WO2021018023A1

WO2021018023A1 - Small molecule glp-1 receptor modulator

Info

Publication number: WO2021018023A1
Application number: PCT/CN2020/104059
Authority: WO
Inventors: 谢方; 于广侠; 张淑娟
Original assignee: 济南泰达领创医药技术有限公司
Priority date: 2019-08-01
Filing date: 2020-07-24
Publication date: 2021-02-04

Abstract

The present invention provides a new compound for modulating a Glucagon-like peptide-1 (GLP-1) receptor, and a synthesis method therefor and a pharmaceutical use thereof. This kind of compound has an obvious agonistic effect on the GLP-1 receptor, but does not have an obvious binding effect on hERG, and therefore, it is indicated that the compound has a low risk of cardiotoxicity. The compound of the present invention can be used alone as a treatment medication for type II diabetes, or be used in combination with a polypeptide GLP-1 receptor agonist such as liraglutide, or be used in combination with the treatment medication for type II diabetes of other mechanisms.

Description

Small molecule GLP-1 receptor modulator

Technical field

The invention belongs to the technical field of medicine, and specifically relates to a small molecule GLP-1 receptor modulator with a new structure, a preparation method and a pharmaceutical application thereof.

Background technique

Diabetes is a metabolic disease characterized by high blood sugar. Due to factors such as insulin secretion defects or decreased physiological function of pancreatic islet B cells, it leads to a long-term high blood sugar state in the body, which in turn leads to chronic diseases of the eyes, kidneys, heart, blood vessels, nerves and other organs and tissues. Damage or dysfunction. According to different pathogenesis, diabetes can be divided into type I diabetes and type II diabetes. Type II diabetes is caused by the decline of pancreatic islet B cell function and insulin resistance, which is the main type of diabetes.

Glucagon-like peptide-1 receptor (GLP-1 receptor) is one of the effective targets for the treatment of type II diabetes. The GLP-1 receptor belongs to the B family of G protein-coupled receptors, with seven transmembrane domains, and its natural agonist ligand is glucagon-like peptide-1 (ie GLP-1). When the GLP-1 receptor is activated, it can further activate downstream signal pathways such as PKA, PI3K, MAPK, and promote the release of insulin and reduce the release of glucagon, thereby reducing blood sugar levels.

The GLP-1 receptor agonists developed in the early stage are GLP-1 analogues, which are all peptides, including liraglutide, semaglutide, etc., through genetic engineering methods and preparation improvements, etc., long-term effects have been developed Polypeptide drugs that can be taken orally, but oral polypeptides are still subject to many disadvantages such as administration time, dosage, and gastrointestinal disturbances. In recent years, researchers have discovered that small molecule compounds can also be used to regulate the function of GLP-1 receptors, and then develop small molecule GLP-1 receptor agonists (Document 1), including some low molecular weight flavonoids and containing Nitrogen heterocyclic compounds and compounds Boc-4 and S4P that mimic the structure of polypeptides, etc. However, the existing small molecule agonists have limited structure types, and the specificity and safety of GLP-1 receptors are not strong, so it is necessary to develop New structure type and specific agonist for GLP-1 receptor.

In 2018, Pfizer Pharmaceuticals reported a series of GLP-1 receptor agonists with a 6-carboxyarylimidazole structure (Document 2). The active compound PF-06882961 is currently in clinical research.

Literature 1: Willard FS, et al. Small molecule drug discovery at the glucagon-like peptide-1 receptor, Journal of Diabetes Research, 2012, 2012: 344-350.

Document 2: WO2018109607A1.

Summary of the invention

The purpose of the present invention is to provide a new structure of small molecule GLP-1 receptor modulator, its preparation method and pharmaceutical application.

Specifically, the present invention provides a small molecule GLP-1 receptor modulator with the following structure, and a pharmaceutically acceptable salt, ester, stereoisomer, ether, prodrug or solvate thereof:

among them,

A is a substituted or unsubstituted aromatic ring, aromatic heterocyclic ring, C ₂ -C ₈ alkenyl group, the substitution is one or more than two, and the substituents may be the same or different;

L is a link chain, selected from -(CH ₂ ) _m -, -CONR ₇ -, -SO ₂ NR ₇ -, -NR ₇ CO-, -NR ₇ SO ₂ -, -(CH ₂ ) _m O-,- O(CH ₂ ) _m -, -(CH ₂ ) _m S-, -S(CH ₂ ) _m -, -(CH ₂ ) _m NR ₇ -, -NR ₇ (CH ₂ ) _m -; said R ₇ It is selected from hydrogen, a C _1-8 straight or branched chain alkyl group or a C _3-8 cycloalkyl group, and the alkyl group is optionally further substituted with one or more halogen, cyano, amino or hydroxy; preferably , R _{7 is} selected from hydrogen, C _1-3 linear alkyl; more preferably, said L is selected from -CH ₂ O-, -CONH-, -CH ₂ NH- or -CH ₂ N(CH ₃ )-.

B is a combined structure of two or more saturated or unsaturated carbocyclic rings, and the carbocyclic ring optionally contains one or more heteroatoms;

m, n are independently selected from 1, 2 or 3;

Z may be the same or different, selected from CH, CR ₉ or N; said R ₉ is halogen, selected from F, Cl, Br, I;

R _{4 is} selected from C _1-4 alkyl or C _1-4 alkoxy. The C _1-4 alkyl or C _1-4 alkoxy is optionally further substituted by one or more halogen or cyano groups, The hydroxyl group is substituted or further substituted by a saturated or unsaturated carbocyclic ring or a heteroatom-containing carbocyclic ring; the saturated or unsaturated carbocyclic ring or a heteroatom-containing carbocyclic ring is optionally further substituted by one or more halogens , Cyano, amino or hydroxyl; preferably, the saturated or unsaturated carbocyclic ring or heteroatom-containing carbocyclic ring is a four-membered ring, a five-membered ring, or a six-membered ring; more preferably, the saturated or The unsaturated carbocyclic ring or the heteroatom-containing carbocyclic ring is a butylene oxide substituent, a pentylene oxide substituent or an oxazole substituent; preferably, R _{4 is} selected from the group consisting of a butylene oxide substituent, pentylene oxide Alkyl substituents or oxazole substituents substituted methyl, ethyl, propyl; more preferably, R _{4 is} selected from methyl substituted with butylene oxide substituents, pentylene oxide substituents or oxazole substituents ；

R ₅ is one or more than two, selected from hydrogen, hydroxy, amino, halogen, cyano, carboxy, tetrazolyl, amide; and the hydroxy, amino, carboxy, tetrazolyl or amide group is optional Is further substituted by one or more hydroxy, hydroxyalkyl, carboxyalkyl, aminoalkyl, and the alkyl is selected from C _1-4 alkyl; preferably, R _{5 is} selected from carboxy, amide, or A hydroxy, hydroxyalkyl, carboxyalkyl, or aminoalkyl substituted amide group; the amide group substituted by a hydroxy, hydroxyalkyl, carboxyalkyl, or aminoalkyl group is, for example, hydroxy, hydroxymethyl, Amido groups substituted by hydroxyethyl, carboxymethyl, and carboxyethyl;

Or R ₅ may form a ring system structure, the ring system structure may further contain one or more nitrogen atoms and/or one or more intra-ring carbonyl groups, and the ring system structure is optionally surrounded by one or more R ₆ substitution, said R _{6 is} selected from hydrogen, hydroxy, amino, halogen, cyano, carboxy, tetrazolyl, amide; and said hydroxy, amino, carboxy, tetrazolyl or amide optionally further Is substituted by one or two or more hydroxy, hydroxyalkyl, carboxyalkyl, aminoalkyl, the alkyl is selected from C _1-4 alkyl; preferably, R ₅ is connected to

Form a ring structure

or

The ring system structure is optionally substituted with one or more R ₆ .

In certain embodiments, L is selected from -(CH ₂ ) _m O-, -SO ₂ NR ₇ -, -(CH ₂ ) _m NR ₇ -, and m is 1; more preferably, R _{7 is} selected from H or methyl;

In certain embodiments, A is one or two or more R ₁ substituted aromatic rings or aromatic heterocycles or C ₂ -C ₈ alkenyl groups, R _{1 is} selected from hydrogen, hydroxyl, amino, halogen, cyano, C _1-4 alkyl, C _1-4 alkoxy, C _1-4 alkyl or C _1-4 alkoxy substituted by one or more halogens; preferably, A is substituted with one or more R ₁ Phenyl

i is selected from an integer of 1-5, such as 1,2,3,4,5; preferably, i is 1 or 2, preferably, R _{1 is} selected from cyano, F, Cl, Br, I, more preferably , A is

The X is halogen, selected from F, Cl, Br, I; preferably, A is

Preferably, R _{1 is} selected from F, Cl, Br, I; in some embodiments, B is selected from the following exemplary structures:

Wherein, Y can be the same or different, and is selected from CH or N; Z is defined as above, R ₂ , R ₃ , R ₈ can be one or more than two, independently selected from hydrogen, hydroxyl, amino, halogen, cyanide Group, C _1-4 alkyl, C _1-4 alkoxy, C _1-4 alkyl or C _1-4 alkoxy substituted with one or more halogens.

Further, B is selected from the following exemplary structures:

The definitions of R ₂ and R ₃ are the same as before;

Further, B is selected from the following exemplary structures:

The definitions of R ₂ and R ₃ are the same as before;

As a specific embodiment, the compound of the present invention has the following structure:

Among them, Y can be the same or different, and is selected from CH or N; Z, R ₂ , R ₃ , R ₄ , R _{5 have} the same definitions as before, (R ₁ ) _i represents that i can be the same or different on the phenyl group substituted by R _1, the same as R ₁ defined before, I is an integer selected from 1-5, for example, 1,2,3,4,5, preferably, i is 1 or 2.

Further, the compound of the present invention has the following structure:

Further, the compound of the present invention has the following structure:

Wherein, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₇ , and i have the same definitions as before, X is selected from -CH ₂ -, -CO- or -SO ₂ -, Y may be the same or different, and From CH or N, Z may be the same or different, and are selected from CH, CR ₉ or N. The R ₉ is halogen, selected from F, Cl, Br, and I.

Wherein, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₇ , and i have the same definitions as before, X is selected from -CH ₂ -, -CO- or -SO ₂ -, Y may be the same or different, and From CH or N.

Wherein, R ₁ , R ₃ , R ₄ , R ₅ , and i are as defined above, Z may be the same or different, and are selected from CH, CR ₉ or N, and R ₉ is halogen, selected from F, Cl, Br, I.

Wherein, R ₁ , R ₂ , R ₃ , R ₄ , and i are as defined above, Y may be the same or different, and are selected from CH or N, and R ₆ is one or more than two, and are selected from hydrogen, hydroxyl, amino, halogen, A cyano group, a carboxyl group, a tetrazolium group, an amide group, and the hydroxyl group, an amino group, a carboxyl group, a tetrazolium group or an amide group is optionally further substituted with one or more hydroxyalkyl groups, carboxyalkyl groups, The alkyl group is selected from C _1-4 alkyl groups;

Wherein, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ and i are as defined above, X is selected from -CH ₂ -, -CO- or -SO ₂ -, Y may be the same or different, and is selected from CH or N , Z may be the same or different, and are selected from CH, CR ₉ or N. The R ₉ is halogen, selected from F, Cl, Br, and I.

Wherein, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₇ , and i are as defined above, X is selected from -CH ₂ -, -CO- or -SO ₂ -, Y may be the same or different, and are selected from CH or N, Z may be the same or different, and are selected from CH, CR ₉ or N, and R ₉ is halogen, selected from F, Cl, Br, and I.

Wherein, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , and i are as defined above, Y may be the same or different, selected from CH or N, Z may be the same or different, selected from CH, CR ₉ or N, so Said R ₉ is halogen, selected from F, Cl, Br, I.

Wherein, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₇ , R ₈ , and i are as defined above, X is selected from -CH ₂ -, -CO- or -SO ₂ -, Y can be the same or different , Selected from CH or N, Z may be the same or different, selected from CH, CR ₉ or N, said R ₉ is halogen, selected from F, Cl, Br, I.

Wherein, R ₁ , R ₄ , R ₅ , R ₇ , and i are as defined above, X is selected from -CH ₂ -, -CO- or -SO ₂ -, Y can be the same or different, and can be selected from CH or N, Z can Same or different, selected from CH, CR ₉ or N, said R ₉ is halogen, selected from F, Cl, Br, I; preferably, R _{1 is} selected from hydrogen, halogen, cyano, C _1-4 alkane Group, C _1-4 alkoxy, C _1-4 alkyl or C _1-4 alkoxy substituted with one or more halogens.

Exemplarily, the compound of the present invention has the following structure:

The general synthesis method of the compound of the present invention is as follows:

Route 1:

Route 2:

Route 3:

Route 4:

The present invention also provides a compound with the following structure, which serves as an intermediate in the preparation of the compound of the present invention:

Wherein, R ₁ , R ₂ , R ₃ , R ₇ , Z, Y, and i are as defined above, Y'is selected from CR ₁₀ or NR ₁₀ , and R _{10 is} selected from H, C1-C8 alkyl, C1 -C4 alkyl carboxyl, C1-C4 alkyl hydroxy; preferably, the R _{10 is} selected from H, methyl, carboxy, carboxymethyl, carboxyethyl.

In a specific embodiment of the present invention, the intermediate has the following structure:

Among them, R ₁ , R ₂ , R ₃ , R ₇ , Z, Y, i, and Y'are as defined above.

Exemplarily, the intermediate described in the present invention has the following structure:

In certain embodiments, the GLP-1 receptor modulator provided by the present invention has the function of activating, enhancing or agonizing the GLP-1 receptor.

In certain embodiments, the present invention provides a medicament comprising a compound of the present invention, a pharmaceutically acceptable salt, hydrolyzable ester, stereoisomer, ether, prodrug, or solvate thereof, and at least one pharmaceutically acceptable carrier combination.

The GLP-1 receptor modulator compound of the present invention, its pharmaceutically acceptable salt, hydrolyzable ester, stereoisomer, ether, prodrug or solvate can be combined with a pharmaceutically acceptable carrier to provide suitable for lactation Pharmaceutical compositions for diseases of animals, preferably humans. The specific carriers used in these pharmaceutical compositions can vary depending on the type of administration required (for example, intravenous, oral, topical, suppository, or parenteral administration type).

In the preparation of oral liquid dosage forms, conventional pharmaceutical media such as water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, etc. can be used for formulation. Similarly, when preparing oral solid dosage forms, carriers such as diluents, granulating agents, lubricants, binders, disintegrating agents and the like can be used.

Embodiments of the present invention also include prodrugs of the compounds of the present invention, which are converted into the compounds of the present invention with pharmacological activity through enzymatic and chemical reactions in vivo. In fact, such prodrugs are functional derivatives of the compounds of the present invention, and conventional methods for selecting and preparing suitable prodrug derivatives are known in the art.

In certain embodiments, the compounds of the present invention provide methods for treating malignant conditions in subjects that medically require activation, enhancement, or agonism of GLP-1 receptors by following a frequency or duration sufficient to provide a beneficial effect to the patient An effective amount of the compound of the invention is administered to the subject for treatment.

The treatment method provided by the present invention includes administering the compound of the present invention alone or in combination with another pharmacologically active agent to a subject or patient suffering from a malignant condition that medically requires the activation, enhancement or agonism of the glucagon-like peptide 1 receptor Or the second drug is to administer the compound of the present invention, the malignant conditions such as type I diabetes, type II diabetes, gestational diabetes, obesity, excessive appetite, insufficient satiety or metabolic disorders.

In certain embodiments, the invention provides a pharmaceutical composition comprising a compound of the invention and a second drug. In certain such embodiments, the second drug is a polypeptide GLP-1 agonist or other type II diabetes treatment drugs, such as DPP-IV inhibitors.

In certain embodiments, the second drug is Exenatide, Semaglutide, Liraglutide, Tasglutide, Albiglutide, Lixisenatide, and/or other insulin modulating peptides.

In certain embodiments, the second drug is a DPPIV inhibitor, such as sitagliptin.

In various combinations, the second drug is a sodium-glucose co-transporter SGLT1 and/or SGLT2 inhibitor; or the second drug is a biguanide hypoglycemic drug, such as metformin; or the second drug is a sulfonylurea hypoglycemic drug , Such as glibenclamide, glipizide, gliclazide and/or glimepiride; or the second drug is a thiazolidinedione hypoglycemic agent, pioglitazone and/or rosiglitazone; or the second drug The drugs are diabetes treatment or adjuvant treatment drugs such as acarbose, miglitol, colesevelam and/or bromocriptine.

Detailed ways

The present invention will be further described in detail below in conjunction with specific embodiments. The following examples are used to understand the method and core idea of the present invention. For those skilled in the art, any possible changes or substitutions made without departing from the concept of the present invention fall within the protection scope of the present invention. Unless otherwise specified, the raw materials and reagents used in the present invention are all chemically pure and above purity.

1. Preparation of exemplary compounds

Compound A

(1) Intermediate preparation

Diisopropylamine (1.84mL, 13.12mmol) was dissolved in tetrahydrofuran (12mL) and the temperature was reduced to -26°C, n-BuLi (2.5M, 5.2mL, 13mmol) was added dropwise within fifteen minutes. The temperature was lowered to -30°C, compound 1a (3.12g, 12.82mmol) was dissolved in tetrahydrofuran (10mL) and then added dropwise. After half an hour of reaction, compound 1 (1.88g, 12.82mmol) was added. Then the temperature was raised to 25°C and stirred for 2.5 hours. Drop HCl solution (6M) into the reaction solution and adjust the pH to 7-8. MTBE extraction, washing with saturated sodium chloride solution (50 mL x 2), and drying. Purification by silica gel column (PE:EA=5:1) gave 3.4 g of yellow oil. Yield: 75.0%. LCMS [M+H] ⁺ =355.1.

Compound 2 (3.4 g, 9.62 mmol) was dissolved in methanol (12 mL) at 43° C., and then added to NaOH solution (4M, 12 mL). The temperature was raised to 50° C. and the mixture was stirred for 40 min. The pH was adjusted to 2.0 by dripping 6M HCl, a white solid was precipitated, and 3.27 g of a white solid was obtained by filtration. Yield: 95.0%. LCMS [M+H] ⁺ =341.1.

Compound 3 (3.27 g, 9.62 mmol) was dissolved in dichloroethane (20 mL), heated to 82° C. and stirred overnight. After spin-drying, 2.99 g of light yellow oil was obtained. LCMS [M+H] ⁺ = 425.1.

Compound 4 (1.2g, 4.05mmol) was dissolved in dioxane (20mL), t-BuOK (0.91g, 8.10mmol) and compound 4a (1.0g, 6.08mmol), Pd ₂ (dba) ₃ ( 0.37g, 0.405mmol), BINAP (0.50g, 0.81mmol). Stir at 110°C for 12h under nitrogen protection. Purify by silica gel column (PE:EA=5:1) and spin dry to obtain 1.12 g of light yellow oil. Yield: 65.0%. LCMS [M+H] ⁺ = 425.1.

Compound 5 (2.12 g, 2.64 mmol) was dissolved in dichloromethane (15 mL), and 5 mL trifluoroacetic acid was added. Stir at room temperature for 1 h. Spin to dryness, re-dissolve in 30mL ethyl acetate and 30mL saturated sodium bicarbonate solution, separate the layers, extract the aqueous phase with ethyl acetate three times (40mL x 3), dry and concentrate to obtain a waxy solid 1.5g, the yield is 90 %. LCMS [M+H] ⁺ = 325.1.

Compound 7 (480 mg, 2.0 mmol) was dissolved in acetonitrile (10 mL), and triethylamine (0.3 g, 3 mmol) and compound 7a (210 mg, 2.4 mmol) were added. Stir at room temperature for 12h under nitrogen protection. It was spin-dried to obtain 618 mg, which was used directly in the next step.

Compound 8 (618 mg, 2.0 mmol) was dissolved in ethanol (20 mL) and water (4 mL), iron powder (0.45 g, 8.10 mmol) and ammonium chloride (0.45 g, 8.1 mmol) were added, and stirred at 80°C for 2 h. Add celite to filter, concentrate, and purify by silica gel column to obtain 560 mg of yellow oil. The yield was 95%. LCMS [M+H] ⁺ = 279.2.

Compound 9 (560 mg, 2 mmol) was dissolved in acetonitrile (20 mL), 9a (450 mg, 3 mmol) and p-toluenesulfonic acid (10 mg) were added, and the mixture was stirred at 70°C for 1 h. It was spin-dried and purified by silica gel column (PE:EA=5:1) to obtain 530 mg of light yellow oil. Yield: 85.0%. LCMS [M+H] ⁺ =337.2.

Compound 6 (80 mg, 0.25 mmol) and compound 10 (108 mg, 0.32 mmol) were dissolved in acetonitrile (6 mL), and K ₂ CO ₃ (104.3 mg, 0.75 mmol), KI (4.15 mg, 0.025 mmol) were added. Stir at 50°C for 2h. Wash with water, extract with ethyl acetate, spin dry, and purify by silica gel column (PE:EA=1:2). 112.0 mg of pale yellow solid was obtained. Yield: 72.0%. LCMS [M+H] ⁺ = 625.2.

(2) Preparation of target compound

Compound 11 (112.0 mg, 0.18 mmol) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (3 mL) was added. Stir at room temperature for 1 h, spin dry, and separate by reversed-phase C-18 column chromatography (formic acid condition) to obtain 23.2 mg of white solid. Yield: 22.75%.

¹ H NMR (400MHz, DMSO) δ 11.04 (s, 1H), 8.27 (s, 1H), 8.00 (d, J = 9.6 Hz, 2H), 7.88-7.74 (m, 4H), 7.64 (d, J ＝8.4Hz,1H), 7.09(d,J＝7.5Hz,1H), 5.09(dd,J＝7.1,2.4Hz,1H), 4.82(dd,J＝15.2,7.2Hz,1H), 4.68(dd ,J=15.2,2.6Hz,1H),4.56–4.45(m,1H),4.43–4.31(m,1H),3.95(d,J=13.5Hz,1H), 3.80(d,J=13.5Hz, 1H), 2.99 (d, J = 10.3 Hz, 1H), 2.87 (d, J = 11.0 Hz, 1H), 2.79-2.68 (m, 1H), 2.65 (d, J = 19.6 Hz, 1H), 2.47- 2.34(m,1H), 2.30-2.09(m,2H), 1.96-1.61(m,4H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₁ H ₃₀ FN ₆ O ₄ 569.61, found 569.23

Compound B

(1) Intermediate preparation

Compound 7 (480 mg, 2.0 mmol) was dissolved in acetonitrile (10 mL), and triethylamine (0.6 g, 6 mmol) and compound 7b (220 mg, 2.4 mmol) were added. Stir at room temperature for 12h under nitrogen protection. Spin to dry to obtain 632 mg, which was used directly in the next step.

Compound 12 (632 mg, 2.0 mmol) was dissolved in ethanol (20 mL) and 4 mL of water, iron powder (0.45 g, 8.10 mmol) and ammonium chloride (0.45 g, 8.1 mmol) were added, and stirred at 80°C for 2 h. Add diatomaceous earth to filter, concentrate, and flash purification to obtain 530 mg of yellow oil with a yield of 88%. LCMS [M+H] ⁺ = 293.2.

Compound 13 (530 mg, 1.9 mmol) was dissolved in acetonitrile (20 mL), and 9a (450 mg, 3 mmol) and p-toluenesulfonic acid (10 mg) were added, and the mixture was stirred at 70°C for 1 h. Spin to dryness and flash purification (PE:EA=5:1) to obtain 530 mg of light yellow oil. Yield: 90.0%. LCMS [M+H] ⁺ =337.2.

Compound 6 (80 mg, 0.25 mmol) and compound 14 (113.75 mg, 0.33 mmol) were dissolved in acetonitrile (6 mL), and K ₂ CO ₃ (104.3 mg, 0.75 mmol), KI (4.15 mg, 0.025 mmol) were added. Stir at 50°C for 2h. Washed with water, extracted with ethyl acetate, spin-dried, and purified by flash (PE:EA=1:3) to obtain 113.68 mg of light yellow solid. Yield: 65.0%. LCMS [M+H] ⁺ = 639.2.

(2) Preparation of target compound

Compound 15 (113.68 mg, 0.18 mmol) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (3 mL) was added. Stir at room temperature for 1 h. After spin-drying, reverse phase C-18 column chromatography (formic acid) to obtain 28.6 mg of white solid. Yield: 27.32%. LCMS [M+H] ⁺ = 583.2.

¹ H NMR (400MHz, DMSO) δ 11.02 (s, 1H), 8.28 (s, 1H), 8.02 (d, J = 9.5 Hz, 2H), 7.83 (t, J = 7.8 Hz, 4H), 7.71 ( s,1H),7.11(s,1H),4.61(d,J=13.8Hz,1H),4.55-4.36(m,1H),4.18(s,1H),3.72(dq,J=68.0,7.1Hz ,3H),3.21-2.75(m,2H),2.29-1.69(m,8H),1.69-1.51(m,1H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₂ H ₃₂ FN ₆ O ₄ 583.64,found 583.24

Compound C

(1) Intermediate preparation

Compound 7 (723 mg, 3.0 mmol) was dissolved in acetonitrile (10 mL), and triethylamine (1.5 g, 15 mmol) and compound 7c (510 mg, 3.0 mmol) were added. Stir at room temperature for 12h under nitrogen protection. Spin to dry to obtain 960 mg, which was used directly in the next step.

Compound 16 (960 mg, 3.13 mmol) was dissolved in EtOH (20 mL), H ₂ O (20 mL), iron powder (0.70 g, 12.52 mmol) and NH ₄ Cl (0.67 g, 12.52 mmol) were added. Stir at 90°C for 4h under nitrogen protection. Filter and spin dry the filtrate. Flash purification (PE:EA=1:1) gave 0.544 g of light yellow oil. Yield: 60.0%. LCMS [M+H] ⁺ = 290.1.

Compound 17 (0.544 g, 1.88 mmol) was dissolved in acetonitrile (10 mL), and p-toluenesulfonic acid monohydrate (17.9 mg, 0.094 mmol) and compound 9a (0.434 g, 2.82 mmol) were added. Stir at 60°C for 1 h under nitrogen protection. Flash purification (PE:EA=5:1), 0.59 g of light yellow oil was obtained. Yield: 90.0%. LCMS [M+H] ⁺ = 348.1.

Compound 6 (80 mg, 0.25 mmol) and compound 18 (114.51 mg, 0.33 mmol) were dissolved in acetonitrile (6 mL), and K ₂ CO ₃ (104.3 mg, 0.75 mmol), KI (4.15 mg, 0.025 mmol) were added. Stir at 50°C for 2h. Spin to dryness and flash purification (PE:EA=1:3) to obtain 101.6 mg of light yellow solid. Yield: 64.0%. LCMS [M+H] ⁺ = 636.2.

(2) Preparation of target compound

Compound 19 (101.6 mg, 0.16 mmol) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (3 mL) was added. Stir at room temperature for 1 h. After spin-drying, reverse phase C-18 column chromatography (formic acid) to obtain 21.6 mg of white solid. Yield: 23.28%.

¹ H NMR(400MHz,DMSO)δ11.04(s,1H),8.32(s,1H),8.29(s,1H),8.01(d,J=9.5Hz,2H),7.86-7.74(m,3H) ), 7.67(d,J=8.4Hz,1H),7.27(s,1H),7.07(d,J=7.5Hz,1H),5.88(s,2H),3.90(s,2H),2.94(s ,2H),2.73-2.58(m,1H),2.20(s,2H),1.80-1.65(m,4H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₁ H ₂₇ FN ₇ O ₄ 580.59,found 580.21

Compound D

(1) Intermediate preparation

Compound 20 (3.0 g, 12.37 mmol) was dissolved in tetrahydrofuran (50 mL), the temperature was reduced to minus 78° C. under N ₂ protection, and n-BuLi was slowly added dropwise. After reacting at minus 78°C for 30 minutes, DMF was added, and the reaction was continued for 30 minutes at elevated temperature. Saturated NH ₄ Cl solution was added dropwise to the reaction solution to quench the reaction, and extracted with ethyl acetate (30 mL x 3). Wash with saturated sodium chloride solution (50 mL) and dry. Flash purification (PE:EA=5:1) to obtain 1.24 g of white solid. Yield: 52.31%. LCMS [M+H] ⁺ = 192.1.

Compound 21 (500 mg, 2.62 mmol) was dissolved in EtOH (10 mL), and CH ₃ CH ₂ NH ₂ HCl (424 mg, 5.24 mmol), AcOH (471.6 mg, 7.86 mmol) and NaBH ₃ CN (330.1 mg, 5.24) were added sequentially mmol). After stirring at 80°C for 1 h, LCMS showed that the compound disappeared, and the main peak LCMS [M+H] ⁺ =221.1. Continue to add Na ₂ CO ₃ (1.39 g, 13.1 mmol) and Boc ₂ O (1.13 g, 5.24 mmol) to the reaction solution, and react at room temperature for 1 hour. Suction filtration, wash the filter cake with methanol, and spin-dry the filtrate. Flash purification (PE:EA=8:1) gave 469.5 mg of pale yellow solid. Yield: 56.0%. LCMS [M+H] ⁺ = 321.1.

Compound 22a (1.11 g, 7.35 mmol) was dissolved in dioxane (20 mL), t-BuOK (1.13 g, 7.35 mmol) and compound 22 (469.5 mg, 1.47 mmol) were added. Stir at 110°C for 2h. Spin to dryness and flash purification (PE:EA=5:1) to obtain 276.2 mg of pale yellow oil. Yield: 43.21%. LCMS [M+H] ⁺ =436.21.

Compound 23 (470 mg, 0.63 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (2 mL) was added. Stir at room temperature for 1 h. Spin to dryness to obtain 395 mg of white solid (hydrochloride). Yield: 98.0%. LCMS [M+H] ⁺ = 336.1.

Compound 24 (60 mg, 0.18 mmol) and compound 10 (78.6 mg, 0.234 mmol) were dissolved in acetonitrile (6 mL), and K ₂ CO ₃ (75.1 mg, 0.54 mmol), KI (3.00 mg, 0.018 mmol) were added. Stir at 50°C for 2h. It was spin-dried and purified by flash (PE:EA=1:3) to obtain 82.3 mg of light yellow solid. Yield: 72.0%. LCMS [M+H] ⁺ = 636.2.

(2) Preparation of target compound

Compound 25 (82.3 mg, 0.130 mmol) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (3 mL) was added. Stir at room temperature for 1 h. After spin-drying, reverse phase C-18 column chromatography (formic acid) to obtain 25.4 mg of white solid. Yield: 28.1%.

¹ H NMR(400MHz,DMSO)δ8.26(d,J=8.9Hz,1H), 8.18(s,1H), 7.94(d,J=9.8Hz,1H), 7.88(d,J=8.2Hz, 1H), 7.82(t,J=7.6Hz,1H),7.80–7.71(m,3H),7.62(d,J=8.4Hz,1H),7.46(dd,J=8.2,1.3Hz,1H), 7.08(d,J＝8.8Hz,1H), 5.64(s,2H), 4.97–4.83(m,1H), 4.66(dd,J＝15.1,8.1Hz,1H),4.50(d,J＝13.3Hz ,1H), 4.30(d,J=5.6Hz,1H),4.05(d,J=13.6Hz,1H), 3.98(dd,J=5.8,3.2Hz,1H), 3.86(t,J=13.0Hz ,2H),3.72(d,J=13.5Hz,1H),2.71-2.57(m,1H),2.21-2.09(m,1H),1.08(t,J＝7.0Hz,3H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₃ H ₃₁ FN ₅ O ₄ 580.63,found 580.23

Compound E

Compound 24 (60 mg, 0.18 mmol) and compound 14 (81.9 mg, 0.24 mmol) were dissolved in acetonitrile (6 mL), and K ₂ CO ₃ (75.06 mg, 0.54 mmol), KI (3.00 mg, 0.018 mmol) were added. Stir at 50°C for 2h. Wash with water, extract with ethyl acetate and spin dry. Flash purification (PE:EA=1:3). 79.4 mg of pale yellow solid was obtained. Yield: 67.5%. LCMS [M+H] ⁺ = 650.1.

Compound 26 (79.4 mg, 0.122 mmol) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (3 mL) was added. Stir at room temperature for 1 h. Spin dry. 86.3 mg (trifluoroacetate) of a white solid was obtained. Reverse phase C-18 column chromatography (formic acid) to obtain 21.5 mg of white solid. Yield: 29.66%.

¹ H NMR (400MHz, DMSO) δ 8.27 (d, J = 8.8 Hz, 1H), 8.13 (d, J = 1.0 Hz, 1H), 7.94 (dd, J = 9.9, 1.4 Hz, 1H), 7.87 ( d,J=8.2Hz,1H),7.82(t,J=7.6Hz,1H),7.79–7.72(m,3H),7.63(d,J=8.4Hz,1H),7.45(dd,J=8.2 ,1.4Hz,1H),7.09(d,J=8.8Hz,1H), 5.65(d,J=13.2Hz,2H), 4.44(dd,J=14.8,2.7Hz,1H), 4.27(dd,J = 14.8, 9.0 Hz, 1H), 4.10–3.95 (m, 2H), 3.90 (d, J = 13.4 Hz, 1H), 3.78 (d, J = 13.5 Hz, 1H), 3.66 (d, J = 13.4 Hz ,1H),3.41–3.35(m,2H), 3.30–3.24(m,2H), 2.69–2.56(m,1H), 1.86(ddd,J=14.8,12.0,7.4Hz,1H),1.67–1.52 (m,1H),1.52–1.38(m,1H),1.26(ddd,J=16.1,12.2,8.2Hz,1H),1.07(t,J=7.1Hz,3H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₄ H ₃₃ FN ₅ O ₄ 594.66,found 594.25

Compound F

Compound 24 (60 mg, 0.18 mmol) and compound 18 (81.2 mg, 0.234 mmol) were dissolved in acetonitrile (6 mL), and K ₂ CO ₃ (75.06 mg, 0.54 mmol), KI (3.00 mg, 0.018 mmol) were added. Stir at 50°C for 2h. Wash with water, extract with ethyl acetate and spin dry. Flash purification (PE:EA=1:3). 81.5 mg of pale yellow solid was obtained. Yield: 70.0%. LCMS [M+H] ⁺ =647.2.

Compound 27 (81.5 mg, 0.126 mmol) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (3 mL) was added. Stir at room temperature for 1 h. It was spin-dried and separated by reverse-phase C-18 column chromatography (formic acid) to obtain 20.3 mg of white solid. Yield: 27.32%. LCMS [M+H] ⁺ =591.2.

¹ H NMR(400MHz,DMSO)δ8.32-8.18(m,3H),7.94(dd,J=9.9,1.4Hz,1H),7.89(d,J=8.1Hz,1H),7.85(dd,J ＝7.1,5.9Hz,1H),7.81(d,J＝7.4Hz,1H),7.74(dd,J＝11.6,5.1Hz,2H),7.52(d,J＝8.0Hz,1H),7.13(d ,J=8.8Hz,2H),5.77(s,2H),5.64(s,2H),4.44-4.31(m,5H),3.01(s,2H),1.23(s,3H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₃ H ₂₈ FN ₆ O ₄ 591.62,found 591.21

Compound G

(1) Intermediate preparation

Dissolve compound 1 (7.5g, 50mmol) in NMP (50mL), add K ₂ CO ₃ (7g, 50mmol), compound 28 (9.3g, 50mmol), then react at 130°C for 10 hours, dilute with water, and ethyl acetate Extraction and purification by column chromatography (PE:EA=5:1), 7.5 g of white solid was obtained, yield: 50.5%.

Compound 29 (1.2g, 4.05mmol) was dissolved in dioxane (20mL), t-BuONa (0.91g, 8.10mmol) and compound 4a (1.0g, 6.08mmol), Pd ₂ (dba) ₃ ( 0.37g, 0.405mmol), BINAP (0.50g, 0.81mmol). Stir at 110°C for 12h under nitrogen protection. Spin dry. Flash purification (PE:EA=5:1) yielded 989 mg of pale yellow oil. Yield: 55.0%. LCMS [M+H] ⁺ = 426.1.

Compound 30 (300 mg, 7.2 mmol) was dissolved in dichloromethane (15 mL), and trifluoroacetic acid (5 mL) was added. Stir at room temperature for 1 h. Rotate to dryness, re-dissolve in 30 mL ethyl acetate and 30 mL saturated sodium bicarbonate solution, separate the liquids, extract the aqueous phase with ethyl acetate, dry, and concentrate to obtain 220 mg waxy solid with a yield of 90%. LCMS [M+H] ⁺ = 326.1.

Compound 31 (40mg, 0.13mmol) and compound 10 (50mg, 0.16mmol) were dissolved in acetonitrile (6mL), K ₂ CO ₃ (104.3mg, 0.75mmol), KI (4.15mg, 0.025mmol), 50°C were added Stir for 2h. Spin to dryness and flash purification (PE:EA=1:3) to obtain 49 mg of pale yellow solid. Yield: 58.0%. LCMS [M+H] ⁺ = 626.2.

(2) Preparation of target compound

Compound 32 (49 mg, 0.15 mmol) was dissolved in dichloromethane (3.0 mL), and trifluoroacetic acid (3.0 mL) was added. Stir at room temperature for 1 h. Reverse phase C-18 column chromatography (formic acid) to obtain 26.5 mg of white solid. Yield: 31.0%.

¹ H NMR (400MHz, DMSO) δ 12.74 (s, 1H), 10.67 (s, 1H), 8.28 (d, J = 1.0 Hz, 1H), 8.00 (d, J = 9.6 Hz, 1H), 7.81 ( dd,J=8.3,1.5Hz,3H),7.66(d,J=8.4Hz,1H),7.58(t,J=8.0Hz,1H),7.44(s,1H),6.59(d,J=8.0 Hz, 1H), 5.11 (dd, J = 7.2, 2.5 Hz, 1H), 4.78 (d, J = 7.2 Hz, 1H), 4.72-4.62 (m, 1H), 4.49 (dd, J = 7.7, 2.0 Hz ,1H), 4.38 (dd, J = 5.9, 3.1 Hz, 1H), 3.98 (d, J = 13.6 Hz, 1H), 3.81 (d, J = 13.6 Hz, 1H), 3.49 (s, 3H), 2.79 –2.64(m,1H),2.55(s,3H),2.43(ddd,J=8.9,4.0,1.7Hz,1H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₀ H ₂₉ FN ₇ O ₄ 570.60,found 570.22

Compound H

Compound 31 (80 mg, 0.25 mmol) and compound 13 (112 mg, 0.32 mmol) were dissolved in acetonitrile (6 mL), and K ₂ CO ₃ (104.3 mg, 0.75 mmol), KI (4.15 mg, 0.025 mmol) were added. Stir at 50°C for 2 hours, wash with water, extract with ethyl acetate, spin dry, and flash purification (PE:EA=1:3) to obtain 96.0 mg of light yellow solid. Yield: 60.0%. LCMS [M+H] ⁺ = 640.2.

Compound 33 (96.0 mg, 0.15 mmol) was dissolved in dichloromethane (3.0 mL), trifluoroacetic acid (3.0 mL) was added, stirred at room temperature for 1 h, and spin-dried to obtain 99 mg of white solid, which was separated by reversed-phase C-18 column chromatography (Formic acid) 20.1 mg of white solid was obtained. Yield: 22.95%.

¹ H NMR (400MHz, DMSO) δ 12.72 (s, 1H), 10.67 (s, 1H), 8.23 (d, J = 1.0 Hz, 1H), 8.00 (d, J = 9.6 Hz, 1H), 7.88- 7.73(m,3H), 7.65(d,J=8.5Hz,1H), 7.58(t,J=8.0Hz,1H),7.45(s,1H), 6.59(d,J=8.0Hz,1H), 4.57(dd,J=14.8,2.8Hz,1H), 4.46(dd,J=14.8,8.2Hz,1H), 4.25(dd,J=7.7,2.7Hz,1H), 4.01(d,J=13.7Hz ,1H),3.79(dt,J=16.1,10.2Hz,2H),3.68–3.58(m,1H),3.48(s,3H),2.55(s,3H),2.06(dt,J=8.1,4.8 Hz, 1H), 1.85 (ddd, J = 21.7, 11.5, 6.6 Hz, 2H), 1.73-1.56 (m, 1H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₁ H ₃₁ FN ₇ O ₄ 584.62,found 584.24

Compound I

Compound 31 (80 mg, 0.25 mmol) and compound 16 (111 mg, 0.32 mmol) were dissolved in acetonitrile (6 mL), and K ₂ CO ₃ (104.3 mg, 0.75 mmol), KI (4.15 mg, 0.025 mmol) were added. Stir at 50°C for 2 hours, wash with water, extract with ethyl acetate, and spin-dry; flash purification (PE:EA=1:3) to obtain 100.2 mg of light yellow solid. LCMS [M+H] ⁺ = 637.2.

Compound 34 (100.2 mg, 0.16 mmol) was dissolved in dichloromethane (3.0 mL), and trifluoroacetic acid (3.0 mL) was added. Stir at room temperature for 1 hour and spin dry to obtain 110 mg of white solid. Reverse phase C-18 column chromatography (formic acid) to obtain 24.13 mg of white solid, yield: 26.0%.

¹ H NMR (400MHz, DMSO) δ 12.68 (s, 1H), 10.66 (s, 1H), 8.30 (d, J = 8.7 Hz, 2H), 8.00 (d, J = 9.7 Hz, 1H), 7.88- 7.74(m,3H),7.69(d,J=8.4Hz,1H),7.58(t,J=8.0Hz,1H),7.44(s,1H),7.24(s,1H),6.58(d,J ＝7.9Hz, 1H), 5.87 (s, 2H), 3.92 (s, 2H), 3.43 (s, 3H), 2.60-2.53 (m, 2H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₀ H ₂₆ FN ₈ O ₄ 581.58,found 581.21

Compound J, K, L, M

(1) Intermediate preparation

In a microwave tube, dissolve compound 4 (0.89g, 3.07mmol) in dioxane (20mL), add 22a (0.7g, 4.6mmol), xantphos (355mg, 0.61mmol), Pd ₂ (dba) ₃ (290 mg, 0.32 mmol) and Cs ₂ CO ₃ (2.5 g, 7.62 mmol). The reaction system was purged with N ₂ purging, heated to 130 ℃ in a microwave reaction apparatus, the reaction 2h. It was spin-dried, column chromatography (PE:EA=2:1) and concentrated to obtain 975 mg of pale yellow oil. Yield: 63.1%. LCMS [M+H] ⁺ =412.5.

Compound 35 (750 mg, 1.82 mmol) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (3 mL) was added. Stir at room temperature for 1 h. It was spin-dried to obtain 700 mg of white solid, which was neutralized with NaHCO ₃ solution, extracted, spin-dried, and flash purified (PE:EA=1:1) to obtain 500 mg of light yellow solid. Yield: 88.34%. LCMS [M+H] ⁺ = 312.2.

Compound 36 (500 mg, 1.61 mmol) and compound 10 (702 mg, 2.09 mmol) were dissolved in acetonitrile (6 mL), and K ₂ CO ₃ (671 mg, 4.83 mmol), KI (26.73 mg, 0.161 mmol) were added. Stir at 50°C for 2 hours, wash with water, extract with ethyl acetate, spin dry, and purify by flash (PE:EA=1:1) to obtain 460 mg of light yellow solid. Yield: 46.74%. LCMS [M+H] ⁺ = 612.2.

Compound 37 (460 mg, 0.753 mmol) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (3 mL) was added. Stir at room temperature for 1 hour and spin dry to obtain 450 mg of white solid. Reverse phase C-18 column chromatography (formic acid) to obtain 320 mg of white solid, yield: 76.56%. LCMS [M+H] ⁺ = 556.2.

Compound 38 (80 mg, 0.14 mmol) was dissolved in DMF (3.0 mL), HATU (60.25 mg, 0.16 mmol), DIPEA (54.2 mg, 0.42 mmol) were added. Stir at room temperature for 10 min, then add tert-butyl glycine (36.7 mg, 0.28 mmol), stir at room temperature for 1 h, wash with water, extract with ethyl acetate, dry, and flash purification (PE:EA=1:2) to obtain 70.0 mg of light yellow solid. Yield: 74.85%. LCMS [M+H] ⁺ = 669.2.

(2) Preparation of target compound

Compound 38 (80 mg, 0.14 mmol) was dissolved in DMF (3.0 mL), HATU (58.5 mg, 0.154 mmol), DIPEA (361.2 mg, 2.8 mmol) were added. Stir at room temperature for 10 min, then add hydroxylamine hydrochloride (96.6 mg, 1.4 mmol), and stir at room temperature for 1 h. It was washed with water, extracted with ethyl acetate, spin-dried, and separated by reverse-phase C-18 column chromatography (formic acid) to obtain 11.8 mg of white solid. Yield: 13.98%.

¹ H NMR(400MHz,DMSO)δ11.25(s,1H),9.99(s,1H),9.07(s,1H), 8.16(d,J=10.4Hz,1H), 7.78(d,J=8.5 Hz, 1H), 7.73–7.65 (m, 2H), 7.56 (dt, J = 20.2, 9.3 Hz, 3H), 6.94 (d, J = 7.3 Hz, 1H), 6.76 (d, J = 8.2 Hz, 1H ), 6.28 (s, 2H), 5.44 (s, 2H), 5.08 (dd, J = 7.1, 2.4 Hz, 1H), 4.88-4.69 (m, 3H), 4.63 (dd, J = 15.3, 2.3 Hz, 1H),4.51(d,J=6.2Hz,1H), 4.37(dd,J=6.0,3.0Hz,1H), 3.74(s,2H), 2.96(s,1H), 2.79–2.71(m,1H ), 2.69(s,3H),2.43-2.28(m,1H),2.07(d,J=28.6Hz,3H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₁ H ₃₅ FN ₇ O ₅ 604.66, found 604.26.

Compound 38 (80 mg, 0.14 mmol) was dissolved in DMF (3.0 mL), HATU (58.5 mg, 0.154 mmol), DIPEA (54.2 mg, 0.42 mmol) were added. Stir at room temperature for 10 min, then add ethanolamine (21.4 mg, 0.35 mmol), and stir at room temperature for 1 h. Washed with water, extracted with ethyl acetate, spin-dried, reversed phase C-18 column chromatography (formic acid) to obtain a white solid 25.6mg, yield: 30.58%.

¹ H NMR(400MHz,DMSO)δ8.47(t,J=5.7Hz,1H), 8.26(s,1H), 7.91(d,J=9.5Hz,1H), 7.86–7.68(m,4H), 6.96(d,J=7.1Hz,1H), 6.80(d,J=8.2Hz,1H), 5.51(s,2H), 5.08(d,J=6.9Hz,1H), 4.78(dd,J=15.5 ,7.1Hz,3H),4.63(d,J=13.0Hz,1H),4.56–4.46(m,1H), 4.38(d,J=2.7Hz,1H),4.33–4.22(m,1H),3.95 -3.54(m,7H), 2.93(s,1H), 2.80-2.58(m,1H), 2.43--2.26(m,1H), 2.22-1.90(m,4H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₃ H ₃₆ FN ₆ O ₄ 599.68, found 599.28.

Compound 38 (80 mg, 0.14 mmol) was dissolved in DMF (3.0 mL), HATU (58.5 mg, 0.154 mmol), DIPEA (54.2 mg, 0.42 mmol) were added. Stir at room temperature for 10 min, then add NH _{3 /} DMF (1M, 0.42 mL, 0.42 mmol), and stir at room temperature for 1 h. It was washed with water, extracted with ethyl acetate, spin-dried, and separated by reverse-phase C-18 column chromatography (formic acid) to obtain 36.1 mg of white solid. Yield: 46.52%.

¹ H NMR(400MHz,DMSO)δ8.30(s,1H),8.01(s,1H),7.91(d,J=9.6Hz,1H), 7.85(dd,J=8.5,1.5Hz,1H), 7.80–7.68(m,4H),7.42(s,1H), 6.96(d,J=7.3Hz,1H), 6.80(d,J=8.2Hz,1H),5.51(s,2H),5.08(dd ,J=7.1,2.4Hz,1H), 4.94–4.72(m,3H), 4.62(dd,J=15.4,2.5Hz,1H), 4.58–4.46(m,1H), 4.37(dt,J=8.9 ,6.0Hz,1H),3.95-3.75(m,4H), 3.32(s,2H), 2.95(s,1H), 2.81–2.64(m,1H), 2.44–2.30(m,1H), 2.15– 1.97(m,4H),1.80-1.61(m,1H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₁ H ₃₂ FN ₆ O ₃ 555.63, found 555.25.

Compound 39 (70 mg, 0.11 mmol) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (3 mL) was added. Stir at room temperature for 1 h and spin dry to obtain 91.5 mg of white solid. Reverse phase C-18 column chromatography (formic acid) to obtain 18.6 mg of white solid. Yield: 27.64%.

¹ H NMR (400MHz, DMSO) δ 12.59 (s, 1H), 10.07 (s, 1H), 8.88 (dd, J = 13.4, 5.9 Hz, 1H), 8.33 (d, J = 23.9 Hz, 1H), 7.97–7.77(m,3H),7.77–7.66(m,3H), 6.94(dd,J=20.0,7.3Hz,1H), 6.80(dd,J=8.2,3.3Hz,1H),5.51(s, 1H), 5.48(d,J=5.4Hz,1H), 5.44–5.25(m,1H), 5.15–5.00(m,1H), 4.98–4.73(m,2H), 4.64(d,J=13.0Hz) ,1H),4.59–4.47(m,1H),4.47–4.34(m,1H),4.17(s,2H),3.99(d,J=4.2Hz,2H),3.76(s,1H),3.62( dd,J=11.0,4.9Hz,1H), 3.10–2.83(m,2H), 2.72(d,J=7.1Hz,1H), 2.41–2.30(m,1H), 2.18(d,J=10.0Hz ,1H),2.02–1.90(m,1H),1.90–1.74(m,1H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₃ H ₃₄ FN ₆ O ₅ 613.66, found 613.25.

Compound N

(1) Intermediate preparation

Compound 21 (700 mg, 3.66 mmol) was dissolved in EtOH (15 mL), and CH ₃ NH ₂ HCl (495 mg, 7.33 mmol), AcOH (660 mg, 10.99 mmol) and NaBH ₃ CN (462 mg, 7.33 mmol) were sequentially added. After stirring at 80°C for 1 h, LCMS showed that the compound disappeared and the main peak was LCMS [M+H] ⁺ = 207.1. Continue to add Na ₂ CO ₃ (1.94 g, 18.32 mmol) and Boc ₂ O (965 mg, 7.33 mmol) to the reaction solution, and react at room temperature for 1 hour. LCMS showed that the main peak was the product. Suction filtration, the filter cake was washed with methanol, spin-dried, and flash purified (PE:EA=8:1) to obtain 620 mg of light yellow solid. Yield: 55.32%. LCMS [M+H] ⁺ = 307.2.

Compound 5 (1.52 g, 10.10 mmol) was dissolved in dioxane (20 mL), t-BuOK (1.13 g, 10.10 mmol) and compound 40 (620 mg, 2.02 mmol) were added. Stir at 110°C for 2h. Spin to dryness and flash purification (PE:EA=5:1) to obtain 334 mg of light yellow oil. Yield: 39.21%. LCMS [M+H] ⁺ = 422.3.

Compound 41 (334 mg, 0.79 mmol) was dissolved in dichloromethane (5 mL), and HCl/dioxane (1 mL) was added. Stir at room temperature for 1 h. Spin to dryness to obtain 228.0 mg of white solid. Yield: 80.41%. LCMS [M+H] ⁺ = 322.3.

Compound 42 (100 mg, 0.28 mmol) and compound 10 (121 mg, 0.36 mmol) were dissolved in acetonitrile (6 mL), and K ₂ CO ₃ (116.8 mg, 0.84 mmol), KI (4.65 mg, 0.028 mmol) were added. Stir at 50°C for 2h. It was washed with water, extracted with ethyl acetate, spin-dried, and purified by flash (PE:EA=1:3) to obtain 113.0 mg of light yellow solid. Yield: 65.0%. LCMS [M+H] ⁺ =622.2.

(2) Preparation of target compound

Compound 43 (113 mg, 0.182 mmol) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (3 mL) was added. Stir at room temperature for 1 h. It was spin-dried to obtain 125 mg of white solid, which was separated by reversed-phase C-18 column chromatography (formic acid) to obtain 32.1 mg of white solid. Yield: 31.06%. LCMS [M+H] ⁺ = 566.2.

¹ H NMR(400MHz,DMSO)δ8.36-8.27(m,2H),7.95(dd,J=13.5,4.7Hz,3H),7.88(dd,J=8.5,1.3Hz,1H),7.84-7.71 (m, 3H), 7.59 (d, J = 8.8 Hz, 1H), 7.18 (d, J = 8.8 Hz, 1H), 5.65 (s, 2H), 4.95 (ddd, J = 14.5, 7.3, 2.8 Hz, 1H), 4.77–4.44(m,5H), 4.36(dd,J=13.7,7.7Hz,1H), 4.23–4.10(m,1H), 2.77(s,3H), 2.71–2.57(m,2H) ,2.29–2.15(m,1H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₂ H ₂₉ FN ₅ O ₄ 566.61, found 566.22.

Compound O

(1) Intermediate preparation

Dissolve compound 1e (0.6g, 2.02mmol) in dioxane (10ml), add K ₂ CO ₃ (0.42g, 3.04mmol) and compound 1f (1.0g, 6.08mmol), Pd ₂ (dba) ₃ (0.18g, 0.202mmol), xantphos (0.50g, 0.81mmol). Stir at 130°C for 1 h under nitrogen protection. Spin to dryness and flash purification (PE:EA=5:1) to obtain 239 mg of light yellow oil. Yield: 27.0%. LCMS [M+1] ⁺ = 437.2.

Compound 2e (220 mg, 0.5 mmol) was dissolved in dichloromethane (3 ml), and trifluoroacetic acid (3 mL) was added. Stir at room temperature for 1 h. Rotate to dryness, re-dissolve in 30 mL ethyl acetate and 30 mL saturated sodium bicarbonate solution, separate liquids, extract the aqueous phase with ethyl acetate, dry and concentrate to obtain 160 mg of waxy solid with a yield of 98%. LCMS [M+1] ⁺ = 327.2.

Compound 3e (110mg, 0.3mmol) and compound 10 (72mg, 0.2mmol, 0.7eq) were dissolved in acetonitrile (6ml), K ₂ CO ₃ (104.3mg, 0.75mmol), KI (4.15mg, 0.025mmol) were added . Stir at 50°C for 2h. Spin to dryness and flash purification (PE:EA=1:3) to obtain 49 mg of pale yellow solid. Yield: 40.0%. LCMS [M+1] ⁺ = 636.3.

Compound 4e (49 mg, 0.15 mmol) was dissolved in dichloromethane (3.0 ml), and trifluoroacetic acid (3.0 mL) was added. Stir at room temperature for 1 h. After concentration, it was separated by reverse phase C-18 column chromatography (formic acid), and 12.5 mg of white solid was obtained after lyophilization. Yield: 31.0%.

¹ H NMR(400MHz,DMSO)δ11.18(s,1H),8.37(s,1H),7.99(td,J=7.7,1.5Hz,1H),7.95-7.88(m,1H),7.78(d ,J=8.5Hz,1H),7.66(dd,J=8.3,5.1Hz,1H),7.46(t,J=8.0Hz,1H),7.40(dd,J=10.0,8.6Hz,1H),7.37 –7.29(m,1H),6.41(d,J=8.4Hz,1H), 6.24(d,J=7.7Hz,1H), 5.04(dd,J=7.1,2.2Hz,1H), 4.80(dd, J = 15.4, 7.1 Hz, 1H), 4.66 (dd, J = 15.3, 2.5 Hz, 1H), 4.50 (dd, J = 13.6, 7.7 Hz, 2H), 4.35 (dt, J = 9.0, 5.9 Hz, 1H) ),3.04(s,6H),2.77–2.64(m,1H),2.41–2.27(m,1H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₂₈ H ₃₀ FN ₆ O ₅ S 581.64, found 581.20.

Compound 1T, 2T

(1) Intermediate synthesis

Compound 1b (1.6g, 5.4mmol) was dissolved in dioxane (20mL), and 1a (1.3g, 10.8mmol), BINAP (670mg, 1.08mmol), Pd ₂ (dba) ₃ (500mg, 0.54 mmol) and Cs ₂ CO ₃ (5.4 g, 16.5 mmol). Heat to 110°C and stir for 12h under nitrogen protection. It was spin-dried, purified by column chromatography (PE:EA=5:1) and concentrated to obtain 1.31 g of light yellow oil. Yield: 63.7%. LCMS [M+H] ⁺ = 382.3.

Compound 2b (1.3g, 3.4mmol) was dissolved in methanol (12mL) at 43°C, and then added to NaOH solution (4M, 12mL), heated to 50°C and stirred for 7h. Extraction with ethyl acetate, drying, concentration, column chromatography (PE:EA=1:1) and concentration to obtain 0.85 g of white solid. Yield: 90.0%. LCMS [M+H] ⁺ = 279.2.

Compound 3b (0.85g, 3.06mmol) was dissolved in DMF (20mL), 3a (840mg, 3.95mmol, 1.3eq) and DIEA (1.1g, 8.5mmol, 2.78eq) were added in sequence, the reaction system was heated to 65℃ and Stir for 5h. Dilute with water, extract with ethyl acetate, wash with saturated brine, dry, concentrate, column chromatography (PE:EA=1:1) and concentrate to obtain 0.83 g of light yellow oil. Yield: 72.0%. LCMS [M+H] ⁺ = 412.2.

Compound 4b (0.83 g, 2.1 mmol) was dissolved in dichloromethane (15 mL), trifluoroacetic acid (5 mL) was added, and the reaction system was stirred at room temperature for 1 h. Concentrate, add ethyl acetate and saturated sodium bicarbonate aqueous solution, separate the layers, extract the aqueous phase with ethyl acetate, wash the organic phase with saturated brine, dry, filter, and concentrate to obtain 0.61 g of waxy solid. Yield: 97.1%. LCMS [M+H] ⁺ = 312.2.

Compounds 5b (80 mg, 0.25 mmol) and 5a (108 mg, 0.32 mmol) were dissolved in acetonitrile (6 mL), and K ₂ CO ₃ (104 mg, 0.75 mmol) and KI (4.15 mg, 0.025 mmol) were added. The reaction system was stirred at 50°C for 2h. Dilute with water, extract with ethyl acetate, concentrate the organic phase, column chromatography (PE:EA=1:2) and concentrate to obtain 112 mg of light yellow solid. Yield: 71.3%. LCMS [M+H] ⁺ = 611.2.

(2) Synthesis of target compound

Compound 6b (112 mg, 0.18 mmol) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (3 mL) was added. Stir at room temperature for 1 h. After concentration, 123 mg of white solid was obtained, and 35 mg of white solid was obtained by reversed-phase C-18 column chromatography (formic acid). Yield: 34.5%.

¹ H NMR (400MHz, MeOD) δ 8.30 (t, J = 7.0 Hz, 1H), 7.94 (dd, J = 8.5, 1.4 Hz, 1H), 7.65 (d, J = 8.5 Hz, 1H), 7.50 ( t,J=7.8Hz,1H),7.48–7.40(m,2H),7.19(t,J=4.0Hz,1H),5.89(dd,J＝17.2,8.0Hz,1H),5.22(ddd,J = 14.6, 7.2, 2.6 Hz, 1H), 4.81 (s, 1H), 4.68 (dd, J = 15.4, 2.6 Hz, 1H), 4.61 (td, J = 7.9, 6.0 Hz, 1H), 4.43 (dt, J = 9.1, 5.9 Hz, 1H), 4.00 (d, J = 13.8 Hz, 1H), 3.88 (d, J = 13.8 Hz, 1H), 3.34 (t, J = 4.8 Hz, 4H), 2.83–2.68 ( m,1H), 2.59--2.43(m,5H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₀ H ₃₁ FN ₇ O ₃ 556.62, found 556.24.

Compound 1T (16 mg, 0.03 mmol) was dissolved in methanol (2 mL), and HCHO aqueous solution (0.2 mL) and sodium cyanoborohydride (10 mg) were sequentially added. Stir at room temperature for 2h. It was concentrated and separated by reversed-phase C-18 column chromatography (formic acid) to obtain 10 mg of white solid. Yield: 60%.

¹ H NMR (400MHz, DMSO) δ 8.39 (s, 1H), 7.98-7.89 (m, 1H), 7.87-7.78 (m, 2H), 7.68-7.59 (m, 1H), 7.45-7.37 (m, 1H), 7.27(dd,J=9.3,6.2Hz,1H), 6.16(d,J=8.1Hz,1H), 6.07(d,J=8.1Hz,1H), 5.07–4.97(m,1H), 4.84-4.82(m,5H), 4.65(dd,J=18.0,5.0Hz,1H),4.56–4.43(m,2H),4.33(dd,J=6.1,3.0Hz,1H), 3.38(d, J = 21.1Hz, 6H), 3.07 (s, 2H), 3.05 (s, 1H), 2.82–2.63 (m, 2H), 2.34 (d, J = 1.7 Hz, 1H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₁ H ₃₄ FN ₇ O ₃ 570.26, found 570.27.

Compound 3T, 4T

(1) Intermediate synthesis

Compound 7b (1.5g, 8.77mmol) was dissolved in dioxane (25mL) and H ₂ O (5mL), and 7a (2.5g, 8.09mmol), Pd(dppf)Cl ₂ (640mg, 0.877mmol) ) And K ₂ CO ₃ (2.4 g, 17.4 mmol). React at 110°C for 5h under N ₂ protection. Concentrate, column chromatography (PE:EA=3:1) and concentrate to obtain 1.1 g of light yellow oil. Yield: 45.6%. LCMS [M+H] ⁺ = 276.2.

Compound 8b (1.1 g, 4.0 mmol) was dissolved in methanol (25 mL), 50% Pd/C (700 mg) was added under N ₂ protection, and stirred at room temperature for 5 h under H ₂ atmosphere. The solid was filtered off with celite, washed with methanol, the filtrate was concentrated, column chromatography (PE:EA=3:1) and then concentrated to obtain 620 mg of a pale yellow oil. Yield: 56.4%. LCMS [M+H] ⁺ = 278.2.

Compound 9b (300 mg, 1.08 mmol) was dissolved in DMF (10 mL), and 9a (230 mg, 1.08 mmol) and DIEA (350 mg, 2.71 mmol) were added sequentially. Stir at 50°C for 3h. Dilute with water, extract with ethyl acetate, wash the organic phase with saturated brine, dry, concentrate, column chromatography (PE:EA=2:1) and concentrate to obtain 210 mg of light yellow solid. Yield: 47.3%. LCMS [M+H] ⁺ = 411.5.

¹ HNMR (400MHz, CDCl3) 7.62 (t, 1H), 7.53 (t, 1H), 7.44 (d, 1H), 7.37 (d, 1H), 6.75 (d, 1H), 6.65 (d, 1H), 5.49 (s, 1H), 4.20 (br s, 2H), 2.81 ((br s, 2H), 2.70 (tt, 1H), 1.82 (d, 2H), 1.67 (d, 2H), 1.49 (s, 9H)

Compound 10b (210 mg, 0.51 mmol) was dissolved in dichloromethane (6 mL), and trifluoroacetic acid (2 mL) was added to the reaction system and stirred at room temperature for 1 h. Concentrate, add ethyl acetate and saturated sodium bicarbonate aqueous solution, separate, extract with ethyl acetate, wash with saturated brine, dry, filter, and concentrate to obtain 130 mg of waxy solid. Yield: 81.8%. LCMS [M+H] ⁺ =311.2.

Compound 11b (60 mg, 0.19 mmol) and 5a (70 mg, 0.21 mmol) were dissolved in acetonitrile (5 mL), and K ₂ CO ₃ (80 mg, 0.58 mmol) and KI (3.21 mg, 0.019 mmol) were added. Stir at 50°C for 2h. Dilute with water, extract with ethyl acetate, concentrate the organic phase, column chromatography (PE:EA=1:2) and concentrate to obtain 80 mg of light yellow solid. Yield: 76.7%. LCMS [M+H] ⁺ = 612.2.

(2) Synthesis of target compound

Compound 12b (80 mg, 0.13 mmol) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (3 mL) was added. Stir at room temperature for 1 h. Concentrated, separated by reversed-phase C-18 column chromatography (formic acid), and lyophilized to obtain 25 mg of white solid. Yield: 34.2%.

¹ H NMR (400MHz, MeOD) δ 8.30 (d, J = 0.9 Hz, 1H), 7.96 (dd, J = 8.5, 1.4 Hz, 1H), 7.68 (d, J = 8.5 Hz, 1H), 7.50 ( d,J=7.8Hz,1H),7.47–7.36(m,1H),7.31(dd,J=8.1,7.4Hz,1H),6.43(d,J=7.2Hz,1H),6.35(d,J =8.2Hz,1H),5.21(ddd,J=14.5,7.1,2.5Hz,1H), 4.79(d,J=7.0Hz,1H), 4.71-4.53(m,3H), 4.42(dt,J= 9.2,5.9Hz,1H),4.19(dd,J=31.8,14.2Hz,2H),3.23-3.15(m,1H),2.81-2.67(m,1H),2.59-2.58(m,2H),2.48 (ddd,J=9.0,4.1,1.8Hz,1H),1.88– 1.72(m,3H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₁ H ₃₂ FN ₆ O ₃ 555.63, found 555.23.

Compound 3T (15 mg, 0.027 mmol, 1.0 eq) was dissolved in methanol (2 mL), and HCHO aqueous solution (0.2 mL) and sodium cyanoborohydride (10 mg) were sequentially added. Stir at room temperature for 2h. Concentration, reverse phase C-18 column chromatography (formic acid) to obtain 8 mg of white solid. Yield: 52.0%.

¹ H NMR(400MHz,DMSO)δ12.67(s,1H), 8.26(t,J=6.0Hz,1H),7.84(dd,J=10.0,1.5Hz,1H),7.80(dd,J=8.4 ,1.5Hz,1H),7.64(d,J=8.4Hz,1H),7.61(dd,J=7.9,1.5Hz,1H),7.52-7.42(m,1H),7.30(t,J=7.7Hz ,1H), 6.49(dd,J=8.3,6.6Hz,2H),5.08(dd,J=7.2,2.5Hz,1H),4.90(s,2H),4.78(dd,J=15.2,7.2Hz, 1H), 4.63 (dd, J = 15.2, 2.7 Hz, 1H), 4.45 (dt, J = 14.0, 7.0 Hz, 1H), 4.37 (dt, J = 9.0, 5.9 Hz, 1H), 3.92 (d, J = 13.5Hz, 1H), 3.74 (d, J = 13.5Hz, 1H), 3.06 (s, 3H), 2.94 (d, J = 10.9Hz, 1H), 2.79 (d, J = 10.6Hz, 1H), 2.72–2.59(m,1H),2.46-2.39(m,2H),2.28–2.06(m,2H),1.72-1.56(m,4H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₂ H ₃₄ FN ₆ O ₃ 569.65, found 569.26.

Compound 5T

(1) Intermediate synthesis

Compound 13b (2g, 11.42mmol) was dissolved in acetonitrile (40mL), and 13a (1.9g, 11.1mmol) and TEA (3.48g, 34.4mmol) were added sequentially. Stir at 50°C for 4h. Concentrate, column chromatography (PE:EA=5:1) and concentrate to obtain 2.2 g of light yellow solid. Yield: 59.9%. LCMS [M+H] ⁺ = 329.4.

In a microwave tube, dissolve compound 14b (1g, 3.07mmol) in dioxane (20mL), add 14a (0.7g, 4.6mmol), xantphos (355mg, 0.61mmol), Pd ₂ (dba) ₃ ( 290 mg, 0.32 mmol) and Cs ₂ CO ₃ (2.5 g, 7.62 mmol). The reaction system was purged with N ₂ purging, heated to 130 ℃ in a microwave reaction apparatus, the reaction 2h. Rotate to dryness, column chromatography (PE:EA=2:1) and concentrate to obtain 690 mg of light yellow oil. Yield: 56.6%. LCMS [M+H] ⁺ =398.2.

Compound 15b (650 mg, 1.64 mmol) was dissolved in tetrahydrofuran (10 mL), and LiOH.H ₂ O (0.5M, 10 mL) was added. Stir at 40°C for 12h. Add 1.0M HCl to adjust pH=4~5, extract with ethyl acetate, concentrate the organic phase, separate by reversed-phase C-18 column chromatography (formic acid), and freeze-dry to obtain 250 mg of white solid. Yield: 41.4%. LCMS [M+H] ⁺ = 370.2.

Compound 16b (100mg, 0.27mmol), HATU (113mg, 0.3mmol), DIPEA (100mg, 0.78mmol) was dissolved in DMF (4mL), stirred at 40°C for 1h, and then added 16a (75mg, 0.27mmol), reaction system Stir at 40°C for 3h. Dilute with water, extract with ethyl acetate, wash the organic phase with saturated brine, dry, filter, and concentrate. Reversed phase C-18 column chromatography (formic acid) was separated and lyophilized to obtain 105 mg of pale yellow solid. Yield: 61.7%. LCMS [M+H] ⁺ =630.2.

Compound 17b (105 mg, 0.17 mmol) was dissolved in HOAc (4 mL). The reaction system was stirred at 70°C for 2h. Concentrated, separated by reversed-phase C-18 column chromatography (formic acid), and lyophilized to obtain 65 mg of yellow solid. Yield: 63.7%. LCMS [M+H] ⁺ = 612.2.

(2) Synthesis of target compound

Compound 18b (30 mg, 0.05 mmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (2 mL) was added. The reaction system was stirred at room temperature for 1 h. Concentrated, separated by reversed-phase C-18 column chromatography (formic acid), and lyophilized to obtain 6 mg of white solid. Yield: 22.1%.

¹ H NMR (400MHz, MeOD) δ 8.23 (d, J = 1.0 Hz, 1H), 7.94 (dd, J = 8.5, 1.5 Hz, 1H), 7.62 (d, J = 8.5 Hz, 1H), 7.59 ( d,J=7.6Hz,1H), 7.54–7.47(m,2H), 7.39(t,J=8.0Hz,1H), 6.25(d,J=8.1Hz,1H), 6.06(d,J=7.8 Hz, 1H), 5.40 (s, 2H), 5.15 (dd, J = 7.3, 2.5 Hz, 1H), 4.70-4.53 (m, 2H), 4.48 (dd, J = 15.6, 2.6 Hz, 1H), 4.41 –4.30(m,1H),4.21(d,J=12.9Hz,2H), 2.94(dd,J=7.3,1.5Hz,2H), 2.81–2.68(m,3H),2.52–2.36(m,1H ), 2.30–2.17 (m, 1H), 1.72 (d, J = 11.1Hz, 2H), 1.30-1.26 (m, 4H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₁ H ₃₁ FN ₅ O ₄ 556.61, found 556.23.

Compound 6T

(1) Intermediate synthesis

In a microwave tube, dissolve compound 14b (0.8g, 2,45mmol) in dioxane (20mL), add 19a (450mg, 3.13mmol), xantphos (290mg, 0.5mmol), Pd ₂ (dba) ₃ (230 mg, 0.25 mmol) and Cs ₂ CO ₃ (2.0 g, 6.1 mmol). The reaction system was purged with N ₂ purging, heated to 130 ℃ in a microwave reaction apparatus, the reaction 2h. Rotate to dryness, column chromatography (PE:EA=2:1) and concentrate to obtain 510 mg of light yellow oil. Yield: 53.3%. LCMS [M+H] ⁺ = 391.2.

Compound 19b (510 mg, 1.31 mmol) was dissolved in tetrahydrofuran (8 mL), and LiOH.H2O aqueous solution (0.5M, 8 mL) was added. Stir at 40°C for 12h. Add 1.0M HCl to adjust pH=4~5, extract with ethyl acetate, concentrate the organic phase, separate by reverse phase C-18 column chromatography (formic acid), and freeze-dry to obtain 210 mg of white solid. Yield: 44.4%. LCMS [M+H] ⁺ = 363.2.

Dissolve compound 20b (100mg, 0.28mmol), HATU (115mg, 0.30mmol), DIPEA (108mg, 0.84mmol) in DMF (4mL), stir at 40℃ for 1h, then add 16a (78mg, 0.28mmol), 40℃ Stir for 3h. It was diluted with water, extracted with ethyl acetate, the organic phase was washed with brine, dried, filtered, concentrated, separated by reverse-phase C-18 column chromatography (formic acid), and lyophilized to obtain 92 mg of light yellow solid. Yield: 53.5%. LCMS [M+H] ⁺ =623.2.

Compound 21b (92 mg, 0.15 mmol) was dissolved in HOAc (3 mL). The reaction system was stirred at 70°C for 2h. Concentrated, separated by reversed-phase C-18 column chromatography (formic acid), and lyophilized to obtain 55 mg of white solid. Yield: 61.8%. LCMS [M+H] ⁺ = 605.3.

Compound 22b (55 mg, 0.05 mmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (2 mL) was added. The reaction system was stirred at room temperature for 1 h. Concentration, reverse phase C-18 column chromatography (formic acid) to obtain 33 mg of white solid. Yield: 66.1%. LCMS [M+H] ⁺ = 549.2.

¹ H NMR (400MHz, MeOD) δ 8.24 (d, J = 1.0 Hz, 1H), 7.94 (dd, J = 8.5, 1.5 Hz, 1H), 7.62 (d, J = 8.4 Hz, 1H), 7.51- 7.41(m,1H),7.37(t,J=8.0Hz,1H), 6.90(ddt,J=8.4,6.2,2.9Hz,2H), 6.24(d,J=8.1Hz,1H),6.01(d ,J=7.8Hz,1H), 5.29(s,2H), 5.15(dd,J=7.3,2.5Hz,1H), 4.68–4.54(m,2H), 4.49(dd,J=15.6,2.6Hz, 1H), 4.37 (dt, J = 9.2, 5.9 Hz, 1H), 4.28 (d, J = 13.1 Hz, 2H), 2.96 (dd, J = 7.3, 1.9 Hz, 2H), 2.86–2.69 (m, 3H ), 2.51–2.40(m,1H), 2.26(ddd,J=11.3,7.6,3.7Hz,1H),1.76(d,J=11.5Hz,2H),1.33(ddd,J=24.8,12.4,3.9 Hz, 2H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₀ H ₃₁ F ₂ N ₄ O ₄ 549.59, found 549.23.

Compound 7T

(1) Intermediate synthesis

In a microwave tube, compound 23b (500 mg, 3.4 mmol) was dissolved in NMP (10 mL), and 14a (513 mg, 3.4 mmol) and Cs ₂ CO ₃ (1.4 g, 4.26 mmol) were sequentially added. Heat to 120°C in a microwave instrument and react for 2h. Dilute with water, extract with ethyl acetate, wash the organic phase with brine, dry, filter, concentrate, column chromatography (PE:EA=5:1) and concentrate to obtain 603 mg of light yellow oil. Yield: 67.6%. LCMS [M+H] ⁺ = 266.3.

Compound 24b (600mg, 2.28mmol) was dissolved in dioxane (10mL) and H ₂ O (2mL), and 7a (705mg, 2.28mmol), Pd(dppf)Cl ₂ (167mg, 0.228mmol) and K ₂ CO ₃ (623 mg, 4.51 mmol). Under N ₂ protection, heat to 110°C and stir for 5 hours. Concentrate, column chromatography (PE:EA=5:1) and concentrate to obtain 560 mg of light brown oil. Yield: 59.9%. LCMS [M+H] ⁺ = 411.2.

Compound 25b (560 mg, 1.37 mmol) was dissolved in ethyl acetate (15 mL), 50% Pd/C (300 mg) was added under N ₂ protection, and stirred at room temperature for 2 h under H ₂ atmosphere. The solid was filtered off with celite, washed with ethyl acetate, the filtrate was concentrated, column chromatography (PE:EA=3:1) and then concentrated to obtain 410 mg of light yellow oil. Yield: 73.2%. LCMS [M+H] ⁺ = 413.2.

Compound 26b (410 mg, 0.996 mmol) was dissolved in dichloromethane (6 mL), trifluoroacetic acid (2 mL) was added, and the mixture was stirred at room temperature for 1 h. Concentrate, add ethyl acetate and saturated sodium bicarbonate aqueous solution, separate the layers, extract the aqueous phase with ethyl acetate, wash with saturated brine, dry, filter, and concentrate to obtain 200 mg of waxy solid. Yield: 64.1%. LCMS [M+H] ⁺ = 313.2.

Compound 27b (60 mg, 0.19 mmol) and 5a (70 mg, 0.21 mmol) were dissolved in acetonitrile (5 mL), and K ₂ CO ₃ (80 mg, 0.58 mmol) and KI (3.21 mg, 0.019 mmol) were added. The reaction system was stirred at 50°C for 2h. Dilute with water, extract with ethyl acetate, concentrate the organic phase, column chromatography (PE:EA=1:2) and concentrate to obtain 80 mg of light yellow solid. Yield: 68.3%. LCMS [M+H] ⁺ = 612.2.

Compound 28b (50 mg, 0.08 mmol) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (2 mL) was added. The reaction system was stirred at room temperature for 1 h. Concentration, reverse phase C-18 column chromatography (formic acid) to obtain 23 mg of white solid. Yield: 51.1%.

¹ H NMR (400MHz, MeOD) δ8.31 (d, J = 0.8 Hz, 1H), 8.08 (d, J = 13.8 Hz, 2H), 7.95 (dd, J = 8.5, 1.5 Hz, 1H), 7.67 ( dd, J = 11.6, 7.9 Hz, 2H), 7.56 (ddd, J = 9.4, 8.8, 1.4 Hz, 2H), 5.54 (s, 2H), 5.25 (dd, J = 7.3, 2.5 Hz, 1H), 4.91 –4.85(m,1H),4.71(dd,J=15.3,2.7Hz,1H),4.66-4.56(m,1H),4.46-4.44(m,1H),4.09-3.95(m,2H),3.17 –3.05(m,1H), 3.00(d,J=11.6Hz,1H), 2.83-2.75(m,2H), 2.57–2.44(m,1H), 2.44–2.26(m,2H), 1.92–1.78 (m,3H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₀ H ₃₀ FN ₆ O ₄ 557.60, found 557.23.

Compound 8T

(1) Intermediate synthesis

In a microwave tube, compound 29b (2 g, 13.6 mmol) was dissolved in NMP (25 mL), and 14a (2.05 g, 13.6 mmol) and Cs ₂ CO ₃ (4.77 g, 14.5 mmol) were sequentially added. The reaction system was heated to 120°C in a microwave oven and reacted for 2 hours. Dilute with water, extract with ethyl acetate, wash the organic phase with brine, dry, filter, concentrate, column chromatography (PE:EA=5:1) and concentrate to obtain 2.4 g of light yellow oil. Yield: 67.0%. LCMS [M+H] ⁺ = 266.3.

Compound 30b (2g, 7.6mmol) was dissolved in dioxane (25mL) and H ₂ O (5mL), and 7a (2.5g, 8.1mmol), Pd(dppf)Cl ₂ (555mg, 0.759mmol) were added sequentially And K ₂ CO ₃ (2.1 g, 15.2 mmol). The reaction system was heated to 110°C under the protection of N ₂ and stirred for 5 hours. Concentrate, column chromatography (PE:EA=5:1) and then concentrate to obtain 1.8 g of light brown oil. Yield: 57.7%. LCMS [M+H] ⁺ = 411.3.

Compound 31b (1.8 g, 4.4 mmol) was dissolved in ethyl acetate (60 mL), 50% Pd/C (1.2 g) was added under N ₂ protection, and the reaction system was stirred at room temperature for 6 h under H ₂ atmosphere. The solid was filtered off with celite, washed with ethyl acetate, the filtrate was concentrated, column chromatography (PE:EA=3:1) and then concentrated to obtain 1.4 g of light yellow oil. Yield: 77.8%. LCMS [M+H] ⁺ = 413.3.

Compound 32b (1.4 g, 3.4 mmol) was dissolved in dichloromethane (20 mL), trifluoroacetic acid (15 mL) was added, and the reaction system was stirred at room temperature for 1 h. Concentrate, add ethyl acetate and saturated sodium bicarbonate aqueous solution, separate the layers, extract the aqueous phase with ethyl acetate, wash the organic phase with brine, dry, filter, and concentrate to obtain 690 mg of light yellow solid. Yield: 65.1%. LCMS [M+H] ⁺ = 313.1.

Compound 33b (200 mg, 0.64 mmol) and 5a (235 mg, 0.70 mmol) were dissolved in acetonitrile (8 mL), and K ₂ CO ₃ (220 mg, 1.59 mmol) and KI (10.6 mg, 0.064 mmol) were added. The reaction system was stirred at 50°C for 3h. Dilute with water, extract with ethyl acetate, concentrate the organic phase, column chromatography (PE:EA=1:2) and concentrate to obtain 250 mg of light yellow solid. Yield: 63.9%. LCMS [M+H] ⁺ = 612.3.

(2) Synthesis of target compound

Compound 34b (150 mg, 0.245 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (5 mL) was added. The reaction system was stirred at room temperature for 1 h. Concentrated, separated by reversed-phase C-18 column chromatography (formic acid), and lyophilized to obtain 70 mg of white solid. Yield: 51.0%.

¹ H NMR (400MHz, MeOD) δ 8.39 (d, J = 5.9 Hz, 1H), 8.31 (d, J = 1.0 Hz, 1H), 7.95 (dd, J = 8.5, 1.5 Hz, 1H), 7.71- 7.63(m,2H),7.57(ddd,J=9.4,8.8,1.4Hz,2H), 6.77(d,J=5.9Hz,1H), 5.58(s,2H), 5.24(dd,J=7.4, 2.5Hz, 1H), 4.88-4.86 (m, 1H), 4.70 (dd, J = 15.4, 2.6 Hz, 1H), 4.62 (td, J = 7.9, 6.0 Hz, 1H), 4.45 (dt, J = 9.2 ,5.9Hz,1H),4.15-4.02(m,2H),3.16(d,J=11.6Hz,1H),3.06(d,J=11.4Hz,1H),2.91-2.71(m,2H),2.58 –2.39(m,3H),2.05–1.85(m,4H)

Compound 9T

Compound 8T (22 mg, 0.040 mmol) was dissolved in DMF (3.0 mL), HATU (16.6 mg, 0.043 mmol), DIPEA (15 mg, 0.116 mmol) were added. Stir at room temperature for 10 min, then add NH _{3 /} DMF (1M, 0.12 mL, 0.12 mmol), and stir at room temperature for 1 h. Dilute with water, extract with ethyl acetate, wash the organic phase with brine, dry, filter, concentrate, and separate by reverse phase C-18 column chromatography (formic acid) to obtain 12 mg of white solid. Yield: 54.8%.

¹ H NMR(400MHz,DMSO)δ8.49(d,J=5.8Hz,1H), 8.20(s,1H), 7.92(d,J=10.0Hz,2H), 7.80-7.68(m,3H), 7.60(d,J=8.4Hz,1H),7.30(s,1H),6.86(d,J=5.8Hz,1H),5.55(s,2H),5.13(dd,J=7.1,3.0Hz,1H ), 4.79 (dd, J = 15.1, 7.2 Hz, 1H), 4.64 (dd, J = 15.1, 3.2 Hz, 1H), 4.56-4.46 (m, 1H), 4.42 (dt, J = 9.0, 6.0 Hz, 1H), 3.95 (d, J = 13.5Hz, 1H), 3.78 (d, J = 13.4Hz, 1H), 3.31 (s, 3H), 2.96 (d, J = 11.2Hz, 1H), 2.84 (d, J = 11.2Hz, 1H), 2.79–2.64 (m, 2H), 2.49–2.39 (m, 1H), 2.32–2.10 (m, 2H), 1.89 (t, J = 11.6 Hz, 2H), 1.84–1.61 (m,2H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₀ H ₃₁ FN ₇ O ₃ 556.61, found 556.27.

Compound 10T

Compound 8T (35 mg, 0.063 mmol) was dissolved in DMF (3.0 mL), HATU (26 mg, 0.068 mmol), DIPEA (25 mg, 0.194 mmol) were added. Stir at room temperature for 10 min, then add tert-butyl glycine (16.5 mg, 0.126 mmol), and stir at room temperature for 1 h. Dilute with water, extract with ethyl acetate, wash the organic phase with brine, dry, filter, concentrate, and separate by reverse phase C-18 column chromatography (formic acid) to obtain 27 mg of white solid. Yield: 64.3%. LCMS [M+H] ⁺ = 669.3.

Compound 35b (27 mg, 0.04 mmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (2 mL) was added. The reaction system was stirred at room temperature for 1 h. Concentrated to obtain 28 mg of white solid. Reverse phase C-18 column chromatography (formic acid) to obtain 16 mg of white solid. Yield: 64.8%.

¹ H NMR(400MHz,DMSO)δ12.25(s,1H), 8.78(t,J=5.8Hz,1H), 8.50(d,J=5.8Hz,1H), 8.20(s,1H), 7.92( d, J = 10.2 Hz, 1H), 7.74 (dd, J = 5.7, 3.2 Hz, 2H), 7.65 (d, J = 8.4 Hz, 1H), 6.87 (d, J = 5.7 Hz, 1H), 5.55 ( s, 2H), 5.14 (dd, J = 7.1, 2.9 Hz, 1H), 4.80 (dd, J = 15.2, 7.2 Hz, 1H), 4.65 (dd, J = 15.1, 3.0 Hz, 1H), 4.56–4.47 (m,1H),4.47–4.37(m,1H),3.97(d,J=5.7Hz,2H), 3.79(d,J=5.0Hz,1H),3.31(s,4H),3.00(s, 2H), 2.79--2.63(m, 2H), 2.26(s, 2H), 1.92(s, 2H), 1.78(s, 2H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₂ H ₃₃ FN ₇ O ₅ 614.65, found 614.25.

Compound 11T

Compounds 33b (35 mg, 0.112 mmol) and 33a (43 mg, 0.123 mmol) were dissolved in acetonitrile (4 mL), and K ₂ CO ₃ (38 mg, 0.275 mmol) and KI (1.9 mg, 0.011 mmol) were added. The reaction system was stirred at 50°C for 2h. Dilute with water, extract with ethyl acetate, concentrate the organic phase, column chromatography (PE:EA=1:2) and concentrate to obtain 36 mg of light yellow oil. Yield: 51.4%. LCMS [M+H] ⁺ = 626.3.

Compound 36b (35 mg, 0.056 mmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (2 mL) was added. The reaction system was stirred at room temperature for 1 h. Concentration, reverse phase C-18 column chromatography (formic acid) to obtain 19 mg of white solid. Yield: 59.7%.

¹ H NMR(400MHz,DMSO)δ12.65(s,1H), 8.49(d,J=5.8Hz,1H), 8.23(d,J=1.0Hz,1H), 7.92(d,J=10.2Hz, 1H), 7.80(dd,J=8.4,1.5Hz,1H),7.77–7.67(m,2H), 7.64(d,J=8.4Hz,1H), 6.86(d,J=5.8Hz,1H), 5.54 (s, 2H), 4.57 (dd, J = 14.8, 3.1 Hz, 1H), 4.48 (dd, J = 14.8, 8.1 Hz, 1H), 4.26 (dd, J = 7.6, 2.9 Hz, 1H), 3.99 (d,J=13.4Hz,1H), 3.82(dd,J=14.8,6.9Hz,1H), 3.73(d,J=13.3Hz,1H), 3.64(td,J=7.7,6.2Hz,1H) , 2.98 (d, J = 10.0 Hz, 1H), 2.82 (d, J = 9.9 Hz, 1H), 2.73 (t, J = 11.4 Hz, 1H), 2.23 (d, J = 37.2 Hz, 2H), 2.13 –1.96(m,1H),1.93-1.77(m,5H),1.75–1.57(m,2H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₁ H ₃₂ FN ₆ O ₄ 571.63, found 571.24.

Compound 12T

(1) Intermediate synthesis

Dissolve trimethyl sulfoxide iodide (22g, 100mmol) in tert-butanol (250mL), add potassium tert-butoxide (11.2g, 100mmol) under N ₂ protection, stir for 30min and add 1c (8.1g, 50mmol) ) The reaction system was stirred at 60°C for 6 hours. Dilute with water, extract, and column chromatography to obtain 7.3g colorless liquid, yield: 90%.

Compound 2c (4g, 22.47mmol) was dissolved in ethyl acetate (60mL), 10% Pd/C (1.2g) was added under N ₂ protection, and the reaction system was stirred at room temperature for 6 h under H ₂ atmosphere. The solid was filtered off with celite, washed with ethyl acetate, the filtrate was concentrated, column chromatography (PE:EA=20:1) and then concentrated to obtain 1.7 g of light yellow oil. Yield: 86.3%.

Compound 3c (1.7g, 19.3mmol) was dissolved in tetrahydrofuran (50mL), and TEA (5.85g, 57.9mmo) and Ms ₂ O (3.7g, 21.3mmol) were added in sequence at 0℃. Stir at room temperature for 3h. Concentrate, column chromatography (PE:EA=20:1) and concentrate to obtain 1.8 g of pale yellow oil. Yield: 56.3%.

Compound 5c (1 g, 4.5 mmol) was dissolved in DMF (15 mL), and 4c (0.82 g, 4.9 mmol), Cs ₂ CO ₃ (1.8 g, 5.5 mmol) and KI (75 mg, 0.45 mmol) were sequentially added. Stir at 60°C for 17h. Dilute with water, extract with ethyl acetate, wash with saturated brine, dry, filter, concentrate, column chromatography (PE:EA=10:1) and concentrate to obtain 910 mg of light yellow oil. Yield: 68.9%.

In a microwave tube, dissolve compound 6c (1.5g, 5mmol) in dioxane (10mL), add phenyl formate (1.32g, 10mmol), xantphos (620mg, 1mmol), Pd(AcO) ₂ (390mg , 0.5mmo) and triethylamine (1mL). The reaction system was purged with N ₂ purging, heated to 130 ℃ in a microwave reaction apparatus, the reaction 2h. Rotate to dryness, column chromatography (PE:EA=4:1) and concentrate to obtain 1.5 g of light yellow solid. Yield: 70.1%.

¹ H NMR (400MHz, CDCl ₃ ): δ9.58(s,1H), 8.01(d,J=7.0Hz,1H), 7.88(s,1H), 7.80(d,J=7.0Hz,1H), 7.66-7.40(m,3H),7.38-7.31(m,2H),5.11-5.03(m,1H)4.71-4.54(m,2H),4.51-4.44(m,1H),4.37-4.29(m, 1H),2.3-2.64(m,1H),2.39-2.29(m,1H)

In a microwave tube, dissolve compound 1b (60mg, 0.202mmol) in dioxane (3mL), add 14a (45mg, 0.298mmol), xantphos (24mg, 0.041mmol), Pd ₂ (dba) ₃ (19mg , 0.021 mmol) and Cs ₂ CO ₃ (166 mg, 0.506 mmol). The reaction system was purged with N ₂ purging, heated to 130 ℃ in a microwave reaction apparatus, the reaction 2h. It was spin-dried, column chromatography (PE:EA=2:1) and concentrated to obtain 50 mg of pale yellow oil. Yield: 60.1%. LCMS [M+H] ⁺ = 413.2.

Compound 37b (100 mg, 0.243 mmol) was dissolved in dichloromethane (4 mL), and trifluoroacetic acid (4 mL) was added. The reaction system was stirred at room temperature for 1 h. Concentrate, column chromatography (PE:EA=1:2) and concentrate to obtain 47 mg of colorless oil. Yield: 62.1%. LCMS [M+H] ⁺ = 413.1.

Compound 38b (45 mg, 0.15 mmol) and compound 7c (78 mg, 0.286 mmol) were dissolved in dichloroethane (5 mL), and the reaction system was stirred at room temperature for 20 min. Then sodium acetate borohydride (36 mg, 0.573 mmol) was added. The reaction system was heated to 30°C and stirred for 24h. Add 0.1M HCl to adjust pH=4～5, and stir for 5min. Extraction with dichloromethane, concentration, column chromatography (PE:EA=1:2) and concentration to obtain 17 mg of colorless oil. Yield: 20.7%. LCMS [M+H] ⁺ = 632.3.

Compound 39b (15 mg, 0.026 mmol) was dissolved in tetrahydrofuran (2 mL), and LiOH.H ₂ O aqueous solution (0.1 M, 1 mL) was added. Stir at 40°C for 12h. Add 0.2M HCl to adjust pH=4～5, extract with ethyl acetate, concentrate, reverse phase C-18 column chromatography (formic acid), freeze-dry to obtain 8mg of white solid. Yield: 54.8%.

¹ H NMR(400MHz,DMSO)δ12.37(s,1H), 8.17(s,1H), 7.88(dd,J=10.0,1.1Hz,1H), 7.70(dd,J=7.9,1.3Hz,1H ), 7.66(d,J=7.3Hz,1H), 7.63(dd,J=8.2,1.4Hz,1H),7.55(d,J=8.3Hz,1H),7.48(t,J=8.0Hz,1H ), 6.50 (s, 1H), 6.34 (d, J = 8.2 Hz, 1H), 6.13 (d, J = 7.8 Hz, 1H), 5.40 (s, 2H), 5.05 (dd, J = 7.2, 2.6 Hz ,1H), 4.72(dd,J=15.3,7.1Hz,1H),4.56(dd,J=15.4,2.5Hz,1H), 4.47(dd,J=10.5,5.0Hz,1H), 4.36(dt, J = 9.0, 5.8 Hz, 1H), 3.84 (d, J = 13.7 Hz, 1H), 3.66 (d, J = 13.7 Hz, 1H), 3.42 (s, 3H), 3.31 (s, 3H), 2.77- 2.60 (m, 2H), 2.47-2.43 (m, 3H), 2.41-2.33 (m, 1H)

Compound 13T

Compound 36 (45 mg, 0.15 mmol) and compound 7c (78 mg, 0.286 mmol) were dissolved in dichloroethane (5 mL), and the reaction system was stirred at room temperature for 20 min. Sodium acetate borohydride (36 mg, 0.573 mmol) was added. The reaction system was heated to 30°C and stirred for 24h. Add 0.1M HCl to adjust pH=4-5, stir for 5min, extract with dichloromethane, concentrate, column chromatography (PE:EA=1:2) and concentrate to obtain 27mg of colorless oil. Yield: 33%. LCMS [M+H] ⁺ = 631.3.

Compound 40b (27 mg, 0.042 mmol) was dissolved in tetrahydrofuran (2 mL), and LiOH.H ₂ O aqueous solution (0.1 M, 1 mL) was added. Stir at 40°C for 12h. Add 0.2M HCl to adjust pH=4~5, extract with ethyl acetate, concentrate, reverse phase C-18 column chromatography (formic acid), and freeze-dry to obtain 8mg of white solid. Yield: 38%. LCMS [M+H] ⁺ = 555.2.

¹ H NMR (400MHz, DMSO) δ 12.42 (s, 1H), 8.17 (s, 1H), 7.89 (d, J = 10.1 Hz, 1H), 7.70 (d, J = 3.0 Hz, 2H), 7.64 ( dd, J = 16.9, 8.8 Hz, 2H), 7.54 (d, J = 8.3 Hz, 1H), 6.89 (d, J = 7.3 Hz, 1H), 6.73 (d, J = 8.2 Hz, 1H), 6.48 ( s, 1H), 5.48 (s, 2H), 5.12–5.01 (m, 1H), 4.73 (dd, J = 15.3, 7.0 Hz, 1H), 4.58 (dd, J = 15.2, 2.3 Hz, 1H), 4.47 (dd, J = 13.6, 7.6 Hz, 1H), 4.36 (dt, J = 9.0, 5.8 Hz, 1H), 3.82 (d, J = 13.7 Hz, 1H), 3.64 (d, J = 13.6 Hz, 1H) ,3.31(s,3H),2.99(d,J=10.8Hz,1H), 2.88(d,J=10.9Hz,1H), 2.75–2.65(m,1H),2.62-2.56(m,1H), 2.48–2.35(m,1H),2.15-2.04(m,2H),1.85-1.58(m,4H)

HRMS(ESI)m/z[M+H] ⁺ calcd for C ₃₂ H ₃₂ FN ₄ O ₄ 555.62, found 555.24.

Compound 14T

Compound 33b (62 mg, 0.20 mmol) and compound 7c (78 mg, 0.286 mmol) were dissolved in dichloroethane (5 mL) and stirred at room temperature for 20 min. Then sodium acetate borohydride (36 mg, 0.573 mmol) was added. Heat to 30°C and stir for 24h. Add 0.1M HCl to adjust pH=4～5, and stir for 5min. Extraction with dichloromethane, concentration, column chromatography (PE:EA=1:2) and concentration to obtain 36 mg of colorless oil. Yield: 30.7%. LCMS [M+H] ⁺ = 632.3.

Compound 41b (26.5 mg, 0.042 mmol) was dissolved in tetrahydrofuran (2 mL), and LiOH.H ₂ O aqueous solution (0.1M, 1 mL) was added. Stir at 40°C for 12h. Add 0.2M HCl to adjust pH=4~5, extract with ethyl acetate, concentrate, reverse phase C-18 column chromatography (formic acid), and freeze-dry to obtain 6.6 mg of white solid. Yield: 28.3%.

¹ H NMR(400MHz,DMSO)δ8.49(d,J=5.8Hz,1H), 8.16(s,1H), 7.92(d,J=10.1Hz,1H), 7.77–7.70(m,2H), 7.63(d,J=8.2Hz,1H), 7.52(d,J=8.2Hz,1H), 6.86(d,J=5.8Hz,1H), 6.46(s,1H), 5.55(s,2H), 5.06 (dt, J = 6.9, 4.2 Hz, 1H), 4.72 (dd, J = 15.3, 7.0 Hz, 1H), 4.57 (dd, J = 15.3, 2.5 Hz, 1H), 4.47 (dt, J = 13.6, 7.0Hz, 1H), 4.36 (dt, J = 9.0, 5.9 Hz, 1H), 3.82 (d, J = 13.7 Hz, 1H), 3.63 (d, J = 13.7 Hz, 1H), 2.96 (d, J = 11.1Hz, 1H), 2.86 (d, J = 10.9Hz, 1H), 2.78–2.59 (m, 2H), 2.47–2.36 (m, 1H), 2.19-2.07 (m, 2H), 2.05–1.94 (m ,1H),1.89(t,J=11.1Hz,2H),1.83-1.66(m,2H)

2. Evaluation of biological activity

The present invention provides a new compound that modulates glucagon-like peptide-1 (GLP-1) receptor, its synthesis method, and its pharmaceutical use. The provided new compound has obvious agonistic effect on GLP-1 receptor, and its EC ₅₀ value reaches the nm level. Moreover, the active compound has no obvious binding effect on hERG, indicating that it has a lower risk of cardiotoxicity. The compound of the present invention can be used alone as a diabetes treatment drug, or combined with a polypeptide GLP-1 receptor agonist such as liraglutide, or combined with drugs for the treatment of type II diabetes with other mechanisms.

1. Evaluation of GLP-1 receptor agonistic activity

The agonistic activity of the compound on the GLP-1 receptor was characterized by the changes in the cAMP content of HEK293 cells that highly expressed the GLP-1 receptor. Use the cAMP detection kit (Cisbio Cat#62AM4PEJ). For the test principle, please refer to the Cisbio CAMP-GS DYNAMIC KIT manual.

Prepare the compound: use the Bravo liquid processing platform, DMSO as the solvent, the starting concentration of the test compound from 100 μm, and the starting concentration of the control compound from 0.5 μm, respectively, with 4-fold gradient dilutions, each with 10 data points. Transfer 100 nL of compound to an OptiPlate-384-well plate and mix at 2000 rpm for 60 sec.

Prepare the cell suspension: quickly thaw the cells in a 37°C water bath, transfer the cell suspension to 10mL HBSS, and place it in a 15mL conical tube, centrifuge at 1000rpm for 5min at room temperature; aspirate the supernatant and flick to loosen the cell clumps. Resuspend the cells in 10mL HBSS; calculate the cell concentration and determine the cell viability; resuspend the cells in buffer at a concentration of 2.0x10 ⁵ /mL.

HTRF cAMP determination of agonist: Use an electronic multi-channel pipette to add 10 μL of cell suspension to the test plate, mix at 1000 rpm for 60 sec, incubate at room temperature for 30 minutes, add 5 μL of detection reagent to each well; cover with a sealing film, Incubate at room temperature for 60 minutes; remove the sealing film, use EnVision (PerkinElmer) to read, calculate the EC ₅₀ value, where +++++ means between 1-2nm, ++++ means between 2-5nm, +++ means Between 5-20nm, ++ means 20-1000nm, + means 1-10μM. As shown in Table 1.

Table 1 Exemplary compounds on GLP-1 agonistic activity EC ₅₀ value (nm)

化合物编号Compound number	活性(EC ₅₀) Activity (EC ₅₀ )	化合物编号Compound number	活性(EC ₅₀) Activity (EC ₅₀ )
AA	++	1T1T	++++++++
BB	++	2T2T	++
CC	++	3T3T	++++++++
DD	++	4T4T	++++
EE	++	5T5T	++++++++
FF	++	6T6T	++++++
GG	++	7T7T	++++++++
HH	++	8T8T	++++++++++
II	++	9T9T	++
JJ	++	10T10T	++
KK	++++	11T11T	++++++
LL	++++	12T12T	++
MM	++++	13T13T	++
NN	++++	14T14T	++
OO	++	To	To

2. Evaluation of hERG binding effect

Human Ether-a-go-go Related Gene (hERG gene) encodes myocardial delayed rectifier potassium channel current. Studies have found that some drugs have an inhibitory effect on hERG potassium channels, which leads to prolonged cardiac QT interval and induces arrhythmia. By detecting the binding capacity of the compound to hERG, the potential risk of cardiotoxicity of the compound can be initially assessed.

By detecting the competitive binding of the compound and [ ³ H]Dofetilide to hERG on CHO cells that highly express the hERG gene, the binding effect of the compound to hERG was evaluated.

The starting concentration of the test compound was 2 mM, the starting concentration of the control compound was 0.2 mM, and DMSO was the solvent, respectively, with 4-fold gradient dilutions, each taking 8 data points. Transfer 1 μL of the compound to the test plate, add 100 μL of hERG/CHO cells, add 100 μL of [ ³ H]Dofetilide, seal the plate, and incubate at room temperature for 1 h. Use 50μL of 0.5% BSA for each well to soak for at least 0.5h at room temperature. After the binding test was completed, the reaction mixture was filtered, washed, and the test plate was dried at 50°C for 1 hour. Subsequently, seal the bottom of the filter plate well, add 50 μL Perkin Elmer Microscint 20 cocktail, and then seal the top of the filter plate well. Use Perkin Elmer MicroBeta2 reader to count ³ H, calculate IC ₅₀ value, as shown in Table 2.

Table 2 IC ₅₀ value (nm) of the binding effect of exemplary compounds on hERG

化合物编号Compound number	活性(IC ₅₀,nm) Activity (IC ₅₀ ,nm)
3T3T	59225922
5T5T	>100000>100000
7T7T	10621062
8T8T	4994549945
dofetilidedofetilide	3.233.23

Claims

A small molecule GLP-1 receptor modulator with the following structure, and its pharmaceutically acceptable salt, ester, stereoisomer, ether, prodrug or solvate:

among them,

A is a substituted or unsubstituted aromatic ring, aromatic heterocyclic ring, C 2 -C 8 alkenyl group, the substitution is one or more than two, and the substituents may be the same or different;

L is a link chain, selected from -(CH 2 ) m -, -CONR 7 -, -SO 2 NR 7 -, -NR 7 CO-, -NR 7 SO 2 -, -(CH 2 ) m O-,- O(CH 2 ) m -, -(CH 2 ) m S-, -S(CH 2 ) m -, -(CH 2 ) m NR 7 -, -NR 7 (CH 2 ) m -; said R 7 Selected from hydrogen, C 1-8 straight or branched chain alkyl or C 3-8 cycloalkyl, and the alkyl is optionally further substituted with one or more halogen, cyano, amino or hydroxy;

m, n are independently selected from 1, 2 or 3;

Preferably, said L is selected from -CH 2 O-, -CONH-, -CH 2 NH- or -CH 2 N(CH 3 )-;

B is a combined structure of two or more saturated or unsaturated carbocyclic rings, and the carbocyclic ring optionally contains one or more heteroatoms;

Z may be the same or different, selected from CH, CR 9 or N; said R 9 is halogen, selected from F, Cl, Br, I;

R 4 is selected from C 1-4 alkyl or C 1-4 alkoxy. The C 1-4 alkyl or C 1-4 alkoxy is optionally further substituted by one or more halogen or cyano groups, The hydroxyl group is substituted or further substituted by a saturated or unsaturated carbocyclic ring or a heteroatom-containing carbocyclic ring; the saturated or unsaturated carbocyclic ring or a heteroatom-containing carbocyclic ring is optionally further substituted by one or more halogens , Cyano, amino or hydroxyl; preferably, the saturated or unsaturated carbocyclic ring or heteroatom-containing carbocyclic ring is a four-membered ring, a five-membered ring, or a six-membered ring; more preferably, the saturated or The unsaturated carbocyclic ring or the heteroatom-containing carbocyclic ring is a butylene oxide substituent, a pentylene oxide substituent or an oxazole substituent; preferably, R 4 is selected from the group consisting of a butylene oxide substituent, pentylene oxide Methyl, ethyl or propyl substituted with alkyl substituents or oxazole substituents; more preferably, R 4 is selected from methyl substituted with butylene oxide substituents, epoxypentane substituents or oxazole substituents ；

R 5 is one or more than two, selected from hydrogen, hydroxy, amino, halogen, cyano, carboxy, tetrazolyl, amide; and the hydroxy, amino, carboxy, tetrazolyl or amide group is optional Is further substituted by one or more hydroxy, hydroxyalkyl, carboxyalkyl, aminoalkyl, and the alkyl is selected from C 1-4 alkyl; preferably, R 5 is selected from carboxy, amide, or A hydroxy, hydroxyalkyl, carboxyalkyl, or aminoalkyl substituted amide group; the amide group substituted by a hydroxy, hydroxyalkyl, carboxyalkyl, or aminoalkyl group is, for example, hydroxy, hydroxymethyl, Amido groups substituted by hydroxyethyl, carboxymethyl, and carboxyethyl;

Or R 5 may form a ring system structure, the ring system structure may further contain one or more nitrogen atoms and/or one or more intra-ring carbonyl groups, and the ring system structure is optionally surrounded by one or more R 6 substitution, said R 6 is selected from hydrogen, hydroxy, amino, halogen, cyano, carboxy or tetrazolyl, amide; and said hydroxy, amino, carboxy, tetrazolyl or amide optionally further Is substituted by one or two or more hydroxy, hydroxyalkyl, carboxyalkyl, aminoalkyl, the alkyl is selected from C 1-4 alkyl; preferably, R5 is connected to
Form a ring structure
or
The ring system structure is optionally substituted with one or more R 6 .
The GLP-1 receptor modulator according to claim 1, its pharmaceutically acceptable salt, ester, stereoisomer, ether, prodrug or solvate, characterized in that:

A is an aromatic ring or aromatic heterocyclic ring substituted by one or more R 1 or a C 2 -C 8 alkenyl group, R 1 is selected from hydrogen, hydroxyl, amino, halogen, cyano, C 1-4 alkyl, C 1-4 alkoxy, C 1-4 alkyl or C 1-4 alkoxy substituted by one or more halogens; preferably, A is phenyl substituted with one or more R 1
i is selected from an integer of 1-5, such as 1,2,3,4,5; preferably, i is 1 or 2, preferably, R 1 is selected from cyano, F, Cl, Br, I, more preferably , A is
The X is halogen, selected from F, Cl, Br, I; preferably, A is
Preferably, R 1 is selected from F, Cl, Br, I;

B has the following structure:

Among them, Y may be the same or different, and are selected from CH or N; R 2 , R 3 , and R 8 are one or more than two, and are selected from hydrogen, hydroxyl, amino, halogen, cyano, C 1-4 alkyl , C 1-4 alkoxy, C 1-4 alkyl or C 1-4 alkoxy substituted by one or more halogens.
The GLP-1 receptor modulator according to claim 2, its pharmaceutically acceptable salt, ester, stereoisomer, ether, prodrug or solvate, characterized in that:

The B is selected from the following structures:
The GLP-1 receptor modulator according to claim 2, its pharmaceutically acceptable salt, ester, stereoisomer, ether, ester, prodrug or solvate, characterized in that:

The B is selected from the following structures:
The GLP-1 receptor modulator according to claim 1, wherein a pharmaceutically acceptable salt, ester, stereoisomer, ether, prodrug or solvate thereof is characterized in that it has the following structure:

Wherein, Y may be the same or different, and is selected from CH or N; (R 1 ) i represents that the phenyl group may be substituted with i same or different R 1 , and i is selected from an integer of 1-5.
The GLP-1 receptor modulator according to claim 5, its pharmaceutically acceptable salt, ester, stereoisomer, ether, prodrug or solvate, characterized in that it has the following structure:
The GLP-1 receptor modulator according to claim 5, its pharmaceutically acceptable salt, ester, stereoisomer, ether, prodrug or solvate, characterized in that it has the following structure:
The GLP-1 receptor modulator according to claim 5, its pharmaceutically acceptable salt, ester, stereoisomer, ether, prodrug or solvate, characterized in that it has the following structure:

Wherein, X is selected from -CH 2 -, -CO- or -SO 2 -, Y can be the same or different, selected from CH or N, Z can be the same or different, selected from CH, CR 9 or N;

Or have the following structure:

Among them, Z can be the same or different, and are selected from CH, CR 9 or N;

Or have the following structure:

Wherein, Y can be the same or different, selected from CH or N, R 6 substituent is one or more, selected from hydrogen, hydroxyl, amino, halogen, cyano, carboxyl, tetrazolyl, amide, and so The hydroxyl group, amino group, carboxyl group, tetrazolium group or amide group is optionally further substituted with one or more hydroxyalkyl groups and carboxyalkyl groups, and the alkyl group is selected from C 1-4 alkyl groups;

Or have the following structure:

Wherein, X is selected from -CH 2 -, -CO- or -SO 2 -, Y can be the same or different, selected from CH or N, Z can be the same or different, selected from CH, CR 9 or N;

Or have the following structure:

Wherein, X is selected from -CH 2 -, -CO- or -SO 2 -, Y can be the same or different, selected from CH or N, Z can be the same or different, selected from CH, CR 9 or N;

Or have the following structure:

Wherein, R 1 , R 2 , R 3 , R 4 , and R 5 are as defined above, Y may be the same or different, selected from CH or N, and Z may be the same or different, selected from CH, CF or N;

Or have the following structure:

Wherein, X is selected from -CH 2 -, -CO- or -SO 2 -, Y can be the same or different, selected from CH or N, Z can be the same or different, selected from CH, CR 9 or N;

Or have the following structure:

Wherein, X is selected from -CH 2 -, -CO- or -SO 2 -, Y may be the same or different, selected from CH or N, and Z may be the same or different, selected from CH, CR 9 or N.
The GLP-1 receptor modulator according to claim 8, which is a pharmaceutically acceptable salt, ester, stereoisomer, ether, ester, prodrug or solvate thereof, characterized in that it has the following structure:
The GLP-1 receptor modulator according to claim 1, its pharmaceutically acceptable salt, ester, stereoisomer, ether, prodrug or solvate, characterized in that it comprises the following compounds:
A pharmaceutical composition, characterized by comprising the GLP-1 receptor modulator according to any one of claims 1-10, and a pharmaceutically acceptable salt, ester, stereoisomer, ether, prodrug or Solvate and at least one pharmaceutically acceptable carrier.
A pharmaceutical composition, characterized by comprising the GLP-1 receptor modulator according to any one of claims 1-10, a pharmaceutically acceptable salt, ester, stereoisomer, ether, prodrug or Solvate, and a second drug, the second drug is a polypeptide GLP-1 receptor agonist or other type II diabetes treatment drugs.
The pharmaceutical composition according to claim 12, wherein the polypeptide GLP-1 receptor agonist is selected from the group consisting of exenatide, semaglutide, liraglutide, tasglutide, a Biglutide, lixisenatide or other insulin-regulating peptides; the second drug of the other type II diabetes treatment drugs is a DPPIV inhibitor, such as sitagliptin; or the second drug is sodium-glucose co-transporter SGLT1 And/or SGLT2 inhibitor; or the second drug is a biguanide hypoglycemic drug, such as metformin; or the second drug is a sulfonylurea hypoglycemic drug, such as glibenclamide, glipizide, gliclazide and / Or glimepiride; or the second drug is a thiazolidinedione hypoglycemic agent, pioglitazone and/or rosiglitazone; or the second drug is acarbose, miglitol, colesevelam and / Or bromocriptine and other diabetes treatment or adjuvant therapy drugs.
The GLP-1 receptor modulator according to any one of claims 1-10, its pharmaceutically acceptable salt, ester, stereoisomer, ether, prodrug or solvate or any one of claims 11-13 An application of the pharmaceutical composition for the preparation of a medicament for the treatment of malignant conditions of a subject that medically requires the activation, enhancement, or stimulus of GLP-1 receptors, characterized in that it is characterized in that the drug An effective amount of the modulator is administered to the subject for treatment at a frequency or duration.
The use according to claim 14, wherein the malignant condition is selected from the group consisting of type I diabetes, type II diabetes, gestational diabetes, obesity, excessive appetite, insufficient satiety or metabolic disorders.
Compounds with the following structure:

Among them, L is a link chain, selected from -(CH 2 ) m -, -CONR 7 -, -SO 2 NR 7 -, -NR 7 CO-, -NR 7 SO 2 -, -(CH 2 ) m O- , -O(CH 2 ) m -, -(CH 2 ) m S-, -S(CH 2 ) m -, -(CH 2 ) m NR 7 -, -NR 7 (CH 2 ) m -; R 7 is selected from hydrogen, C 1-8 linear or branched alkyl or C 3-8 cycloalkyl, and the alkyl is optionally further substituted with one or more halogen, cyano, amino or hydroxy ; Preferably, said L is selected from -CH 2 O-, -CONH-, -CH 2 NH- or -CH 2 N(CH 3 )-;

R 1 is selected from hydrogen, hydroxy, amino, halo, cyano, C 1-4 alkyl, C 1-4 alkoxy, halo-substituted by one or more C 1-4 alkyl or C 1-4 alkoxy Oxy; i is selected from an integer of 1-5; preferably, i is 1 or 2;

m is selected from 1, 2 or 3;

Z may be the same or different, selected from CH, CR 9 or N; said R 9 is halogen, selected from F, Cl, Br, I;

Y can be the same or different, selected from CH or N; R 2 and R 3 are one or more than two, selected from hydrogen, hydroxyl, amino, halogen, cyano, C 1-4 alkyl, C 1-4 Alkoxy, C 1-4 alkyl or C 1-4 alkoxy substituted by one or more halogens;

Y'is selected from CR 10 or NR 10 , said R 10 is selected from H, C1-C8 alkyl, C1-C4 alkyl carboxyl, C1-C4 alkyl hydroxy; preferably, said R 10 Selected from H, methyl, carboxyl, carboxymethyl, carboxyethyl.
The compound of claim 16, wherein the compound has the following structure:
The compound of claim 16, wherein the compound has the following structure: