Trisubstituted tetrahydroisoquinoline amide derivatives and medical applications thereof

Abstract

The application discloses a tri-substituted tetrahydroisoquinoline amide derivative with a novel structure or a pharmaceutically acceptable salt thereof, which is a compound shown in formula I. The compound has a strong inhibiting capacity on immune cell migration, the compound itself has high permeability and good water solubility, and can be used as an immune cell migration inhibitor and used for developing a medicine for treating ophthalmic diseases such as dry eye.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical chemistry. More specifically, it relates to a trisubstituted tetrahydroisoquinoline amide derivative or its pharmaceutically acceptable salt or hydrate, as well as deuterated or other isotopically substituted compounds, their preparation methods, and their application in treating ophthalmic diseases such as dry eye. Background Technology

[0002] Dry eye, formerly known as keratoconjunctivitis sicca or dry eye syndrome (DED), is a diagnosed condition characterized by dry eyes due to reduced tear production or excessive tear evaporation. It is a common ocular surface disease that significantly impacts vision and quality of life; severe dry eye can have a quality of life almost comparable to dialysis or severe angina. The incidence of dry eye is as high as 93% among frequent users of video terminals and as high as 90% among contact lens wearers. Dry eye is becoming another major national eye health problem after myopia. The dry eye drug market is projected to grow significantly to $6.7 billion by 2030, at a CAGR of 28.4%.

[0003] Dry eye is a chronic ocular surface disease caused by multiple factors, which may be accompanied by ocular surface inflammation, tissue damage, and neurosensory abnormalities, resulting in various ocular discomfort symptoms and / or visual dysfunction. The clinical manifestations of dry eye are highly heterogeneous; no single symptom or set of symptoms can be specifically used to define dry eye. Dry eye presents with various signs, but these signs vary among patients due to factors such as etiology, severity, and disease course. Studies show that the objective signs of dry eye have a low correlation with and are often inconsistent with the primary symptoms. Currently, the most widely accepted characteristics of dry eye are inflammatory response and reduced tear secretion, along with elevated levels of cytokines in the tears.

[0004] Dry eye can be treated by addressing the underlying cause and symptoms, depending on its type and severity. Currently, medications for dry eye mainly fall into two categories: those that lubricate the ocular surface and promote repair, and anti-inflammatory drugs. In Europe and the United States, the main drugs approved for treating dry eye are cyclosporine and lifitigrast. Cyclosporine is an immunosuppressant, belonging to the calcineurin inhibitor class. Its eye drops are a white, milky liquid (with a cationic / positively charged surface), but long-term use may damage the autoimmune system, and the cationic / positively charged milky liquid is not suitable for long-term storage. Lifitigrast is an immune cell migration inhibitor, achieving its therapeutic effect by preventing immune cells from entering the site of inflammation. However, this drug is highly lipophilic, and its metabolites can easily cause irritation and other adverse reactions. The incidence of treatment-oriented adverse events (TEAEs) during lifitigrast ocular treatment is as high as 53.6%, with 8.2% of subjects withdrawing from the trial. Common reasons for withdrawal include discomfort at the instillation site, irritation or pain at the instillation site, increased tearing, decreased visual acuity, and blurred vision.

[0005] LFA-1 (lymphocyte function-associated antigen 1) is a cell adhesion molecule widely expressed in the immune system. Its binding to its ligand ICAM-1 (intercellular adhesion molecule 1) plays a crucial role in the migration, activation, and inflammatory responses of immune cells such as T cells. In the pathogenesis of dry eye syndrome, T cell infiltration and activation are key factors leading to ocular inflammation and damage to the ocular surface tissues.

[0006] Lifitigrast, the first approved LFA-1 antagonist, treats dry eye by preventing T-cell adhesion to ICAM-1, inhibiting cytokine secretion, and ultimately preventing inflammation. It was approved by the US FDA in July 2016, with CAS number 1025967-78-5, and its structural formula is [structural formula missing]. The patent is disclosed in Chinese patent CN 1902195B. In addition, VVN001, an LFA-1 antagonist currently in Phase III clinical trials, has a similar mechanism of action to ristatin. It mainly targets the structure of ristatin by modifying it. While retaining the right-side methanesulfonylphenyl structure of ristatin, the tetrahydroisoquinoline structure in the parent nucleus is modified into a monocyclic (CN 109134533B), or benzo[a]pentacyclic bicyclic (CN 110483437B), or tricyclic (CN109721605B) structure.

[0007] Given the significant and unmet clinical need for dry eye medications, developing a new generation of LFA-1 antagonists with efficacy equivalent to or even superior to existing products, and improving upon their irritation and other defects, is a technical challenge that urgently needs to be addressed by those skilled in the art. Summary of the Invention

[0008] The technical problem to be solved by the present invention is to provide a novel trisubstituted tetrahydroisoquinoline amide derivative that can be used as a new generation of LFA-1 antagonist to treat dry eye syndrome, thereby overcoming the shortcomings of existing dry eye drugs, such as limited efficacy, inability to simultaneously improve and treat dry eye symptoms, or high TEAEs.

[0009] To solve the above-mentioned technical problems, the present invention provides the following technical solution:

[0010] A trisubstituted tetrahydroisoquinoline amide derivative, which is a compound, isomer, or pharmaceutically acceptable salt thereof represented by Formula I:

[0011]

[0012] Wherein, Y is selected from carbonyl, C 1-6 Alkyl or CR2R3;

[0013] X1 and X2 are each independently selected from halogens and C. 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, Halogenated C 1-6 Alkoxy, hydroxy, nitro, cyano, amino, C 1-6 alkylamine group, C 1-6 Alkyl, amide, or sulfonamide groups;

[0014] X3 is selected from hydrogen, halogen, substituted or unsubstituted C. 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, hydroxy, nitro, cyano, amino, substituted or unsubstituted C 1-6 Alkylamine group, substituted or unsubstituted C 1-6 Alkyl, amide, or sulfonamide groups; when C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 alkylamine group, C 1-6 When an alkyl acyl group has substituents, it can be substituted by at least one of the following groups: deuterium, halogen;

[0015] R1 is selected from substituted or unsubstituted C. 6-14 aryl, substituted or unsubstituted 5-14 heteroaryl, substituted or unsubstituted C 3-8 Cycloalkyl, substituted or unsubstituted 3-10 membered heterocyclic groups; when C 6-14 Aryl, 5-14 heteroaryl, C 3-8 When cycloalkyl or 3-10 membered heterocyclic groups have substituents, they can be monosubstituted or polysubstituted by one of the following groups, or polysubstituted by multiple of the following groups: deuterium, hydroxyl, halogen, C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylamine group, -OR';

[0016] R2 and R3 are each independently selected from hydrogen and C. 1-6 Alkyl or halogenated C 1-6 alkyl;

[0017] R' is selected from C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 2-6 alkenyl or C 1-6 Alkyl group;

[0018] Furthermore, when R1 is not a substituted or unsubstituted benzodioxolane, X3 is not hydrogen;

[0019] The heterocyclic group or heteroaryl group contains at least one heteroatom, which is selected from N, O or S.

[0020] Preferably, in Formula I, R1 is arbitrarily selected from one of the following structures:

[0021] in,

[0022] R4 is selected from hydrogen, deuterium, hydroxyl, halogen, and C. 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylamine group or -OR';

[0023] R' is selected from C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 2-6 alkenyl or C 1-6 Alkyl group;

[0024] R5 is selected from hydrogen, deuterium, or C. 1-6 alkyl or alkylyl, C 2-6 Enyl or halogenated C 2-6 Enyl group;

[0025] R6 and R7 are each independently selected from hydrogen, deuterium, halogens, and carbon. 1-6 Alkyl or halogenated C 1-6 alkyl;

[0026] n is selected from 0, 1, 2 or 3.

[0027] Preferably, in Formula I, the halogen is selected from fluorine, chlorine, bromine, iodine, and more preferably fluorine or chlorine.

[0028] Preferably, it is a compound represented by Formula II, an isomer, or a pharmaceutically acceptable salt thereof:

[0029]

[0030] Wherein, Y is selected from carbonyl or -CH2- group;

[0031] X1 and X2 are each independently selected from halogens and C. 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, Halogenated C 1-6 Alkoxy, nitro, cyano, amino, or sulfonamide groups;

[0032] X3 is selected from hydrogen, halogen, substituted or unsubstituted C. 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, hydroxy, nitro, cyano, amino, substituted or unsubstituted C 1-6 Alkylamine group, substituted or unsubstituted C 1-6 Alkyl, amide, or sulfonamide groups; when C 1-6 Alkyl, C1-6 Alkoxy, C 1-6 alkylamine group, C 1-6 When an alkyl acyl group has substituents, it can be substituted by at least one of the following groups: deuterium, halogen;

[0033] R4 is selected from hydrogen, deuterium, hydroxyl, halogen, and C. 1-6 Alkyl or halogenated C 1-6 alkyl;

[0034] R6 and R7 are each independently selected from hydrogen, deuterium, halogens, and carbon. 1-6 Alkyl or halogenated C 1-6 alkyl;

[0035] n is selected from 0, 1, 2 or 3.

[0036] Preferably, in Formula II, X1 and X2 are each independently selected from chlorine.

[0037] Preferably, in Formula II, when X1 and X2 are each chlorine, X3 is selected from hydrogen, nitro, cyano, halogen, acetyl, methoxy, amino or substituted amino, sulfonamide, and further preferably from hydrogen, nitro, methoxy, chlorine, bromine, fluorine and sulfonamide.

[0038] Preferably, in Formula II, R4 is selected from hydrogen.

[0039] Preferably, in Formula II, R6 and R7 are selected from hydrogen and fluorine.

[0040] Preferably, it is a compound represented by Formula II-1, an isomer, or a pharmaceutically acceptable salt thereof:

[0041]

[0042] Wherein, Y is selected from carbonyl or -CH2- group;

[0043] X3 is selected from hydrogen, nitro, methoxy, chlorine, bromine, fluorine, or sulfonamide groups;

[0044] R6 and R7 are each independently selected from hydrogen, halogen, or C. 1-6 Alkyl group; preferably derived from hydrogen or fluorine.

[0045] More preferably, in the compound, isomer or pharmaceutically acceptable salt of Formula II-1, Y is selected from carbonyl; X3 is selected from hydrogen, nitro, methoxy, chlorine, bromine or fluorine; R6 and R7 are each independently selected from hydrogen or fluorine.

[0046] Preferably, it is a compound represented by Formula III, an isomer, or a pharmaceutically acceptable salt thereof:

[0047]

[0048] Where n is selected from 0, 1, 2 or 3;

[0049] X1 and X2 are each independently selected from halogens and C. 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, Halogenated C 1-6 Alkoxy, nitro, cyano, amino, or sulfonamide groups;

[0050] X3 is selected from halogenated, substituted, or unsubstituted C. 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, hydroxy, nitro, cyano, amino, substituted or unsubstituted C 1-6 Alkylamine group, substituted or unsubstituted C 1-6 Alkyl, amide, or sulfonamide groups; when C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 alkylamine group, C 1-6 When an alkyl acyl group has substituents, it can be substituted by at least one of the following groups: deuterium, halogen;

[0051] R4 is selected from hydrogen, deuterium, hydroxyl, halogen, and C. 1-6 Alkyl, Halogenated C 1-6 Alkyl or C 1-6 Alkyl group.

[0052] Preferably, in Formula III, X1 and X2 are each independently selected from chlorine.

[0053] Preferably, when X1 and X2 are both chlorine in Formula III, X3 is selected from nitro, cyano, halogen, acetyl, methoxy, amino or substituted amino, sulfonamide, and further preferably from nitro, methoxy, chlorine, bromine, fluorine or sulfonamide.

[0054] Preferably, in Formula III, R4 is selected from hydroxyl, halogen, methyl, difluoromethyl, trifluoromethyl, and methoxy.

[0055] Preferably, it is a compound represented by Formula III-1, an isomer thereof, or a pharmaceutically acceptable salt thereof:

[0056]

[0057] Wherein, X3 is selected from nitro, cyano, halogen, acetyl, methoxy, amino or substituted amino, sulfonamide, and further preferably from nitro, methoxy, chlorine, bromine, fluorine or sulfonamide.

[0058] R4 is selected from hydroxyl, halogen, methyl, difluoromethyl, trifluoromethyl, and methoxy.

[0059] n is selected from 1 or 2.

[0060] More preferably, in the compound, isomer or pharmaceutically acceptable salt of formula III-1, X3 is selected from chlorine, bromine or fluorine; R4 is selected from hydroxyl, halogen, methyl, difluoromethyl or trifluoromethyl; and n is selected from 1.

[0061] Preferably, it is a compound represented by Formula IV, an isomer, or a pharmaceutically acceptable salt thereof:

[0062]

[0063] Among them, X1 and X2 are each independently selected from halogens and C. 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, Halogenated C 1-6 Alkoxy, nitro, cyano, amino, or sulfonamide groups;

[0064] X3 is selected from halogenated, substituted, or unsubstituted C. 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, hydroxy, nitro, cyano, amino, substituted or unsubstituted C 1-6 Alkylamine group, substituted or unsubstituted C 1-6 Alkyl, amide, or sulfonamide groups; when C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 alkylamine group, C 1-6 When an alkyl acyl group has substituents, it can be substituted by at least one of the following groups: deuterium, halogen;

[0065] R1 is selected from

[0066] R4 is selected from hydrogen, deuterium, hydroxyl, halogen, and C. 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylamine group or -OR';

[0067] R' is selected from C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 2-6 alkenyl or C 1-6 Alkyl group;

[0068] R5 is selected from hydrogen, deuterium, or C. 1-6 alkyl;

[0069] n is selected from 0, 1, 2 or 3.

[0070] Preferably, in Formula IV, X1 and X2 are each independently selected from chlorine.

[0071] Preferably, when X1 and X2 are both chlorine in Formula IV, X3 is selected from nitro, cyano, halogen, acetyl, methoxy, amino or substituted amino, sulfonamide, and further preferably from nitro, methoxy, chlorine, bromine, fluorine and sulfonamide.

[0072] Preferably, in Formula IV, R4 is selected from hydrogen, hydroxyl, halogen, methyl, difluoromethyl, trifluoromethyl, and methoxy.

[0073] Preferably, it is a compound represented by Formula IV-1, an isomer thereof, or a pharmaceutically acceptable salt thereof:

[0074]

[0075] X3 is selected from nitro, methoxy, chlorine, bromine, fluorine and sulfonamide groups;

[0076] R4 is selected from hydrogen, deuterium, hydroxyl group, or -OR'; R' is selected from C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 2-6 alkenyl or C 1-6 Alkyl group;

[0077] Furthermore, R4 is preferably derived from hydrogen, hydroxyl, halogen, methyl, difluoromethyl, trifluoromethyl, or methoxy.

[0078] More preferably, in the compound, isomer or pharmaceutically acceptable salt of Formula IV-1, X3 is selected from methoxy, fluorine or sulfonamide groups; R4 is selected from hydrogen or deuterium.

[0079] Preferably, it is a compound represented by Formula IV-2, an isomer thereof, or a pharmaceutically acceptable salt thereof:

[0080]

[0081] X3 is selected from nitro, methoxy, chlorine, bromine, fluorine and sulfonamide groups;

[0082] R1 is selected from

[0083] R4 is selected from hydrogen, deuterium, hydroxyl, halogen, and C. 1-6 Alkyl, Halogenated C 1-6 Alkyl or C 1-6 Alkoxy;

[0084] R5 is selected from hydrogen, deuterium, or C. 1-6 alkyl;

[0085] n is selected from 0, 1, or 2.

[0086] The present invention also provides a trisubstituted tetrahydroisoquinoline amide derivative, comprising compounds, isomers or pharmaceutically acceptable salts thereof numbered * to * and * to * having the following structures:

[0087]

[0088]

Terminology Definition

[0089] Unless otherwise stated, the following terms and phrases as used herein are intended to have the following meanings. A particular term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient.

[0090] The "compound" described in this invention includes, but is not limited to, compounds in the following forms: free base, stereoisomer, geometric isomer, tautomer, isotope, pharmaceutically acceptable salt, solvate, hydrate, prodrug (ester), etc.

[0091] The "compound" described in this invention can be asymmetric, for example, having one or more stereoisomers. Unless otherwise stated, all stereoisomers include, for example, enantiomers and diastereomers. Compounds containing asymmetric carbon atoms in this invention can be isolated in optically active pure form or in racemic form. Optically active pure form can be obtained by resolution of racemic mixtures, synthesis using chiral starting materials or chiral reagents.

[0092] In this invention, "isomer" refers to stereoisomers or tautomers unless otherwise specified. Unless otherwise specified, the term "stereoisomer" refers to compounds having the same chemical structure but with different spatial arrangements of atoms or groups. Stereoisomers include, but are not limited to, enantiomers, diastereomers, conformational isomers (rotational isomers), geometric isomers (cis / trans) isomers, and transisomers. Any mixture of stereoisomers obtained can be separated into pure or substantially pure geometric isomers, enantiomers, and diastereomers based on differences in the physicochemical properties of the components, for example by chromatography and / or fractional crystallization. Unless otherwise specified, the term "tautomer" refers to structural isomers with different energies that can interconvert through a low energy barrier. If tautomerism is possible (e.g., in solution), chemical equilibrium of the tautomers can be achieved. For example, proton tautomers (also known as proton transfer tautomers) include interconversions via proton transfer, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions via the rearrangement of some bonding electrons.

[0093] In this invention, "isotope" refers to a compound of this invention, unless otherwise specified, existing in an isotopically traced or enriched form, containing one or more atoms whose atomic weight or mass number differs from the atomic weight or mass number of the most abundant atoms found in nature. Isotopes can be radioactive or non-radioactive. Commonly used isotopes for isotopic labeling include hydrogen isotopes, including but not limited to... 2 H and 3 H; Carbon isotopes: including but not limited to 13 C and 14 C; Chlorine isotopes: including but not limited to 35 Cl and 37 Cl; Fluorine isotopes: including but not limited to 18 F; Iodine isotopes: including but not limited to 123 I and 125 I; Nitrogen isotopes: including but not limited to 13 N and 15 N; oxygen isotopes: including but not limited to 15 O、 17 O and 18 O; sulfur isotopes: including but not limited to 35 S. These isotope-labeled compounds can be used to study the distribution of pharmaceutical molecules in tissues, especially 3 H and 13 C, because they are easy to label and convenient to detect, are more widely used. Some heavy isotopes, such as deuterium (… 2 Substitution with H can enhance metabolic stability and prolong the half-life, thereby achieving the goal of reducing dosage and providing therapeutic advantages. Isotope-labeled compounds are generally synthesized from labeled starting materials using known synthetic techniques, just like non-isotope-labeled compounds.

[0094] In this invention, "pharmaceutically acceptable salt" refers to the salt of the compounds of this invention, which are compounds with specific substituents discovered in this invention, and are compatible with 2-acetoxybenzoic acid, 2-hydroxyethanesulfonic acid, acetic acid, ascorbic acid, benzenesulfonic acid, benzoic acid, bicarbonate, carbonic acid, citric acid, edetate, ethanedisulfonic acid, ethanesulfonic acid, fumaric acid, glucohepose, gluconic acid, glutamic acid, glycolic acid, hydrobromic acid, hydrochloric acid, hydroiodide, hydroxynaphthalene, hydroxyethanesulfonic acid, lactic acid, lactose, and dodecyl sulfonic acid. A base addition salt can be obtained by contacting a compound in its neutral form with a sufficient amount of base in a pure solution or a suitable inert solvent when it contains a relatively acidic functional group, such as maleic acid, malic acid, mandelic acid, methanesulfonic acid, nitric acid, oxalic acid, dihydroxynaphthyl acid, pantothenic acid, phenylacetic acid, phosphoric acid, polygalacturonic acid, propionic acid, salicylic acid, stearic acid, acetic acid, succinic acid, aminosulfonic acid, p-aminobenzenesulfonic acid, sulfuric acid, tannin, tartaric acid, and p-toluenesulfonic acid. Pharmaceutically acceptable base addition salts include, but are not limited to, sodium, potassium, calcium, magnesium, ammonium, or organic amine salts. Examples include alkali metal salts, alkaline earth metal salts, other metal salts, inorganic base salts, organic base salts, inorganic acid salts, lower alkyl sulfonates, aryl sulfonates, organic acid salts, and amino acid salts.

[0095] In this invention, "prodrug" refers to a prodrug of the compound of this invention that readily undergoes a chemical change under physiological conditions to be converted into the compound of this invention. Furthermore, prodrugs can be converted into the compound of this invention in the in vivo environment through chemical or biochemical methods.

[0096] The terms used in this article have the following meanings:

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

[0098] The term "alkyl" refers to a straight-chain or branched saturated hydrocarbon group composed of carbon and hydrogen atoms, such as C... 1-6 Alkyl groups, including but not limited to methyl, ethyl, propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, sec-butyl, and tert-butyl), pentyl (including n-pentyl, isopentyl, and neopentyl), and hexyl (n-hexyl, 2-methylpentyl, 3-methylpentyl, 2,3-dimethylbutyl, and 2,2-dimethylbutyl).

[0099] The term "alkenyl" refers to an unsaturated aliphatic hydrocarbon group consisting of a straight or branched chain of carbon and hydrogen atoms, having at least one double bond. Alkenyl groups can contain 2-20 carbon atoms, preferably 2-10 carbon atoms (i.e., C2H2O). 2-10 Alkenyl), further preferably 2-8 carbon atoms (i.e., C14-C2 ... 2-8 Alkenyl), more preferably 2-6 carbon atoms (i.e., C14-C2 ... 2-6 alkenyl), 2-5 carbon atoms (i.e., C) 2-5alkenyl), 2-4 carbon atoms (i.e., C) 2-4 alkenyl), 2-3 carbon atoms (i.e., C) 2-3 Alkenyl), 2 carbon atoms (i.e., C2 alkenyl), for example "C 2-6 "Alkenyl" refers to a group that is alkenyl and has 2 to 6 carbon atoms in its carbon chain (i.e., 2, 3, 4, 5, or 6). Non-limiting examples of alkenyl groups include, but are not limited to, vinyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 1-methyl-1-propenyl, 2-methyl-1-propenyl, 1,3-butadien-1-yl, 1,3-butadien-2-yl, etc.

[0100] The term "cycloalkyl" refers to a monocyclic alkyl group composed of carbon and hydrogen atoms, such as C1. 3-8 Cycloalkyl groups, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0101] The term "alkoxy" refers to a straight-chain or branched alkyl group linked by an oxygen atom, such as C... 1-6 Alkoxy groups, including but not limited to methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), butoxy (including n-butoxy, isobutoxy, sec-butoxy, and tert-butoxy), pentoxy (including n-pentoxy, isopentoxy, and neopentoxy), and hexoxy (n-hexoxy, 2-methylpentoxy, 3-methylpentoxy, 2,3-dimethylbutoxy, and 2,2-dimethylbutoxy).

[0102] The term "alkylamine" refers to an open-chain alkyl group containing a nitrogen atom, such as C... 1-6 Alkylamine groups, including but not limited to methylamino, ethylamino, isopropylamino, dimethylamino, methylethylamino, diethylamino, etc.

[0103] The term "alkanoyl" refers to a straight-chain or branched alkyl group linked by an acyl group, i.e., -alkyl-CO-, such as C 1-6 Alkyl groups, including but not limited to formyl, acetyl, propionyl (including n-propionyl and isopropionyl), butyryl (including n-butyryl, isobutyryl, sec-butyryl, and tert-butyryl), valeryl (including n-valeryl, isovaleryl, and neovaleryl), and hexanoyl (n-hexanoyl, 2-methylvaleryl, 3-methylvaleryl, 2,3-dimethylbutyryl, and 2,2-dimethylbutyryl).

[0104] The term "aryl" refers to a monocyclic or fused polycyclic group with 6-16 carbon atoms, possessing a fully conjugated π-electron system, including but not limited to phenyl, naphthyl, anthracene, etc., with phenyl being preferred.

[0105] The term "heterocyclic group" refers to a saturated or partially unsaturated monocyclic or polycyclic (e.g., spirocyclic, bridged, etc.) group containing 3-20 ring atoms, and having a non-aromatic structure. The polycyclic group may consist entirely of non-aromatic rings, or at least one ring may be aromatic while the rest are non-aromatic. The aforementioned 3-20 ring atoms contain one or more (e.g., 2, 3, 4 or more) heteroatoms, with the remainder being carbon atoms selected from one or more of N, O, and S. The aforementioned monocyclic or polycyclic group may include the same or different heteroatoms in one or more rings, and the number of heteroatoms may be one or more. Non-limiting examples of "heterocyclic group" include, but are not limited to, azirropropyl, oxadiropropyl, thioherropropyl, azirrobutyl, oxadirobutyl, thioherrobutyl, furanyl, piperidinyl, piperazinyl, morpholinyl, etc.

[0106] The term "heteroaryl" refers to an aromatic monocyclic or polycyclic (e.g., fused ring) group containing 5-16 ring atoms, wherein the aforementioned 5-16 ring atoms contain one or more (e.g., 2, 3, 4 or more) heteroatoms, and the remainder are carbon atoms selected from one or more of N, O, and S. The aforementioned monocyclic or polycyclic group may include the same or different heteroatoms in one or more rings, and the number of heteroatoms may be one or more. The aforementioned "heteroaryl" preferably contains 5-14, 5-12, 5-10, or 5-8 ring atoms, more preferably 5-6 ring atoms. Non-limiting examples of "heteroaryl" include, but are not limited to, tetrahydrofuranyl, thiophenyl, oxazolyl, thiazolyl, pyrroleyl, pyrazolyl, imidazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, quinolinyl, indolyl, benzofuranyl, benzothiophenyl, benzimidazolyl, benzopyridyl, benzopyrimidinyl, benzopyrazinyl, etc.

[0107] The method for preparing the above-mentioned compound provided by the present invention involves the following steps, but is not limited to these steps:

[0108] Synthesis Method 1:

[0109] The reaction route is as follows:

[0110]

[0111] Where Y is selected from carbonyl, then X is selected from hydroxyl, pentafluorophenoxy, halogen, or The radical; when Y is selected from -CH2-, then X is selected from halogen or sulfonate; Pro1 is Me, Et, iPr, t-Bu or Bn; R1, X1, X2, X3 are defined as described in claim 1, and R1 is further preferably as described in claim 2.

[0112] The reaction process is as follows:

[0113] 1) Preparation of starting material A for the reaction: When Y is carbonyl (i.e. -CO-), formula A is the product of aryl and / or heteroaryl carboxylic acids and / or carboxyl groups after activation; when Y is methylene (i.e. -CH2-), A is a benzyl active compound of aryl and / or heteroaryl groups.

[0114] The main step in this reaction is to first prepare the left-wing fragment. When Y = carbonyl and X = hydroxyl, it is an aryl or heteroaryl carboxylic acid. The carboxyl group can be activated to form a sulfonate, pentafluorophenyl ester, or acyl chloride, or reacted with ethyl chloroformate or isobutyl ester to generate a mixed anhydride, which then further undergoes a condensation reaction with the amino group of the central nucleus, a "trisubstituted tetrahydroisoquinoline (carboxylic acid) compound." Alternatively, this condensation reaction can be performed directly with the central nucleus using aryl or heteroaryl carboxylic acids in the presence of condensing agents such as DCC, DIC, EDCI, HOBt, HBTU, and TBTU.

[0115] When Y = carbon (i.e. -CH2-), A is an aryl or heteroaryl benzyl active compound, including but not limited to benzyl halides, benzyl sulfonates, or other easily leaving mixed esters.

[0116] 2) Preparation of the starting material B-acid: A protected tetrahydroisoquinoline (carboxylic acid) compound is deprotected under acidic conditions or acidified after deprotection to obtain the key starting material B-acid; wherein the acid radical can be an inorganic acid or an organic acid such as HCl, HBr, HI or CF3COOH; (see Synthesis Method II and Examples below for details)

[0117] This step primarily yields the central core, a trisubstituted tetrahydroisoquinoline (carboxylic acid) compound. To facilitate the preservation of the intermediate, it is typically prepared as a hydrochloride salt. During the subsequent reaction, the amino group of tetrahydroisoquinoline (THIQ) is neutralized in situ and further reacted. Since the protecting group is mainly a conventional amino protecting group, the removal of the protecting group can be done under appropriate acidic conditions.

[0118] 3) A and B-acids undergo a condensation reaction to generate the key intermediate product C;

[0119] The main purpose of this condensation reaction is to generate the amide structure on the left wing. If B is in the form of hydrochloride, the condensation can be carried out in situ after neutralization under alkaline conditions. If A is an unactivated carboxylic acid, it can also react directly with the central nucleus in the presence of condensing agents such as DCC, DIC, EDCI, HOBt, HBTU, TBTU, and HATU.

[0120] 4) Preparation of starting material D: The carboxyl group of m-methylsulfonyl L-phenylalanine was protected to obtain compound D;

[0121] This protection reaction is a conventional reaction for protecting carboxyl groups. Pro1 includes, but is not limited to, Me, Et, iPr, t-Bu, and Bn, as long as the protecting group can be successfully removed subsequently.

[0122] 5) In the presence of a condensing agent, the above intermediate products C and D are subjected to a condensation reaction to obtain intermediate product E;

[0123] The main purpose of the condensation reaction between C and D is to generate the right-wing amide structure. Since this condensation reaction is a direct condensation between a carboxyl group and an amino group, the condensing agents used include, but are not limited to, DCC, DIC, EDCI, HOBt, HBTU, TBTU, and HATU. A mild condensation reaction can be carried out under alkaline conditions. The preferred process is as follows: intermediate products C and D are mixed and reacted in a DIPEA (or TEA) + HATU system with stirring for 2-8 hours. The reaction is then quenched with an acid solution, followed by extraction and rotary evaporation. The molar ratio of intermediate products C to D is 1:1-3.

[0124] 6) After removing the protecting group Pro1 from intermediate product E, the target product is obtained.

[0125] The final step is to remove the protecting group on the right-hand carboxyl group under mild conditions, while avoiding the hydrolysis of the amide bond. Any method can be used to convert one of the ester groups to a carboxyl group, such as conventional acidic or basic hydrolysis. A preferred synthetic method involves adding a polar solvent, water, and an alkali (such as hydroxide, or TEA+LiBr) to intermediate E, reacting at 10-35°C for 0.1-10 hours, quenching with an inorganic or organic acid, and then extracting and drying. The molar / mass ratio of intermediate E to the alkali is generally 1:0.1-0.5. If acidic hydrolysis is used, such as selective ester hydrolysis in formic acid, the temperature can be 20-70°C.

[0126] Of course, the final product will undergo a series of purification processes such as recrystallization. Therefore, adding corresponding purification processes does not mean exceeding the scope of this patent.

[0127] The first general synthesis method provided by this invention mainly involves first constructing the left-wing active intermediate, and then completing the condensation reaction of the right wing to obtain the target compound.

[0128] The present invention also provides the following second synthesis method, which mainly involves first constructing the right-wing active intermediate, and then completing the condensation reaction of the left wing to obtain the target compound.

[0129] Synthesis Method Two:

[0130] The reaction route is as follows:

[0131]

[0132] Wherein, Pro2 is H or other amine protecting groups (Bn, Cbz, Boc, Fmoc, Alloc, Teoc, Tos, Tfa, Trt, Dmb, PMB, Me, Et, etc.) and their salts; the definitions of the remaining substituents are the same as in Synthesis Method 1.

[0133] The reaction process is as follows:

[0134] 1) Mix IM4 and another starting material D in a polar solvent, add an organic base and a condensing agent to carry out a condensation reaction to obtain the key intermediate G;

[0135] 2) Selectively remove the protecting group Pro2 of G under acidic conditions to obtain the key intermediate H; wherein the acid radical can be an inorganic acid or an organic acid such as HCl, HBr, HI or CF3COOH.

[0136] 3) The above-mentioned key intermediate H is further condensed with compound A under alkaline conditions to obtain intermediate E;

[0137] 4) Remove the protecting group Pro1 from intermediate E to obtain the target product formula (I).

[0138] This invention also provides the key starting material IM4 in the above-mentioned synthesis method (II) and its preparation method:

[0139] The reaction route is as follows:

[0140]

[0141] The definitions of each substituent are the same as in Synthesis Method 1.

[0142] Starting with trisubstituted benzene (SM), a trisubstituted isoquinoline intermediate (IM1) was obtained through formylation, condensation, reduction, Ts protection, and ring closure. The trisubstituted isoquinoline intermediate (IM1) was then reduced to a trisubstituted tetrahydroisoquinoline intermediate (IM2). After amino protection, the trisubstituted tetrahydroisoquinoline intermediate (IM2) underwent a carboxylation reaction to obtain the aforementioned protected trisubstituted tetrahydroisoquinoline (carboxylic acid) compound (IM4). The protecting group of IM4 was removed under acidic conditions to obtain compound B-acid.

[0143] The present invention also provides a pharmaceutical composition comprising at least one compound as described above or a pharmaceutically acceptable salt thereof as an active ingredient, and at least one or more pharmaceutically acceptable carriers.

[0144] The term "pharmaceutical composition" as used in this invention refers to a formulation comprising one or more compounds of the invention or salts thereof, and a carrier, in commonly accepted terms in the art, for delivering a bioactive compound to an organism (e.g., a human). The purpose of a pharmaceutical composition is to facilitate drug delivery to an organism.

[0145] The routes of administration of the compounds described in this invention or their pharmaceutically acceptable salts or hydrates, as well as deuterated or other isotopically substituted compounds or their pharmaceutical compositions, include, but are not limited to, oral, rectal, transmucosal, enteral administration, or local, transdermal, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, and intravenous administration.

[0146] Preferably, the route of administration is intraocular (eye drops).

[0147] The present invention also provides the use of the preparation of the compounds or pharmaceutical compositions described above in immune cell migration inhibitors.

[0148] The present invention also provides the use of the preparation of the compound or pharmaceutical composition as described above in the prevention or treatment of diseases associated with LFA-1 activity.

[0149] Preferably, the diseases mentioned above include, but are not limited to, dry eye syndrome and T-cell inflammatory response diseases.

[0150] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0151] This invention, based on the target design of LFA-1 antagonists, has developed a series of novel multi-substituted tetrahydroisoquinoline amide derivatives. Related biological experiments have shown that most compounds exhibit strong permeability and good water solubility, possessing significant LFA-1 antagonistic activity. Some compounds even demonstrate superior irritation-reducing effects compared to positive control drugs, significantly reducing adverse reactions and demonstrating significant clinical application value. Furthermore, the synthetic routes provided by this invention are novel, safe, environmentally friendly, and have good production feasibility. Detailed Implementation

[0152] The following are specific embodiments of the present invention, which further describe the technical solution of the present invention. However, the scope of protection of the present invention is not limited to these embodiments. All changes or equivalent substitutions that do not depart from the concept of the present invention are included within the scope of protection of the present invention.

[0153] Furthermore, all operations involving easily oxidized or hydrolyzed raw materials are performed under nitrogen protection. Unless otherwise stated, the raw materials used in this invention are commercially available and can be used directly without further purification.

[0154] All reaction raw materials and common intermediates involved in the embodiments of the present invention can be obtained commercially or by self-production. The preparation process of raw materials and common intermediates that need to be self-produced is described in detail below.

[0155] I. Preparation of the common intermediates IM4 and β-acid

[0156]

[0157] 1) Starting with trisubstituted benzene (SM), the trisubstituted isoquinoline intermediate (IM1) was obtained after formylation, condensation, reduction, Ts protection and cyclization.

[0158] The process from SM to IM1 actually involves five chemical reaction steps, including formylation, condensation, reduction, Ts protection, and cyclization. These unit reactions are all performed under classic organic synthesis conditions. Through the combination of these five unit reactions, the trisubstituted isoquinoline ring intermediate (IM1) required by this invention was successfully constructed. Due to the differences in the functional groups X1, X2, and X3, some classical protection and deprotection measures for these functional groups may be involved in certain stages of the above unit reactions. However, these measures can all be completed at this stage and therefore do not exceed the scope of protection of this patent.

[0159] A typical reaction equation from SM to IM1 is shown below:

[0160]

[0161] 2) After reduction, the trisubstituted isoquinoline intermediate (IM1) yields the trisubstituted tetrahydroisoquinoline intermediate (IM2);

[0162] IM1 is hydrogenated to reduce the pyridine ring of the isoquinoline ring, yielding the trisubstituted tetrahydroisoquinoline intermediate (IM2). Reducing agents that can be used include, but are not limited to, NaBH4, KBH4, or their partially substituted derivatives such as sodium cyanoborohydride, lithium aluminum hydride, etc. Other suitable reducing agents for this type of reaction can also be used, such as Pd / C, hydrogen, etc.

[0163] 3) The trisubstituted tetrahydroisoquinoline intermediate (IM2) is protected with an amino group to obtain intermediate IM3, which is then further reacted with a carboxyl group to obtain the trisubstituted tetrahydroisoquinoline (carboxylic acid) compound (IM4) with a protecting group.

[0164] The carboxylation (addition of a carboxyl group) reaction involves the insertion of CO2, therefore the amino group of the trisubstituted tetrahydroisoquinoline intermediate (IM2) needs to be protected beforehand (to obtain IM3). The protecting group Pro2 involved is hydrogen or other amino protecting groups (Bn, Cbz, Boc, Fmoc, Alloc, Teoc, Tos, Tfa, Trt, Dmb, PMB, Me, Et groups) and their salts. The above amino protection reaction can be carried out under alkaline conditions: add the chloride or anhydride of the corresponding protecting group (such as Boc anhydride) (1-3 eq) to the intermediate IM2 (1.0 eq), and then stir until the starting material spot IM2 disappears on the TLC, thus obtaining IM3.

[0165] After IM3 is mixed and reacted with butyllithium at a low temperature (generally below -20℃), dry ice is added (or CO2 gas is introduced), and the reaction continues until carboxylation is complete. Then, the reaction solution is acidified to obtain a trisubstituted tetrahydroisoquinoline (carboxylic acid) compound (IM4) with a protecting group. The molar ratio of IM3 to butyllithium is (1:1) to (1:4) (i.e., butyllithium should be in excess, but generally not exceeding 4.0 eq). This reaction can also be activated by adding tetramethylethylenediamine (TMEDA), hexamethylphosphoramide (HMPA), 1,4-diazabicyclo[2.2.2]octane (DABCO), or 2,2,6,6-tetramethylpiperidine.

[0166] 4) The protecting group of a trisubstituted tetrahydroisoquinoline (carboxylic acid) compound (IM4) is deprotected under acidic conditions or acidified after deprotection to give the salt of intermediate product B (B-acid). The anion of B-acid can be a conventional inorganic or organic acid, including but not limited to HCl, HBr, HI, or CF3COOH.

[0167] II. Preparation of the common intermediate β-acid compound

[0168] When X1 = X2 = Cl, X3 is nitro (B1.1), methoxy (B1.2), halogen (Cl: B1.3; Br: B1.4; F: B1.5), or sulfonamide (B1.6), the specific codes and structures are shown in Table 1 and Table 1b below.

[0169]

[0170] The anions of β-acids include, but are not limited to, inorganic or organic acids such as HCl, HBr, HI, or CF3COOH.

[0171] Table 1: Codes of specific intermediates for intermediate series B compounds

[0172] <![CDATA[X1 / X2 / X3 / Pro2]]> SM IM1 IM2 IM3 IM4 B-acid <![CDATA[Cl / Cl / NO2 / Boc]]> SM1.1 IM1.1 IM2.1 IM3.1 IM4.1 B1.1 Cl / Cl / OMe / Bn SM1.2 IM1.2 IM2.2 IM3.2 IM4.2 B1.2 Cl / Cl / Cl / Boc SM1.3 IM1.3 IM2.3 IM3.3 IM4.3 B1.3 Cl / Cl / Br / Trt SM1.4 IM1.4 IM2.4 IM3.4 IM4.4 B1.4 Cl / Cl / F / Trt SM1.5 IM1.5 IM2.5 IM3.5 IM4.5 B1.5 <![CDATA[Cl / Cl / SO2NH2 / Boc]]> SM1.6 IM1.6 IM2.6 IM3.6 IM4.6 B1.6

[0173] Table 1b: Codes and structural formulas of intermediate B-series compounds

[0174]

[0175] Example (1) Preparation of intermediate B1.3:

[0176] The synthetic route for intermediate B1.3 is as follows:

[0177]

[0178] 1. Starting with SM1.3, after formylation, condensation, reduction, Ts protection, and ring closure, a trisubstituted isoquinoline intermediate (IM1.3) is obtained. A typical reaction equation from SM1.3 to IM1.3 is shown below:

[0179]

[0180] 1) Formylation: To a frozen (<-10℃) tetrahydrofuran solution containing 2,2,6,6-tetramethylpiperidine (1-2 eq) and butyllithium (1-2 eq), SM1.3 (1.0 eq) of THF solution and DMF (1-10 eq) were added dropwise, followed by slow heating to 0℃. After the reaction was complete, the mixture was quenched with NH4Cl aqueous solution, extracted with EtOAc, and the combined organic phases were washed with brine, dried with Na2SO4, concentrated under vacuum, and the residue was purified by silica gel column chromatography (petroleum ether / EtOAc = 50 / 1 to 10 / 1) to give a yellow oily formylated intermediate SM1.3.1 (yield ~85%).

[0181] 2) Condensation: The formylated intermediate SM1.3.1 (1.0 eq) was dissolved in toluene by stirring. Aminoacetaldehyde dimethyl acetal (or diethyl acetal) (1–3 eq) was added, and the mixture was heated under reflux to dehydrate until the starting material was almost completely eliminated. The solvent was evaporated to dryness to obtain a brown liquid condensed imine intermediate SM1.3.2, which can be used in the next step without further purification.

[0182] 3) Reduction: NaBH4 (0.5–2.0 eq) was added in portions to a stirred methanol solution of crude imine SM1.3.2 (1.0 eq) at 0 °C, and the mixture was stirred at room temperature for 4 hours until the reaction was complete. The reaction was quenched with ice water, and the solvent was evaporated. The residue was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, and evaporated to dryness. Purification was performed by column chromatography using an ethyl acetate / n-hexane (7:3) mixture to give the yellow liquid compound SM1.3.3 (two-step yield approximately 84%).

[0183] 4) Ts protection: DMAP (0.01–0.2 eq) and triethylamine (1–3 eq) were added to a stirred dichloromethane solution of the reduced product SM1.3.3 (1.0 eq), followed by the addition of p-toluenesulfonyl chloride (1–2 eq) at 0 °C. The reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was quenched with ice water and extracted with dichloromethane. The combined organic layers were washed with water and brine, dried over sodium sulfate, and concentrated to dryness. Purification was performed by column chromatography using an ethyl acetate / n-hexane (1:3) mixture to give a white, foamy SM1.3.4 (approximately 87% yield).

[0184] LC-MS m / z = 497, [M+H+NH3] + .

[0185] 5) Cyclation: At 0°C, a dichloromethane solution of the above-mentioned Ts-protected compound SM1.3.4 (1.0 eq) was added to a stirred suspension of AlCl3 (2–10 eq) in dichloromethane. The reaction mixture was stirred at room temperature for approximately 16 hours until complete. The reaction mixture was poured into ice water and extracted with dichloromethane (500 mL). The combined organic layers were washed successively with saturated NaHCO3 solution, brine, and dried over sodium sulfate, and then concentrated to dryness. Purification was performed by column chromatography using an ethyl acetate / n-hexane (2:8) mixture to give a white to brownish-white solid as the desired trisubstituted isoquinoline IM1.3 (yield approximately 47%).

[0186] LC-MS m / z = 232, [M+H] + .

[0187] 2. Trisubstituted isoquinoline IM1.3 was reduced to prepare trisubstituted tetrahydroisoquinoline IM2.3.

[0188] At 0 °C, NaBH4 (1.5–5 eq) was added in portions to an acetic acid solution of trisubstituted isoquinoline IM1.3 (1.0 eq), and the reaction mixture was stirred at room temperature until complete. The reaction mixture was quenched with saturated sodium bicarbonate solution. The mixture was extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to dryness to give the desired compound IM2.3, which is a brown to brown oily or semi-solid substance that can be used without further purification.

[0189] 1 H NMR (400MHz, DMSO-d6) δ7.50 (s, 1H), 3.82 (s, 2H), 2.96-3.10 (t, J = 8.5Hz, 2H), 2.73-2.79 (t, J = 8.5Hz, 2H).

[0190] LC-MS m / z = 236.0, [M+H] +.

[0191] 3. Preparation of IM3.3 by protecting the 3-amino group of trisubstituted tetrahydroisoquinoline IM2.

[0192] While stirring at 0 °C, DIPEA (1.0–3.0 eq) and (Boc)₂O (1.0–2.5 eq) were added to a THF solution of IM₂.₃ (1.0 eq). The reaction mixture was stirred at room temperature until the reaction was complete. The reaction mixture was concentrated to dryness under reduced pressure, and the residue was dissolved in ethyl acetate. The combined organic layers were washed with water, then with brine, dried over anhydrous sodium sulfate, and the solution was concentrated to dryness. Purification was performed by column chromatography using an ethyl acetate / petroleum ether (1:10) mixture to give the Boc-protected compound IM₃.₃ (two-step yield approximately 83%).

[0193] 1 H NMR (400MHz, DMSO-d6) δ7.32 (d, 1H), 4.59 (s, 2H), 3.66 (t, J = 5.52Hz, 2H), 2.80 (t, J = 5.77Hz, 2H), 1.49 (s, 9H).

[0194] LC-MS m / z = 353, [M+H+NH3] + .

[0195] 4. Preparation of compound IM4.3 by carboxylation of compound IM3.3.

[0196] Under nitrogen protection, a THF solution of IM3.3 (1.0 eq) was added to the reaction flask. The temperature was lowered to below -40°C, and TMEDA (1–4 eq) and n-BuLi (1–4 eq) were added. After the reaction, dry ice was added (or CO2 was introduced), and the reaction was allowed to proceed at low temperature for a period of time before being allowed to return to the room temperature until carboxylation was complete. The reaction solution was quenched with ammonium chloride or citric acid solution, extracted with ethyl acetate, and the combined organic phases were concentrated to dryness to obtain the carboxylated product IM4.3 with the protecting group. The subsequent deprotection reaction can be carried out directly without separation.

[0197] 5. Compound B1.3 was prepared by removing the protecting group from compound IM4.3.

[0198] The above-mentioned IM4.3 was dissolved in an appropriate amount of dichloromethane, and trifluoroacetic acid (1-20 eq) was added. The Boc protecting group was removed by stirring at room temperature. The reaction solution was concentrated to dryness and then dissolved in water. The solution was neutralized with sodium bicarbonate aqueous solution and then extracted with dichloromethane. After drying with anhydrous sodium sulfate, the solution was filtered. The filtrate was acidified with hydrochloric acid and then filtered or concentrated to dryness to obtain B1.3 (the hydrochloride salt of compound B). The yield of the two steps was 75%.

[0199] 1H NMR (400MHz, DMSO-d6) δ 10.08 (br, 2H), 4.29 (s, 2H), 3.35 (t, J = 5.87Hz, 2H), 2.95 (t, J = 5.87Hz, 2H).

[0200] LC-MS m / z = 280.1, [M+H] + .

[0201] The above deprotection reaction can also be carried out under acetone and concentrated hydrochloric acid conditions. After the reaction solution is concentrated to dryness, it can be pulped with acetone or filtered to obtain B1.3.

[0202] Example (II) Preparation of intermediate B1.5:

[0203] When X1 = X2 = Cl and X3 = F, the intermediate is B1.5. The synthesis method can be the same as the preparation method of the intermediate B1.3. Pro2 can be a Trt group.

[0204] The synthetic route for intermediate B1.5 is as follows:

[0205]

[0206] 1) IM1.5 was obtained by referring to the preparation method of IM1.3.

[0207] Following the synthetic route of intermediate B1.3, the preparation method of IM1.3 was used, but only SM1.3 was replaced with SM1.5. The rest of the method was the same, and compound IM1.5 was prepared.

[0208] 2) IM2.5 was obtained by referring to the preparation method of IM2.3.

[0209] Following the synthetic route of intermediate B1.3, the preparation method of IM2.3 was used, but only IM1.3 was replaced with IM1.5. The rest of the method was the same, and compound IM2.5 was prepared.

[0210] 3) Preparation of IM3.5 by Trt protection reaction.

[0211] Compound IM2.5 (1.0 eq) was dissolved in THF, and DIPEA (1–3 eq) was added with stirring, followed by TRT-Cl (1–3 eq). The reaction was monitored by TLC until the starting material spot disappeared. The reaction mixture was filtered to remove the precipitate (which was washed three times with THF). The filtrate was concentrated under reduced pressure to obtain the crude product, which was then dispersed in EA and water. After freezing to 5°C, the mixture was filtered to obtain a white granular solid IM3.5 with an HPLC purity greater than 90% and a yield of approximately 85%.

[0212] 1H NMR (400MHz, DMSO-d6) δ7.14~7.52 (m, 16H), 3.46 (s, 2H), 2.90 (t, J = 5.92Hz, 2H), 2.51 (t, J = 5.91Hz, 2H).

[0213] The solvent for the above reaction can be replaced with dichloromethane or other solvents that can dissolve Trt-Cl. DIPEA can also be replaced with other organic bases. The order of adding the reactants is not limited. If the entire reaction is protected with nitrogen, the production of N-oxides can be reduced, and the appearance of the product can be improved. Slurrying EA helps to remove byproduct impurities.

[0214] 4) Preparation of IM4.5 by carboxylation reaction

[0215] THF solvent and 2,2,6,6-tetramethylpiperidine (1–4 eq) were added to the reaction flask, and the mixture was purged with nitrogen and cooled to approximately -70°C. Then, n-BuLi (1–4 eq) was added dropwise to the system, and the mixture was stirred at -70°C for 30 min after the addition was complete. Next, 1.0 eq of IM3.5 THF solution was added dropwise, and the mixture was stirred for 1 h after the addition was complete. Then, dry ice (1–10 eq) was added to the reaction system, and the mixture was kept at -60°C for 1 h, and then cooled to -10°C. The reaction solution was quenched with ice water, and the pH was adjusted to 1 with hydrochloric acid. A large amount of white solid precipitated out. The solid was filtered out and dried to obtain IM4.5 (TLC purity approximately 95%, DCM:MeOH:HOAc mixed eluent, IM4.5 product Rf approximately 0.3), yield approximately 88%.

[0216] 5) Preparation of B1.5 by removing protecting groups:

[0217] Add acetone to IM4.5 (1.0 eq) and stir until the material is dispersed. Add concentrated hydrochloric acid (approximately 0.5–2 w / w) and stir at room temperature to reflux temperature until deprotection is complete. Filter, and slurry the filter cake with acetone and dry to obtain a white to pink solid B1.5, which is approximately one molecule of hydrochloride, with a yield of approximately 75%. Mp 280–283℃.

[0218] 1 H NMR (400MHz, DMSO-d6) δ 10.02 (br, 2H), 4.33 (s, 2H), 3.39 (t, J = 8.00Hz, 2H), 3.00 (t, J = 8.00Hz, 2H).

[0219] LC-MS m / z = 264, [M+H] + .

[0220] Example (3) Preparation of intermediate B1.2:

[0221] The synthetic route for intermediate B1.2 is as follows:

[0222] When X1 = X2 = Cl and X3 is methoxy (B1.2), the synthesis method can be the same as the preparation method of the intermediate B1.3 mentioned above, and Pro2 can be Bn-based.

[0223]

[0224] 1) IM1.2 was obtained by referring to the preparation method of IM1.3.

[0225] Following the synthetic route of intermediate B1.3, the preparation method of IM1.3 was used, but only SM1.3 was replaced with SM1.2. The rest of the method was the same, and compound IM1.2 was prepared.

[0226] 2) IM2.2 was obtained by referring to the preparation method of IM2.3.

[0227] Following the synthetic route of intermediate B1.3, the preparation method of IM2.3 was used, but only IM1.3 was replaced with IM1.2. The rest of the method was the same, and compound IM2.2 was prepared.

[0228] 3) Preparation of IM3.2 by Bn protection reaction.

[0229] DMF (or DMSO) solvent, potassium carbonate (1–5 eq), and IM2.2 (1.0 eq) were added to the reaction flask. While stirring, benzyl bromide (1–2 eq) was added dropwise at approximately 0°C. The reaction mixture was stirred until TLC showed complete reaction of the starting materials. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined and washed with saturated brine. The organic phase was concentrated under reduced pressure to dryness to obtain a brownish-yellow oily substance IM3.2, with a yield of 80%.

[0230] 1 H NMR (400MHz, CDCl3) δ7.58(s,1H),7.34-7.45(m,5H),3.83(s,3H),3.67(s,2H),3.53(s,2H),2.74-2.83(m,4H).

[0231] 4) Preparation of IM4.2 by carboxylation reaction:

[0232] THF solvent, TMEDA (1–4 eq), and IM3.2 (1.0 eq) were added to the reaction flask. After purging with nitrogen, the mixture was cooled to approximately -70°C. Then, n-BuLi (1–4 eq) was added dropwise to the system. After the addition was complete, the mixture was stirred at -70°C for 30 min. CO2 was then introduced into the system, and the temperature was maintained at -60°C. The mixture was then stirred at -60°C for 1 h, and then allowed to naturally return to 0°C. Ammonium chloride solution was added to adjust the pH to 6, and the mixture was extracted with ethyl acetate. The organic phases were combined, and a suitable amount of concentrated hydrochloric acid was added. The mixture was stirred at 10°C until crystallization was complete. The mixture was filtered, and the filter cake was dried to obtain a white to off-white solid, IM4.2, with a yield of 79%.

[0233] 1 H NMR (400MHz, DMSO-d6) δ10.05(br,1H),7.32-7.47(m,5H),4.43(s,3H),4.38(s,2H),4.29(s,2H),3.07-3.51(m,4H).

[0234] 5) Preparation of B1.2 by removing protecting groups:

[0235] IM4.2 (1.0 eq), 5% Pd / C (10% w / w), 12N HCl (1-2 w / w), and MeOH (5-10 w / w) were added sequentially to the hydrogenation reactor; the reactor was purged with nitrogen three times, followed by purging with hydrogen three times; the temperature was raised to 50°C, and the hydrogen pressure was maintained at 1-5 atm for about 20 h; TLC showed that the reaction was complete; the mixture was filtered, and the filter cake was washed with methanol; the filtrate was concentrated to dryness under reduced pressure; the mixture was dispersed in methanol and water, slurried, filtered, and dried to obtain a white to off-white B1.2 solid with a yield of 90%.

[0236] LC-MS m / z = 276, [M+H] + .

[0237] Example (4) Preparation of intermediate B1.6:

[0238] Refer to Table 1: Codes for specific intermediates in the B series, with B1.6 representing the specific compounds.

[0239] When X1 = X2 = Cl and X3 is SO2NH2 (B1.6), the synthesis method can be the same as the preparation method of the intermediate B1.3 mentioned above, and Pro2 can be a Boc group or a Trt group.

[0240]

[0241] The yields of each step of the reaction are as follows: SM1.6 was prepared into an isoquinoline ring via formylation, condensation, reduction, Ts protection, and ring closure (combined yield approximately 35%). IM1.6 was further reduced to yield a crude IM2.6 with a higher yield of 95%, and the yield of the Boc protection step was approximately 90% (the yield of the Tlt group protection step was approximately 94%). The carboxylation and deprotection reactions could be carried out consecutively to give B1.6 with a yield of approximately 72%.

[0242] LC-MS m / z = 325, [M+H] + .

[0243] Example (5) Preparation of intermediate B1.4:

[0244] Refer to Table 1: Codes of specific intermediates in the B series. B1.4 represents a specific compound, X3 is Br (B1.4). The synthesis method can be the same as that for intermediate B1.5 mentioned above. Pro2 can be a Trt group.

[0245]

[0246] Example (VI) Preparation of intermediate B1.1:

[0247] Refer to Table 1: Codes of specific intermediates in the B series of compounds. B1.1 represents a specific compound, X3 is NO2 (B1.1). The synthesis method can be the same as the preparation method of B1.3 above. Pro2 can be a Boc group.

[0248]

[0249] Example 1: Synthesis of compound FR111 (X3 = NO2; B-acid = B1.1)

[0250] The synthesis route is as follows:

[0251]

[0252] 1) Benzofuran-2-carboxylic acid (SM2, 1.0 eq) was treated in toluene with oxaloyl chloride (1-2 eq) and a catalytic amount of DMF, followed by the addition of pentafluorophenol (1-1.5 eq) and DIPEA (1-10 eq). The reaction was continued until no raw material was observed on TLC. The reaction was quenched with water and separated. The organic phase was washed once with sodium bicarbonate aqueous solution and once with water. The solution was concentrated under reduced pressure to a viscous state to obtain a light brown oily substance (or semi-solid) as the pentafluorophenyl ester intermediate.

[0253] 2) Dissolve the above pentafluorophenyl ester in acetonitrile (or other suitable solvents such as DCM for amine ester exchange reactions). Then add B1.1 (0.8–1.0 eq) and DIPEA (1–10 eq), and continue the reaction until no B1.1 raw material is visible on the TLC plate (THF:methanol:ammonia = 8:1:1). If necessary, the reaction can be warmed to promote complete reaction. After the reaction is complete, quench the reaction with dilute hydrochloric acid, cool to room temperature with stirring, and filter to obtain a light brown solid C111 (i.e., C11, X3 = NO2), with an HPLC purity of approximately 98% and a yield of approximately 75%.

[0254] 3) Referring to the literature, (S)-2-amino-3-(3-(methylsulfonyl)phenyl)propionic acid (SM1 or its HCl, HBr, HI, CF3COOH salt) (1.0 eq) was dissolved in tert-butyl acetate. Perchloric acid (1-3 eq) was added at 0°C, and the mixture was kept at 25°C for 6-7 hours. After the reaction was complete, water (50 L) was added and the organic layer was separated. The pH of the aqueous layer was adjusted to alkaline, and the solution was extracted with dichloromethane to obtain a dichloromethane solution containing tert-butyl (S)-2-amino-3-(3-(methylsulfonyl)phenyl)propionic acid (D11) for later use.

[0255] 4) Add the C111 intermediate (1.0 eq) and triethylamine (1-5 eq) from 2) above to DMF (5-10 w / w) at approximately 30°C and stir until homogeneous. Add HATU (1-2 eq), then heat to 45°C and maintain for 1-2 hours. After the reaction is complete, cool the reaction to 30°C and add the dichloromethane solution containing D11 (1-2 eq) from 3) above. Maintain the resulting reaction solution at 30°C for 5-6 hours until TLC shows complete reaction. After coupling is complete, add water (5-10 w / w) and separate the organic layer. Wash the obtained organic layer with 5% sodium carbonate solution, then with dilute hydrochloric acid and 5% brine. Concentrate the organic layer to obtain the E111 intermediate (i.e., E11, X3 = NO2), with an HPLC purity of approximately 97% and a yield of approximately 92%.

[0256] 5) Dilute the E111 intermediate with formic acid (5-10 w / w) and maintain at 60°C for 6-7 hours to complete hydrolysis. Add the reactants to chilled water, maintain at 0°C for 2 hours, and filter. The resulting wet solid, after drying, yields the FR111 compound as an off-white solid, with a yield of 87.5%.

[0257] LC-MS m / z = 660, [M+H] + .

[0258] The above reaction can also be simplified by treating C111 in DCM with oxalyl chloride and a catalytic amount of DMF to obtain the acyl chloride compound of C111. SM1 (or its HCl, HBr, HI, or CF3COOH salt) is dissolved in acetonitrile, treated with DIPEA, and then the above-mentioned acyl chloride compound of C111 is slowly added, and the reaction is stirred until TLC analysis confirms the completion of the reaction. Adding water to precipitate the solid and drying under reduced pressure can also yield the FR111 compound, with a yield of approximately 65%.

[0259] The above reaction can also be carried out by first activating C111 with HATU and DIPEA, and then directly adding SM1 (or its HCl, HBr, HI, or CF3COOH salts) to perform a condensation amidation reaction. After the reaction is complete, the reaction is quenched with dilute hydrochloric acid, extracted with EA, and concentrated to dryness under reduced pressure. Then, the compound is purified by column chromatography using AcOH / EA = 3 / 97 to obtain a white to off-white solid FR111 compound with a yield of about 75%.

[0260] Example 2: Synthesis of compound FR112 (X3 = OMe; IM4.2; β-acid = B1.2)

[0261]

[0262] Referring to the synthesis method of compound FR111 in Example 1, B1.2 of this Example 1 was used instead of B1.1 of Example 1, and the remaining preparation steps were the same as in Example 1, to obtain the target compound FR112.

[0263] The β-acid used is B1.2, which is an HCl salt. Other β-acids include, but are not limited to, HBr, HI, or CF3COOH. Following steps 1) and 2) of Example 1, C112 (i.e., C11, X3 = OMe) was prepared with an HPLC purity of approximately 98% and a yield of approximately 71%.

[0264] Continuing with steps 3) and 4) of Example 1, E112 (i.e., E11, X3 = OMe) was prepared with an HPLC purity of approximately 96% and a yield of approximately 83%.

[0265] The final step, deprotection, was performed under the same acidic conditions as step 5) in Example 1 to dissociate the ester protecting group, yielding compound FR112 with a yield of 91%.

[0266] LC-MS m / z = 645, [M+H] + .

[0267] Example 3: Preparation of FR113 (X3 = Cl; IM4.3; β-acid = B1.3)

[0268]

[0269] Referring to the synthesis method of compound FR111 in Example 1, B1.3 of this Example 1 was used instead of B1.1 of Example 1, and the remaining preparation steps were the same as in Example 1, to obtain the target compound FR113.

[0270] The β-acid used was B1.3. Following steps 1) and 2) of Example 1, C113 (i.e., C11, X3 = Cl) was prepared with an HPLC purity of approximately 95% and a yield of approximately 89%.

[0271] Continuing with steps 3) and 4) of Example 1, E113 (i.e., E11, X3 = Cl) was prepared with an HPLC purity of approximately 98% and a yield of approximately 80%.

[0272] The final step, deprotection, was performed under the same acidic conditions as step 5) of Example 1 to dissociate the ester protecting group, yielding compound FR113 with a yield of 87%.

[0273] LC-MS m / z = 649, [M+H] + .

[0274] Example 4: Preparation of FR114 (X3 = Br; IM4.4; β-acid = B1.4)

[0275]

[0276] Referring to the synthesis method of compound FR111 in Example 1, B1.4 of this Example 1 was used instead of B1.1 of Example 1, and the remaining preparation steps were the same as in Example 1, to obtain the target compound FR114.

[0277] The β-acid used was B1.4. Following steps 1) and 2) of Example 1, C114 (i.e., C11, X3 = Br) was prepared with an HPLC purity of approximately 96% and a yield of approximately 81%.

[0278] Continuing with steps 3) and 4) of Example 1, E114 (i.e., E11, X3 = Br) was prepared with an HPLC purity of approximately 96% and a yield of approximately 87%.

[0279] The final step, deprotection, was performed under the same acidic conditions as step 5) in Example 1 to dissociate the ester protecting group, yielding compound FR114 with a yield of 89%.

[0280] Example 5: Preparation of FR115 (X3 = F; IM4.5; B-acid = B1.5)

[0281]

[0282] Referring to the synthesis method of compound FR111 in Example 1, B1.5 of this Example 1 was used instead of B1.1 of Example 1, and the remaining preparation steps were the same as in Example 1, to obtain the target compound FR115.

[0283] The B-acid used was B1.5. Following steps 1) and 2) of Example 1, C115 (i.e., C11, X3 = F) was prepared with an HPLC purity of approximately 96.5% and a yield of approximately 90%.

[0284] Continuing with steps 3) and 4) of Example 1, E115 (i.e., E11, X3 = F) was prepared with an HPLC purity of approximately 96% and a yield of approximately 87%.

[0285] The final step, deprotection, was performed under the same acidic conditions as step 5) in Example 1 to dissociate the ester protecting group, yielding compound FR115 with a yield of 87% and an HPLC purity of 98.5%.

[0286] LC-MS m / z = 633, [M+H] + .

[0287] Example 6: Preparation of FR116 (X3 = SO2NH2; IM4.6; β-acid = B1.6)

[0288] Referring to the synthesis method of compound FR111 in Example 1, B1.6 of this Example 1 was used instead of B1.1 of Example 1, and the remaining preparation steps were the same as in Example 1, to obtain the target compound FR116.

[0289] The β-acid used was B1.6. Following steps 1) and 2) of Example 1, C116 (i.e., C11, X3 = SO2NH2) was prepared with an HPLC purity of approximately 98.5% and a yield of approximately 92%.

[0290] Continuing with steps 3) and 4) of Example 1, E116 (i.e., E11, X3 = SO2NH2) was prepared with an HPLC purity of approximately 96.5% and a yield of approximately 81%.

[0291] The final step, deprotection, was performed under the same acidic conditions as step 5) in Example 1 to dissociate the ester protecting group, yielding compound FR116 with a yield of 79% and an HPLC purity of 99.0%.

[0292] LC-MS m / z = 694, [M+H] + .

[0293] The preparation route of the target compound FR116 is as follows:

[0294]

[0295] Examples 1B to 6B represent another preparation method for FR111 to FR116, namely the aforementioned synthesis method two. This method mainly involves first constructing the right-wing active intermediate and then completing the left-wing condensation reaction to obtain the target compound.

[0296] The general preparation reaction route is shown in the following formula, and the corresponding codes of each substituent and protecting group, as well as intermediates and final products, are shown in Tables 2 and 2b.

[0297]

[0298] Table 2: Corresponding codes for substituents and protecting groups, intermediates and final products in the FR11 series

[0299] <![CDATA[X3 / Pro2]]> IM4 <![CDATA[Pro1 / D]]> intermediate G Acid intermediate H <![CDATA[NO2 / Boc]]> IM4.1 Bn / D10 G101 HCl H1011 OMe / Bn IM4.2 t-Bu / D11 G112 no H1120 Cl / Boc IM4.3 Bn / D10 G103 HCl H1031 Br / Trt IM4.4 Bn / D10 G104 HCl H1041 F / Trt IM4.5 Bn / D10 G105 HCl H1051 <![CDATA[SO2NH2 / Boc]]> IM4.6 Bn / D10 G106 HCl H1061

[0300] Table 2b: Structural formulas and corresponding intermediate codes for FR11 series products

[0301]

[0302] Example 1B: Preparation of FR111 (X3 = NO2; IM4.1; G101; H1011; E101)

[0303]

[0304] 1) Dissolve 1.0 eq of IM4.1 (X3 = NO2, Pro2 = Boc) in DMF, add benzyl (S)-2-amino-3-(3-(methanesulfonyl)phenyl)propionate (D10, Pro1 = Bn) or its hydrochloride (1-3 eq), then add DIPEA (2-10 eq) and HATU (1-3 eq). Stir at room temperature for 10-20 h until the IM4.1 reaction is complete. After routine post-treatment, the reaction solution is subjected to column chromatography to obtain G101 in 70% yield.

[0305] G101 was treated with a dioxane solution of HCl (4N, excess) for about 2 hours to remove the Boc protecting group of the amine group. The precipitate was precipitated by adding a poor solvent such as diethyl ether. After filtration and drying, H1011 (X3 = NO2, Pro1 = Bn, HCl salt) was obtained with a yield of 92%.

[0306] LC-MS m / z = 606, [M+H] + .

[0307] 2) Prepare benzofuran-2-carboxylic acid (SM2, 1-1.5 eq) into acyl chloride using thionyl chloride for later use; or treat toluene with oxalyl chloride (1-2 eq) and a catalytic amount of DMF, stir for about 5 hours until a clear solution is obtained, remove the solvent under reduced pressure and dissolve in DCM to obtain a DCM solution of acyl chloride, which is then kept under nitrogen or argon protection for later use.

[0308] The above-mentioned H1011 (1.0 eq) was dissolved in DCM and cooled to 0-5°C. DIPEA (1-10 eq) was added, followed by the slow addition of the above-mentioned acyl chloride DCM solution, ensuring the reaction temperature did not exceed 5°C. After the addition was complete, the mixture was stirred at 5°C for several hours until the reaction was complete. The reaction solution was quenched with ammonium chloride solution, and the DCM organic layer was separated. The organic layer was then washed with NaHCO3 solution, washed with water, and concentrated. Intermediate E101 was obtained in a quantitative yield.

[0309] 3) Debenzylation protection of intermediate E101 to prepare FR111. The reaction can be carried out by hydrolysis to remove benzyl under acidic (HCl) or alkaline (NaOH) conditions, or by hydrogenation to remove benzyl using transfer hydrogenation (such as HCOOH-TEA or ammonium formate).

[0310] Under nitrogen protection, TEA (2-10 eq) was added to a methanol:THF (2:1 to 10:1) mixture. The mixture was cooled to 4°C in an ice bath, and formic acid (2-25 eq) (formic acid:TEA = 5:1 to 1:5) was slowly added. Then, 10% palladium / carbon and intermediate E101 (1.0 eq) were added, and the mixture was stirred at room temperature until the transfer hydrogenation debenzylation reaction was complete.

[0311] The reaction solution was first filtered to recover the palladium / carbon catalyst. The filtrate was poured into an ice-water mixture, the pH was adjusted to acidic, and the solution was extracted with EA. The organic layer was dried with anhydrous sodium sulfate and then evaporated to dryness. The residue was subjected to column chromatography to obtain FR111 in 82% yield.

[0312] LC-MS m / z = 660, [M+H] + .

[0313] Example 2B: Preparation of FR112 (X3 = OMe; IM4.2; G112; H1120; E112)

[0314]

[0315] 1) Dissolve 1.0 eq of IM4.2 (X3 = OMe, Pro2 = Bn) in DMF, add tert-butyl (S)-2-amino-3-(3-(methanesulfonyl)phenyl)propionate (D11, Pro1 = tBu) (1-3 eq), then add DIPEA (2-10 eq) and HATU (1-3 eq). Stir at room temperature until the IM4.2 reaction is complete. After routine post-treatment, the reaction solution is separated by column chromatography, yielding G112 in 78% yield.

[0316] LC-MS m / z = 647, [M+H] + .

[0317] 2) Add G112 (1.0 eq), 5% Pd / C (5%–10% w / w), 12N HCl (0.5–2 w / w), and MeOH (5–10 w / w) sequentially to the hydrogenation reactor; purge with nitrogen three times, then purge with hydrogen three times; heat to 60°C, maintain hydrogen pressure at 1–5 atm, and react for approximately 20 h; TLC shows complete reaction; filter, wash filter cake with methanol; adjust pH of filtrate to alkaline, extract twice with EA, combine organic layers, wash with water, and then concentrate the organic layer to dryness under reduced pressure to obtain H1120, HPLC >98%, yield 93%.

[0318] LC-MS m / z = 557, [M+H] + .

[0319] 3) Benzofuran-2-carboxylic acid (SM2, 1-1.5 eq) was pretreated in toluene with oxalyl chloride (1-2 eq) and a catalytic amount of DMF. The mixture was stirred until a clear solution was obtained. The solvent was removed under reduced pressure and then dissolved in DCM to prepare a DCM solution of A acyl chloride. The solution was then stored under nitrogen or argon protection.

[0320] Dissolve the above H1120 (1.0 eq) in DCM, cool to 0-5°C, add DIPEA (1-10 eq), and then slowly add the above A acyl chloride DCM solution, ensuring the reaction temperature does not exceed 5°C. After the addition is complete, allow the reaction temperature to return to room temperature and continue stirring until the reaction is complete. Quench the reaction solution with water and separate the DCM organic layer. Wash the organic layer with NaHCO3 solution, wash with water, and concentrate to obtain intermediate E112, which can be directly used for the next reaction without purification.

[0321] Using the same deprotection method as in Example 2, the tert-butyl protecting group of E112 was removed under acidic conditions to prepare compound FR112, with a yield of approximately 90%.

[0322] LC-MS m / z = 645, [M+H] + .

[0323] Example 3B: Preparation of FR113 (X3 = Cl; IM4.3; G103; H1031; E103)

[0324]

[0325] Referring to the synthesis method of compound FR111 in Example 1B, IM4.3 of this example is used instead of IM4.1 of Example 1B, and the remaining preparation steps are the same as in Example 1B, to obtain the target compound FR113.

[0326] The yields for each step are as follows:

[0327] 1) IM4.3 was reacted with D10 to prepare G103, and then the Boc protecting group was removed under acidic conditions to obtain H1031, with a yield of 86%.

[0328] LC-MS m / z = 595, [M+H] + .

[0329] 2) H1031 is condensed with A to obtain E103 intermediate, which can be directly reacted in the next step without purification.

[0330] 3) The intermediate E103 was debenzylated by transfer hydrogenation in the presence of a palladium / carbon catalyst to obtain FR113 with a yield of 87%.

[0331] LC-MS m / z = 649, [M+H] + .

[0332] Example 4B: Preparation of FR114 (X3 = Br; IM4.4; G104; H1041; E104)

[0333]

[0334] Referring to the synthesis method of compound FR111 in Example 1B, IM4.4 of this example is used instead of IM4.1 of Example 1B, and the remaining preparation steps are the same as in Example 1B, to obtain the target compound FR114.

[0335] The yields for each step are as follows:

[0336] 1) Using DMF as solvent, compound IM4.4 (1.0 eq) was first stirred with TEA (2-10 eq) and HATU (1-3 eq) for 10-120 min, and then D10 (1-2 eq) was added. The mixture was stirred at room temperature for about 18 hours until the IM4.4 spot on the TLC plate disappeared. While stirring, the reaction solution was slowly poured into ice water, and the pH was adjusted to alkaline. The precipitate was then filtered, and the product, intermediate G104, was obtained in 75% yield.

[0337] 2) Intermediate G104 was treated with excess HCl / dioxane solution (4N, 10:1 (w / w)) until TLC showed complete removal of the Trt protecting group. Then MTBE was added to precipitate the product, which was filtered and washed with MTBE to obtain compound H1041 with a yield of about 94%.

[0338] LC-MS m / z = 639, [M+H] + .

[0339] 3) Same as step 2) of Example 1B, H1041 is condensed with the pre-activated A acyl chloride compound to obtain intermediate E104, which can be directly reacted in the next step without purification.

[0340] 4) Similar to step 3) of Example 1B, the intermediate E104 was subjected to a debenzylation reaction using a palladium / carbon-ammonium formate transfer hydrogenation system. The reaction was carried out under nitrogen protection in a mixed solution of methanol:THF (2:1 to 10:1), with 10% palladium / carbon (1-10 w / w) and 5-25 eq of ammonium formate. After the reaction, the palladium / carbon catalyst was first recovered by filtration. The filtrate was poured into an ice-water mixture, the pH was adjusted to acidic, and the mixture was extracted with EA. The organic layer was dried with anhydrous sodium sulfate and then evaporated to dryness. The residue was subjected to column chromatography to obtain FR114 in 76% yield.

[0341] LC-MS m / z = 694, [M+H] + .

[0342] Example 5B: Preparation of FR115 (X3 = F; IM4.5; G105; H1051; E105)

[0343]

[0344] Referring to the synthesis method of compound FR114 in Example 4B, IM4.5 of this example was used instead of IM4.4 of Example 4B, and the remaining preparation steps were the same as in Example 4B, to obtain the target compound FR115.

[0345] The starting materials and intermediates for each step are as follows:

[0346] IM4.5 (X3 = F; Pro2 = Trt), D10 (Pro1 = Bn), G105 (X3 = F; Pro2 = Trt; Pro1 = Bn), H1051 (HCl salt; X3 = F; Pro1 = Bn), E105 (X3 = F; Pro1 = Bn). The finished product is FR115 (X3 = F). The overall yield of FR115 prepared from IM4.5 is 65%.

[0347] Example 6B: Preparation of FR116 (X3 = SO2NH2; IM4.6; G106; H1061; E106)

[0348]

[0349] Referring to the synthesis method of compound FR111 in Example 1B, IM4.6 of this example is used instead of IM4.1 of Example 1B, and the remaining preparation steps are the same as in Example 1B, to obtain the target compound FR116.

[0350] The starting materials and intermediates for each step are as follows:

[0351] IM4.6(X3=SO2NH2; Pro2=Boc), D10(Pro1=Bn), G106(X3=SO2NH2; Pro2=Boc; Pro1=Bn), H1061

[0352] (HCl salt; X3 = SO2NH2; Pro1 = Bn), E106 (X3 = SO2NH2; Pro1 = Bn). The finished product is FR116 (X3 = SO2NH2). The overall yield of FR116 prepared from IM4.6 is 60%.

[0353] Example 7: Synthesis of compound FR121 (X3 = NO2; IM4.1; β-acid = B1.1)

[0354]

[0355] Referring to the synthesis method of compound FR111 in Example 1, the starting material benzofuran-2-carboxylic acid (SM2, 1.0 eq) of Example 1 was replaced with benzofuran-6-carboxylic acid (SM3, 1.0 eq) of this example, and the remaining preparation steps were the same as in Example 1, to obtain the target compound FR121.

[0356] The intermediates obtained and their yields are as follows:

[0357] 1) Benzofuran-6-carboxylic acid (SM3) was activated by the carboxyl group and then reacted with B-Acid (B1.1, X1=X2=Cl, X3=NO2) to prepare C121 (X3=NO2), with an HPLC purity of about 97.5% and a yield of 80%.

[0358] LC-MS m / z = 435, [M+H] + .

[0359] 2) Referring to steps 3) and 4) of Example 1, C121 and D11 were condensed under HATU and TEA conditions to prepare E121 (X3 = NO2), with an HPLC purity of about 97% and a yield of 83%.

[0360] 3) E121 was deprotected by removing the tert-butyl ester protecting group under acidic conditions to obtain FR121, with a yield of 76%.

[0361] LC-MS m / z = 660, [M+H] + .

[0362] Example 8: Synthesis of compound FR122 (X3 = OMe; B-acid = B1.2)

[0363]

[0364] Following the same preparation method as in Example 7, except that B-acid was replaced with B1.2 (X3 = OMe), compound FR122 (X3 = OMe) was isolated in an overall yield of 56% after each reaction step.

[0365] Example 9: Synthesis of compound FR123 (X3 = Cl; B-acid = B1.3)

[0366]

[0367] Following the same preparation method as in Example 7, except that B-acid was replaced with B1.3 (X3 = Cl), compound FR123 was isolated with an overall yield of 67% after each reaction step.

[0368] LC-MS m / z = 649, [M+H] + .

[0369] Example 10: Synthesis of compound FR124 (X3 = Br; B-acid = B1.4)

[0370]

[0371] Following the same preparation method as in Example 7, except that B-acid was replaced with B1.4 (X3 = Br), compound FR124 was isolated in an overall yield of 59% after each reaction step.

[0372] LC-MS m / z = 694, [M+H] + .

[0373] Example 11: Synthesis of compound FR125 (X3 = F; B-acid = B1.5)

[0374]

[0375] The preparation method of Example 7 is the same as that of Example 7, except that B-acid is replaced with B1.5 (X3 = F).

[0376] 1) Benzofuran-6-carboxylic acid (SM3) was activated by the carboxyl group and then reacted with B-Acid (B1.5, X1=X2=Cl, X3=F) to prepare C125 (X3=F), with an HPLC purity of about 98.9% and a yield of 67%.

[0377] 1 H NMR (400MHz, DMSO-d6) δ8.13 (s, 1H), 7.76-7.77 (m, 2H), 7.37-7.40 (dd, J = 4.00Hz, 1H), 7.05-7.06 (d, J=4.00Hz, 1H), 4.79 (bs, 2H), 3.71 (bs, 2H), 2.87-2.90 (m, 2H).

[0378] LC-MS m / z = 408, [M+H] + .

[0379] 2) Referring to step 2 of Example 7, C125 and D11 were condensed under HATU and TEA conditions. After post-processing and concentration, E125 (X3 = F) was prepared in quantitative yield with an HPLC purity of approximately 97.1%.

[0380] 3) E125 was deprotected by removing the tert-butyl ester protecting group under acidic conditions to obtain FR125, with an HPLC yield of 98.7% and a recovery rate of 71%.

[0381] FR125: 1 H NMR (400MHz, DMSO-d6) δ12.96 (br, 1H), 9.10-9.12 (d, J = 8.00Hz, 1H), 8.12-8.1 4(m,1H),7.86-7.87(m,1H),7.74-7.79(m,3H),7.66-7.68(m,1H),7.56-7.59(t ,J=8.00Hz,1H),7.34-7.36(d,J=8.00Hz,1H),7.05-7.06(m,1H),4.77-4.83(bm ,3H),3.68(br,2H),3.29-3.34(bm,4H),3.00-3.06(m,1H),2.79-2.82(bm,2H).

[0382] LC-MS m / z = 633, [M+H] + .

[0383] Example 12: Synthesis of compound FR126 (X3 = SO2NH2; B-acid = B1.6)

[0384]

[0385] The preparation method of Example 7 is the same as that of Example 7, except that B-acid is replaced with B1.6 (X3 = SO2NH2).

[0386] After each reaction step, compound FR126 (X3 = SO2NH2) was isolated in an overall yield of 73%.

[0387] LC-MS m / z = 694, [M+H] + .

[0388] Examples 13 to 31: Synthesis of compounds corresponding to the FR13 series to FR31 series, respectively.

[0389] In Examples 1-6 and Examples 7-12, compounds FR111-FR116 (FR11 series) and FR121-FR126 (FR12 series) containing benzofuran-2-carboxylic acid (SM2) and benzofuran-6-carboxylic acid (SM3) as the left wings were prepared, respectively. When different aryl and / or heteroaryl carboxylic acids are changed, compounds of the FR13-FR31 series can be prepared by the same method.

[0390] The general synthetic method is shown in the following formula, and the compounds corresponding to the specific embodiments are shown in Table 3 below:

[0391]

[0392] Table 3: Compounds of the FR13-FR31 series corresponding to Examples 13-31

[0393]

[0394]

[0395]

[0396] The proton NMR data for compound FR290 are as follows:

[0397] 1H NMR (600MHz, DMSO-d6) δ12.86(s,1H),9.02(d,J=8.3Hz,1H),7.86(d,J=1.8Hz,1H),7.77(d t,J=7.8,1.5Hz,1H),7.67(dt,J=7.7,1.4Hz,1H),7.57(t,J=7.7Hz,1H),7.28-7.43(m,1H), 6.88-7.07(m,3H),6.09(s,2H),4.78(ddd,J=10.4,8.3,4.7Hz,1H),4.68(s,2H),3.67(m,2 H),3.02 / 3.29(dd,J=14.2,4.7Hz / dd,J=14.2,10.4Hz,2H),3.15(s,3H),2.71-2.78(m,2H).

[0398] The proton NMR data for compound FR300 are as follows:

[0399] 1 H NMR(600MHz,DMSO-d6)δ12.86(s,1H),9.00(d,J=8.3Hz,1H),7.86(t,J=1.7Hz,1H),7 .77(dt,J=7.8,1.5Hz,1H),7.67(d,J=7.7Hz,1H),7.52-7.61(m,2H),7.50(d,J=8.2H z,1H),7.16-7.47(m,2H),4.78(ddd,J=10.2,8.3,4.7Hz,1H),4.50-4.75(m,2H),3.4 7-3.97(m,2H),2.98-3.07 / 3.25-3.30(m / m,2H),3.15(s,3H),2.76(t,J=6.3Hz,2H).

[0400] Examples 32 to 36: Synthesis of compounds corresponding to the FR32 series to FR36 series, respectively.

[0401] Examples 1 to 31 above respectively prepared aryl or heteroaryl carboxylic acids where Y = carbonyl. As compounds of the left wing FR11 to FR31 series. When Y = methylene (i.e. -CH2-), A2 is an aryl or heteroaryl benzyl active compound, including but not limited to benzyl halides, benzyl sulfonates or other easily leaving mixed esters.

[0402] The general synthesis method is shown in the following formula:

[0403]

[0404] Amine or heteroaryl benzyl chloride (or benzyl bromide, benzyl sulfonate) is amination reaction with B-acid under basic conditions to prepare the corresponding intermediate K; then it is condensed with D11 to prepare intermediate M; M is hydrolyzed under acidic or basic conditions to obtain the corresponding FR32-36 series compounds. Specific examples of the corresponding compounds are shown in Table 4 below:

[0405] Table 4: Compounds of the FR32-FR36 series corresponding to Examples 32-36

[0406]

[0407]

[0408] For a more detailed explanation, taking the preparation of FR355 using A2 = piperonyl chloride (A2-5) as an example, the following formula is shown:

[0409]

[0410] 1) Preparation of intermediate K355:

[0411] 1.1) Add DMF (or DMSO) solvent, potassium carbonate (2-10 eq), and B1.5 (1.0 eq) to the reaction flask. While stirring, add piperonyl chloride (or bromine, or Ts ester) (1-5 eq) dropwise at around 0°C. Heat and stir until TLC shows complete reaction of the starting materials. Add water to the reaction solution and extract with ethyl acetate. Combine the organic phases and wash with saturated brine. Concentrate the organic phase under reduced pressure to dryness to obtain K355 (X3 = Cl, R1 is piperonyl), yield 50%.

[0412] 1.2) K355 can also be prepared by a similar method to steps 3) and 4) of Example (III) (preparation of IM4.2) (i.e., first protecting IM2.2 with an N-alkyl group, then carboxylating it to obtain IM4.2). First, N-alkylation is applied to obtain IM355, which is then carboxylated. As shown in the following formula:

[0413]

[0414] Referring to the carboxylation procedure in step 3) of Example (III), dissolve IM2.5 (1.0 eq) in DMF (or DMSO) solvent, add potassium carbonate (1-5 eq), and dropwise add piperonyl chloride (or bromine, or Ts ester) (1-2 eq) while stirring and controlling the temperature at around 0°C. Continue stirring until TLC shows complete reaction of the starting material. Add water to the reaction solution and extract with ethyl acetate. Combine the organic phases and wash with saturated brine. Concentrate the organic phase under reduced pressure to dryness to obtain IM355 (X3 = Cl, R1 is piperonyl), with a yield of 80%.

[0415] Referring to the carboxylation procedure in step 4) of Example (III), IM355 (1.0 eq) reacted with n-BuLi (1-4 eq) dropwise under the action of TMEDA (1-4 eq). After the addition was complete, the mixture was stirred at -70°C for 30 min. CO2 was introduced into the system (or dry ice was added), and the reaction was controlled at -60°C. After stirring for 1 h, the mixture was allowed to cool naturally to 0°C. The reaction was quenched with an aqueous solution, and the pH was adjusted to 6. The mixture was extracted with ethyl acetate, and the organic phases were combined. An appropriate amount of concentrated hydrochloric acid was added, and the mixture was stirred at 10°C until crystallization was complete. The mixture was filtered, and the filter cake was dried to obtain a white to off-white solid (K355 or its hydrochloride), with a yield of 70%.

[0416] 2) Preparation of intermediate M355:

[0417] K355 intermediate (1.0 eq) and triethylamine (1-5 eq) were added to DMF (5-10 w / w) at approximately 30°C and stirred until homogeneous. HATU (1-2 eq) was added, followed by heating to 45°C and maintaining the temperature for 1-2 hours. After the reaction was complete, the reaction mixture was cooled to 30°C, and a pre-prepared dichloromethane solution containing D11 (1-2 eq) was added. The reaction mixture was maintained at 30°C for 5-6 hours until TLC showed complete reaction. After coupling, water (5-10 w / w) was added, and the organic layer was separated. The resulting organic layer was washed with 5% sodium carbonate solution, followed by washing with dilute hydrochloric acid and 5% brine. The organic layer was concentrated to obtain M355 intermediate in approximately 81% yield.

[0418] 3) Preparation of FR355:

[0419] The protection of M355 tert-butyl ester can be removed by acidic or alkaline hydrolysis. To avoid racemization of the chiral center, hydrolysis with formic acid or NaOH is more suitable for removing the tert-butyl ester protecting group.

[0420] Formic acid (5-10 w / w) was used to dissolve the M355 intermediate, and the mixture was kept at 60°C for 6-7 hours to complete the hydrolysis. The reactants were then added to chilled water, kept at 0°C for 2 hours, and filtered. The resulting wet solid was dried to give the FR355 compound in 80% yield.

[0421] LC-MS m / z = 623, [M+H] + .

[0422] Example 37: Cell Adhesion Inhibition Experiment

[0423] T-cell adhesion assays were performed using the human T-lymphocyte line HuT 78 (ATCC TIB-161). Goat anti-Hu IgG (Fc) was diluted to 2 μg / mL in PBS and spread at 50 μL / well in 96-well plates for 1 hour at 37°C. The plates were washed with PBS and blocked with 1% BSA in PBS at room temperature for 1 hour. 5-domain ICAM-Ig was diluted to 100 ng / mL in PBS and then added at 50 μL / well to the O / N plate at 4°C. HuT 78 cells were centrifuged at 100 g, and cell pellets were incubated in a 5% CO2 incubator at 37°C with 5 mM EDTA for approximately 5 minutes. Cells were washed and centrifuged in a mixture of 0.14 M NaCl, 0.02 M Hepes, 0.2% glucose, and 0.1 mM MnCl2 (assay buffer). Cells were resuspended in the assay buffer to a final volume of 3.0 x 10⁻⁶. 6 cells / mL.

[0424] The test compound and positive control (Lifitegrast) were diluted to a final concentration of 2-fold in assay buffer and pre-incubated with HuT 78 cells for 30 minutes at room temperature. 100 μL / well of cells and compound were added to the plate and incubated for 1 hour at room temperature. Total fluorescence intensity was measured using a fluorometer (ex: 485; em: 530; cutoff: 530). The plate was washed once with assay buffer, and fluorescence intensity was measured again using a fluorometer (ex: 485; em: 530; cutoff: 530). The results were plotted as an inhibition-concentration curve, and the IC50 was calculated using standard methods. 50 .

[0425] Table 5: Cell adhesion inhibition assay IC50 50 value

[0426] Example serial number <![CDATA[IC 50 (μM)]]> Example serial number <![CDATA[IC 50 (μM)]]> 1 FR111 * 7 FR121 * 2 FR112 * 8 FR122 ** 3 FR113 * 9 FR123 * 4 FR114 * 10 FR124 ** 5 FR115 * 11 FR125 *** 6 FR116 * 12 FR126 *** 13 FR134 ** 13 FR135 ** 25 FR255 * 25 FR256 * 28 FR285 ** 28 FR286 * 29 FR290 ** 30 FR300 ** 29 FR295 *** 30 FR305 *** 33 FR335 ** 35 FR355 ** 36 FR365 ** Yang Shen Lifitegrast **

[0427] [Note] The average value of three retests was taken. "***": 0.001~0.010μM; "**": 0.011~0.100μM; "*": 0.101~1.0μM

[0428] Example 38: Effect of experimental compounds on Hut-78 cell viability

[0429] The cell adhesion inhibition IC of each compound measured in Example 37 50 Values ​​were used to determine the effect of each compound on the viability of Hut-78 cells at this concentration.

[0430] After culturing Hut-78 cells to a certain cell mass in low-adhesion culture flasks, resuspend them in culture medium at the required density (e.g., 3000-5000 cells / well). Add 50 μL of cell suspension to each well of a 96-well plate. Prepare a 2x solution of the target small molecule at the concentration required for the cell adhesion inhibition experiment. Add 50 μL of the target small molecule solution to each well. After adding all the solution, gently shake the plate to mix the drug and cell suspension. Incubate at 37°C for 1 hour in a 5% CO2 incubator. Centrifuge the 96-well plate at 100g. The cells will aggregate at the bottom of the plate; discard the supernatant. Add 100-150 μL of CTG working solution to each well and incubate at 37°C in the dark with gentle horizontal shaking for 5-10 minutes. Remove the incubated cell culture plate, transfer the liquid from each well to a black-walled 96-well plate, and place it in a microplate reader to detect the chemiluminescence value and calculate the viability of each experimental group.

[0431] Table 6. Effects of each compound on the viability of Hut-78 cells at specific concentrations

[0432] Example serial number viability of Hut-78 cells Example serial number viability of Hut-78 cells 1 FR111 ++ 7 FR121 ++ 2 FR112 ++ 8 FR122 ++ 3 FR113 ++ 9 FR123 ++ 4 FR114 ++ 10 FR124 ++ 5 FR115 +++ 11 FR125 +++ 6 FR116 +++ 12 FR126 +++ 13 FR134 +++ 13 FR135 +++ 25 FR255 ++ 25 FR256 ++ 28 FR285 ++ 28 FR286 ++ 29 FR290 +++ 29 FR295 +++ 30 FR300 +++ 30 FR305 +++ 33 FR335 ++ 35 FR355 ++ 36 FR365 +++ Yang Shen Lifitegrast ++

[0433] [Note] The specific concentrations mentioned in Table 6 refer to the IC50 concentrations of each compound that inhibit cell adhesion, as measured in Example 37. 50 concentration.

[0434] "+++": 85%–100%; "++": 70%–84%; "+": less than 70%.

[0435] Example 39: Schirmer Tear Test (STT) of Experimental Compounds

[0436] The Schirmer tear test (STT) was used to study tear production as the primary endpoint. Preliminary assessments were performed in unanesthetized dogs before initial drug administration and after 4 weeks of continuous TID administration. The test was performed before any topical medication was instilled. One Schirmer tear test strip was used per study eye for each test. The strip was placed in the lower palpebral conjunctival sac, at the junction of the medial two-thirds and lateral one-third of the lower eyelid margin, for 60 seconds, and the length of the wetted portion below the groove of the strip was recorded (in millimeters).

[0437] Observe and record the number of blinks 0-5 minutes after the instillation to compare the irritation level.

[0438] Table 7. STT test results and irritation comparison results for some compounds.

[0439] Example serial number 4-week STT results* Comparison of blink counts** 8 FR122 + 50 11 FR125 ++ 30 12 FR126 ++ 38 25 FR256 - 65 28 FR286 -- 87 29 FR290 + 42 29 FR295 ++ 48 30 FR300 + 51 30 FR305 ++ 53 Yang Shen Lifitegrast 0 55

[0440] [Note*]: Refers to the STT result after 4 weeks of continuous TID administration; the STT result of Yangshen is used as the baseline, and the definition is as follows:

[0441] "0" indicates an increase of less than 10% (excluding 10%) or a decrease of less than 10% (excluding 10%) compared to Yangcan.

[0442] "+" indicates an improvement of 10% to 30% compared to Yangcan; "++" indicates an improvement of 31% to 60% compared to Yangcan.

[0443] "-" indicates a reduction of 10% to 30% compared to Yangshen; "--" indicates a reduction of 31% to 60% compared to Yangshen.

[0444] [Note**]: This refers to the average number of blinks within 5 minutes after each instillation on the last day of 4 consecutive weeks of TID administration.

[0445] STT results showed that, compared with Lifitegrast, most of the compounds of this invention had higher Schirmer test values ​​after 4 weeks of treatment, indicating better efficacy; this also suggests that these compounds can directly or indirectly promote the secretory function of the lacrimal gland.

Claims

1. A trisubstituted tetrahydroisoquinoline amide derivative, which is a compound, isomer, or pharmaceutically acceptable salt thereof represented by Formula I: in, Y is selected from carbonyl group, C 1-6 Alkyl or CR2R3; X1 and X2 are each independently selected from halogens and C. 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, Halogenated C 1-6 Alkoxy, hydroxy, nitro, cyano, amino, C 1-6 alkylamine group, C 1-6 Alkyl, amide, or sulfonamide groups; X3 is selected from hydrogen, halogen, substituted or unsubstituted C. 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, hydroxy, nitro, cyano, amino, substituted or unsubstituted C 1-6 Alkylamine group, substituted or unsubstituted C 1-6 Alkyl, amide, or sulfonamide groups; when the C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 alkylamine group, C 1-6 When an alkyl acyl group has substituents, it can be substituted by at least one of the following groups: deuterium, halogen; R1 is selected from substituted or unsubstituted C. 6-14 aryl, substituted or unsubstituted 5-14 heteroaryl, substituted or unsubstituted C 3-8 Cycloalkyl, substituted or unsubstituted 3-10 membered heterocyclic groups; when the C 6-14 Aryl, 5-14 heteroaryl, C 3-8 When cycloalkyl or 3-10 membered heterocyclic groups have substituents, they can be monosubstituted or polysubstituted by one of the following groups, or polysubstituted by multiple of the following groups: deuterium, hydroxyl, halogen, C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylamine group, -OR'; R2 and R3 are each independently selected from hydrogen and C. 1-6 Alkyl or halogenated C 1-6 alkyl; R' is selected from C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 2-6 alkenyl or C 1-6 Alkyl group; Furthermore, when R1 is not a substituted or unsubstituted benzodioxolane, X3 is not hydrogen; The heterocyclic group or heteroaryl group contains at least one heteroatom, which is selected from N, O or S; The halogen is selected from fluorine, chlorine, bromine, and iodine, with fluorine or chlorine being preferred.

2. The trisubstituted tetrahydroisoquinoline amide derivative according to claim 1, characterized in that: In Equation I, R1 is arbitrarily selected from one of the following structures: in, R4 is selected from hydrogen, deuterium, hydroxyl, halogen, and C. 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylamine group or -OR'; R' is selected from C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 2-6 alkenyl or C 1-6 Alkyl group; R5 is selected from hydrogen, deuterium, and C. 1-6 alkyl or alkylyl, C 2-6 Enyl or halogenated C 2-6 Enyl group; R6 and R7 are each independently selected from hydrogen, deuterium, halogens, and carbon. 1-6 Alkyl or halogenated C 1-6 alkyl; n is selected from 0, 1, 2 or 3.

3. The trisubstituted tetrahydroisoquinoline amide derivative according to any one of claims 1 to 2, characterized in that: The compound represented by Formula II, its isomer, or a pharmaceutically acceptable salt thereof: Wherein, Y is selected from carbonyl or -CH2- group; X1 and X2 are each independently selected from halogens and C. 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, Halogenated C 1-6 Alkyl, nitro, cyano, amino, or sulfonamide groups; X1 and X2 are preferably both chlorine; X3 is selected from hydrogen, halogen, substituted or unsubstituted C. 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, hydroxy, nitro, cyano, amino, substituted or unsubstituted C 1-6 Alkylamine group, substituted or unsubstituted C 1-6 Alkyl, amide, or sulfonamide groups; when the C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 alkylamine group, C 1-6 When the alkyl acyl group has substituents, it can be substituted by at least one of the following groups: deuterium, halogen; when X1 and X2 are preferably chlorine, X3 is preferably hydrogen, nitro, cyano, halogen, acetyl, methoxy, amino or substituted amino, sulfonamide, and further X3 is preferably hydrogen, nitro, methoxy, chlorine, bromine, fluorine or sulfonamide. R4 is selected from hydrogen, deuterium, hydroxyl, halogen, and C. 1-6 Alkyl or halogenated C 1-6 Alkyl; preferably hydrogen; R6 and R7 are each independently selected from hydrogen, deuterium, halogens, and carbon. 1-6 Alkyl or halogenated C 1-6 Alkyl groups; preferably derived from hydrogen or fluorine; n is selected from 0, 1, 2 or 3.

4. The trisubstituted tetrahydroisoquinoline amide derivative according to any one of claims 1 to 2, characterized in that: The compound represented by Formula III, its isomer, or a pharmaceutically acceptable salt thereof: Among them, X1 and X2 are each independently selected from halogens and C. 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, Halogenated C 1-6 Alkyl, nitro, cyano, amino, or sulfonamide groups; X1 and X2 are preferably both chlorine; X3 is selected from halogenated, substituted, or unsubstituted C. 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, hydroxy, nitro, cyano, amino, substituted or unsubstituted C 1-6 Alkylamine group, substituted or unsubstituted C 1-6 Alkyl, amide, or sulfonamide groups; when the C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 alkylamine group, C 1-6 When the alkanoyl group has substituents, it can be substituted by at least one of the following groups: deuterium, halogen; when X1 and X2 are preferably chlorine, X3 is preferably from nitro, cyano, halogen, acetyl, methoxy, amino or substituted amino, sulfonamide, and further X3 is preferably from nitro, methoxy, chlorine, bromine, fluorine or sulfonamide. R4 is selected from hydrogen, deuterium, hydroxyl, halogen, and C. 1-6 Alkyl, Halogenated C 1-6 Alkyl or C 1-6 Alkyl group; preferably hydroxyl, halogen, methyl, difluoromethyl, trifluoromethyl, or methoxy. n is selected from 0, 1, 2 or 3, preferably 1 or 2.

5. The trisubstituted tetrahydroisoquinoline amide derivative according to any one of claims 1 to 2, characterized in that: The compound represented by Formula IV, its isomer, or a pharmaceutically acceptable salt thereof: Among them, X1 and X2 are each independently selected from halogens and C. 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, Halogenated C 1-6 Alkyl, nitro, cyano, amino, or sulfonamide groups; X1 and X2 are preferably both chlorine; X3 is selected from halogenated, substituted, or unsubstituted C. 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, hydroxy, nitro, cyano, amino, substituted or unsubstituted C 1-6 Alkylamine group, substituted or unsubstituted C 1-6 Alkyl, amide, or sulfonamide groups; when the C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 alkylamine group, C 1-6 When the alkanoyl group has substituents, it can be substituted by at least one of the following groups: deuterium, halogen; when X1 and X2 are preferably chlorine, X3 is preferably from nitro, cyano, halogen, acetyl, methoxy, amino or substituted amino, sulfonamide, and further X3 is preferably from nitro, methoxy, chlorine, bromine, fluorine or sulfonamide. R1 is selected from R4 is selected from hydrogen, deuterium, hydroxyl, halogen, and C. 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylamine group or -OR'; R' is selected from C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 2-6 alkenyl or C 1-6 Alkyl group; R4 is preferably selected from hydrogen, hydroxyl, halogen, methyl, difluoromethyl, trifluoromethyl, or methoxy. R5 is selected from hydrogen, methyl, formyl, acetyl, acryloyl, or 2-fluoroacryloyl; n is selected from 0, 1, 2 or 3; preferably 0, 1 or 2.

6. A trisubstituted tetrahydroisoquinoline amide derivative, selected from the following characteristic compounds or their isomers, or pharmaceutically acceptable salts thereof:

7. The method for preparing the trisubstituted tetrahydroisoquinoline amide derivative according to any one of claims 1 to 6, characterized in that, The following synthetic route (I) is included: Where Y is selected from carbonyl, then X is selected from hydroxyl, pentafluorophenoxy, halogen, The radical; when Y is selected from -CH2-, then X is selected from halogens or sulfonates; Pro1 = Me,Et,iPr,t-Bu,Bn; R1, X1, X2, X3 are defined as described in any one of claims 1 to 5; 1) Compound A and trisubstituted tetrahydroisoquinoline carboxylic acid compound B-acid undergo a condensation reaction under alkaline conditions to generate the key intermediate C; 2) The carboxyl group protected by tert-butyl, benzyl or other easily removable alkyl groups, m-methylsulfonyl L-phenylalanine (compound D) is reacted with the above key intermediate C in the presence of a condensing agent to obtain intermediate E; the preferred reaction conditions are: after mixing C and D, stirring the reaction in a DIPEA (or TEA) + HATU system for 2-8 hours, quenching the reaction with acid solution, and extracting and drying to obtain intermediate E, wherein the molar ratio of intermediate C to D is 1:(1-3); 3) After removing the protecting group Pro1 from intermediate product E, the target product (I) is obtained. The preferred reaction conditions are: add intermediate product E to a polar solvent, water, and an alkaline agent (such as hydroxide, or TEA+LiBr, etc.), react at a temperature of 10-35℃ for 0.1-10 hours, quench with an inorganic or organic acid, and then extract and evaporate to dryness.

8. The preparation method according to claim 7, characterized in that, The key starting material B-acid in the synthetic route (I) has the structural formula shown in formula B-acid. The acid radical can be an inorganic acid or an organic acid such as HCl, HBr, HI, or CF3COOH; The key intermediate C has the structural formula shown in equation C. The key intermediate E has the structural formula shown in Equation E. The definitions of X1, X2, and X3 are as described in any one of claims 1 to 5.

9. The method for preparing the trisubstituted tetrahydroisoquinoline amide derivative according to any one of claims 1 to 6, characterized in that, The following synthetic route (II) is included: Where Y is selected from carbonyl, and X is selected from hydroxyl, pentafluorophenoxy, halogen, The group is defined as follows: when Y is selected from -CH2- group, X is selected from halogen or sulfonate ester; Pro1 = Me, Et, iPr, t-Bu, Bn; Pro2 is an amino protecting group (Bn, Cbz, Boc, Fmoc, Alloc, Teoc, Tos, Tfa, Trt, Dmb, PMB, Me, Et group) and its salt; R1, X1, X2, X3 are defined as described in any one of claims 1 to 5; 1) Compound IM4 and another starting material compound D were mixed and dissolved in a polar solvent, and an organic base and a condensing agent were added to carry out a condensation reaction to obtain the key intermediate G; 2) Selectively remove the protecting group Pro2 of G under acidic conditions to obtain the key intermediate H, wherein the acid radical can be an inorganic acid or an organic acid such as HCl, HBr, HI or CF3COOH. 3) The above-mentioned key intermediate H is further condensed with compound A under alkaline conditions to obtain intermediate E; 4) Remove the protecting group Pro1 from intermediate E to obtain the target product formula (I).

10. The preparation method according to claim 9, characterized in that, The starting material IM4 of the synthetic route (II) has the structural formula shown in formula IM4: The key intermediate G has the following structure: The key intermediate H has the structural formula shown in equation H: The key intermediate E has the following structure:

11. The preparation method according to claim 9 or 10, characterized in that, The preparation methods of the compounds IM4 and B-acid are as follows: Starting with trisubstituted benzene (SM), a trisubstituted isoquinoline intermediate (IM1) was obtained through formylation, condensation, reduction, Ts protection, and ring closure. The trisubstituted isoquinoline intermediate (IM1) was then reduced to a trisubstituted tetrahydroisoquinoline intermediate (IM2). After amino protection, the trisubstituted tetrahydroisoquinoline intermediate (IM2) underwent a carboxylation reaction to obtain the aforementioned protected trisubstituted tetrahydroisoquinoline (carboxylic acid) compound (IM4). The protecting group of IM4 was removed under acidic conditions to obtain compound B-acid.

12. A pharmaceutical composition comprising at least one active ingredient as described in any one of claims 1 to 11 and at least one pharmaceutically acceptable carrier or excipient.

13. Use of a compound prepared according to any one of claims 1 to 11 or the pharmaceutical composition of claim 12 in an immune cell migration inhibitor.

14. Use of a compound prepared according to any one of claims 1 to 11 or a pharmaceutical composition according to claim 12 in the prevention or treatment of diseases associated with LFA-1 activity.

15. The use according to claim 15, characterized in that, The diseases mentioned include, but are not limited to, dry eye syndrome and T-cell inflammatory response diseases.

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