind to melanin is an element that must be considered in the development of ocular drugs.

[0200] Reagents:

[0201] Melanin was derived from Sepia officinalis melanin, purchased from Sigma Aldrich (St Louis, MO).

[0202] Experimental procedure:

[0203] First, a 1 mg / ml melanin suspension was prepared using PBS (pH 7.4). As a washing step of the matrix, the suspended particles were centrifuged at 3000 rpm for 5 minutes. The obtained particles were resuspended in fresh PBS buffer and heated to 37°C under continuous shaking and incubated for 1 hour. 2 μl of the test compound stock solution (0.1 mM) was accurately pipetted into 98 μl of the melanin suspension (1 mg / ml) so that the final incubation concentration was 2 μM. Similarly, 2 μl of the stock solution (0.1 mM) was accurately pipetted into 98 μl of PBS buffer (containing 0.1% BSA) as a blank control sample. The samples were incubated at 37°C for 2 hours in a shaker (80-90 rpm). Then, centrifugation was performed at room temperature. 50 μl of supernatant was pipetted into 50 μl of precipitation agent (containing internal standard). After all the samples were precipitated, the supernatant was centrifuged and diluted with water (if necessary) before injection for analysis. The high binding compounds Sunitinib, Chloroquine and the low binding compound Celecoxib were determined as positive controls. An LC-MS / MS analysis method was established to determine the concentration of the compounds, and the ratio of the peak areas of the analyte / internal standard (Aanalyte / AIS) in the determined samples was calibrated as the replacement concentration of the sample for data processing. The binding rate (F b ) of the compound in the melanin was calculated according to the following formula:

[0204] C test is the drug concentration of the compound in the melanin incubation sample or the peak area ratio of the test compound to the internal standard. control is the drug concentration of the compound in the blank control sample or the peak area ratio of the test compound to the internal standard. The percentage of free drug (F u %) = 100 - F b .

[0205] The binding rate of the compound to melanin is shown in the following table, with the compound PAN90806 (commercial product) as a positive control.

[0206] Table 2 Experimental results

[0207] The results in Table 2 show that the compounds of the present application increase the percentage of free drug and reduce the binding rate to melanin, can interact with the receptors in the ocular tissues and increase the bioavailability. In comparison, the percentage of free drug of PAN90806 is low.

[0208] Example 5: Maximal tolerated dose (MITD) of eye drop administration of the compound

[0209] The maximum tolerated dose (MTD) is generally defined as the dose that produces an "acceptable" level of toxicity during a particular study, or if exceeded, would place the animal at "unacceptable" risk of toxicity, and is sometimes referred to as the "minimum toxic dose". We used male Dutch belted rabbits to evaluate the maximum tolerated dose of compound eye drops.

[0210] Experimental procedure:

[0211] Male Dutch belted rabbits (body weight 1.5 to 2.5 kg) were purchased from a qualified supplier. The test protocol designed for the animals in this test was implemented after being approved by the Institutional Animal Care and Use Committee (IACUC). Any operation on experimental animals was in accordance with the relevant requirements of the IACUC test operation.

[0212] Healthy animals with normal eyes (no trauma, hyperemia, swelling, etc.) were selected for the experiment. Rabbits received bilateral eye drops (50 microliters were dropped into the lower eyelid). Randomized grouping, 2 per group. Eye drops were given 3 times a day, 6±0.5 hours apart, for 3 consecutive days. Before administration, the state of the administration preparation was checked to ensure the uniformity of the preparation by vortexing, stirring or shaking. The eye condition of the animals was observed before each administration, including but not limited to eye color, material around the eye, whether the conjunctiva was swollen, eye exudate, secretion, cornea, pupil reflex, etc.

[0213] Experimental results:

[0214] One rabbit had a slight redness of the left eyelid on the second day of eye drop administration of the positive compound PAN90806 (175 micrograms / eye). On the third day of eye drop administration, one rabbit had a slight redness of the left and right eyelids, and another rabbit had a redness of the left eyelid, white secretion in the eye, redness of the right eyelid, and yellow secretion in the eye. The maximum tolerated dose (MTD) of PAN90806 on the third day of eye drop administration was 100 micrograms / eye.

[0215] T078 and T116 were administered eye drops at a dose of 1000 micrograms / eye for three consecutive days, and the results of the toxicity risk assessment showed that no abnormalities were observed in the eyes of all rabbits. Therefore, the maximum tolerated dose (MTD) of T078 and T116 on the fourth day of eye drop administration was 1000 micrograms / eye or higher.

[0216] In addition, other compounds described in the present application (such as those shown in general formula IV) also have similar properties to compounds T078 and T116 (the preparation and activity testing of these compounds are described in patent application PCT / CN2023 / 142499).

[0217] Example 6: Preparation of a salt of compound T116

[0218] Compound T116 is a free base with a pKa value of 7.4. The following acids were selected to prepare its salt forms: fumaric acid, succinic acid, hemimellaric acid, L-tartaric acid.

[0219] 1. Experimental method

[0220] About 25 mg of compound T116 (prepared in Example 2) was weighed into a 2 mL glass vial with 0.5 equivalent of acid and methanol was added.

[0221] The resulting sample was allowed to suspend at 50°C for 2 hours, then allowed to cool to 25°C naturally, and was allowed to suspend at 25°C for at least 48 hours.

[0222] The resulting suspension was centrifuged at 14000 rpm through a 0.45 μm nylon filter membrane, and the resulting solid was vacuum dried at 50°C for 2 hours to obtain a solid.

[0223] The resulting solid was subjected to 1 H-NMR analysis to confirm the formation of the salt. The H-NMR spectra of the hemifumarate salt, hemisuccinate salt, and hemi-L-tartrate salt are shown in Figures 1-3, respectively, which show that the molar ratio of the free base to the acid is 1:0.5. 1

[0224] 2. Experimental results

[0225] The hemifumarate salt, hemisuccinate salt, hemimellarate salt, and hemi-L-tartrate salt of compound T116 were prepared, and each was dissolved in methanol to obtain a clear solution.

[0226] The solubility of the free base and its salts of compound T116 in PBS buffer at pH 7.4 (final pH) is shown below. The solubility of the resulting salts was significantly improved relative to the free base.

[0227] Table 3 Solubility

[0228] In addition, the water content of the resulting hemifumarate salt solid was 0.5 wt%, and the hygroscopicity was low (0.7% water absorption from 40% to 80% RH) (the water content of the free base solid was 2.7 wt%, and the water absorption from 40% to 80% RH was 1.8%).

[0229] The solid stability, solubility, and hygroscopicity of the salts of compound T116 were investigated, taking the hemisuccinate salt as an example.

[0230] Example 7: Investigation of solid stability

[0231] 1. Experimental method

[0232] ​Open containers containing the hemi-succinate salt (prepared in Example 6) were placed at 25°C / 92.5% RH and 40°C / 75% RH for 1 week and 4 weeks, respectively. Closed containers containing the hemi-succinate salt were placed at 60°C for 1 week and 4 weeks. The samples under these conditions were tested by HPLC and observed for color change.

[0233] 2. Experimental Results

[0234] The experimental results are shown in the following table.

[0235] Table 4. Results of Solid Stability Investigation

[0236] Note: A: no color change; B: slight color change; C: moderate color change; D: severe color change

[0237] From the above table, it can be seen that the hemi-succinate salt of compound T116 showed good physical and chemical stability under the above accelerated stability conditions for 1 week and 4 weeks (even better than the mono-succinate salt of compound T116, which deliquesced under 25°C / 92.5% RH).

[0238] Example 8: Solubility Investigation

[0239] Into a 2 ml glass bottle, 22.6 mg of the hemi-succinate salt (prepared in Example 6) (equivalent to 20 mg of anhydrous free state) was weighed. 1 ml of solvent was added. The pH of the ES1-ES3 samples was adjusted to 6.0, 6.5 and 7.0, respectively, using 2M KOH solution or phosphoric acid solution. The time for adjusting the pH was about 30 minutes. The resulting suspensions were stirred at 25°C at 400 rpm for 2 hours and 24 hours. Subsequently, the resulting suspensions were centrifuged at 25°C at 14,000 rpm for 5 minutes. The solubility of the supernatant was tested by HPLC, and the pH of the supernatant was measured by a pH meter.

[0240] 2. Experimental Results

[0241] The experimental results are shown in the following table.

[0242] Table 5. Results of Solubility Investigation

[0243] Note: " / / ": not carried out.

[0244] Example 9: Hygroscopicity Investigation

[0245] 1. Experimental Method

[0246] The water absorption and dehydration behavior of the hemi-succinate salt (prepared in Example 6) was investigated by DVS testing at 25°C. The experimental conditions were as follows:

[0247] Instrument: Intrinsic

[0248] Total gas flow: 200 seem

[0249] Temperature: 25°C

[0250] Solvent: water

[0251] Method: Cycle: 40-0-95-0-40% RH

[0252] Step: 10% RH

[0253] Equilibrium: 0.002 dm / dt (% / min)

[0254] Minimum dm / dt maintenance time: 60 min

[0255] Maximum dm / dt maintenance time: 360 min

[0256] Sample size: ~5-50 mg

[0257] 2. Experimental results

[0258] The experimental results are shown in the following table.

[0259] Table 6. Adsorption and desorption test results of water

[0260] Note: " / / ": no test was performed

[0261] No or almost no hygroscopicity: hygroscopic weight gain less than 0.2%

[0262] Slightly hygroscopic: hygroscopic weight gain less than 2% but not less than 0.2%

[0263] Hygroscopic: hygroscopic weight gain less than 15% but not less than 2%

[0264] Extremely hygroscopic: hygroscopic weight gain not less than 15%

[0265] Deliquescent: absorbs sufficient moisture to form a liquid

[0266] Water absorption amount = water absorption amount at specific RH (80% to 95%) - water absorption amount at 40% RH

[0267] From the results in the above table, the hemi-succinate is almost non-hygroscopic, and it adsorbs about 0.1% moisture at 40% RH to 80% RH at 25°C.

[0268] It can be seen that the salt prepared in the present application exhibits good stability, solubility, and almost no hygroscopicity.

[0269] Example 10: Preparation of a salt of compound T078

[0270] 1. Experimental method

[0271] About 40 mg of compound T078 (prepared in Example 1) and 0.5 equivalent of acid were placed in a 2 mL glass bottle, and acetone was added.

[0272] The resulting mixture was stirred at 50°C for 2 hours, and then stirred at 25°C for 48 hours.

[0273] The resulting suspension was centrifuged at 14000 rpm. It was dried in vacuum at 50°C for 3 hours to obtain a solid.

[0274] The resulting solid was subjected to H-NMR analysis to confirm the formation of the salt. 1 The H-NMR spectra of the hemi-citrate salt, hemi-fumarate salt, hemi-succinate salt, hemi-L-tartrate salt, and hemi-adipate salt are shown in FIGS. 4-8, respectively, which show that the molar ratio of the free base to citric acid, fumaric acid, L-tartaric acid, and adipic acid is 1:0.5. 1 The H-NMR spectra of the hemi-citrate salt, hemi-fumarate salt, hemi-succinate salt, hemi-L-tartrate salt, and hemi-adipate salt are shown in FIGS. 4-8, respectively, which show that the molar ratio of the free base to citric acid, fumaric acid, L-tartaric acid, and adipic acid is 1:0.5.

[0275] 2. Experimental results

[0276] The hemi-sulfate salt, hemi-citrate salt, hemi-fumarate salt, hemi-succinate salt, hemi-L-tartrate salt, and hemi-adipate salt of compound T078 were prepared, and each of them was dissolved in methanol to obtain a clear solution.

[0277] The above-described only the preferred embodiments of the present application, and not to limit the present application, any modification, equivalent replacement, etc. made within the spirit and principles of the present application, should be included in the scope of protection of the present application.

[0278] The foregoing examples and methods described in the present application can vary based on the ability, experience, and preference of a person skilled in the art.

[0279] The order of the steps of the method in the present application does not constitute any limitation on the order of the steps of the method.

Claims

1. A salt form of a compound, said salt form being selected from: hemisulfates, hemicitrates, hemifumarates, hemisuccinates, hemi-L-tartrates, and hemiadipates, said compound having the following structure: in, R 1b Selected from: C1-C6 haloalkyl, cyano, nitro, azide, -OR 101 -C(O)R 101 -C(O)OR 101 -NHC(O)OR 101 -OC(O)R 101 -NHSO2R 101 -SO2NR 101 R 102 -NHC(O)R 101 -C(O)NR 101 R 102 -NR 101 R 102 -SR 101 -S(O)2R 101 -SO3H, -(C0-C6 alkylene)-(phenyl), -(C0-C6 alkylene)-(4-10 membered heterocyclic group), -(C0-C6 alkylene)-(C3-C 10 cycloalkyl); R 1c Selected from: H, F, C1-C6 alkyl, cyano, nitro, azide, -OR 101 -C(O)R 101 -C(O)OR 101 -NHC(O)OR 101 -OC(O)R 101 -NHSO2R 101 -SO2NR 101 R 102 -NHC(O)R 101 -C(O)NR 101 R 102 -NR 101 R 102 -SR 101 -S(O)2R 101 -SO3H, -(C0-C6 alkylene)-(phenyl), -(C0-C6 alkylene)-(4-10 membered heterocyclic group), -(C0-C6 alkylene)-(C3-C 10 cycloalkyl); R 1a R 1d R 1e Independently selected from: H, C1-C6 alkyl, halogen, C1-C6 haloalkyl, cyano, nitro, azide, -OR 101 -C(O)R 101 -C(O)OR 101 -NHC(O)OR 101 -OC(O)R 101 -NHSO2R 101 -SO2NR 101 R 102 -NHC(O)R 101 -C(O)NR 101 R 102 -NR 101 R 102 -SR 101 -S(O)2R 101 -SO3H, -(C0-C6 alkylene)-(phenyl), -(C0-C6 alkylene)-(4-10 membered heterocyclic group), -(C0-C6 alkylene)-(C3-C 10 cycloalkyl); Each R 101 and R 102 Independently selected from: H, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxy-substituted alkyl, C1-C6 alkoxy-substituted alkyl, C1-C6 amino-substituted alkyl, C1-C6 alkylamino-substituted alkyl, C3-C 10 cycloalkyl, C4-C 10 Cycloalkyl, substituted or unsubstituted 4-10 membered heterocyclic groups, phenyl, benzyl, C1-C 10 Silyl-substituted alkyl, C1-C 10 Silyl group; L2 is C0-C 10 Alkylene, wherein one or more H atoms in the alkylene are optionally and independently substituted with the following groups: C1-C6 alkyl, halogen, cyano, nitro, trifluoromethyl, trifluoromethoxy, azide, hydroxyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C4-C 10 cycloalkylalkyl; R0 is selected from: H, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne, halogen, cyano, nitro, trifluoromethyl, trifluoromethoxy, azide, -OR 001 -C(O)R 001 -C(O)OR 001 -NR 002 C(O)OR 001 -OC(O)R 001 -NR 002 SO2R 001 -SO2NR 001 R 002 -NR 002 C(O)R 001 -C(O)NR 001 R 002 -NR 001 R 002 -S(O) i R 002 Where i is an integer from 0 to 2, -SO3H, -NR 002 (CR 003 R 004 ) t OR 001 , t is an integer from 1 to 5; Among them, ring E is C6-C 10 Aromatic rings or 4-10 quintone heterocyclic rings; R2 is selected from: H, =O, Cl-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne, halogen, cyano, nitro, trifluoromethyl, trifluoromethoxy, azide, -OR 201 -C(O)R 201 -C(O)OR 201 -NR 202 C(O)OR 201 -OC(O)R 201 -NR 202 SO2R 201 -SO2NR 201 R 202 -NR 202 C(O)R 201 -C(O)NR 201 R 202 -NR 201 R 202 -S(O) i R 201 Where i is an integer from 0 to 2, -SO3H, -(C0-C6 alkylene)-(C6-C 10 aryl), -SO2-(C0-C6 alkylene)-(C6-C 10 aryl), -S-(C0-C6 alkylene)-(C6-C 10 aryl), -O-(C0-C6 alkylene)-(C6-C 10 Aryl), -(C0-C6 alkylene)-(4-10 heterocyclic), -SO2-(C0-C6 alkylene)-(4-10 heterocyclic), -S-(C0-C6 alkylene)-(4-10 heterocyclic), -O-(C0-C6 alkylene)-(4-10 heterocyclic), -(C0-C6 alkylene)-(C3-C 10 cycloalkyl), -SO2-(C0-C6 alkylene)-(C3-C 10 cycloalkyl), -S-(C0-C6 alkylene)-(C3-C 10 cycloalkyl), -O-(C0-C6 alkylene)-(C3-C 10 cycloalkyl); wherein, the C1-C 10 Alkyl, C6-C 10 The aryl and 4-10 membered heterocyclic groups are optionally substituted with groups selected from the following: halogen, cyano, nitro, azide, -OR', -C(O)R', -C(O)OR', -OC(O)R', -NR'C(O)OR”, -NR'SO2R”, -SO2NR'R”, -NR'C(O)R”, -C(O)NR'R”, -NR'R”, -SR', -SOR', -SO2R', -SO3H, C3-C 10 Cycloalkyl, phenyl, 4-10 membered heterocyclic groups; n is an integer from 1 to 5 (e.g., 1, 2, 3, 4, 5, if the valence allows); Each R 001 To R 004 Independently selected from: H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkylalkyl, substituted or unsubstituted heterocyclic group, substituted or unsubstituted heterocyclic alkyl, C1-C 10 Silyl group, wherein the alkyl, cycloalkyl, or heterocyclic group is optionally substituted with a group selected from the following: halogen, cyano, nitro, azide, -OR', -C(O)R', -C(O)OR', -OC(O)R', -NR'C(O)OR”, -NR'SO2R”, -SO2NR'R”, -NR'C(O)R”, -C(O)NR'R”, -NR'R”, -SR', -SOR', -SO2R', -SO3H, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Silyl, C3-C 10 Cycloalkyl, phenyl, 4-10 membered heterocyclic groups; Each R 201 To R 204 Independently selected from: H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkylalkyl, substituted or unsubstituted heterocyclic group, substituted or unsubstituted heterocyclic alkyl, C1-C 10 Silyl group, wherein the alkyl, cycloalkyl, or heterocyclic group is optionally substituted with a group selected from the following: halogen, cyano, nitro, azide, -OR', -C(O)R', -C(O)OR', -OC(O)R', -NR'C(O)OR”, -NR'SO2R”, -SO2NR'R”, -NR'C(O)R”, -C(O)NR'R”, -NR'R”, -SR', -SOR', -SO2R', -SO3H, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Silyl, C3-C 10 Cycloalkyl, phenyl, 4-10 membered heterocyclic groups; R' and R" are independently selected from: H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkylalkyl, substituted or unsubstituted heterocyclic group, substituted or unsubstituted heterocyclic alkyl; Preferably, the molar ratio of the compound to the acid is 1:0.4-0.6, more preferably 1:0.5; Preferably, the salt form is amorphous or crystalline.

2. The salt form as described in claim 1, characterized in that, R 1b Selected from: -CF3, -CHF2, -CH2F Cyano, Nitro, Azide, -OH Preferably, R 1a R 1e Independently selected from: H, halogens, C1-C3 alkyl groups; Preferably, R 1c Selected from: H, F; Preferably, R 1d Selected from: H, halogens, C1-C3 alkyl groups.

3. The salt form as described in claim 1, characterized in that, The compound has the following structure: Preferably, R 101 Selected from: H, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxysubstituted alkyl, C3-C6 cycloalkyl, C4-C 10 Cycloalkylalkyl.

4. The salt form according to any one of claims 1-3, characterized in that, L2 is a C0-C6 alkylene, such as a C2-C6 straight-chain alkylene.

5. The salt form according to any one of claims 1-4, characterized in that, R0 is Among them, ring E is a 4-10 membered heterocycle, especially a 4-8 membered saturated nitrogen-containing heterocycle; or, R0 is -NR 001 R 002 , where R 001 and R 002 Independently selected from: H, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxy-substituted alkyl, C1-C6 alkoxy-substituted alkyl, C1-C6 amino-substituted alkyl, C1-C6 alkylamino-substituted alkyl, C3-C6 cycloalkyl, C4-C 10 cycloalkylalkyl; or, R0 is selected from: H, cyano, -O(CO-C) 10 Alkyl), -O(C1-C) 10 silyl), -S(CO-C) 10 Alkyl), -C(O)(C0-C 10 Alkyl), -C(O)O(C0-C) 10 Alkyl), -OC(O)(C0-C 10 Alkyl), -N(C0-C) 10 Alkyl)SO2(C0-C 10 Alkyl), -SO2N(C0-C) 10 Alkyl) (C0-C 10 Alkyl), -N(C0-C) 10 alkyl)C(O)(C0-C 10 Alkyl), -C(O)N(C0-C) 10 Alkyl) (C0-C 10 Alkyl), -SO2(C0-C) 10 Alkyl), wherein the alkyl group is optionally substituted with a group selected from the following: halogen, cyano, hydroxy, amino, C1-C6 alkoxy, C1-C6 alkylamine, C3-C6 cycloalkyl.

6. The salt form as described in claim 5, characterized in that, Part of R2 is selected from: H, halogen, cyano, nitro, azide, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxy-substituted alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C4-C 10 Cycloalkylalkyl, -C(O)(C1-C6 alkyl), -C(O)O(C1-C6 alkyl); Preferably, R2 is selected from: H, methyl, ethyl, n-propyl, isopropyl, -CF3, -CHF2, -CH2F, F, Cl, Br, I, cyano, nitro, azide, hydroxyl, methoxy, ethoxy More preferably, R0 is selected from:

7. The salt form as described in claim 5, characterized in that, R0 is selected from: H, cyano, -OH -COOH, 8. The salt form as described in claim 1, characterized in that, The compound is selected from the following structures:

9. A pharmaceutical composition comprising the salt form according to any one of claims 1-8, and one or more pharmaceutically acceptable excipients; Preferably, the pharmaceutical composition is an ophthalmic preparation, preferably selected from: eye drops, ophthalmic ointments, and ophthalmic gels.

10. The use of the salt form according to any one of claims 1-8 in the preparation of a medicament for the prevention and / or treatment of diseases related to protein tyrosine kinases; Preferably, the tyrosine kinase is VEGFR, and more preferably one or more of VEGFR1, VEGFR2, and VEGFR3; More preferably, the disease is selected from: tumors, autoimmune diseases, inflammatory diseases, diseases related to pathogen infection, neurodegenerative diseases, cardiovascular diseases, metabolic diseases, fibrotic diseases, and eye diseases; Preferably, the tumor is selected from: breast cancer, lung cancer, colorectal cancer, kidney cancer, liver cancer, pancreatic cancer, ovarian cancer, prostate cancer, glioblastoma, glioma, unrelated myeloid metaplasia, mesothelioma, sarcoma, myelodysplastic syndrome, and hematologic malignancies, and more preferably from: renal cell carcinoma, hepatocellular carcinoma, colorectal cancer, lung cancer, gastric cancer, and leukemia; Preferably, the ocular disease is selected from: diabetic retinopathy, age-related macular degeneration (AMD), pathological choroidal neovascularization (CNV) of any pathological mechanism, pathological retinal neovascularization of any pathological mechanism, uveitis, retinal vein occlusion, ocular trauma, surgical edema, surgical neovascularization, cystoid macular edema, ocular ischemia, retinopathy of prematurity, Coats' disease, sickle cell retinopathy and / or neovascularizing glaucoma, retinoblastoma, preferably from: diabetic retinopathy, age-related macular degeneration (AMD), and Coats' disease.

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