Pan KRAS protein degradation agent as well as preparation method and application thereof

Abstract

The invention relates to a Pan KRAS protein degradation agent as well as a preparation method and application thereof. Specifically, the compound provided by the invention has a structure as shown in a formula I. The invention also discloses a preparation method of the compound and application of the compound in prevention and / or treatment of KRAS mutation related diseases.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of pharmaceutical chemistry, in particular to a Pan KRAS protein degrader, a preparation method and an application thereof. BACKGROUND

[0002] KRAS is a small GTPase, which is one of the important members of the RAS family of oncogenes. As a molecular switch, KRAS can transmit upstream signals of various tyrosine kinases to downstream pathways by cycling between inactive (GDP-bound) and active (GTP-bound) states, and plays an important role in cell proliferation, survival, migration and metabolism. KRAS mutations include many different site mutations, of which the most common are G12C, G12D and G12V mutations, as well as G12A, G12S, G12R, etc. KRAS mutations are found in a variety of malignant tumors, the most common of which include non-small cell lung cancer (G12C), colon cancer (G12D and G12V), pancreatic cancer (G12D and G12V), etc. Moreover, studies on the molecular mechanism show that the genetic mutation of KRAS makes it continuously active, mediates the activation of downstream signaling pathways, including the PI3K-AKT-mTOR and RAF-MEK-ERK pathways, and the cell will uncontrollably grow and divide, eventually leading to the occurrence, development and invasion and metastasis of cancer.

[0003] PROTAC is a heterobifunctional molecule composed of a Protein of interest (POI) ligand, a linker and an E3 ligase ligand. After entering the cell, the Protein of interest (POI) ligand in the molecule can specifically bind to the corresponding target protein, and the other end can recruit E3 ligase to form a POI-Linker-E3 ligase ternary complex, in which the E3 ligase can mediate the ubiquitination of POI by E2 ubiquitin-conjugating enzyme. The POI labeled with ubiquitin is recognized by the proteasome and degraded. This process does not require the target protein ligand to occupy the active site for a long time, and only a short-lived ternary complex is needed to transiently complete the ubiquitination of the target protein and cause degradation, which is conducive to overcoming difficult drug targets. And PROTAC can be recycled in the cell, similar to the biological mechanism of a catalyst, and only a low dose is needed to play a role, which is safer. PROTAC molecules directly degrade target proteins, overcoming the drug resistance problems of protein overexpression and mutations caused by traditional small molecules.

[0004] According to statistics, in about 20% of human tumor cells, RAS genes have mutations. Among them, KRAS is the most common mutant oncogene. At present, only 14% of cancers have marketed targeted drugs, and KRAS-driven cancers account for 11.6%, but only G12C mutations have made a breakthrough.

[0005] Pan-KRAS inhibitors do not need to develop drugs for each specific mutation, and the degradation agent modified by the PROTAC technology has greater clinical value. SUMMARY

[0006] The purpose of the present application is to provide a pan KRAS degradation compound which can effectively degrade KRAS protein and inhibit the proliferation of KRAS related cells, and can prevent and / or treat KRAS mutation related diseases.

[0007] In a first aspect, the present application provides a compound of formula I, or a pharmaceutically acceptable salt thereof, wherein

[0008]

[0009] wherein is a single bond or a double bond as valence allows;

[0010] X is selected from N or CH;

[0011] R1 is selected from C6-C18 aryl or 5-18 membered heteroaryl, which C6-C18 aryl or 5-18 membered heteroaryl is optionally substituted with one or more Ra, which Ra is independently selected from C1-C4 alkyl, C1-C4 alkoxy, halogen, C2-C4 alkynyl, hydroxyl;

[0012] R2 is selected from H, fluorine, chlorine, bromine, cyano;

[0013] R3 is selected from 5-18 membered heterocyclyl, which heterocyclyl can be monocyclic or bicyclic forming a bridged ring or a spiro ring, which heterocyclyl contains 1-3 heteroatoms, which heteroatoms are O, S and N, and which heterocyclyl is optionally substituted with one or more R b , which R b is independently selected from C1-C4 alkyl, C1-C4 alkoxy, carbonyl;

[0014] Each A is independently O or N;

[0015] Each n is independently 0, 1 or 2;

[0016] R4 is selected from the group consisting of H, D and C1-C4 alkyl;

[0017] C is selected from 5-10 membered heterocyclyl, which heterocyclyl can be monocyclic or bicyclic forming a spiro ring or a bridged ring, which heterocyclyl contains 1-3 heteroatoms, which heteroatoms are O, S and N, and which heterocyclyl is optionally substituted with one or more R c , which R c is independently selected from C1-C4 alkyl, C1-4 alkoxy, halogen;

[0018] The L is a divalent linker, and the L is selected from:

[0019]

[0020] F1, F2, F3, and F4 are selected from O, NH, or are not present;

[0021] n1, n2, n3 and n4 are selected from 0, 1, 2, 3 or 4;

[0022] G1 is independently selected from the following groups: C3-C10 cyclic aliphatic rings, C6-C10 cyclic aromatic rings, or 5-10 cyclic heterocyclic aromatic rings;

[0023] The adipose ring is a helical ring or bridge ring composed of a single ring or a double ring; the selected G1 can have 0, 1, 2 or 3 R... g The R that was replaced g Selected from the following group: C1-4 alkyl, C1-C4 oxoalkyl, halogen, C3-C10 cycloalkyl; the selected 5-10 membered heteroaryl ring contains 1-3 heteroatoms selected from O, N, and S;

[0024] The E is selected from the protein-binding fragment represented by formula III:

[0025]

[0026] R5 is selected from methyl and H;

[0027] R6 is selected from methyl and hydroxymethyl;

[0028] R7 is selected from methyl and H;

[0029] R8 is selected from -NH,

[0030] In another preferred embodiment, R1 is phenyl or naphthalene ring; Ra is independently selected from C1-C4 alkyl, halogen, or hydroxyl groups; and R2 is F.

[0031] In another preferred embodiment, R3 is selected from the following group:

[0032]

[0033] and their respective chiral isomers.

[0034] In another preferred embodiment, R3 is

[0035] In another preferred embodiment, the compound has the structure described in Formula I-1:

[0036]

[0037] The remaining substituents are as described above.

[0038] In another preferred embodiment, C is selected from the group consisting of:

[0039]

[0040] In another preferred embodiment, the C is In another preferred embodiment, the R4is hydrogen, and n is 1. In another preferred embodiment, the A is O.

[0041] In another preferred embodiment, the L is selected from the group consisting of:

[0042]

[0043] wherein n1, n2, n3, and n4are defined as described above.

[0044] In another preferred embodiment, the compound is selected from the group consisting of:

[0045]

[0046]

[0047]

[0048]

[0049]

[0050]

[0051]

[0052]

[0053]

[0054] In a second aspect, the present application provides a pharmaceutical composition comprising:

[0055] (1) a compound according to the first aspect of the present application, a tautomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof; and

[0056] (2) a pharmaceutically acceptable carrier.

[0057] In a third aspect, the present application provides the use of a compound, a tautomer, a racemate, an enantiomer, a diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof according to the first aspect of the present application, or a pharmaceutical composition according to the second aspect of the present application, for the preparation of a medicament for preventing, treating and / or ameliorating a disease mediated by or dependent on KRAS protein.

[0058] In another preferred embodiment, the disease is a tumor.

[0059] In another preferred embodiment, the tumor is selected from the group consisting of breast cancer, multiple myeloma, bladder cancer, endometrial cancer, gastric cancer, cervical cancer, rhabdomyosarcoma, non-small cell lung cancer, small cell lung cancer, pleomorphic lung cancer, ovarian cancer, esophageal cancer, melanoma, colorectal cancer, hepatocellular carcinoma, head and neck tumor, cholangiocellular carcinoma, myelodysplastic syndrome, malignant glioma, prostate cancer, thyroid cancer, schwannoma, lung squamous cell carcinoma, keratosis, synovial sarcoma, skin cancer, pancreatic cancer, testicular cancer or liposarcoma.

[0060] It should be understood that, within the scope of the present application, each of the technical features described above and each of the technical features specifically described hereinafter (such as in the examples) can be combined with each other to form a new or preferred technical solution. Due to the limited space, they will not be listed one by one here. DETAILED DESCRIPTION

[0061] The present inventors have made extensive and in-depth research, and through a large number of screening and testing, provided a compound that can be used as a PanKRAS protein degradation agent. The compound is a heterobifunctional molecule composed of a POI ligand, a linker and an E3 ligase ligand, directly targets KRAS protein, degrades target protein, inhibits cell proliferation, and some compounds exhibit KRAS mutation selectivity, which can prevent and / or treat KRAS mutation related diseases such as tumors, etc. On this basis, the present application is completed.

[0062] Terminology

[0063] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

[0064] As used herein, the term "about," when used in reference to a particular recited numerical value, means that the value can vary from the recited value by not more than 1%. For example, as used herein, the expression "about 100" includes all values between 99 and 101 (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0065] As used herein, the term "containing" or "including" can be open, semi-closed and closed. In other words, the term also includes "consisting essentially of" or "consisting of".

[0066] As used herein, the term "room temperature" or "ambient temperature" means a temperature of 4-40°C, preferably 25±5°C.

[0067] The term "halogen" means fluorine, chlorine, bromine or iodine.

[0068] The term "C1-C4 alkyl" means a straight or branched chain alkyl group including 1-4 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl or different butyl, pentyl or hexyl isomers, etc.

[0069] The term "C1-C4 alkyl" means a straight or branched chain alkyl group including 1-4 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl or different butyl, pentyl or hexyl isomers, etc.

[0070] "C3-C6 aliphatic ring" means a cyclic alkyl group having 3-6 carbon atoms in the ring, including, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.

[0071] "5-18 membered heterocyclic group" means a ring group having 5-18 atoms including 1, 2 or 3 heteroatoms selected from N, O, S, for example:

[0072] "C6-C18 aryl" means a polyunsaturated (usually aromatic) hydrocarbon group having from 6-18 carbon atoms, which can be a single ring or multiple rings that are fused together or linked covalently (up to three rings). Examples of aryl groups include: phenyl.

[0073] "Aromatic heterocycle" or "heteroaryl" have the same meaning, referring to a heteroaromatic group containing one to several heteroatoms. For example "C3-C 10 Heteroaryl" means an aromatic heterocycle having 1-4 heteroatoms selected from oxygen, sulfur and nitrogen, and 3-10 carbon atoms. Non-limiting examples include: furanyl, thienyl, pyridyl, pyrazolyl, pyrrolyl, N-alkyl pyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, etc. The heteroaryl group can be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring attached to the parent structure together is a heteroaryl ring.

[0074] Typical examples of the Pan KRAS protein degrader represented by formula (I) of the present application are shown in Table 1, and the present application is not limited thereto.

[0075] Unless otherwise stated, the structural formulae described herein are intended to include all isomeric forms (e.g., enantiomeric, diastereomeric, and geometric (or conformational) isomers): for example, the R, S configurations for asymmetric centers, the (Z), (E) isomers for double bonds, and the like. Thus, individual stereochemical isomers and mixtures of their enantiomers, diastereomers, or geometric (or conformational) isomers, where possible, of the present compounds, are within the scope of the application.

[0076] As used herein, the term "tautomer" means structural isomers that differ in energy by a low energy barrier, and thus interconvert. For example, prototropic tautomers (i.e., proton shift) include interconversion by proton migration, such as 1H-indazole and 2H-indazole. Valence tautomers include interconversion by reorganization of some of the bonding electrons.

[0077] As used herein, the term "solvate" means a complex of a compound of the present application with solvent molecules in specific stoichiometric ratios.

[0078] As used herein, the term "hydrate" means a complex of a compound of the present application with water.

[0079] Active ingredient

[0080] In the present application, an active ingredient capable of effectively degrading Pan KRAS protein is provided. The active ingredient is a compound represented by general formula (I), and the active ingredient can effectively prevent, treat, and / or alleviate Pan KRAS protein-related diseases.

[0081] Experiments show that the active ingredient of the present application can effectively degrade Pan KRAS protein, thereby preventing, treating, and / or alleviating Pan KRAS protein-related diseases.

[0082] It should be understood that the active ingredient of the present application includes a compound represented by general formula (I), or a pharmaceutically acceptable salt thereof, or a prodrug thereof. It should be understood that the active ingredient of the present application also includes a crystalline form of the compound of general formula (I), an amorphous compound, and a deuterated compound, and the like.

[0083] The term "pharmaceutically acceptable salt" refers to those salts of the compounds of the present application which are suitable for use as medicaments. Pharmaceutically acceptable salts include inorganic and organic salts. One preferred class of salts are those formed from the compounds of the present application and acids. Suitable acids for salt formation include, but are not limited to, hydrochloric, hydrobromic, hydrofluoric, sulfuric, nitric, phosphoric, and the like inorganic acids; formic, acetic, trifluoroacetic, propionic, oxalic, malonic, succinic, fumaric, maleic, lactic, malic, tartaric, citric, picric, benzoic, methanesulfonic, ethanesulfonic, p-toluenesulfonic, benzenesulfonic, naphthalenesulfonic, and the like organic acids; and amino acids such as proline, phenylalanine, aspartic acid, glutamic acid. Another preferred class of salts are those formed from the compounds of the present application and bases, such as alkali metal salts (e.g., sodium or potassium salts), alkaline earth metal salts (e.g., magnesium or calcium salts), ammonium salts (e.g., lower alkylammonium salts and other pharmaceutically acceptable amine salts), such as methylamine salts, ethylamine salts, propylamine salts, dimethylamine salts, trimethylamine salts, diethylamine salts, triethylamine salts, t-butylamine salts, ethylenediamine salts, hydroxyethylamine salts, dihydroxyethylamine salts, trihydroxyethylamine salts, and amine salts of morpholine, piperazine, lysine, respectively.

[0084] Pharmaceutical compositions and methods of administration

[0085] Since the compounds of the present application have excellent Pan KRAS protein degradation activity, the compounds of the present application and various crystal forms thereof, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present application as the main active ingredient can be used for preventing, treating and / or alleviating Pan KRAS protein related diseases, such as treating cancer.

[0086] The pharmaceutical composition of the present application comprises a safe and effective amount of the compound of the present application and a pharmaceutically acceptable excipient or carrier. Among them, "safe and effective amount" refers to the amount of the compound sufficient to significantly improve the condition without causing serious side effects. Generally, the pharmaceutical composition contains 1-2000 mg of the compound of the present application per dose, more preferably 10-200 mg of the compound of the present application per dose. Preferably, the "dose" is a capsule or a tablet.

[0087] "Pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid filler substances or gel materials, which are suitable for human use and which are of sufficient purity and sufficiently low toxicity. By "compatible" it is meant that the components of the composition are capable of being commingled with the compounds of the application, and with each other, in the dosage form with no interaction that significantly affects the efficacy of the compounds. Examples of suitable pharmaceutically acceptable carriers are celluloses and their derivatives (e.g., sodium carboxymethylcellulose, ethylcellulose sodium, cellulose acetate, etc.), gelatin, talc, solid lubricants (e.g., stearic acid, magnesium stearate), calcium sulfate, vegetable oils (e.g., soybean oil, sesame oil, peanut oil, olive oil, etc.), polyhydric alcohols (e.g., propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (e.g., lecithin), wetting agents (e.g., sodium lauryl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, and the like.

[0088] The mode of administration of the compounds or pharmaceutical compositions of the present application is not narrowly critical and representative modes of administration include, but are not limited to, oral, parenteral (intravenous, intramuscular or subcutaneous).

[0089] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dosage forms, the active compound is admixed with at least one inert excipient (or carrier) such as sodium citrate or dicalcium phosphate, or with such other ingredients as binders, (a) fillers or extenders, e.g., starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, e.g., hydroxymethylcellulose, alginic acid, gelatin, polyvinylpyrrolidone, sucrose, and acacia; (c) humectants, e.g., glycerol; (d) disintegrating agents, e.g., agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solution retarders, e.g., paraffin; (f) absorption accelerators, e.g., quaternary ammonium compounds; (g) wetting agents, e.g., cetyl alcohol and glycerol monostearate; (h) absorbents, e.g., kaolin and bentonite clay; and (i) lubricants, e.g., talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, buffering agents can be included.

[0090] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings and other materials well known in the art. They can contain opacifying agents, and can also be of such composition that they release the active compound or compounds in a certain part of the intestinal tract in a delayed manner. Examples of embedding compositions that can be used are polymeric substances and waxes. The active compounds can also be in micro-encapsulated form, if appropriate, with one or more of the above-mentioned excipients.

[0091] ​Liquid dosage forms for oral administration include pharmaceutically-acceptable emulsions, solutions, suspensions, syrups, or elixirs. In addition to the active compounds, the liquid dosage forms can contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers, as, for example, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3- butylene glycol, dimethylformamide, and the like, or mixtures thereof.

[0092] Besides such inert diluents, the composition can also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0093] Suspensions, in addition to the active compounds, can contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, agar-agar, or mixtures thereof, and the like.

[0094] Compositions for parenteral injection can contain physiologically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols and suitable mixtures thereof.

[0095] The compounds of the present application can be administered alone or in combination with other pharmaceutically acceptable therapeutic agents.

[0096] In combination therapy, the pharmaceutical composition further comprises one or more (2, 3, 4, or more) other pharmaceutically acceptable therapeutic agents. One or more (2, 3, 4, or more) of the other pharmaceutically acceptable therapeutic agents can be used simultaneously, separately or sequentially with the compounds of the present application to prevent, treat and / or ameliorate FGFR-mediated diseases.

[0097] The pharmaceutical composition is used in a safe and effective amount of the compounds of the present application for a mammal (e.g., human) in need of treatment, wherein the dosage is administered in a pharmaceutically effective amount, and the daily dosage is generally 1-2000 mg, preferably 20-500 mg, for a 60 kg body weight. Of course, the specific dose will also take into account a number of factors such as the route of administration, the patient's health status, and the like, which are within the skill of the skilled practitioner.

[0098] Abbreviations

[0099]

[0100]

[0101] Advantages of the present application

[0102] According to the embodiments of the present application, the present application provides KRAS protein degradation agents with novel structures and significant activities, which can be used for effectively treating diseases and disorders related to KRAS protein.

[0103] Some compounds in the present application exhibit selectivity for KRAS mutations, which can reduce the occurrence of adverse reactions while exerting therapeutic effects.

[0104] The present application will be further described in conjunction with specific implementations. It should be understood that these examples are only used to illustrate the present application and not to limit the scope of the present application. The experimental methods in the following examples, if not specified, are generally carried out according to conventional conditions, such as the conditions described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the conditions recommended by the manufacturer. Unless otherwise specified, percentages and parts are calculated by weight.

[0105] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary skill in the art. In addition, any method and material similar or equivalent to those described herein can be used in the present application. The preferred methods and materials described herein are only used as examples.

[0106] Synthesis of general intermediates

[0107] First step:

[0108]

[0109] Under the protection of nitrogen, drop DIPEA (31.0 g, 240 mmol, 3.0 eq) into a solution of raw material C-1 (15.1 g, 60 mmol, 1.0 eq) and C-0 (9.0 g, 60 mmol, 1.0 eq) in dichloromethane (200 mL) at -40℃, and maintain the temperature not more than -40℃; after drop completion, the obtained dark brown reaction solution is naturally raised to room temperature 26℃ and stirred for 3 hours; pour the reaction solution into an ice water mixture (100 g), stir for 10 minutes, remove the aqueous phase, and concentrate the organic phase under reduced pressure. The residue is purified by slurry with methyl tert-butyl ether: petroleum ether = 1:10 (100 mL) to obtain the light yellow solid product C-2 (21 g).

[0110] MS (ESI, m / z): 331.3 [M+H]+.

[0111] 1H NMR (600MHz, CDCl3) δ9.08 (s, 1H), 4.59 (d, J = 13.2Hz, 1H), 4.44 (d, J = 13.6Hz, 1H), 3.37 (dd, J = 21.8, 12.8 Hz,2H),2.12(dddd,J=16.2,12.1,8.1,4.2Hz,1H),1.92(d,J=13.8Hz,1H),1.85-1.68(m,3H),1.38(s,3H)

[0112] Step Two:

[0113]

[0114] Compound C-2 (3.5 g, 10.57 mmol) was dissolved in dimethyl sulfoxide (10 mL) at 20-25 °C, followed by the addition of potassium fluoride (5.85 g, 100.57 mmol, 10 eq). The reaction was heated to 120 °C and stirred for 1 hour. The reaction was monitored by TLC. After the reaction was complete, the reaction solution was cooled to room temperature, 50 mL of water was added, and the mixture was extracted with ethyl acetate (50 mL * 4). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 1) to give compound 2 (yellow solid, 2 g, yield 60%).

[0115] Step 3:

[0116]

[0117] Compound C-3 (2 g, 6.37 mmol) was dissolved in 1,4-dioxane (40 ml) and water (8 ml) under stirring at 20–25 °C. Then, compound C-4 (4.6 g, 12.74 mmol, 2 eq), potassium phosphate (4.07 g, 19.11 mmol, 3 eq), and Pd Cata CXium A Pd G3 (466.11 mg, 0.64 mmol, 0.1 eq) were added sequentially. The reaction mixture was heated to 100 °C and stirred for 3 hours. The reaction was monitored by TLC. After cooling to room temperature, 100 ml of water was added, followed by extraction with ethyl acetate (30 ml x 3). The combined organic phases were extracted with saturated brine (30 ml x 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (developing solvent: petroleum ether / ethyl acetate = 1 / 1) to give compound C-5 (yellow solid, 700 mg, yield 21%).

[0118] Example 1 (077)

[0119] Step 1 : Synthesis of compound 1-2

[0120]

[0121] To a solution of compound 1-1 (10.4 g, 0.1 mmol, 1.0 eq) in THF (100 mL) was added NaH (4.0 g, 0.1 mmol, 60% content, 1.0 eq) in portions under ice water bath 0 °C and nitrogen protection, after stirring for 30 minutes, the raw material ethyl bromoacetate (32.4 g, 0.15 mmol, 1.5 eq) was added at once, and the reaction mixture was naturally raised to room temperature, then heated to 55 °C and stirred for 4 hours. The reaction mixture was poured into dilute hydrochloric acid aqueous solution (100 mL, 0.5 M) to quench, and MTBE (100 mL) was added to extract, and the combined organic phase was concentrated under reduced pressure, and the residue was purified by flash silica gel column (ethyl acetate: petroleum ether = 0-10% gradient elution) to obtain the product 1-2 (7.1 g) as colorless oil.

[0122] 1 H NMR (300 MHz, CDC13) δ 4.23 (q, J = 7.1 Hz, 2H), 4.07 (s, 2H), 3.57 (t, J = 6.1 Hz, 2H), 3.48 (t, J = 6.7 Hz, 2H), 2.06-1.94 (m, 2H), 1.78 (dq, J = 9.6, 6.3 Hz, 2H), 1.30 (t, J = 7.1 Hz, 3H).

[0123] Step 2: Synthesis of compound 1-3

[0124]

[0125] To a solution of compound 1-2 (2.0 g, 8.4 mmol, 1.0 eq) and L-prolinol (0.85 g, 8.4 mmol, 1.0 eq) in MeCN was added potassium carbonate (2.3 g, 16.7 mmol, 2.0 eq) and potassium iodide (260 mg, 1.6 mmol, 0.2 eq), and the resulting mixture was stirred at room temperature 22 °C-31 °C for 16 hours. The reaction mixture was filtered and washed with dichloromethane (20 mL), and the combined organic phase was concentrated under reduced pressure, and the crude product was purified by flash silica gel column (methanol:dichloromethane = 0-15%, gradient elution with the addition of 0.1% ammonia) to obtain the product 1-3 (2.1 g) as light brown oil.

[0126] MS (ESI, m / z): 260.4 [M+H] + .

[0127] 1H NMR (600 MHz, Chloroform-d) δ 4.24 (q, J = 7.1 Hz, 2H), 4.08 (s, 2H), 3.70 (d, J = 9.4 Hz, 1H), 3.56 (tt, J = 9.1, 4.5 Hz, 2H), 3.52 - 3.42 (m, 2H), 3.30 (s, 1H), 2.81 (d, J = 78.0 Hz, 2H), 2.41 (d, J = 46.5 Hz, 2H), 1.98 - 1.88 (m, 1H), 1.83 (d, J = 13.0 Hz, 3H), 1.76 - 1.61 (m, 4H), 1.31 (t, J = 7.1 Hz, 3H).

[0128] Third Step: Synthesis of compound 1-4

[0129]

[0130] To a solution of starting material 1-3 (820 mg, 3.2 mmol, 1.2 eq) and compound C-2 (870 mg, 2.6 mmol, 1.0 eq) in THF (10 mL) was added dropwise LiHMDS THF solution (3.2 g, 3.2 mmol, 1 mol / L, 1.2 eq) under ice water bath 0 °C and nitrogen protection. The resulting light brown reaction solution was stirred for 30 min, then naturally raised to room temperature 32 °C and stirred for 4 h. The reaction mixture was quenched by pouring into aqueous citric acid solution (3 mL, 0.5 mol / L), extracted with ethyl acetate:methanol = 10:1 (30 mL*3), and the combined organic phase was concentrated under reduced pressure. The residue was purified by column chromatography, eluted with methanol:dichloromethane = 0-10%, to give the foam paper solid product 1-4 (1.1 g).

[0131] MS (ESI, m / z): 554.5 [M+H] + .

[0132] Fourth Step: Synthesis of compound 1-5

[0133]

[0134] To a solution of compound 1-4 (1.1 g, 2.0 mmol, 1.0 eq) and C-4 (950 mg, 2.6 mmol, 1.3 eq) in dioxane and water (15 mL, 5:1) was added potassium phosphate (1.3 g, 6.1 mmol), after three times nitrogen replacement, Pd CataCXium A Pd G3 (150 mg, 0.2 mmol, 0.1 eq) was added, the resulting light brown solution was heated to 80 °C and stirred for 6 h. To the reaction mixture was added water (5 mL) and ethyl acetate (10 mL*3) was extracted, the combined organic phase was concentrated under reduced pressure, the residue was purified by flash silica gel (methanol:dichloromethane 0~10% gradient elution) to give the product 1-5 (810 mg) as a light yellow foamy solid.

[0135] MS (ESI, m / z): 752.8 [M+H] + .

[0136] Step 5: Synthesis of compound 1-6

[0137]

[0138] To a solution of compound 1-5 (150 mg, 0.2 mmol, 1.0 eq) in acetonitrile (2 mL) was added Et3N (61 mg, 0.6 mmol, 3.0 eq), water (11 mg, 0.6 mmol, 3.0 eq) and lithium bromide (180 mg, 2 mmol, 10.0 eq) sequentially at room temperature 20-27 °C, the reaction mixture was stirred at 27 °C~33 °C for 16 h. To the reaction mixture was added aqueous citric acid (0.5 mol / L) to pH~5, ethyl acetate (5 mL*3) was added to extract, the combined organic phase was concentrated under reduced pressure, the residue was purified by flash silica gel column to give 1-6 (120 mg) as a light yellow foamy solid.

[0139] MS (ESI, m / z): 724.5 [M+H] + .

[0140] Step 6: Synthesis of compound 1-7

[0141]

[0142] To a solution of compound 1-6 (610 mg, 0.84 mmol, 1.0 eq) and C-6 (410 mg, 0.92 mmol, 1.1 eq) in DMF (10 mL) was added DIPEA (330 mg, 2.54 mmol, 3.0 eq) and HATU (440 mg, 1.16 mmol, 1.4 eq), the resulting light brown reaction solution was stirred at room temperature 29 °C for 2 h. To the reaction solution was added water (15 mL) and saturated brine (10 mL) and stirred for 5 min, then added ethyl acetate (10 mL*3) and extracted, the combined organic phase was concentrated under reduced pressure, the residue was purified by column chromatography, eluted with methanol: dichloromethane = 0-10%, to give the product 1-7 (820 mg) as a light yellow foamy solid.

[0143] MS (ESI, m / z): 1150.6 [M+H]+.

[0144] Seventh step: synthesis of example 1

[0145]

[0146] To a solution of compound 1-7 (820 mg, 0.71 mmol, 1.0 eq) in dichloromethane (10 mL) was added HCl-dioxane solution (2.8 mL, 2.8 mmol, 4 mol / L), the reaction solution was stirred at room temperature 29 °C for 1 h. Then concentrated under reduced pressure, the residue was purified by column chromatography pre-HPLC to give example 1 (345 mg, light yellow solid).

[0147] MS (ESI, m / z): 1106.7 [M+H] + .

[0148] 1H NMR (300 MHz, MeOD) δ 9.31 - 9.21 (m, 1H), 8.86 (d, J = 3.9 Hz, 1H), 7.78 - 7.56 (m, 2H), 7.39 (d, J = 10.2 Hz, 4H), 7.31 (d, J = 2.6 Hz, 1H), 7.25 (t, J = 9.4 Hz, 1H), 7.07 (s, 1H), 5.03 - 4.94 (m, 1H), 4.74 - 4.48 (m, 5H), 4.37 (dd, J = 21.6, 13.9 Hz, 2H), 4.11 - 3.91 (m, 3H), 3.86 - 3.69 (m, 3H), 3.68 - 3.37 (m, 6H), 3.24 (s, 2H), 2.40 (d, J = 9.6 Hz, 2H), 2.28 - 2.05 (m, 6H), 2.00 - 1.66 (m, 9H), 1.59 - 1.44 (m, 4H), 1.29 (t, J = 4.8 Hz, 3H), 1.00 (d, J = 5.2 Hz, 9H), 0.81 (q, J = 7.3 Hz, 3H).

[0149] Example 2 (078)

[0150] First Step: Synthesis of compound 2-2

[0151]

[0152] To a solution of starting material 2-1 (2.0 g, 8.4 mmol, 1.0 eq) and L-prolinol (0.85 g, 8.4 mmol, 1.0 eq) in MeCN (10 ml) was added potassium carbonate (2.3 g, 16.7 mmol, 2.0 eq) and potassium iodide (260 mg, 1.6 mmol, 0.2 eq) and the resulting mixture was stirred at room temperature 22 °C to 31 °C for 16 hours. The reaction mixture was filtered, washed with dichloromethane (20 mL), and the combined organic phases were concentrated under reduced pressure. The residue was purified by column chromatography (methanol:dichloromethane 0-15% gradient elution with 0.1% aqueous ammonia) to give the product 2-2 (2.1 g) as a light brown oil. MS (ESI, m / z): 260.4 [M+H] + .

[0153] The following steps were identical to Example 1 to give Example 2, see Table 1 for characterization spectra.

[0154] Example 3 (079)

[0155] First Step: Synthesis of compound 3-2

[0156]

[0157] Compound 3-1 (3 g, 21.58 mmol) was dissolved in acetonitrile (15 ml) at 20-25 °C, and then ethyl acrylate (4.3 g, 43.16 mmol, 2 eq) and cesium carbonate (716.8 mg, 2.2 mmol, 0.1 eq) were added. The reaction was stirred at 20-25 °C for 16 hours, and the reaction was monitored by TLC. After the reaction was completed, the reaction was concentrated under reduced pressure to obtain a crude product. The crude product was purified by flash column chromatography (eluent: dichloromethane / methanol = 10 / 1) to obtain compound 3-2 (colorless oil, 1.8 g, yield 35%).

[0158] 1 H NMR (300 MHz, CDCl3) δ 4.16 (q, J = 7.1 Hz, 2H), 3.72 (t, J = 6.3 Hz, 2H), 3.57 (t, J = 5.8 Hz, 2H), 3.48 (t, J = 6.5 Hz, 2H), 2.56 (t, J = 6.3 Hz, 2H), 2.14-2.02 (m, 2H), 1.27 (t, J = 7.1 Hz, 3H).

[0159] Second step: synthesis of compound 3-3

[0160]

[0161] Compound 3-2 (1.60 g, 6.69 mmol, 1.2 eq) was dissolved in N,N-dimethylformamide (20 ml) at 20-25 °C, and then L-prolinol (563.63 mg, 5.58 mmol, 1 eq), potassium carbonate (2.30 g, 16.73 mmol, 3 eq) and potassium iodide (185.09 mg, 1.12 mmol, 0.2 eq) were added in sequence. The reaction was heated to 90 °C and stirred for 5 hours. The reaction was monitored by TLC, and after the reaction was completed, the reaction system was cooled to 25 °C, 200 ml of water was added to the reaction, and then extracted with ethyl acetate (50 ml*3). The organic phase was washed with saturated brine (30 ml*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by flash column chromatography (eluent: dichloromethane / methanol = 10 / 1) to obtain compound 3-3 (yellow oil, 750 mg, yield 52%).

[0162] MS (ESI, m / z): 260.4 [M+H] + .

[0163] 1H NMR (300 MHz, CDC13) δ 4.15 (q, J = 7.1 Hz, 2H), 3.69 (td, J = 6.5, 1.4 Hz, 2H), 3.63 (dd, J = 10.8, 3.6 Hz, 1H), 3.51 (td, J = 6.3, 3.5 Hz, 2H), 3.38 (dd, J = 10.9, 2.9 Hz, 1H), 3.18 (dt, J = 9.2, 4.6 Hz, 1H), 2.92-2.82 (m, 1H), 2.64-2.48 (m, 3H), 2.39-2.19 (m, 2H), 1.93-1.67 (m, 6H), 1.26 (t, J = 7.1 Hz, 3H).

[0164] Third Step: Synthesis of compound 3-4

[0165]

[0166] Compound C-5 (133.75 mg, 0.52 mmol, 1.2 eq) was dissolved in tetrahydrofuran (5 ml) at 20-25 °C, then compound 3-3 (220 mg, 0.43 mmol, 1 eq) was added. After the reaction solution was protected by nitrogen, LiNMDS (0.52 ml, 1 mol / L, 1.2 eq) was slowly added dropwise to the reaction solution at 0-5 °C, and the reaction solution was slowly raised to 25 °C and stirred for 1 hour. The reaction was monitored by TLC. After the reaction was completed, saturated aqueous ammonium chloride solution (10 ml) was added to the reaction solution, which was then extracted with dichloromethane / methanol mixed solvent (10 / 1, 10 ml*3). The organic phase was combined and washed with saturated brine (30 ml*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by flash silica gel column (eluent: dichloromethane / methanol = 10 / 1) to obtain compound 3-4 (yellow solid, 130 mg, yield 40%).

[0167] MS (ESI, m / z): 752.6 [M+H] +

[0168] Fourth Step: Synthesis of compound 3-5

[0169]

[0170] Compound 3-4 (130 mg, 0.17 mmol) was dissolved in acetonitrile (2 ml) at 20-25 °C, then lithium bromide (130.30 mg, 1.70 mmol, 10 eq), triethylamine (51.61 mg, 0.51 mmol, 3 eq) and water (9.18 mg, 0.51 mmol, 3 eq) were added successively. The reaction was stirred at 15 °C for 16 hours. The reaction process was monitored by TLC. After the reaction was completed, 10% citric acid aqueous solution was slowly added to the reaction solution to pH ~ 4, then extracted with dichloromethane / methanol mixed solvent (10 / 1, 10 ml*3). The organic phase was combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give compound 3-5 (50 mg, white solid, yield 41%).

[0171] Step 5: Synthesis of compound 3-6

[0172]

[0173] Compound 3-5 (50 mg, 0.07 mmol) was dissolved in N,N-dimethylformamide (1.5 ml) at 20-25 °C, then C-6 (46.68 mg, 0.11 mmol, 1.5 eq), HATU (39.93 mg, 0.11 mmol, 1.5 eq) and N,N-diisopropyl ethylamine (27.14 mg, 0.21 mmol, 3 eq) were added. The reaction was stirred at 20-25 °C for 3 hours. The reaction process was monitored by LC-MS. After the reaction was completed, 5 ml of water was added to the reaction solution, then extracted with ethyl acetate (5 ml*5), the organic phase was combined and extracted with saturated brine (30 ml*2), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 10 / 1) to give compound 3-6 (yellow solid, 30 mg, yield 37%).

[0174] MS (ESI, m / z): 1150.6 [M+H] +

[0175] Step 6: Synthesis of Example 3

[0176]

[0177] Compound 3-6 (30 mg, 0.03 mmol) was dissolved in dichloromethane (1 ml) at 20-25 °C, then hydrochloric acid dioxane solution (4 mol / L, 0.5 ml) was added, and the reaction was stirred for 1 hour. The reaction process was monitored by LCMS. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain the crude product. The crude product was purified by a reversed-phase chromatographic column to obtain compound Example 3 (white solid, 2.1 mg, formate, yield 6%).

[0178] MS (ESI, m / z): 1106.6 [M+H] +

[0179] 1 H NMR (300 MHz, DMSO-d6) δ 9.22 (s, 1H), 8.98 (s, 1H), 8.36 (d, J = 7.7 Hz, 1H), 8.24 (s, 1H), 7.80 (d, J = 9.2 Hz, 1H), 7.77-7.73 (m, 1H), 7.46-7.41 (m, 2H), 7.40-7.31 (m, 4H), 7.04 (d, J = 2.5 Hz, 1H), 4.97-4.84 (m, 2H), 4.74 (s, 2H), 4.51 (d, J = 9.2 Hz, 2H), 4.46-4.35 (m, 3H), 4.16-4.04 (m, 3H), 3.67-3.52 (m, 7H), 2.45 (s, 3H), 2.21-1.98 (m, 8H), 1.76-1.59 (m, 8H), 1.36 (d, J = 7.0 Hz, 3H), 1.24 (s, 1H), 1.16 (d, J = 7.1 Hz, 3H), 0.89 (s, 9H), 0.73 (t, J = 6.5 Hz, 3H).

[0180] Example 4 (080)

[0181] First step: synthesis of compound 4-2

[0182]

[0183] Compound 4-2 (yellow oil, 1.4 g, yield 46%) was obtained by the reaction of 4-1 (2.5 g, 11.61 mmol) with bromoethanol (1.73 g, 13.93 mmol, 1.2 eq), potassium carbonate (4.80 g, 24.83 mmol, 3 eq) and potassium iodide (385.50 mg, 2.32 mmol, 0.2 eq) in N,N-dimethylformamide (20 ml) at 20-25 °C for 5 h under nitrogen. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was cooled to 20 °C and 30 ml of water was added to the reaction mixture, which was then extracted with ethyl acetate (20 ml*3). The organic phase was washed with saturated brine (20 ml*2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product. The crude product was purified by flash column chromatography (eluent: dichloromethane / methanol = 10 / 1) to obtain compound 4-2 (yellow oil, 1.4 g, yield 46%).

[0184] MS (ESI, m / z): 260.4 [M+H] + .

[0185] 1 H NMR (300 MHz, CDC13) δ 3.70 - 3.52 (m, 3H), 3.44 (dd, J = 10.0, 6.1 Hz, 1H), 3.14 (dt, J = 9.5, 4.8 Hz, 1H), 3.02 (ddd, J = 12.5, 9.1, 4.9 Hz, 1H), 2.70 (dq, J = 8.3, 5.6 Hz, 1H), 2.56 (dt, J = 12.5, 3.7 Hz, 1H), 2.31 (dt, J = 9.0, 8.0 Hz, 1H), 1.95 - 1.81 (m, 1H), 1.80 - 1.67 (m, 2H), 1.65 - 1.53 (m, 1H), 0.90 (s, 9H), 0.06 (s, 6H).

[0186] Second Step: Synthesis of compound 4-3

[0187]

[0188] Compound 4-2 (900 mg, 3.47 mmol) was dissolved in acetonitrile (8 ml) at 25 °C, then ethyl acrylate (1.74 g, 17.35 mmol, 5 eq) and cesium carbonate (113.06 mg, 0.35 mmol, 0.1 eq) were added. The reaction was heated to 70 °C and stirred for 8 hours. The reaction was monitored by TLC. After the reaction was completed, the reaction was cooled to 20 °C, and the reaction was concentrated under reduced pressure to obtain a crude product. The crude product was purified by flash silica gel column (eluent: dichloromethane / methanol = 10 / 1) to obtain compound 4-3 (yellow oil, 0.6 g, yield 48%).

[0189] MS (ESI, m / z): 360.2 [M+H] + .

[0190] Third step: synthesis of example 4

[0191]

[0192] Compound 4-3 (500 mg, 1.39 mmol) was dissolved in hydrochloric acid dioxane solution (5 ml, 4 mol / L) at 25 °C, and the reaction was stirred at 25 °C for 2 hours. The reaction was monitored by liquid chromatography-mass spectrometry. After the reaction was completed, the reaction was cooled to room temperature, and saturated sodium bicarbonate aqueous solution was slowly added to the reaction until the pH was about 8, and then extracted with dichloromethane (20 ml*3). The organic phase was combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 4-4 (yellow oil, 350 mg, yield 100%).

[0193] MS (ESI, m / z): 246.3 [M+H] + .

[0194] The following steps were the same as example 3 to obtain example 4, and the characterization data are shown in Table 1.

[0195] Example 5 (081)

[0196] First step: synthesis of compound 5-3 & 5-4

[0197]

[0198] Dimethyl sulfoxide (1.50 g, 19.26 mmol, 2 eq) was dissolved in dichloromethane (20 ml) at 20 °C. The reaction was replaced by nitrogen and cooled to -60 to -70 °C. Oxalyl chloride (1.80 g, 14.45 mmol, 1.5 eq) was slowly added to the reaction. The reaction was stirred at this temperature range for 0.5 h. Compound 5-1 (1.20 g, 9.63 mmol) was slowly added to the reaction and the reaction was stirred for 40 min. Then triethylamine (4.90 g, 48.15 mmol, 5 eq) was added to the reaction and the reaction was slowly warmed to 15 °C and stirred for 1 h. Compound 5-2 (4.96 g, 11.56 mmol, 1.2 eq) and potassium tert-butoxide (2.2 g, 19.26 mmol, 2 eq) were dissolved in THF (50 ml) and slowly added to the reaction at -60 to -70 °C. The reaction was slowly warmed to 15 °C and stirred for 2 h. The reaction was monitored by TLC. The reaction was cooled to room temperature and 50 ml of water was added to the reaction. The reaction was extracted with ethyl acetate (50 ml*3) and the organic phase was extracted with saturated brine (30 ml*2). The organic phase was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 3 / 1) to give compound 5-3 (yellow oil, 800 mg, yield 44%) and compound 5-4 (yellow oil, 300 mg, yield 16%).

[0199] 1 H NMR (300 MHz, CDC13) δ 6.95 (dt, J = 15.7, 4.3 Hz, 1H), 6.11 (dt, J = 15.7, 2.0 Hz, 1H), 4.26-4.19 (m, 4H), 3.78-3.73 (m, 2H), 3.68-3.63 (m, 2H), 1.30 (t, J = 7.1 Hz, 3H).

[0200] 1 H NMR (300 MHz, CDC13) δ 6.39 (dt, J = 11.7, 4.9 Hz, 1H), 5.84 (dt, J = 11.7, 2.4 Hz, 1H), 4.66 (dd, J = 4.9, 2.4 Hz, 2H), 4.18 (q, J = 7.1 Hz, 2H), 3.78-3.72 (m, 2H), 3.69-3.61 (m, 2H), 1.30 (t, J = 7.1 Hz, 3H).

[0201] Second step: synthesis of compound 5-5

[0202]

[0203] A mixture of compound 5-3 and 5-4 (750 mg, 3.89 mmol, 1.1 eq) was dissolved in N,N-dimethylformamide (6 ml) at 25 °C, then L-prolinol (357.90 mg, 3.54 mmol), potassium carbonate (1.47 g, 10.62 mmol, 3 eq) and potassium iodide (117.53 mg, 0.71 mmol, 0.2 eq) were added successively, and the reaction was stirred at 90 °C for 4 h. The reaction was monitored by LCMS. After the reaction was completed, the reaction system was cooled to 25 °C, 100 ml of water was added to the reaction solution, and then extracted with ethyl acetate (50 ml*3). The organic phase was combined and washed with saturated brine (30 ml*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 10 / 1) to obtain compound 5-5 (yellow oil, 400 mg, yield 44%).

[0204] MS (ESI, m / z): 258.3 [M+H] + .

[0205] Third step: synthesis of compound 5-6

[0206]

[0207] Pd / C (50 mg, 10% content) was added to a 100 ml single-necked flask, then wetted with a small amount of methanol (3 ml), and compound 5-4 (250 mg, 0.97 mmol) was added to the reaction solution, and methanol (10 ml) was added. After replacing hydrogen, the reaction mixture was stirred at 20 °C for 1 h. The reaction process was monitored by TLC. After the reaction was completed, the reaction solution was filtered through diatomite, the filter cake was rinsed with methanol (10 ml*3), and the combined filtrate was concentrated under reduced pressure to obtain compound 5-6 (yellow oil, 200 mg, yield 75%).

[0208] MS (ESI, m / z): 260.4 [M+H] + .

[0209] The following steps were the same as Example 3 to obtain Example 5, and the characterization spectrum was analyzed in Table 1.

[0210] Example 6(082)

[0211] First step: synthesis of compound 6-2

[0212]

[0213] To a mixture of compound 6-1 (1 g, 0.008 mol, 1 eq) and ethyl bromoacetate (1.3 g, 0.008 mol, 1 eq) in tetrahydrofuran (10 mL) at 0 °C under nitrogen protection, NaH (322 mg, 0.008 mol, content 60% 1 eq) was added in batches, and after the addition was completed, it was slowly warmed to room temperature and stirred for 2 hours. To the reaction system, saturated aqueous ammonium chloride solution (2 mL) was added dropwise to quench, then water (30 mL) was added, and extracted with ethyl acetate (20 mL x 3). The organic phase was combined, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated in vacuo, and the residue was purified by flash silica gel column (ethyl acetate / petroleum ether 0-100% gradient elution) to give compound 6-2 (925 mg, purity 80%, yield 54.6%) as colorless oil.

[0214] MS (ESI, m / z): 211.1 [M+H] + .

[0215] 1 H NMR (300 MHz, CDCl3) δ 4.15 (q, J = 7.1 Hz, 2H), 4.09 (s, 2H), 3.84-3.62 (m, 6H), 3.62-3.50 (m, 2H), 1.27-1.15 (m, 3H).

[0216] Second step: synthesis of compound 6-3

[0217]

[0218] Compound 6-2 (631 mg, 3 mmol, 1 eq), L-prolinol (364 mg, 3.60 mol, 1.2 eq), potassium carbonate (1.2 g, 9.00 mmol, 3 eq) and potassium iodide (100 mg, 0.60 mmol, 0.2 eq) were added to DMF (10 mL), then heated to 90 °C and stirred for 16 h. After the reaction was completed, the reaction system was poured into water (50 mL) and extracted with ethyl acetate (40 mL x 3), and all the organic phases were combined, washed with saturated brine (30 mL x 2), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated in vacuo. The residue was purified by flash silica gel column (methanol / dichloromethane 0-10% gradient elution) to give compound 6-3 (230 mg, purity 80%, yield 27.9%) as anhydrous oil.

[0219] MS (ESI, m / z): 276.3 [M+H] + .

[0220] The following steps were the same as Example 3 to give Example 6, and the characterization spectrum was analyzed in Table 1.

[0221] Example 7(083)

[0222] First Step: Synthesis of compound 7-2

[0223]

[0224] To a solution of compound 7-1 (3 g, 0.017 mol, 1 eq) and ethyl bromoacetate (2.96 g, 0.017 mol, 1 eq) in tetrahydrofuran (30 mL) was added NaH (714 mg, 0.017 mol, 1 eq, purity 60%) in portions at 0 °C under nitrogen protection, and the reaction was slowly warmed to room temperature and stirred for 2 hours. The reaction was quenched by dropwise addition of saturated aqueous ammonium chloride solution (5 mL), then water (100 mL) was added, and the mixture was extracted with ethyl acetate (80 mL x 3). The organic phases were combined, dried over anhydrous magnesium sulfate, and filtered. The filtrate was concentrated in vacuo, and the crude product was purified by flash silica gel column (eluted with ethyl acetate / petroleum ether 0-100% gradient) to give compound 7-2 (1.73 g, purity 75%, yield 40%) as colorless oil.

[0225] MS (ESI, m / z): 255.2 [M+H] + .

[0226] 1 H NMR (300 MHz, CDCl3) δ 4.30-4.02 (m, 4H), 3.73-3.51 (m, 12H), 1.22 (t, J = 7.2 Hz, 3H).

[0227] Second Step: Synthesis of compound 7-3

[0228]

[0229] Compound 7-2 (1.7 g, 0.006 mol, 1 eq), L-prolinol (0.7 g, 0.007 mol, 1.2 eq), potassium carbonate (2.8 g, 0.02 mmol, 3 eq) and potassium iodide (0.2 g, 0.001 mmol, 0.2 eq) were added to DMF (15 mL) and heated to 90 °C and stirred for 16 hours.

[0230] The reaction was poured into water (75 mL) and extracted with ethyl acetate (60 mL x 3), and all organic phases were combined, washed with saturated brine (50 mL x 2), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated in vacuo, and the crude product was purified by flash silica gel column (eluted with methanol / dichloromethane 0-10% gradient) to give compound 7-3 (304 mg, purity 80%, yield 15.7%) as colorless oil.

[0231] MS (ESI, m / z): 320.4 [M+H] + .

[0232] The following steps are the same as Example 3 to obtain Example 7, see Table 1 for characterization spectrum analysis.

[0233] Example 8(084)

[0234] First step: synthesis of compound 8-1

[0235]

[0236] Bromoethanol (10 g, 1.0 eq.) and ethyl bromoacetate (7.48 g, 1.0 eq.) were dissolved in 100 ml of anhydrous tetrahydrofuran, cooled to 0°C, and NaH (2.4 g, 1.0 eq.) was added in three batches. After the addition was completed, the temperature was gradually increased to room temperature (15°C), and after 3 hours the reaction was completed. The reaction solution was added to 100 ml of saturated aqueous ammonium chloride solution at 0°C to quench, and extracted with ethyl acetate (200 ml*3). The organic phase was combined and dried over anhydrous sodium sulfate, and concentrated to obtain 4.97 g of colorless oil.

[0237] 1 H NMR (300 MHz, CDCl3) δ 4.22-4.12 (m, 2H), 4.09 (s, 2H), 3.83 (t, J = 6.2 Hz, 2H), 3.45 (t, J = 6.2 Hz, 2H), 1.23 (t, J = 7.1 Hz, 3H).

[0238] Second step: synthesis of compound 8-2

[0239]

[0240] Compound 8-1 (2.03 g, 1.0 eq.) and L-prolinol (1.167 g, 1.2 eq.) were dissolved in 20 ml of N,N-dimethylformamide, and potassium iodide (1.595 g, 1.0 eq.) and potassium carbonate (2.65 g, 2.0 eq.) were added under nitrogen protection. The temperature was increased to 90°C and stirred for 4 hours. The reaction solution was cooled to room temperature, 10 ml of water was added, and extracted with a mixture of dichloromethane and methanol (50 ml*3, 5:1). The organic phase was combined and dried over anhydrous sodium sulfate, and concentrated to obtain 3.2 g of crude product. Purification was performed on a flash silica gel column (DCM / MeOH: 2%-10% gradient elution), and 0.97 g of yellow oil was obtained.

[0241] 1H NMR (300 MHz, CDC13) δ 4.23-4.11 (m, 2H), 4.11-3.92 (m, 3H), 3.67-3.56 (m, 3H), 3.45-3.34 (m, 1H), 3.28-3.19 (m, 1H), 3.08-2.95 (m, 1H), 2.79-2.68 (m, 1H), 2.67-2.51 (m, 1H), 2.47-2.22 (m, 1H), 1.89-1.65 (m, 4H), 1.23 (t, J = 7.2 Hz, 3H).

[0242] The following steps are the same as Example 3 to obtain Example 8, and the characterization spectrum is shown in Table 1.

[0243] Example 9 (086)

[0244]

[0245] NaH (519.36 mg, 12.98 mmol, purity 60%, 1.2 eq) and tetrahydrofuran (10 mL) were placed in a 100 mL three-necked flask, replaced with nitrogen for three times, and cooled to -5-0 °C. Compound 9-2 (1.5 g, 10.8 mmol, 1 eq) tetrahydrofuran solution (5 mL) was slowly added to the reaction solution at -5-0 °C with a syringe, and the temperature was kept at -5-0 °C for 0.5 h. Ethyl bromoacetate (2.2 g, 13.0 mmol, 1.2 eq) was slowly added to the reaction solution at a constant temperature, and the reaction was slowly raised to 15 °C for 3 h. The reaction system was cooled to 0 °C, and saturated ammonium chloride solution (10 mL) was slowly added to the reaction solution at -5-0 °C for quenching, and then raised to room temperature. Ethyl acetate (20 mL*3) was used for extraction, and the organic phase was combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by a flash silica gel column (eluent: petroleum ether / ethyl acetate = 3 / 1) to obtain compound 9-3 (1 g, yield 42%).

[0246] 1 H NMR (300 MHz, CDC13) δ 4.22 (q, J = 7.1 Hz, 2H), 4.15 (s, 2H), 3.76-3.70 (m, 2H), 3.69-3.59 (m, 6H), 2.04 (p, J = 6.2 Hz, 2H), 1.29 (t, J = 7.1 Hz, 3H).

[0247] Step 3: Synthesis of compound 9-4

[0248]

[0249] Compound 9-3 (1 g, 4.4 mmol, 1.2 eq) was dissolved in DMF (5 mL) at 25 °C, then compound L-prolinol (375 mg, 3.7 mmol, 1 eq), potassium carbonate (1.54 g, 11.1 mmol, 3.0 eq) and potassium iodide (123.2 mg, 0.74 mmol, 0.2 eq) were added successively, and the reaction was heated to 90 °C and stirred for 4 h. Water (15 mL) was added to the reaction solution, which was extracted with ethyl acetate (15 mL*3), and the combined organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column (eluent: dichloromethane / methanol = 10 / 1) to obtain compound 9-4 (yellow oil, 300 mg, yield 28%). MS (ESI, m / z): 290.2 [M+H] + .

[0250] The following steps were the same as Example 3 to obtain Example 9, and the characterization spectrum is shown in Table 1.

[0251] Example 10(088)

[0252] First step: synthesis of compound 10-2

[0253]

[0254] To a solution of compound 10-1 (3 g, 18 mmol, 1 eq) in tetrahydrofuran / water (20 mL / 10 mL) was added 10% sodium hydroxide solution (7.6 mL, 0.018 mol, 1 eq), followed by the addition of Boc2O (5.96 g, 27 mmol, 1.5 eq) in portions, and the mixture was stirred at room temperature for 2 h after the addition was completed. The tetrahydrofuran was concentrated under vacuum, the pH was adjusted to ~2 using 10% sodium bisulfate solution, and the mixture was extracted with ethyl acetate (15 mL x 3), and the organic phase was combined and washed with water and saturated brine once, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under vacuum to obtain a crude product compound 10-2 (4 g), which was used directly in the next step.

[0255] 1 H NMR (300 MHz, DMSO-d6) δ 4.47 (dd, J = 9.2, 5.1 Hz, 1H), 4.17 (s, 1H), 3.82 (d, J = 12.6 Hz, 1H), 3.68 (d, J = 7.3 Hz, 3H), 2.89 (q, J = 13.7, 12.7 Hz, 1H), 2.45 (d, J = 13.1 Hz, 1H), 1.47 (s, 9H).

[0256] Second step: synthesis of compound 10-3

[0257]

[0258] Control 0-5 ℃ and nitrogen protection, in compound 10-2 (2 g, 7.5 mmol, 1 eq) tetrahydrofuran (20 mL) solution in batches lithium aluminum hydride (286 mg, 8.2 mol, 1.1 eq) was added, after the addition was completed, slowly to room temperature stirring 2 hours. The temperature was reduced to 0 ℃, the reaction was added dropwise water (0.3 mL), 15% sodium hydroxide solution (0.3 mL) and water (0.9 mL) in turn, then added anhydrous sodium sulfate stirring and filtration, the filtrate was concentrated in vacuum, the residue was purified by flash silica gel column (methanol / dichloromethane 0-10% gradient elution) to give colorless oil compound 10-3 (1.05 g, purity 90%).

[0259] 1 H NMR (300 MHz, CDCl3) δ 4.10 (s, 1H), 3.89-3.49 (m, J = 12.8, 11.3 Hz, 4H), 3.41 (s, 1H), 2.51-2.31 (m, 1H), 1.41 (s, 9H).

[0260] Third step: synthesis of compound 10-4

[0261]

[0262] At 25 ℃, to compound 10-3 (1.05 g) in dichloromethane (10 mL) solution was added hydrogen chloride dioxane solution (4 mol / L, 5 mL), stirring for 2 hours. The reaction was concentrated in vacuum to give crude compound 10-4 (1 g) was directly used in the next step reaction.

[0263] MS (ESI, m / z): 138.1 [M+H] + .

[0264] Fourth step: synthesis of compound 10-5

[0265]

[0266] Compound 10-4 (606 mg, 4.4 mmol, 1 eq), 1-2 (1.2 g, 4.86 mol, 1.1 eq), potassium carbonate (1.2 g, 13.3 mmol, 3 eq) and potassium iodide (146 mg, 0.88 mmol, 0.2 eq) were dissolved in DMF (10 mL), heated to 80 °C and stirred for 16 h. The reaction system was poured into water (50 mL) and extracted with ethyl acetate (40 mL x 3). All organic phases were combined, washed with saturated brine (30 mL*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was purified by flash silica gel column (gradient elution with methanol / dichloromethane 0-10%) to obtain compound 10-5 (490 mg, purity 85%, yield 38%) as an anhydrous oil.

[0267] MS (ESI, m / z): 296.2 [M+H] + .

[0268] The following steps were the same as Example 3 to obtain Example 10. See Table 1 for characterization spectrum analysis.

[0269] Example 11(090)

[0270] First step: synthesis of compound 11-2

[0271]

[0272] Compound 11-1 (900 mg, 3.53 mmol) was dissolved in tetrahydrofuran (10 ml) at 25 °C and under stirring conditions, and cooled to 0-5 °C under nitrogen replacement. BH3-THF solution (7.06 mmol, 1 mol / L, 2 eq) was slowly added to the reaction solution. The reaction was slowly raised to 25 °C and stirred for 2 h. The reaction progress was monitored by liquid chromatography-mass spectrometry (LCMS), and after the reaction was completed, the reaction was cooled to 0-5 °C, 10 ml of methanol was slowly added to the reaction solution, and then the reaction solution was heated to 65 °C and stirred for 2 h. The reaction solution was concentrated under reduced pressure to obtain compound 11-2 (colorless oil, 800 mg, yield 94%).

[0273] MS (ESI, m / z): 186.3 [M-56+H] + .

[0274] 1 H NMR (600 MHz, Chloroform-d) δ 3.92 (dd, J = 8.0, 3.0 Hz, 1H), 3.79 (dd, J = 10.8, 3.2 Hz, 1H), 3.76-3.67 (m, 2H), 3.51-3.46 (m, 1H), 3.41 (d, J = 11.4 Hz, 1H), 1.46 (s, 9H), 1.04 (s, 3H), 0.94 (s, 3H).

[0275] Second Step: Synthesis of compound 11-3

[0276]

[0277] Compound 11-2 (500 mg, 2.07 mmol) was dissolved in dichloromethane (30 ml) at 25 °C under stirring, then hydrochloric acid dioxane solution (3 ml, 4 mol / L) was added, the reaction solution was stirred at 25 °C for 2 hours. The reaction progress was monitored by thin layer chromatography. The reaction solution was concentrated under reduced pressure to obtain compound 11-3 (colorless oil, 440 mg, yield 100%).

[0278] Third Step: Synthesis of compound 11-4

[0279]

[0280] Compound 11-3 (440 mg, 2.48 mmol) was dissolved in N,N-dimethylformamide (5 ml) at 25 °C under stirring, then compound 1-2 (652.74 mg, 2.73 mmol, 1.2 eq), potassium carbonate (1.03 g, 7.44 mmol, 3 eq) and potassium iodide (83.00 mg, 0.50 mmol, 2 eq) were added in turn. The reaction was heated to 90 °C and stirred for 4 hours. The reaction was monitored by liquid chromatography-mass spectrometry, after the reaction was completed, the reaction system was cooled to 25 °C, 20 ml of water was added to the reaction solution, then extracted with ethyl acetate (20 ml*3), the organic phase was combined and extracted with saturated brine (30 ml*2), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (developing agent: dichloromethane / methanol = 10 / 1) to obtain compound 11-4 (yellow oil, 210 mg, yield 28%).

[0281] MS (ESI, m / z): 300.5 [M+H] + .

[0282] 1 H NMR (600 MHz, CDCl3) δ 4.22 (t, J = 7.1 Hz, 2H), 4.07 (s, 2H), 3.69-3.59 (m, 2H), 3.58-3.52 (m, 2H), 2.69 (s, 2H), 2.58 (s, 0H), 2.39-2.33 (m, 1H), 1.77-1.55 (m, 3H), 1.46-1.41 (m, 1H), 1.37-1.32 (m, 1H), 1.30 (t, J = 7.1 Hz, 3H), 1.27 (s, 1H), 1.05 (s, 3H), 0.96 (s, 3H).

[0283] The following steps are the same as Example 3 to obtain Example 11, see Table 1 for characterization spectrum analysis.

[0284] Example 12(089)

[0285] First step: synthesis of compound 12-2

[0286]

[0287] To a solution of triphenylphosphine bromide (5.4 g, 15 mmol, 1.5 eq) in THF (25 mL) was added t-BuOK tetrahydrofuran solution (16 mL, 16 mmol, 1.6 eq, 1 mol / L) dropwise under ice water bath 0 °C and nitrogen protection. After stirring for 30 min, a light yellow mixture was obtained, and compound 12-1 (2.4 g, 10 mmol, 1.0 eq) was added to it, and stirring was continued for 4 h. To the reaction mixture was added saturated aqueous ammonium chloride solution to quench, and ethyl acetate was extracted. The aqueous phase was removed, and the organic phase was combined and concentrated under reduced pressure. The residue was purified by flash silica gel column to obtain the product 12-2 (2.4 g) as a light yellow oil.

[0288] MS (ESI, m / z): 242.3 [(M+H)] + .

[0289] Second step: synthesis of compound 12-3

[0290]

[0291] To a solution of compound 12-2 in THF (20 mL) was added TMSCF3 (2.2 g, 15.5 mmol, 2.5 eq) and sodium iodide (1.9 g, 12.4 mmol, 2.0 eq) to obtain a light yellow mixture which was heated to 70 °C and stirred for 3 h. To the reaction mixture was added water (10 mL) to quench, and ethyl acetate (20 mL*2) was extracted. The aqueous phase was removed, and the organic phase was combined and concentrated under reduced pressure. The residue was purified by column chromatography to obtain the product 12-3 (1.4 g, yield 78%) as a light yellow oil.

[0292] MS (ESI, m / z): 292.4 [(M+H)] + .

[0293] Third step: synthesis of compound 12-4

[0294]

[0295] To a solution of compound 12-3 (1.4 g, 4.8 mmol, 1.0 eq) in THF (20 mL) was added lithium aluminum hydride (180 mg, 4.8 mmol, 1.0 eq) portionwise under ice water bath at 0 °C with nitrogen protection, the reaction mixture was allowed to naturally rise to room temperature 28 °C and stirred for another 2 hours. To the reaction solution was added water (180 mg), 10% NaOH aqueous solution (180 mg) and water (540 mg) successively to quench the reaction, stirred for 10 minutes, filtered, THF (10 mL) was added to wash the solid twice, the filtrate was mixed and concentrated under reduced pressure, the residue was purified by column chromatography to obtain compound 12-4 (900 mg).

[0296] MS (ESI, m / z): 264.3 [(M+H)] + .

[0297] Fourth step: synthesis of compound 12-5

[0298]

[0299] To a solution of compound 12-4 (900 mg, 3.4 mmol) in dichloromethane (5 mL) was added HCl-dioxane solution (2.0 mL, 8.0 mmol, 4 mol / L) under temperature control at 20-25 °C, the reaction mixture was stirred for 2 hours. To the reaction solution was concentrated under reduced pressure to obtain compound 12-5 (650 mg).

[0300] MS (ESI, m / z): 164.3 [(M+H)] + .

[0301] Fifth step: synthesis of example 12

[0302]

[0303] To a solution of compound 12-5 (500 mg, 2.5 mmol, 1.0 eq) and 1-2 (630 mg, 2.6 mmol, 1.1 eq) in DMF (5 mL) was added potassium carbonate (1.1 g, 8.0 mmol, 3.0 eq) and potassium iodide (80 mg, 0.5 mmol, 0.2 eq), the resulting mixture was stirred at 90 °C for 4 hours. The reaction mixture was filtered, washed with dichloromethane (20 mL), the combined organic phase was concentrated under reduced pressure, the residue was purified by flash silica gel column elution (methanol:dichloromethane = 0-15%, with the addition of 0.1% ammonia water) to obtain the product 12-6 (640 mg, yield 79%) as a light brown oil.

[0304] MS (ESI, m / z): 322.4 [M+H]+.

[0305] 1H NMR (300 MHz, Chloroform-d) δ 4.22 (q, J = 7.1 Hz, 2H), 4.06 (s, 2H), 3.76-3.65 (m, 1H), 3.61-3.50 (m, 2H), 3.46-3.18 (m, 1H), 3.05-2.63 (m, 3H), 2.49-2.09 (m, 3H), 1.85 (ddt, J = 13.2, 9.8, 5.0 Hz, 1H), 1.78-1.52 (m, 4H), 1.38-1.15 (m, 5H).

[0306] 19 F NMR (282 MHz, Chloroform-d) δ -135.19 - -139.76 (m).

[0307] The following steps are the same as Example 3 to obtain Example 12, see Table 1 for characterization spectrum analysis.

[0308] Example 13 (085)

[0309] First step: synthesis of compound 13-2

[0310]

[0311] To a solution of starting material 13-1 (15.2 g, 100 mmol, 1.0 eq) and ethyl acrylate (12 g, 120 mmol, 1.2 eq) in MeCN (100 mL) was added benzyltrimethylammonium hydroxide (4.2 g, 10 mmol, 40% aqueous solution, 0.1 eq) in portions under ice water bath 5 °C and nitrogen protection. The reaction solution was allowed to naturally rise to room temperature and stirred for 12 hours. The reaction mixture was poured into water (200 mL), extracted with ethyl acetate (50 mL*2), the combined organic phase was concentrated under reduced pressure, and the residue was purified by flash column chromatography (ethyl acetate: petroleum ether = 0-30% gradient elution) to obtain the product 13-2 (10.6 g) as colorless oil.

[0312] 1 H NMR (300 MHz, Chloroform-d) δ 7.39-7.27 (m, 5H), 4.57 (s, 2H), 4.15 (q, J = 7.1 Hz, 2H), 3.77 (t, J = 6.5 Hz, 2H), 3.69-3.58 (m, 4H), 2.60 (t, J = 6.5 Hz, 2H), 1.26 (t, J = 7.1 Hz, 3H).

[0313] Second step: synthesis of compound 13-3

[0314]

[0315] To a solution of intermediate 13-2 (10.6 g, 42.0 mmol, 1.0 eq) in THF (100 mL) was added lithium aluminum hydride (1.8 g, 47.4 mmol, 1 eq) in portions under ice water bath 0 °C and nitrogen protection. The reaction mixture was allowed to warm to room temperature naturally and stirred for 3 hours. After the reaction was completed, water (1.8 g), NaOH (1.8 g, 10% aqueous solution) and water (5.4 g) were added to the reaction mixture in turn, stirred for 10 minutes, filtered, THF (50 mL*2) was added to wash the solid, and the combined organic phase was concentrated under reduced pressure to obtain the crude compound 13-3 (8.8 g) which was directly used in the next step.

[0316] 1 H NMR (400 MHz, CDCl3) δ 7.40-7.29 (m, 5H), 4.60 (s, 2H), 4.23 (q, J = 7.1 Hz, 2H), 4.08 (d, J = 2.2 Hz, 2H), 3.81-3.54 (m, 8H), 2.00-1.90 (m, 2H), 1.33-1.28 (m, 3H).

[0317] Third step: synthesis of compound 13-4

[0318]

[0319] To a solution of intermediate 13-3 (8.8 g, 41.8 mmol, 1.0 eq) in THF (100 mL) was added NaH (2.5 g, 62.5 mmol, 60% content, 1.5 eq) in portions under ice water bath 0 °C and nitrogen protection. After stirring for 30 minutes, ethyl bromoacetate (8.4 g, 50.3 mmol, 1.2 eq) was added, the reaction mixture was allowed to warm to room temperature naturally, and then heated to 55 °C and stirred for 4 hours. The reaction mixture was poured into dilute aqueous hydrochloric acid solution (100 mL, 0.5 M) to quench, and MTBE (100 mL) was added to extract, the combined organic phase was concentrated under reduced pressure, and the residue was purified by flash column chromatography (ethyl acetate: petroleum ether = 0-10% gradient elution) to obtain the product 13-4 (10 g) as colorless oil.

[0320] 1 H NMR (400 MHz, CDCl3) δ 7.40-7.29 (m, 5H), 4.60 (s, 2H), 4.23 (q, J = 7.1 Hz, 2H), 4.08 (d, J = 2.2 Hz, 2H), 3.81-3.54 (m, 8H), 2.00-1.90 (m, 2H), 1.33-1.28 (m, 3H).

[0321] Fourth step: synthesis of compound 13-5

[0322]

[0323] To a solution of intermediate 13-4 (7.1 g, 24 mmol, 1.0 eq) in ethanol was added Pd / C (1.0 g, 5% w / w) under nitrogen protection at room temperature 20-25 °C, the mixture was stirred under hydrogen atmosphere for 16 hours. After the reaction was completed, the catalyst was filtered and washed with methanol (50 mL), and the filtrate was concentrated under reduced pressure to give compound 13-5 (4.5 g) as a colorless oil, which was used directly in the next step.

[0324] MS (ESI, m / z): 211.3 [M+H] + .

[0325] Step 5: Synthesis of compound 13-6

[0326]

[0327] To a solution of intermediate 13-5 (206 mg, 1.0 mmol, 1.0 eq) in pyridine (2 mL) was added p-toluenesulfonyl chloride (210 mg, 1.1 mmol, 1.1 eq) under nitrogen protection at room temperature 20-25 °C, the resulting light yellow reaction solution was stirred for 2 hours, quenched by adding water (2 mL), extracted with ethyl acetate (2 mL*2), the organic phase was combined and concentrated under reduced pressure, the residue was purified by flash column chromatography to give product 13-6 (220 mg, yield 61%) as a colorless oil.

[0328] MS (ESI, m / z): 361.3 [M+H] +

[0329] 1 H NMR (600 MHz, CDC13) δ 7.82 (d, J = 8.2 Hz, 2H), 7.36 (d, J = 8.0 Hz, 2H), 4.23 (dq, J = 13.2, 6.8 Hz, 2H), 4.17 (q, J = 4.2, 3.7 Hz, 2H), 4.06 (s, 2H), 3.63 (q, J = 5.1 Hz, 2H), 3.58 (t, J = 6.3 Hz, 2H), 3.53 (t, J = 6.3 Hz, 2H), 2.47 (s, 3H), 1.84 (p, J = 6.3 Hz, 2H), 1.31 (t, J = 7.1 Hz, 3H).

[0330] Step 6: Synthesis of compound 13-7

[0331]

[0332] To a solution of intermediate 13-6 (220 mg, 0.6 mmol, 1.0 eq) and L-prolinol (62 g, 0.61 mmol, 1.0 eq) in DMF (2 mL) was added potassium carbonate (250 mg, 1.6 mmol, 3.0 eq) and potassium iodide (20 mg, 0.1 mmol, 0.2 eq), and the reaction mixture was stirred at room temperature 22 °C ~ 31 °C for 16 hours. The reaction mixture was filtered, washed with dichloromethane (20 mL), and the combined organic phases were concentrated under reduced pressure. The residue was purified by flash column chromatography (methanol:dichloromethane = 0 ~ 15%, gradient elution with the addition of 0.1% aqueous ammonia) to give the product 13-7 (130 mg) as a light brown oil.

[0333] MS (ESI, m / z): 290.4 [M+H] + .

[0334] 1 H NMR (600 MHz, Chloroform-d) δ 4.23 (q, J = 7.1 Hz, 2H), 4.08 (s, 2H), 3.68 - 3.55 (m, 9H), 3.48 - 3.39 (m, 1H), 3.30 (s, 1H), 3.08 - 2.99 (m, 1H), 2.81 (s, 1H), 2.71 - 2.64 (m, 1H), 2.52 - 2.44 (m, 1H), 1.91 (tt, J = 10.1, 4.9 Hz, 4H), 1.84 - 1.70 (m, 4H), 1.30 (t, J = 7.1 Hz, 3H).

[0335] The following steps were the same as in Example 3 to give Example 13. See Table 1 for characterization of the spectra.

[0336] Example 14 (091)

[0337] First step: synthesis of compound 14-1

[0338]

[0339] To a solution of intermediate 14-1 (1 g, 4.1 mmol, 1.0 eq) in THF (10 mL) was added BH3-THF solution (8.2 mL, 8.2 mmol, 2 eq, 1 mol / L) dropwise under ice water bath 0 °C and nitrogen protection, then the reaction solution was slowly warmed to room temperature and stirred for 2 hours. The reaction solution was cooled to 0 °C, methanol (5 mL) was slowly added, and then heated to 60 °C and stirred for 1 hour to quench. The reaction solution was concentrated, dissolved in dichloromethane, washed with water and saturated brine once respectively, and the organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to give white solid compound 14-2 (937 mg, crude), which was directly used in the next step.

[0340] MS (ESI, m / z): 214.2 [M+H] + .

[0341] Second step: synthesis of compound 14-2

[0342]

[0343] To a solution of intermediate 14-2 (1 g) in dichloromethane (10 mL) was added hydrogen chloride (10 mL, 4 mol / L solution in dioxane) at room temperature 25 °C, the resulting reaction solution was stirred at 25 °C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain white solid compound 14-3 (613 mg), which was directly used in the next step reaction.

[0344] MS (ESI, m / z): 114.2 [(M+H)] + .

[0345] Third step: synthesis of compound 14-3

[0346]

[0347] To a solution of intermediate 14-3 (613 mg, 5.4 mmol, 1 eq) in DMF (5 mL) was added compound 1-2 (1.2 g, 5.4 mmol, 1.0 eq), potassium carbonate (2 g, 14.7 mmol, 3 eq) and potassium iodide (43 mg, 1 mmol, 2 eq) successively at room temperature 25 °C, the resulting mixture was heated to 90 °C and stirred for 3 hours. After cooling to 25 °C, water (20 mL) was added to the reaction solution, which was extracted with ethyl acetate (20 mL*3), the organic phase was washed with saturated brine (30 mL*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash silica gel column (eluent: dichloromethane / methanol = 10 / 1) to obtain compound 14-4 (375 mg, yield 28%).

[0348] MS (ESI, m / z): 272.2 [M+H] + .

[0349] The following steps were the same as Example 3 to obtain Example 14, and the characterization spectrum was analyzed in Table 1.

[0350] Example 15(096)

[0351] First step: synthesis of compound 15-2

[0352]

[0353] Control 0-5 ℃, to compound 15-1 (1 g, 4.1 mmol, 1 eq) of tetrahydrofuran (10 mL) solution dropwise BH3-THF (, 8.2 mL, 8.2 mmol, 1 mol / L, 2 eq), after dropwise addition slowly to room temperature after stirring for 2 hours. The reaction was monitored by liquid chromatography-mass spectrometry, after the reaction was completed, the system was reduced to 0 ℃, slowly dropwise methanol (5 mL) reaction to 60 ℃ stirring 1 hour quenching. Concentration, then dissolved with dichloromethane, the obtained organic phase was washed with water and saturated brine once. Dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated in vacuum to obtain white solid compound 15-2 crude (1 g), which can be directly used in the next step reaction.

[0354] MS (ESI, m / z): 228.1 [M+H] + .

[0355] Second step: synthesis of compound 15-3

[0356]

[0357] At 25 ℃, to the dichloromethane solution (10 mL) of intermediate 15-2 (1 g) was added HCl dioxane solution (4 mol / L, 10 mL). The reaction liquid was stirred at 25 ℃ for 2 hours. The reaction was monitored by liquid chromatography-mass spectrometry, after the reaction was completed, the solution was vacuum concentrated to obtain white solid compound 15-3 crude (630 mg), which can be directly used in the next step reaction.

[0358] MS (ESI, m / z): 128.2 [(M+H)] + .

[0359] Third step: synthesis of compound 15-4

[0360]

[0361] Compound 15-3 (256 mg, 2.016 mmol, 1.6 eq), compound 1-2 (300 mg, 1.26 mol, 1 eq), potassium carbonate (521 mg, 3.78 mmol, 3 eq) and potassium iodide (42 mg, 0.252 mmol, 0.2 eq) were added to DMF (5 mL), heated to 90 ℃ and stirred for 3 h, the reaction was monitored by liquid chromatography-mass spectrometry. After the reaction was completed, the reaction system was poured into water (20 mL), extracted with ethyl acetate (15 mL x 3), combined all organic phases, washed with saturated brine (15 mL x 2), dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated in vacuum. The residue was purified by flash silica gel column (methanol / dichloromethane 0-10% gradient elution) to obtain compound 15-4 (119 mg, purity 80%, yield 33%) in yellow oil.

[0362] MS (ESI, m / z): 286.4 [M+H] + .

[0363] The following steps are the same as Example 3 to obtain Example 15, see Table 1 for characterization of the spectrum.

[0364] Example 16 (098)

[0365] Step 1: synthesis of compound 16-2

[0366]

[0367] Compound 16-1 (700 mg, 2.93 mmol, 2 eq) was dissolved in acetonitrile (15 ml) at 25 °C, then compound 1-2 (570 mg, 1.46 mmol, 1 eq) and potassium carbonate (1.6 g, 5.8 mmol, 4 eq) were added. The reaction was stirred at 25 °C for 16 hours. The reaction was monitored by TLC. After the reaction was completed, 200 ml of water was added to the reaction solution, extracted with ethyl acetate (50 ml*3), the organic phase was combined and washed with saturated brine (30 ml*2), the mixed organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 10 / 1) to obtain compound 16-2 (colorless oil, 880 mg, yield 62%).

[0368] The following steps are the same as Example 1 to obtain Example 16, see Table 1 for characterization of the spectrum.

[0369] Example 17 (099)

[0370] Step 1: synthesis of compound 17-2

[0371]

[0372] Lithium aluminum hydride (4.2 g, 112 mmol, 1.3 eq) was added to tetrahydrofuran (200 ml) at 0 °C under nitrogen protection, then compound 17-1 solid (15 g, 86.1 mmol, 1 eq) was slowly added. The reaction was stirred at 25 °C for 2 hours. The reaction was monitored by TLC. After the reaction was completed, the temperature was lowered to 0 °C, 5 ml of water, 5 ml of 10% sodium hydroxide aqueous solution and 15 ml of water were slowly added to the reaction solution in turn, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 5 / 1) to obtain compound 17-2 (colorless oil, 12 g, yield 96%).

[0373] Step 2: Synthesis of compound 17-3

[0374]

[0375] Compound 17-2 (5 g, 34.2 mmol, 1 eq) was dissolved in tetrahydrofuran (80 ml) at 25 °C under stirring, and then sodium hydride (1.5 g, 37.6 mmol, 1.1 eq) was slowly added after cooling to 0 °C. The reaction solution was stirred at 0 °C for 0.5 h, and then compound 3 was added. The reaction solution was slowly warmed to room temperature and stirred at 25 °C for 1 h. The reaction was monitored by TLC. After the reaction was completed, the reaction system was cooled to 25 °C, 100 ml of saturated aqueous ammonium chloride solution was added to the reaction solution, and then extracted with ethyl acetate (50 ml*3). The organic phase was combined and extracted with saturated brine (30 ml*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 1) to obtain compound 17-3 (yellow oil, 3 g, yield 38%).

[0376] Step 3: Synthesis of compound 17-4

[0377]

[0378] Compound 17-4 (3 g, 12.9 mmol, 1 eq) was dissolved in acetone (20 ml) and water (1 ml) at 25 °C, and then p-toluenesulfonic acid monohydrate (245 mg, 1.29 mmol, 0.1 eq) was added. The reaction solution was stirred at 25 °C for 1 h. The reaction was monitored by TLC. Saturated aqueous sodium carbonate solution was added to the reaction solution to adjust the pH to about 8, and then extracted with ethyl acetate (30 ml*3). The organic phase was combined and extracted with saturated brine (30 ml*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 17-5 (yellow oil, 2.24 g, crude).

[0379] Step 4: Synthesis of compound 17-5

[0380]

[0381] Compound 17-5 (2.24 g, 12 mmol, 1 eq) was dissolved in 1,2-dichloroethane (20 ml) under stirring at 25 °C, then L-prolinol (1.35 g, 13.2 mmol, 1.1 eq) was added, and acetic acid was added dropwise, and the pH of the reaction solution was adjusted to 5. After stirring for 30 min, NaBH(OAc)3(6.3 g, 30 mmol, 3.0 eq) was added, and the reaction was stirred at 25 °C for 1 h. The reaction process was monitored by TLC. After the reaction was completed, 100 ml of water was added to the reaction solution, which was extracted with dichloromethane / methanol = 10 / 1 (30 ml*3). The organic phase was combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 10 / 1) to obtain compound 17-6 (yellow oil, 2.1 g, yield 65%).

[0382] The following steps were the same as in Example 1 to obtain Example 17, and the characterization spectrum is shown in Table 1.

[0383] Example 18(097)

[0384] First step: synthesis of compound 18-2

[0385]

[0386] Under ice water bath at 0 °C and nitrogen protection, acetic acid (0.6 g, 10 mmol, 1.0 eq) was added to a dichloromethane solution of raw material 18-1 (2.1 g, 10 mmol, 1.0 eq) and L-prolinol (1.0 g, 10 mmol, 1.0 eq), and stirred for 30 min, then NaBH(OAc)3(6.3 g, 30 mmol, 3.0 eq) was added, and the obtained white mixture was stirred for 4 h. The pH of the reaction solution was adjusted to ~7 by adding a protective sodium bicarbonate aqueous solution, the water phase was removed, and the organic phase was concentrated under reduced pressure. The residue 18-2 (2.3 g, 80% yield) was directly used in the next step reaction.

[0387] 1 H NMR (600 MHz, CDC13) δ 7.37 (d, J = 7.9 Hz, 2H), 7.29 (d, J = 8.1 Hz, 2H), 4.30 (d, J = 13.1 Hz, 1H), 3.80 (d, J = 13.1 Hz, 1H), 3.77-3.72 (m, 2H), 3.71 (s, 3H), 3.64 (s, 2H), 3.29-3.13 (m, 2H), 2.70-2.60 (m, 1H), 2.01-1.94 (m, 1H), 1.93-1.77 (m, 3H).

[0388] The following steps were the same as Example 3 to obtain Example 18, and the characterization spectrum was analyzed as shown in Table 1.

[0389] Example 19 (100)

[0390] First step: synthesis of compound 19-2

[0391]

[0392] Under ice water bath 0°C and nitrogen protection, to the raw material 19-1 (2.1 g, 10 mmol, 1.0 eq) and L-prolinol (1.0 g, 10 mmol, 1.0 eq) in dichloromethane solution, acetic acid (0.6 g, 10 mmol, 1.0 eq) was added, after stirring for 30 minutes, NaBH(OAc)3(6.3 g, 30 mmol, 3.0 eq) was added, and the reaction mixture was stirred for 4 hours. To the reaction solution, sodium bicarbonate aqueous solution was added to adjust pH ~ 7, the water phase was removed, the organic phase was concentrated under reduced pressure, and the residue 19-2 (2.7 g, yield 90%) was directly used in the next step reaction.

[0393] 1 H NMR (600 MHz, CDCl3) δ 7.36 (d, J = 8.6 Hz, 2H), 7.00 (dd, J = 45.1, 7.4 Hz, 2H), 6.91 (d, J = 8.6 Hz, 2H), 4.63 (s, 2H), 4.36-4.23 (m, 3H), 3.89 (d, J = 13.1 Hz, 1H), 3.80-3.72 (m, 2H), 3.30 (dddd, J = 15.3, 9.0, 6.5, 3.9 Hz, 2H), 2.76-2.68 (m, 1H), 2.05-1.90 (m, 5H), 1.84 (tq, J = 12.1, 5.6, 4.5 Hz, 2H), 1.31 (t, J = 7.1 Hz, 3H).

[0394] The following steps were the same as Example 1 to obtain Example 19, and the characterization spectrum was analyzed as shown in Table 1.

[0395] Example 20 (101)

[0396] First step: synthesis of compound 20-3

[0397]

[0398] To a solution of compound 20-1 (280 mg, 1.8 mol, 1 eq) in tetrahydrofuran (5 mL) was added NaBH3(OAC)3 (667 mg, 3.1 mmol, 2 eq), compound 20-2 (300 mg, 2.4 mmol, 1.5 eq) and acetic acid (0.5 mL, 7.8 mmol, 5 eq) sequentially under ice water bath 0 °C and nitrogen protection. The mixture was allowed to warm to room temperature 27 °C and stirred for 16 hours. The reaction mixture was poured into water (20 mL) and extracted with ethyl acetate (8 mL*3). The organic phase was combined, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel, methanol / dichloromethane 0-10% gradient) to give compound 20-3 (249 mg, purity 85%, yield 55%) as brown oil.

[0399] MS (ESI, m / z): 290.2 [M+H] + .

[0400] The following steps were the same as Example 1 to give Example 20. See Table 1 for characterization of the spectra.

[0401] Example 21 (102)

[0402] First step: synthesis of compound 21-2

[0403]

[0404] To a solution of compound 21-1 (1.0 g, 4.1 mmol, 1 eq) in tetrahydrofuran (10 mL) was added BH3-THF solution (8.2 mL, 8.2 mmol, 2 eq, 1 mol / L) dropwise under ice water bath 0 °C and nitrogen protection. The reaction mixture was allowed to warm to room temperature 26 °C and stirred for 2 hours. The reaction mixture was cooled to 0 °C and methanol (5 mL) was added dropwise. The reaction mixture was heated to 60 °C and stirred for 1 hour. The reaction mixture was concentrated under reduced pressure. The residue was dissolved in dichloromethane (10 mL) and washed with water (10 mL) and saturated brine (10 mL) sequentially. The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to give compound 21-2 (1 g, crude) as white solid, which was used directly in the next step.

[0405] MS (ESI, m / z): 228.1 [M+H] + .

[0406] Second step: synthesis of compound 21-3

[0407]

[0408] To a solution of compound 21-2 (1 g, crude) in dichloromethane (10 mL) was added HC1 dioxane solution (10 mL, 4 mol / L) at room temperature 25 °C, the reaction mixture was stirred for 2 hours. The reaction solution was concentrated under reduced pressure to obtain white solid compound 21-3 (630 mg, crude), which was directly used in the next step reaction.

[0409] MS (ESI, m / z): 128.2 [(M+H)] + .

[0410] Third step: synthesis of compound 21-4

[0411]

[0412] To a solution of compound 21-3 (347 mg, 2.1 mmol, 1.0 eq) and compound 1-2 (560 mg, 2.3 mol, 1 eq) in MeCN (5 mL) was added potassium carbonate (585 mg, 4.3 mmol, 3 eq) and potassium iodide (70 mg, 0.21 mmol, 0.2 eq), the reaction mixture was heated to 90 °C and stirred for 3 hours. Water (5 mL) was added to the reaction system, and ethyl acetate (10 mL*3) was added to extract, the organic phase was combined and washed with saturated brine (15 mL*2), dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by flash column chromatography (silica gel, 0-10% gradient of methanol / dichloromethane) to obtain compound 21-4 (305 mg, yield 50%) as yellow oil.

[0413] MS (ESI, m / z): 286.4 [M+H] + .

[0414] The following steps were the same as example 3 to obtain example 21, and the characterization spectrum was analyzed in table 1.

[0415] Example 22 (105)

[0416] First step: synthesis of compound 22-1

[0417]

[0418] Compound 17-2 (9 g, 61 mmol, 1 eq) was dissolved in tetrahydrofuran (80 mL) under stirring at room temperature 25 °C, and then the system was cooled to 0 °C, NaH (2.6 g, 68 mmol, 1.1 eq) was slowly added, the mixture was stirred for 0.5 h, then compound 3-bromopropynyl (7.5 g, 68 mmol, 1.1 eq) was added, the reaction solution was slowly raised to room temperature 25 °C, and stirred for 1 h. The reaction solution was quenched by adding saturated aqueous ammonium chloride solution (100 mL), and then extracted with ethyl acetate (50 mL*3), the organic phase was washed with saturated brine (30 mL*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column (eluent: petroleum ether / ethyl acetate = 1 / 1) to obtain compound 22-1 (8.46 g, yield 74%) in yellow oil.

[0419] Second step: synthesis of compound 22-2

[0420]

[0421] Compound 22-1 (8.46 g, 45.9 mmol, 1.0 eq) was dissolved in a mixed solution of acetone (80 mL) and water (4 mL) under stirring at room temperature 25 °C, then p-toluenesulfonic acid monohydrate (873 mg, 4.6 mmol, 0.1 eq) was added, and the obtained compound was stirred for 1 h. The pH of the reaction solution was adjusted to about 8 by adding saturated aqueous sodium carbonate solution, and then extracted with ethyl acetate (100 mL*3), the organic phase was extracted with saturated brine (100 mL*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 22-2 (6.25 g, crude product was used directly in the next step) in yellow oil.

[0422] Third step: synthesis of compound 22-3

[0423]

[0424] Compound 22-2 (6.25 g, 45.2 mmol, 1 eq) was dissolved in 1,2-dichloroethane (60 mL) under stirring at room temperature 25 °C, then L-prolinol (5.0 g, 49.7 mmol, 1.1 eq) was added, acetic acid was added dropwise to adjust the pH to about 5, then sodium borohydride acetate (1.9 g, 90.4 mmol, 2 eq) was added, and the reaction mixture was stirred for 1 h. The reaction solution was added with aqueous sodium carbonate solution (200 mL), and then extracted with dichloromethane / methanol mixed solvent (10 / 1, 50 mL*3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column (eluent: dichloromethane / methanol = 10 / 1) to obtain compound 22-3 (2.5 g, yield 23%) in yellow oil.

[0425] Fourth step: synthesis of compound 22-4

[0426]

[0427] After compound 22-3 (2.5 g, 7.7 mmol, 1 eq) was dissolved in 1,4-dioxane (20 mL) under stirring at 25 °C, compound C-2 (1.73 g, 7.7 mmol, 1 eq), cesium carbonate (7.5 g, 23.1 mmol, 3 eq) and cuprous cyanide (206 mg, 2.3 mmol, 0.3 eq) were added in turn, and the mixture was heated to 85 °C and stirred for 12 h. After cooling, water (100 mL) was added to the reaction solution, which was extracted with ethyl acetate (50 mL*5). The organic phase was combined and washed with saturated brine (30 mL*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column (eluent: dichloromethane / methanol = 10 / 1) to obtain compound 22-4 (2.1 g, yield 26%) as a yellow solid.

[0428] MS (ESI, m / z): 716.7 [M+H] + .

[0429] Fifth step: synthesis of compound 22-5

[0430]

[0431] Compound 22-4 (500 mg, 0.96 mmol, 1 eq) was dissolved in ethyl acetate / water (5 mL / 5 mL) under nitrogen protection at room temperature 25 °C, and then compound 22-9 (270 mg, 1.16 mmol, 1.2 eq), anhydrous copper sulfate (77 mg, 0.48 mmol, 0.5 eq) and sodium ascorbate (191 mg, 0.96 mmol, 1 eq) were added. The resulting mixture was stirred for 2 h. The reaction solution was filtered, extracted with ethyl acetate (20 mL*3), and the organic phase was combined and washed with saturated brine (20 mL*2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column (eluent: dichloromethane / methanol = 10 / 1) to obtain compound 22-5 (250 m, yield 34%) as a yellow oil.

[0432] MS (ESI, m / z): 751.7 [M+H] + .

[0433] Sixth step: synthesis of compound 22-6

[0434]

[0435] To a solution of compound 22-5 (250 mg, 0.33 mmol, 1 eq) and compound C-4 (240 mg, 0.66 mmol, 2 eq) in 1,4-dioxane (10 mL) and water (0.5 mL) was added potassium phosphate (212 mg, 1.00 mmol, 3 eq) under nitrogen protection at room temperature 25 °C, replaced with nitrogen for three times, added catalyst Pd Cata CXium A Pd G3((24 mg, 0.03 mmol, 0.1 eq), the mixture was heated to 95 °C and stirred for 12 hours. After cooling, water (50 mL) was added to the reaction solution, extracted with ethyl acetate (30 mL*5), the organic phase was washed with saturated brine (30 mL*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column (eluent: dichloromethane / methanol = 10 / 1) to obtain compound 22-6 (150 mg, yield 47.4%) as a yellow solid.

[0436] MS (ESI, m / z): 949.7 [M+H] + .

[0437] Seventh step: synthesis of compound 22-7

[0438]

[0439] Compound 22-6 (150 mg, 0.158 mmol, 1 eq) was dissolved in acetonitrile (5 mL) at room temperature 25 °C, and then lithium bromide (273 mg, 3.16 mmol, 20 eq), triethylamine (47.8 mg, 0.474 mmol, 3 eq) and water (8.5 mg, 0.474 mmol, 3 eq) were added in sequence. The reaction mixture was stirred for 12 hours. To the reaction solution was slowly added 10% aqueous citric acid solution to pH ~ 4, and then extracted with dichloromethane / methanol = 10 / 1 (10 mL*3). The organic phase was combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 22-7 (140 mg) as a white solid.

[0440] MS (ESI, m / z): 859.6 [M+H] + .

[0441] Eighth step: synthesis of compound 22-8

[0442]

[0443] Compound 22-7 (140 mg, 0.16 mmol, 1 eq) was dissolved in dichloromethane (3 mL) at 25 °C under stirring, then compound C-6 (81 mg, 0.24 mmol, 1.5 eq), HATU (68.6 mg, 0.24 mmol, 1.5 eq) and N-methylimidazole (66.8 mg, 0.85 mmol, 3 eq) were added successively. The reaction mixture was stirred for 2 h. Water (15 mL) was added to the reaction solution, which was then extracted with ethyl acetate (15 mL*5). The organic phase was combined and washed with saturated brine (30 mL*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column (elution development agent: dichloromethane / methanol = 10 / 1) to obtain compound 22-8 (100 mg, yield 52%) as a yellow solid.

[0444] MS (ESI, m / z): 1172.6 [M+H] + .

[0445] Ninth step: synthesis of example 22

[0446]

[0447] Compound 22-8 (100 mg, 0.17 mmol) was dissolved in dichloromethane (1 mL) at 25 °C, and a hydrochloric acid dioxane solution (0.5 mL, 4 mol / L) was added. The reaction mixture was stirred for 1 h. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by pre-HPLC to obtain example 22 (6.5 mg, white solid formate, yield 6.7%).

[0448] MS (ESI, m / z): 1128.7 [M+H] + .

[0449] 1H NMR (600 MHz, Methanol-d4) δ 9.29 (d, J = 6.0 Hz, 1H), 8.92 (d, J = 12 Hz, 1H), 8.20 - 8.06 (m, 1H), 7.75 - 7.65 (m, 1H), 7.52 - 7.37 (m, 4H), 7.36 - 7.23 (m, 2H), 7.14 - 7.06 (m, 1H), 5.37 - 5.02 (m, 3H), 4.70 - 4.31 (m, 9H), 4.01 (s, 1H), 3.93 - 3.78 (m, 2H), 3.67 - 3.44 (m, 5H), 2.60 - 2.50 (m, 2H), 2.45 - 2.35 (m, 2H), 2.32 - 2.03 (m, 9H), 1.93 - 1.85 (m, 1H), 1.88 (s, 1H), 1.85 - 1.75 (m, 2H), 1.68 (d, J = 6.0 Hz, 7H), 1.55 - 1.47 (m, 2H), 1.33 - 1.29 (m, 4H), 1.17 - 1.07 (m, 3H), 0.86 - 0.79 (m, 3H), 0.78 - 0.71 (m, 3H).

[0450] Example 23 (106)

[0451] First step:

[0452]

[0453] After compound 22-7 (50 mg, 0.06 mmol, 1 eq) was dissolved in dichloromethane (3 mL) under stirring at 25 °C, compound 23-1 (40 mg, 0.12 mmol, 2 eq), HATU (31 mg, 0.12 mmol, 2 eq) and N-methylimidazole (25 mg, 0.3 mmol, 5 eq) were added successively. The resulting mixture was stirred for 2 hours. Water (15 mL) was added to the reaction solution, which was extracted with ethyl acetate (15 mL*5). The organic phase was washed with saturated brine (30 mL*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column (eluent: dichloromethane / methanol = 10 / 1) to obtain compound 23-2 (50 mg, yield 72%) as a yellow solid.

[0454] MS (ESI, m / z): 1188.6 [M+H] + .

[0455] Second step: synthesis of compound 23

[0456]

[0457] Compound 23-2 (50 mg, 0.04 mmol) was dissolved in dichloromethane (1 mL) at 25 °C, then HCl dioxane solution (0.5 mL, 4 mol / L) was added, stirred for 1 hour. The reaction solution was directly concentrated under reduced pressure, and the obtained crude product was purified by Yongpre-HPLC to obtain white solid compound 23 (9 mg, HCOOH salt, yield 18.7%).

[0458] MS (ESI, m / z): 1144.5 [M+H] + .

[0459] 1 H NMR (600 MHz, DMSO-d6) δ 10.16 (d, J = 162 Hz, 1H), 9.35-9.25 (m, 1H), 9.25-8.85 (m, 6H), 8.57-8.31 (m, 1H), 8.19-8.08 (m, 1H), 7.79-7.70 (m, 1H), 7.50-7.41 (m, 1H), 7.40-7.32 (m, 3H), 7.31-7.03 (m, 2H), 5.41-5.27 (m, 1H), 5.02-4.73 (m, 2H), 4.65-4.48 (m, 6H), 4.46-4.19 (m, 2H), 4.17-4.04 (m, 1H), 3.91-3.72 (m, 2H), 3.70-3.53 (m, 5H), 3.10-3.00 (m, 2H), 2.45-2.4 (m, 4H), 2.34-2.30 (m, 2H), 2.23 (s, 2H), 2.15-2.10 (m, 2H), 1.98-1.78 (m, 5H), 1.82-1.62 (m, 4H), 1.54 (d, J = 6.0 Hz, 6H), 1.24 (s, 1H), 1.20-1.14 (m, 3H), 1.06-0.95 (m, 3H), 0.86-0.84 (m, 1H), 0.80-0.73 (m, 3H), 0.69-0.53 (m, 3H).

[0460] Example 24 (107)

[0461] First Step: Synthesis of compound 24-2

[0462]

[0463] To compound 24-1 (790 mg, 3.3 mmol, 1.2 eq) and compound 1-2 (450 mg, 2.75 mmol, 1 eq) in acetonitrile (10 mL) was added potassium carbonate (1.1 g, 8.3 mmol, 3 eq) and potassium iodide (46 mg, 0.3 mmol, 0.1 eq) at 25 °C under stirring. The reaction was stirred at 80 °C for 4 h. After the reaction was cooled to room temperature, water (200 mL) was added and the mixture was extracted with ethyl acetate (50 mL*3). The organic phase was combined and washed with saturated brine (30 mL*2), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (dichloromethane / methanol = 10 / 1) to give compound 24-2 (340 mg, yield 43%) as yellow oil. MS (ESI, m / z): 286.4 [M+H] + .

[0464] 1 H NMR (600 MHz, Chloroform-d) δ 4.24 (q, J = 7.1 Hz, 2H), 4.08 (s, 2H), 3.77 - 3.69 (m, 1H), 3.62 - 3.46 (m, 4H), 3.01 - 2.81 (m, 3H), 2.59 (d, J = 9.3 Hz, 1H), 2.44 (s, 1H), 1.98 (dd, J = 12.6, 8.6 Hz, 1H), 1.73 (dt, J = 13.0, 6.5 Hz, 2H), 1.65 (s, 3H), 1.31 (t, J = 7.1 Hz, 3H), 0.66 - 0.51 (m, 4H).

[0465] Second step: synthesis of compound 24-3

[0466]

[0467] Compound 24-2 (300 mg, 1.1 mmol, 1.5 eq) was dissolved in 1,4-dioxane (5 mL) at 25 °C under stirring, and then compound C-2 (232 mg, 0.7 mmol, 1 eq), cesium carbonate (685 mg, 2.1 mmol, 3 eq) and cuprous cyanide (19 mg, 0.21 mmol, 0.3 eq) were added successively. The reaction was heated to 80 °C and stirred for 12 h. The reaction was cooled to 25 °C, and then water (200 mL) was added to the reaction solution, which was extracted with ethyl acetate (50 mL*3). The organic phase was washed with saturated brine (30 mL*2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: dichloromethane / methanol = 10 / 1) to give compound 24-3 (170 mg, yield 42%) as a yellow oil. MS (ESI, m / z): 580.5 [M+H] + .

[0468] The following steps were the same as in Example 1 to give Example 24. The structure confirmation information is shown in Table 1.

[0469] Example 25(092)

[0470] First step: synthesis of compound 25-2

[0471]

[0472] Compound 25-1 (18 g, 2.0 mol, 1 eq) and imidazole (16.3 g, 2.4 mol, 1.2 eq) were dissolved in dichloromethane (200 mL) under ice water bath at 0 °C and nitrogen protection, and then TBSCl (30 g, 2.0 mol, 1 eq) was added in batches. After the addition was completed, the temperature was slowly increased to room temperature, and the mixture was stirred for 16 h. The reaction was quenched by adding ice water (100 mL) dropwise, and then dichloromethane (100 mL) was added for extraction. The organic phase was combined and washed with saturated brine (200 mL), dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The residue was directly used in the next step.

[0473] Second step: synthesis of compound 25-3

[0474]

[0475] Compound 25-2 (10 g, 48 mmol, 1 eq) and trichloroethylene (7.6 g, 58 mol, 1.2 eq) were dissolved in tetrahydrofuran (100 mL) under ice water bath at 0 °C and nitrogen protection, and then NaH (1.2 g, 48 mmol, 1 eq) was added in batches. After the addition was completed, the temperature was slowly increased to room temperature, and the mixture was stirred for 16 h. The reaction was quenched by adding ice water (100 mL) dropwise, and then dichloromethane (100 mL) was added for extraction. The organic phase was combined and washed with saturated brine (200 mL), dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The residue was directly used in the next step.

[0476] (2.35 g, 58 mol, 1 eq, 40% purity), and the temperature was slowly raised to room temperature after the addition was completed and stirred for 16 hours. Ice water (20 mL) was added dropwise to quench the reaction at low temperature, and then water (300 mL) was added, and extracted with ethyl acetate (300 mL*3). The organic phase was combined and washed with saturated brine (200 mL), dried over anhydrous magnesium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash silica gel column chromatography (eluted with 0-10% ethyl acetate / petroleum ether gradient) to give compound 25-3 (9.8 g, purity 80%, yield 67%) as a colorless oil.

[0477] 1 H NMR (600 MHz, DMSO-d6) δ 6.09 (s, 1H), 4.05 (t, J = 6.5 Hz, 2H), 3.65-3.57 (m, 2H), 1.68 (dq, J = 8.2, 6.5 Hz, 2H), 1.58 (dq, J = 9.6, 6.4 Hz, 2H), 0.87 (d, J = 3.3 Hz, 9H), 0.03 (d, J = 6.9 Hz, 6H).

[0478] Third step: synthesis of compound 25-4

[0479]

[0480] To a solution of compound 25-3 (5 g, 0.017 mol, 1 eq) in tetrahydrofuran (50 mL) was slowly added dropwise n-butyllithium (2.5 M in tetrahydrofuran, 16.7 mL) at -70 °C under nitrogen protection. After the addition was completed, the temperature was slowly raised to room temperature and stirred for 3 hours. Saturated aqueous ammonium chloride solution (10 mL) was added dropwise to quench the reaction, and then ice water (150 mL) was added, and extracted with ethyl acetate (150 mL*3). The organic phase was combined, dried over anhydrous magnesium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash silica gel column chromatography (eluted with 0-100% ethyl acetate / petroleum ether gradient) to give compound 25-4 (3.88 g, purity 70%, yield 70%) as a colorless oil.

[0481] 1 H NMR (600 MHz, DMSO-d6) δ 4.06 (dt, J = 60.0, 6.6 Hz, 2H), 3.60 (q, J = 5.8, 5.4 Hz, 2H), 2.35 (s, 1H), 1.77-1.57 (m, 2H), 1.55-1.47 (m, 2H), 0.87 (d, J = 4.0 Hz, 9H), 0.04 (d, J = 3.4 Hz, 6H).

[0482] Fourth step: synthesis of compound 25-6

[0483]

[0484] Compound 25-4 (1.8 g, 7.9 mmol, 1.2 eq) was dissolved in ethyl acetate / water (10 mL / 10 mL) at 25 °C under nitrogen protection, then compound 22-9 (1.53 g, 6.6 mmol, 1 eq), anhydrous copper sulfate (524 mg, 3.3 mmol, 0.5 eq) and sodium ascorbate (1.3 g, 6.6 mmol, 1 eq) were added, and the resulting mixture was stirred for 2 hours. The reaction mixture was filtered, extracted with ethyl acetate (20 mL*3), and the organic phase was combined and washed with saturated brine (20 mL*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (eluted with a gradient of 0-100% ethyl acetate / petroleum ether) to give compound 25-6 (750 mg, purity 80%, yield 30%) as a brown oil.

[0485] MS (ESI, m / z): 462.4 [M+H] + .

[0486] Step 5: Synthesis of compound 25-7

[0487]

[0488] To a solution of compound 25-6 (700 mg) in dichloromethane (7 mL) was added a solution of HCl in dioxane (7 mL, 4 mol / L) at 25 °C, and the resulting mixture was stirred for 2 hours. The reaction was concentrated under vacuum to give crude compound 25-7 (1 g), which was used directly in the next step.

[0489] MS (ESI, m / z): 348.4 [M+H] + .

[0490] Step 6: Synthesis of compound 25-8

[0491]

[0492] Compound 25-7 (1 g, crude) was dissolved in dichloromethane (7 mL), then p-toluenesulfonyl chloride (1.2 g, 6.6 mmol, 1.2 eq), triethylamine (1.2 mL, 8.6 mmol, 1.6 eq) and 4-dimethylaminopyridine (1.2 g, 9.8 mmol, 1.8 eq) were added in turn, and the resulting mixture was stirred at 25 °C for 2 hours. The reaction was concentrated under reduced pressure, and the residue was purified by flash column chromatography on silica gel (eluted with a gradient of 0-100% ethyl acetate / petroleum ether) to give compound 25-8 (1.2 g, purity 80%, yield 80%) as a white solid.

[0493] (347 mg, 1.82 mmol, 1.2 eq) and triethylamine (0.85 mL, 6.068 mmol, 4 eq) and the resulting reaction was stirred for 16 h. The reaction was poured into water (10 mL) and washed twice, then the organic phase was combined, dried over anhydrous magnesium sulfate and filtered, and the filtrate was concentrated in vacuo to give the crude product which was purified using flash silica gel column (0-100% ethyl acetate / petroleum ether gradient elution) to give compound 25-8 (439 mg, purity 90%, yield 67%) as yellow oil.

[0494] MS (ESI, m / z): 502.2 [M+H] + .

[0495] Seventh step: synthesis of compound 25-9

[0496]

[0497] Compound 25-8 (976 mg, 1.95 mmol, 1 eq), compound L-prolinol (220 mg, 1.95 mol, 1 eq), potassium carbonate (806 mg, 5.84 mmol, 3 eq) and potassium iodide (65 mg, 0.389 mmol, 0.2 eq) were added into DMF (10 mL), the resulting mixture was heated to 90 °C and stirred for 3 h, then the reaction was poured into water (50 mL) and extracted with ethyl acetate (40 mL*3), the organic phase was combined and washed with saturated brine (30 mL*2), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated in vacuo. The residue was purified by flash silica gel column (0-30% methanol / dichloromethane gradient elution) to give compound 25-9 (410 mg, purity 85%, yield 49%) as yellow oil.

[0498] MS (ESI, m / z): 431.4 [M+H] + .

[0499] Eighth step: synthesis of compound 25-10

[0500]

[0501] Compound C-2 (141 mg, 0.43 mol, 1 eq) was dissolved in 1,4-dioxane (4 mL) under nitrogen atmosphere, compound 25-9 (360 mg, 0.83 mmol, 2 eq), cesium carbonate (418 mg, 1.28 mmol, 3 eq) and copper cyanide (15 mg, 0.08 mmol, 0.3 eq) were added successively, the resulting mixture was heated to 80 °C and stirred for 4 h. The reaction mixture was directly concentrated under reduced pressure, the residue was purified by flash silica gel column (0-10% methanol / dichloromethane gradient elution) to give compound 25-10 (95 mg, 85% purity, 31% yield) as brown semi-solid. MS (ESI, m / z): 663.4 [M+H] + .

[0502] Ninth step: synthesis of compound 25-11

[0503]

[0504] Compound 25-10 (85 mg, 0.13 mol, 1 eq) was dissolved in 1,4-dioxane (1 mL) and water (0.2 mL), compound C-4 (115 mg, 0.32 mmol, 2 eq) and potassium phosphate (81 mg, 0.384 mmol, 3 eq) were added, Pd Cata CXium A Pd G3 (10 mg, 0.013 mmol, 0.1 eq) was added under nitrogen atmosphere, the resulting mixture was stirred at 90 °C for 2 h. The reaction was concentrated under reduced pressure, the residue was purified by flash column (0-10% methanol / dichloromethane gradient elution) to give compound 25-11 (62 mg, 80% purity, 55% yield) as brown solid.

[0505] MS (ESI, m / z): 861.7 [M+H] + .

[0506] Tenth step: synthesis of compound 25-12

[0507]

[0508] Compound 25-11 (62 mg, 0.07 mmol, 1 eq) was dissolved in acetonitrile (2 mL), lithium bromide (94 mg, 1.1 mmol, 15 eq), triethylamine (30 μL, 0.22 mmol, 3 eq) and water (4 μL, 0.22 mmol, 3 eq) were added, the resulting mixture was stirred at room temperature for 24 hours. Water (10 mL) was added to the reaction, 2M citric acid was added to adjust the pH of the system to weakly acidic, ethyl acetate (8 mL*3) was added to extract, the organic phase was combined, dried over anhydrous magnesium sulfate and filtered, the filtrate was concentrated under vacuum to obtain brown solid compound 25-12 (50 mg), which was directly used in the next step reaction.

[0509] MS (ESI, m / z): 833.41 [M+H] + .

[0510] Eleventh step: synthesis of compound 25-14

[0511]

[0512] Compound 25-12 (40 mg, 0.05 mmol, 1 eq), N,N,N',N'-tetramethylchloroformamidium hexafluorophosphate (27 mg, 0.10 mmol, 2 eq) and N-methylimidazole (12 mg, 0.14 mmol, 3 eq) were first dissolved in dimethyl sulfoxide (1 mL), after stirring for 20 minutes, compound C-6 (46 mg, 0.10 mmol, 2 eq) was added, the resulting mixture was stirred at room temperature for 2 hours. The reaction was poured into water (5 mL), extracted with ethyl acetate (5 mL*3), the organic phase was combined, dried over anhydrous magnesium sulfate and filtered, the filtrate was concentrated under vacuum, and the residue was purified by flash column chromatography (0-10% gradient of methanol / dichloromethane) to obtain brown solid compound 25-14 (25 mg, purity 70%, yield 45%).

[0513] MS (ESI, m / z): 1146.7 [(M+H)] + .

[0514] Twelfth step: synthesis of example 25

[0515]

[0516] To a solution of compound 25-14 (25 mg) in dichloromethane (0.5 mL) was added HCl dioxane solution (0.5 mL, 4 mol / L) at 25 °C, the resulting mixture was stirred for 2 hours. The reaction mixture was directly concentrated under vacuum, and the residue was purified by pre-HPLC to obtain example 25 (2 mg, purity 95%, yellow solid).

[0517] MS (ESI, m / z): 1102.6 [M+H] + .

[0518] Example 26 (093)

[0519] First Step: Synthesis of compound 26-2

[0520]

[0521] To a solution of compound 26-2 (50 mg) in dichloromethane (1 mL) was added hydrogen chloride (1 mL, 1M in dioxane) at room temperature 25 °C. The resulting mixture was stirred for 2 hours. The reaction mixture was directly concentrated in vacuum. The residue was purified by pre-HPLC to give Example 26 (10 mg, yellow solid, purity 85%).

[0522] MS (ESI, m / z): 1162.7 [(M+H)] + .

[0523] Second Step: Synthesis of Example 26

[0524]

[0525] To a solution of compound 26-2 (50 mg) in dichloromethane (1 mL) was added hydrogen chloride (1 mL, 1M in dioxane) at room temperature 25 °C. The resulting mixture was stirred for 2 hours. The reaction mixture was directly concentrated in vacuum. The residue was purified by pre-HPLC to give Example 26 (10 mg, yellow solid, purity 85%).

[0526] MS (ESI, m / z): 1118.7 [M+H] + .

[0527] 1H NMR (600 MHz, DMSO-d6) δ 10.00 (s, 1H), 9.29 (t, J = 2.8 Hz, 1H), 8.99 (d, J = 7.4 Hz, 1H), 8.48 (d, J = 7.6 Hz, 1H), 7.77 (dd, J = 9.1, 6.0 Hz, 1H), 7.69 (d, J = 3.3 Hz, 1H), 7.44 (dd, J = 8.2, 2.5 Hz, 2H), 7.40 - 7.33 (m, 4H), 7.04 (t, J = 2.6 Hz, 1H), 5.16 (dd, J = 44.0, 10.4 Hz, 2H), 4.95 - 4.81 (m, 2H), 4.78 - 4.70 (m, 2H), 4.66 (s, 1H), 4.58 - 4.48 (m, 1H), 4.44 (t, J = 8.1 Hz, 1H), 4.40 - 4.26 (m, 2H), 4.16 - 4.08 (m, 3H), 3.95 (s, 1H), 3.78 - 3.73 (m, 1H), 3.64 (d, J = 5.4 Hz, 3H), 3.17 (s, 1H), 2.45 (d, J = 13.4 Hz, 4H), 2.34 (s, 1H), 2.26 (d, J = 7.9 Hz, 1H), 2.17 - 1.93 (m, 6H), 1.84 (q, J = 7.5 Hz, 2H), 1.82 - 1.72 (m, 4H), 1.67 (s, 1H), 1.52 (d, J = 7.0 Hz, 2H), 1.24 (s, 1H), 1.20 - 1.14 (m, 4H), 1.01 (dd, J = 27.1, 6.6 Hz, 3H), 0.74 (dt, J = 14.5, 7.4 Hz, 3H), 0.67 (dd, J = 14.3, 6.5 Hz, 3H).

[0528] Example 27 (094)

[0529] First Step: Synthesis of compound 27-2

[0530]

[0531] To a solution of compound 27-1 (7.1 g, 65.0 mmol, 2 eq) in acetonitrile (100 mL) was added compound 4-bromobutanol (7.0 g, 32.5 mmol, 1 eq), potassium carbonate (13.5 g, 97.5 mmol, 3 eq) and potassium iodide (1.1 g, 6.5 mmol, 0.2 eq) successively at room temperature 25 °C with stirring, and the resulting mixture was heated to 80 °C and stirred for 5 h. The reaction system was cooled to 25 °C, water (100 mL) was added, and then extracted with ethyl acetate (50 mL*3). The organic phase was combined and washed with saturated brine (30 mL*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (elution developing agent: dichloromethane / methanol = 10 / 1) to obtain compound 27-2 (6.3 g, yield 67%).

[0532] MS (ESI, m / z): 288.5 [M+H] +

[0533] Second step: synthesis of compound 27-3

[0534]

[0535] Sodium hydride (2.6 g, 65.8 mmol, 3 eq) was added to tetrahydrofuran (50 mL) under an ice water bath 0-5 °C and nitrogen protection, and then compound 27-2 (6.3 g, 21.9 mmol, 1 eq) was added dropwise while maintaining 0-5 °C and stirring for 1 h. Finally, compound bromopropargyl (7.8 g, 65.8 mmol, 3 eq) was added, and the temperature was allowed to rise to room temperature 25 °C and stirred for 16 h. The reaction system was cooled to 0-5 °C, water (100 mL) was added, and then extracted with ethyl acetate (50 mL*3). The organic phase was combined and extracted with saturated brine (30 mL*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (elution developing agent: petroleum ether / ethyl acetate = 1 / 1) to obtain compound 27-3 (2.9 g, yield 41%) as a yellow oil.

[0536] MS (ESI, m / z): 326.5 [M+H] + .

[0537] Third step: synthesis of compound 27-4

[0538]

[0539] Compound 27-3 (2.4 g, 7.37 mmol) was dissolved in dichloromethane (5 mL) at 25 °C, and a hydrochloric acid dioxane solution (5 mL, 1 mol / L) was added. After stirring for 1 h, the reaction solution was concentrated under reduced pressure to obtain compound 27-4 (1.4 g, yield 89%) in the form of a red oil.

[0540] MS (ESI, m / z): 212.3 [M+H] + .

[0541] Fourth step: synthesis of compound 27-5

[0542]

[0543] Compound C-2 (1.0 g, 4.73 mmol, 1.6 eq) and compound 27-4 (979.8 mg, 2.96 mmol, 1 eq) were dissolved in dioxane (20 mL) at 25 °C under stirring, and cesium carbonate (2.9 g, 8.9 mmol, 3 eq) and cuprous cyanide (79.7 mg, 0.9 mmol, 0.3 eq) were added in sequence. After nitrogen replacement for three times, the temperature was raised to 80 °C and stirring was performed for 16 h. Saturated aqueous ammonium chloride solution (5 mL) was added to the reaction solution, and extraction was performed with ethyl acetate (15 mL*3). The combined organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. Purification was performed on a silica gel column (eluent: dichloromethane / methanol = 10 / 1) to obtain compound 27-5 (670 mg, yield 45%) in the form of a yellow oil.

[0544] MS (ESI, m / z): 506.4 [M+H] + .

[0545] Fifth step: synthesis of compound 27-6

[0546]

[0547] To a solution of compound 27-5 (700 mg, 1.4 mmol, 1 eq) in ethyl acetate / water (5 mL / 5 mL) was added compound 22-9 (387.2 mg, 1.7 mmol, 1.2 eq), anhydrous copper sulfate (110 mg, 0.7 mmol, 0.5 eq) and sodium ascorbate (273 mg, 1.4 mmol, 1 eq) at 25 °C under nitrogen protection. The reaction system was stirred for 2 hours. After the reaction was completed, diatomite was added to filter, and the aqueous phase was extracted with ethyl acetate (20 mL*3). The organic phase was washed with saturated brine (20 mL*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. Purification was performed by silica gel column (elution solvent: dichloromethane / methanol = 10 / 1) to obtain compound 27-6 (600 mg, yield 59%) in yellow oil.

[0548] MS (ESI, m / z): 739.7 [M+H] + .

[0549] Step six: synthesis of compound 27-7

[0550]

[0551] To a solution of compound 27-6 (500 mg, 0.68 mmol, 1 eq) and compound C-4 (367.4 mg, 1.02 mmol, 1.5 eq) in dioxane / water (10 mL / 2 mL) was added potassium phosphate (432.5 mg, 2.0 mmol, 3 eq) at room temperature 25 °C, and a catalyst Pd Cata CXium A Pd G3 (51 mg, 0.07 mmol, 0.1 eq) was added after nitrogen replacement for three times. The resulting mixture was heated to 100 °C and stirred for 16 hours. After the reaction liquid was cooled to room temperature, water (20 mL) was added and extracted with ethyl acetate (20 mL*3). The organic phase was combined and washed with saturated brine (20 mL*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column (eluent: dichloromethane / methanol = 10 / 1) to obtain compound 27-7 (60 mg, yield 10%) in yellow oil.

[0552] MS (ESI, m / z): 847.6 [M+H] + .

[0553] Step seven: synthesis of compound 27-8

[0554]

[0555] To a solution of compound 27-7 (50 mg, 0.06 mmol, 1 eq) and compound C-6 (39.8 mg, 0.12 mmol, 2 eq) in dichloromethane (1 mL) was added HATU (33.7 mg, 0.12 mmol, 2 eq) and N-methylimidazole (14.78 mg, 0.18 mmol, 3 eq) at room temperature 25 °C. The reaction mixture was stirred for 2 hours. To the reaction solution was added water (5 mL), and then extracted with dichloromethane / methanol mixed solvent (5 mL*3, 10 / 1), and the combined organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column (eluent: dichloromethane / methanol = 10 / 1) to obtain compound 27-8 (100 mg, crude) as a yellow oil.

[0556] MS (ESI, m / z): 1160.7 [M+H] + .

[0557] Eighth step: synthesis of example 27

[0558]

[0559] To a solution of compound 27-8 (100 mg, crude) in dichloromethane (1 mL) was added hydrochloric acid dioxane solution (0.5 mL, 4 mol / L) at room temperature 25 °C. The reaction mixture was stirred for 1 hour. To the reaction solution was slowly added saturated aqueous sodium bicarbonate solution until the pH of the reaction solution was about 8, and then extracted with dichloromethane / methanol mixed solvent (10 mL*5, 10 / 1). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by pre-HPLC to obtain compound example 27 (15.2 mg, HCOOH salt, yield 23%) as a yellow solid.

[0560] MS (ESI, m / z): 1116.8 [M+H] + .

[0561] 1H NMR (600 MHz, DMSO-d6) δ 9.99 (s, 1H), 9.52 (s, 1H), 9.29 (s, 1H), 8.99-8.97 (m, 1H), 8.39 (d, J = 7.8 Hz, 1H), 8.10 (s, 1H), 7.77 (dd, J = 9.1, 6.0 Hz, 1H), 7.49-7.43 (m, 1H), 7.40-7.34 (m, 4H), 7.30-7.25 (m, 2H), 7.20 (s, 1H), 7.12 (s, 1H), 7.03 (s, 2H), 5.36 (d, J = 9.1 Hz, 1H), 5.00-4.91 (m, 1H), 4.84-4.80 (m, 1H), 4.68 (d, J = 11.9 Hz, 1H), 4.52-4.43 (m, 4H), 4.28 (s, 1H), 4.18-4.02 (m, 3H), 3.90 (s, 2H), 3.75-3.58 (m, 8H), 3.13 (s, 2H), 2.37-2.08 (m, 4H), 2.01 (t, J = 9.0 Hz, 2H), 1.80-1.60 (m, 7H), 1.46 (dd, J = 19.3, 7.0 Hz, 2H), 1.33 (d, J = 7.0 Hz, 2H), 1.25 (s, 3H), 1.18-1.14 (m, 3H), 0.98 (d, J = 6.7 Hz, 2H), 0.85 (q, J = 6.0 Hz, 1H), 0.73 (dt, J = 12.8, 7.4 Hz, 3H), 0.65-0.58 (m, 3H).

[0562] Example 28 (095)

[0563] First Step: Synthesis of compound 28-1

[0564]

[0565] To a solution of compound 27-7 (40 mg, 0.05 mmol, 1 eq) and compound 23-1 (34.7 mg, 0.10 mmol, 2 eq) in DMSO (1 mL) were added HATU (28.1 mg, 0.10 mmol, 2 eq) and N-methylimidazole (12.3 mg, 0.15 mmol, 3 eq) successively at room temperature 25 °C. The reaction mixture was stirred for 2 h. Water (5 mL) was added to the reaction solution, which was then extracted with a dichloromethane / methanol mixed solution (10 / 1, 5 mL*3). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column (eluent: dichloromethane / methanol = 10 / 1) to obtain compound 28-1 (50 mg, crude) as a yellow oil.

[0566] MS (ESI, m / z): 1176.6 [M+H] + .

[0567] Second Step: Synthesis of Example 28

[0568]

[0569] To the solution of compound 28-1 (50 mg, crude) in dichloromethane (1 mL) was added hydrochloric acid dioxane solution (0.5 mL, 4 mol / L) at room temperature 25 ℃, the reaction mixture was stirred for 1 hour. To the reaction solution was slowly added saturated aqueous sodium bicarbonate solution until the pH of the reaction solution was about 8, then extracted with dichloromethane / methanol=10 / 1 (10 mL*5), the organic phase was combined and dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, the residue was purified by pre-HPLC to obtain example 28 (5.6 mg, HCOOH salt, yellow solid, yield 10%).

[0570] MS (ESI, m / z): 1132.6 [M+H] + .

[0571] 1H NMR (600 MHz, DMSO-d6) δ 10.03 (s, 1H), 9.29 (s, 1H), 8.99 (s, 1H), 8.48 (dd, J = 7.9, 3.0 Hz, 1H), 8.14 (d, J = 2.8 Hz, 1H), 7.77 (dd, J = 9.0, 6.0 Hz, 1H), 7.46 - 7.42 (m, 2H), 7.40 - 7.32 (m, 4H), 7.24 (s, 1H), 7.16 (s, 1H), 7.07 (s, 1H), 7.04 (s, 1H), 5.29 (dd, J = 10.1, 2.1 Hz, 1H), 4.83 (q, J = 6.6 Hz, 1H), 4.78 - 4.60 (m, 3H), 4.50 (q, J = 4.8, 3.9 Hz, 2H), 4.43 (t, J = 8.2 Hz, 1H), 4.18 - 4.06 (m, 1H), 3.92 (s, 2H), 3.76 (dd, J = 10.9, 4.2 Hz, 1H), 3.68 - 3.52 (m, 9H), 3.17 (s, 3H), 3.15 - 3.08 (m, 3H), 2.46 (s, 3H), 2.33 (s, 1H), 2.24 (s, 1H), 1.93 (s, 2H), 1.82 - 1.63 (m, 7H), 1.61 - 1.54 (m, 2H), 1.24 (s, 2H), 1.20 - 1.15 (m, 4H), 1.04 (d, J = 6.6 Hz, 3H), 0.87 - 0.84 (m, 1H), 0.74 (dt, J = 14.6, 7.4 Hz, 3H), 0.64 (d, J = 6.5 Hz, 3H).

[0572] Example 29 (108)

[0573] First Step: Synthesis of compound 29-2

[0574]

[0575] After replacing nitrogen three times, DIPEA (3.3 g, 25.8 mmol, 3.0 eq) and T3P (4 g, 2 eq, 50% in EtOAc) were added to a solution of raw material 29-1 (1 g, 8.6 mmol, 1.0 eq) and 4-1 (2.78 g, 12.9 mmol, 1.5 eq) in DMF (10 mL) in turn, and the resulting light yellow reaction solution was stirred at room temperature 29°C for 12 hours. Water (150 mL) was added to the reaction solution, followed by extraction with ethyl acetate (15 mL*3), and the combined organic phase was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (methanol: dichloromethane = 0-5% gradient elution) to obtain the product 29-2 (800 mg, yield 30%) as a light yellow oil. MS (ESI, m / z): 314.5 [M+H]+ .

[0576] Second Step: Synthesis of compound 29-3

[0577]

[0578] To a solution of compound 29-2 (800 mg, 2.55 mmol, 1.0 eq) in THF (4 mL) was added 1 mol / L BH3-THF solution (12.7 mL, 12.7 mmol, 5 eq) at 0 °C under nitrogen protection, the reaction was warmed to 65 °C and stirred for 3 hours, the reaction was cooled to 30 °C, 10 mL of methanol was added to the reaction, and the reaction was quenched by stirring at 65 °C for 2 hours, then the reaction was concentrated under reduced pressure, and the residue was purified by silica gel column (methanol: dichloromethane = 0-10% gradient elution) to give the product 29-3 (500 mg, yield: 65%) as a light yellow oil.

[0579] MS (ESI, m / z): 300.5 [M+H] + .

[0580] Third Step: Synthesis of compound 29-4

[0581]

[0582] To a solution of compound 29-3 (200 mg, 0.67 mmol, 1.0 eq) in THF (3 mL) was added NaH (80 mg, 2 mmol, 60% purity, 3.0 eq) at 0 °C under nitrogen protection, after stirring for 30 minutes, ethyl bromoacetate (122 mg, 0.73 mmol, 1.1 eq) was added, and the reaction mixture was naturally warmed to room temperature and stirred for 4 hours. To the reaction was added dilute hydrochloric acid aqueous solution (1 mL, 0.5 M) to quench, and EtOAc (20 mL) was added to extract, and the organic phase was concentrated under reduced pressure, and the residue was purified by silica gel column (ethyl acetate: petroleum ether = 0-40% gradient elution) to give the product 29-4 (60 mg, yield: 58%) as a light yellow oil. MS (ESI, m / z): 386.5 [M+H] + .

[0583] Fourth Step: Synthesis of compound 29-5

[0584]

[0585] To a solution of compound 29-4 (150 mg, 0.39 mmol) in dichloromethane (1.5 mL) was added HC1 in dioxane (1.5 mL, 4 mol / L), and stirred for 2 hours. The reaction was concentrated under reduced pressure, and the residue was purified by column chromatography (eluted with methanol:dichloromethane (0-5%) gradient) to give the product 29-5 (80 mg, yield: 78%) as a light yellow oil.

[0586] MS (ESI, m / z): 272.4 [M+H]+. + .

[0587] Fifth step: synthesis of compound 29-6

[0588]

[0589] To a solution of compound 29-5 (50 mg, 0.18 mmol, 1 eq) and compound C-2 (61 mg, 0.18 mmol, 1 eq) in dioxane (2 mL) was added CsCO3(180 mg, 0.55 mmol, 3 eq) and CuCN (4.9 mg, 0.05 mmol, 0.3 eq), and the reaction was stirred at 90 °C for 12 hours under nitrogen. The reaction was cooled to room temperature, and water (10 mL) was added. The reaction was extracted with ethyl acetate (10 mL*3), and the organic phase was washed with saturated brine (10 mL*2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (eluted with methanol:dichloromethane = 0-5%) to give 7 (50 mg, yield 47%) as a light yellow solid.

[0590] MS (ESI, m / z): 566.4 [M+H]+.

[0591] The following steps were the same as Example 1 to give Example 29, and the characterization spectrum is shown in Table 1.

[0592] Example 30 (103)

[0593] First step: synthesis of compound 30-2

[0594]

[0595] To a solution of compound 30-1 (2.1 g, 10 mmol, 1.0 eq) and L-prolinol (1.0 g, 10 mmol, 1.0 eq) in dichloromethane was added acetic acid (0.6 g, 10 mmol, 1.0 eq) under ice water bath 0 °C and nitrogen protection, after stirring for 30 min, NaBH(OAc)3(6.3 g, 30 mmol, 3.0 eq) was added, and the resulting white mixture was stirred for 4 h. To the reaction solution was added a saturated aqueous sodium bicarbonate solution to adjust pH ~ 7, the aqueous phase was removed, and the organic phase was concentrated under reduced pressure. The residue 30-2 (2.7 g, yield 90%) was directly used in the next step reaction.

[0596] 1H NMR (600 MHz, CDCl3) δ 7.50 (d, J = 8.1 Hz, 2H), 7.37 (d, J = 8.1 Hz, 2H), 5.75 (s, 3H), 4.29 (d, J = 13.2 Hz, 1H), 3.81-3.66 (m, 4H), 3.22 (ddd, J = 10.5, 6.8, 3.4 Hz, 1H), 3.17-3.07 (m, 2H), 2.64-2.55 (m, 1H), 2.07-1.95 (m, 4H), 1.86 (dddd, J = 28.2, 16.3, 11.3, 7.2, 3.8 Hz, 3H), 1.25 (t, J = 7.0 Hz, 1H).

[0597] Second step: synthesis of compound 30-3

[0598]

[0599] To a solution of compound 30-2 (600 mg, 2.8 mmol, 1.0 eq) and compound 22-9 (650 mg, 2.8 mmol, 1.0 eq) in ethyl acetate / water = 1:1 (10 mL) was added copper sulfate pentahydrate (350 mg, 1.4 mmol, 1.4 eq) and sodium ascorbate (560 mg, 2.8 mmol, 1.0 eq) at room temperature 27 °C, and the resulting compound was stirred for 3 h. To the reaction solution was added saturated aqueous sodium bicarbonate solution (10 mL) and ethyl acetate (10 mL) and stirred for 10 min, the aqueous phase was removed, and the organic phase was concentrated under reduced pressure. The residue was purified by column chromatography to obtain compound 30-3 (0.9 g, yield 72%) as a light yellow oil. MS (ESI, m / z): 449.4 [M+H]+.

[0600] Third step: synthesis of compound 30-4

[0601]

[0602] To a solution of starting material 30-3 (800 mg, 1.8 mmol, 1.5 eq) and compound C-2 (400 mg, 0.3 mmol, 1.0 eq) in 1,4-dioxane (10 mL) was added cesium carbonate (1.75 g, 5.4 mmol, 3.0 eq) and cuprous cyanide (32 mg, 0.4 mmol, 0.4 eq) under nitrogen at room temperature 27 °C. The resulting mixture was heated to 90 °C and stirred for 12 h. The reaction mixture was directly concentrated under reduced pressure, and the residue was purified by column chromatography to give compound 30-4 (210 mg) as a light yellow foam.

[0603] MS (ESI, m / z): 743.7 [M+H]+.

[0604] Fourth step: synthesis of compound 30-5

[0605]

[0606] To a solution of starting material 30-4 (210 mg, 0.28 mmol, 1.0 eq) and C-4 (150 mg, 0.42 mmol, 1.3 eq) in dioxane: water = 5:1 (5 mL) was added potassium phosphate (180 mg, 0.85 mmol, 3.0 eq). After nitrogen was replaced for three times, Pd Cata CXium A Pd G3 (30 mg, 0.04 mmol, 0.1 eq) was added. The resulting light brown reaction mixture was heated to 80 °C and stirred for 6 h. Water (5 mL) was added to the reaction mixture, and ethyl acetate (10 mL*3) was extracted. The combined organic phase was concentrated under reduced pressure, and the residue was purified by column chromatography on silica gel (methanol:dichloromethane = 0-10% gradient elution) to give product 30-5 (200 mg) as a light yellow foam. MS (ESI, m / z): 941.7 [M+H]+.

[0607] Fifth step: synthesis of compound 30-6

[0608]

[0609] To a solution of compound 30-5 (200 mg, 0.2 mmol, 1.0 eq) in MeCN (2 mL) were added Et3N (61 mg, 0.6 mmol, 3.0 eq), water (11 mg, 0.6 mmol, 3.0 eq) and lithium bromide (180 mg, 2 mmol, 10.0 eq) successively at room temperature 27 °C. The resulting mixture was stirred at 27 °C to 33 °C for 16 hours. To the reaction mixture was added aqueous citric acid (0.5 mol / L) to pH ~ 5, and extracted with ethyl acetate (5 mL*3), and the combined organic phase was concentrated under reduced pressure. The residue was purified by column chromatography to give compound 30-6 (120 mg) as a light yellow foamy solid. MS (ESI, m / z): 851.7 [M+H]+.

[0610] Sixth step: synthesis of compound 30-7

[0611]

[0612] After three times of nitrogen replacement, to a solution of compound 30-6 (85 mg, 0.1 mmol, 1.0 eq) and C-6 (36.5 mg, 0.11 mmol, 1.1 eq) in DMF (2 mL) were added N-methylimidazole (25 mg, 0.3 mmol, 3.0 eq) and TCFH (42 mg, 0.15 mmol, 1.4 eq), and the resulting light brown reaction solution was stirred at room temperature 29 °C for 2 hours. To the reaction mixture was added water (5 mL) and saturated brine (5 mL) and stirred for 5 minutes, followed by extraction with ethyl acetate (10 mL*3), and the combined organic phase was concentrated under reduced pressure. The residue was purified by column chromatography (methanol: dichloromethane 0-10% gradient elution) to give product 30-7 (110 mg) as a light yellow foamy solid. MS (ESI, m / z): 1164.7 [M+H]+.

[0613] Seventh step: synthesis of example 30

[0614]

[0615] To a solution of compound 30-7 (110 mg, 0.1 mmol, 1.0 eq) in dichloromethane (2 mL) was added HCl-dioxane solution (0.5 mL, 4 mol / L), and the resulting light brown reaction solution was stirred at room temperature 29 °C for 1 hour. The reaction mixture was concentrated under reduced pressure, and the residue was purified by column chromatography pre-HPLC to give example 30 (11 mg light yellow solid). MS (ESI, m / z): 1120.7 [M+H]+. 1H NMR (600 MHz, DMSO-d6) δ 10.01 (s, 1H), 9.24 (d, J = 11.8 Hz, 1H), 8.99 (s, 1H), 8.76 (s, 1H), 8.53 (d, J = 7.4 Hz, 1H), 7.92 (s, 2H), 7.80 - 7.73 (m, 1H), 7.68 (s, 1H), 7.45 (d, J = 8.0 Hz, 2H), 7.40 - 7.32 (m, 4H), 7.31 - 7.19 (m, 1H), 7.07 (s, 1H), 5.39 (dd, J = 35.7, 9.1 Hz, 1H), 5.21 (s, 1H), 4.93 (q, J = 6.9 Hz, 1H), 4.77 (d, J = 9.8 Hz, 1H), 4.74 - 4.50 (m, 2H), 4.49 - 4.21 (m, 5H), 4.08 (dd, J = 32.0, 12.6 Hz, 2H), 3.85 - 3.66 (m, 2H), 3.65 - 3.45 (m, 2H), 2.45 (d, J = 14.1 Hz, 4H), 2.40 - 2.30 (m, 2H), 2.21 - 1.83 (m, 7H), 1.83 - 1.59 (m, 6H), 1.38 (d, J = 6.9 Hz, 3H), 1.25 - 1.12 (m, 4H), 1.07 (dd, J = 29.5, 5.8 Hz, 4H), 0.77 - 0.70 (m, 6H).

[0616] Example 31(0104)

[0617] First Step: Synthesis of compound 31-1

[0618]

[0619] To a solution of compound 30-6 (75 mg, 0.09 mmol, 1.0 eq) and 23-1 (40 mg, 0.11 mmol, 1.1 eq) in DMSO (2 mL) was added NMM (22 mg, 0.3 mmol, 3.0 eq) and TCFH (40 mg, 0.14 mmol, 1.6 eq), after nitrogen replacement for three times, the resulting light brown reaction solution was stirred at room temperature 29 °C for 2 hours. Water (5 mL) and saturated brine (5 mL) were added to the reaction solution and stirred for 5 minutes, then extracted with ethyl acetate (10 mL*3), the combined organic phase was concentrated under reduced pressure, the residue was purified by column chromatography, gradient elution with methanol: dichloromethane (0-10%), to obtain light yellow foam solid product 27-1 (70 mg). MS (ESI, m / z): 1180.7 [M+H]+.

[0620] Second Step: Synthesis of Example 31

[0621]

[0622] To the solution of compound 31-1 (70 mg, 0.06 mmol, 1.0 eq) in dichloromethane (2 mL) was added HCl-dioxane solution (0.5 mL, 2 mmol, 4 mol / L), the resulting light brown reaction solution was stirred at room temperature 29 °C for 1 hour. The reaction solution was concentrated under reduced pressure, the residue was purified by column chromatography pre-HPLC to give example 31 (light yellow solid 10 mg). MS (ESI, m / z): 1136.7 [M+H]+.

[0623] 1 H NMR (600 MHz, DMSO-d6) δ 10.01 (s, 1H), 9.24 (d, J = 11.8 Hz, 1H), 8.99 (s, 1H), 8.76 (s, 1H), 8.53 (d, J = 7.4 Hz, 1H), 7.92 (s, 2H), 7.80 - 7.73 (m, 1H), 7.68 (s, 1H), 7.45 (d, J = 8.0 Hz, 2H), 7.40 - 7.32 (m, 4H), 7.31 - 7.19 (m, 1H), 7.07 (s, 1H), 5.39 (dd, J = 35.7, 9.1 Hz, 1H), 5.21 (s, 1H), 4.93 (q, J = 6.9 Hz, 1H), 4.77 (d, J = 9.8 Hz, 1H), 4.74 - 4.50 (m, 2H), 4.49 - 4.21 (m, 5H), 4.08 (dd, J = 32.0, 12.6 Hz, 2H), 3.85 - 3.66 (m, 2H), 3.65 - 3.45 (m, 2H), 2.45 (d, J = 14.1 Hz, 4H), 2.40 - 2.30 (m, 2H), 2.21 - 1.83 (m, 7H), 1.83 - 1.59 (m, 6H), 1.38 (d, J = 6.9 Hz, 3H), 1.25 - 1.12 (m, 4H), 1.07 (dd, J = 29.5, 5.8 Hz, 4H), 0.77 - 0.70 (m, 6H).

[0624] Table 1 Partial compound structure and spectral analysis control list

[0625]

[0626]

[0627]

[0628]

[0629]

[0630]

[0631]

[0632]

[0633] Bioactivity experiment

[0634] 1. Protein degradation experiment:

[0635] The Western Blot method was used to detect the degradation of KRAS G12D mutant and KRAS G12C protein by the test substance.

[0636] 1.1 Cell information

[0637] Serial No. Cell No. Mutation Type Final Cells / 25 μl / well 1 ASPC-1 KRAS G12D mutation 12000.00 2 NCI-H358 KRAS G12C mutation 12000.00 3 NCI-H727 KRAS G12V mutation 12000.00

[0638] 1.2 Reagent information:

[0639]

[0640] 1.3 Instrument information:

[0641]

[0642] 1.4 Preparation and storage method of test drug:

[0643] 1) 10 mM stock solution was prepared using DMSO.

[0644] 2) The test substance stock solution was sequentially diluted with DMSO to obtain 3000 μΜ, 1000 μΜ, 300 μΜ, 100 μΜ, 30 μΜ and 10 μΜ dilutions.

[0645] 3) The diluted compounds were added to the cell well plate to obtain a final concentration of 10000 nΜ, 3000 nΜ, 1000 nΜ, 300 nΜ, 100 nΜ, 30 nΜ and 10 nM working solution, and the DMSO concentration was 0.1%.

[0646] 4) The solubility of the test sample was observed visually, and all concentrations were completely dissolved without visible precipitate.

[0647] 5) The test drug stock solution and working solution were stored at -20°C.

[0648] 1.5 Cell culture and treatment

[0649] 1) ASPC-1 and NCI-H358 cells were inoculated into 12-well plates and cultured overnight at 37°C, 5% CO2.

[0650] 2) When the cells grow to 80% coverage, add the test substance, and incubate at 37°C, 5% CO2 for 48 hours.

[0651] 3) Collect the cells, digest the cells with trypsin, centrifuge at 1000 x g and wash twice with ice-cold PBS, and store in a -80°C refrigerator.

[0652] 1.6 Cell lysis and protein extraction

[0653] 1) Thaw the cell mass on ice, add pre-cooled cell lysis solution (0.1 mL / 10 6 cells), and use a pipette to blow multiple times until the cells are loose.

[0654] 2) Lysate on ice for 30 minutes, shake every 10 minutes (20000 rpm, 2 seconds x 3).

[0655] 3) Centrifuge at 12000 rpm at 4°C for 15 minutes.

[0656] 4) Collect the supernatant and transfer it to a pre-cooled EP tube, determine the protein concentration in the lysis solution by the BCA method, and adjust the protein concentration of all samples to be consistent with the lysis solution.

[0657] 1.7 Western blot immunodetection

[0658] 1) Take an appropriate amount of sample, add 4x Laemmli Sample buffer, and incubate in a 100°C metal bath for 10 minutes.

[0659] 2) When the sample is cooled to room temperature, centrifuge at 1000 rpm for 5 minutes.

[0660] 3) Use 4-12% SDS-PAGE precast gel for electrophoresis, load 10 μg, and electrophorese at 120V for 10 min, and transfer at 150V.

[0661] 4) Wet transfer at 300mA for 1 hour.

[0662] 5) Use 5% BCA solution for blocking, and block at room temperature for 1 hour.

[0663] 6) Primary antibody incubation at 4°C overnight, dilution ratio KRAS 1:1000, RAS-G12D 1:1000, GAPDH 1:8000. TBST wash 3 times, 5 minutes each time.

[0664] 7) Secondary antibody (Anti-Mouse IgG, CST, 7076) incubation at room temperature for 1 hour, dilution ratio 1:5000.

[0665] 8) TBST wash 3 times, 5 minutes each time.

[0666] 9) ECL method color development, imaging system imaging.

[0667] 1.8 Data analysis

[0668] 1) Quantify the bands using Image Lab Touch software in the ChemiDoc imaging system.

[0669] 2) The gray value of the target protein of the control group (DMSO treatment) is divided by the gray value of the internal reference protein, and the drug treatment group is normalized (R = Target intensity / GAPDH intensity, R DMSO as 100%)

[0670] Inhibition rate calculation:

[0671] Degradation% = (R DMSO -R cpd ) / R DMSO ×100%

[0672] The data were analyzed using GraphPad Prism, and the test results and conclusions

[0673] 1.9 Test results

[0674]

[0675]

[0676] Note: - indicates no activity or undetected

[0677] The experimental results show that each compound of the present application exhibits pan-KRAS inhibitory activity, and has good inhibitory activity against G12C mutant, G12D mutant and G12V mutant.

[0678] 2. Cell inhibition test

[0679] The CTG method was used to detect the inhibitory activity of the test drug on the proliferation of ASPC-1 and H358 cells.

[0680] 2.1 Reagents, consumables and instruments

[0681]

[0682]

[0683] 2.2 Test instruments

[0684] Name Manufacturer Model No. CO2 Incubator Thermo 3111 Biological Safety Cabinet Thermo 3189 Electric Pipettor INTEGRA 155015 Centrifuge Thermo / Xiangyi 75009740 / L535R Cell Counter LUNA II T13001 Multifunctional Microplate Reader Biotech 3000D

[0685] 2.3 Experimental steps:

[0686] 2.3.1 Drug Dissolution

[0687] Dissolve to 10 mM stock concentration in DMSO according to compound information.

[0688] 2.3.2 Drug Dilution

[0689] Dilute the stock solution to get working solutions with concentrations of 10 μM, 3 μM, 1 μM, 0.3 μM, 0.1 μM, 0.03 μM, 0.01 μM, 0.003 μM, 0.001 μM respectively. The control is STS with a concentration of 1 μM.

[0690] 2.3.3 Cell Plating

[0691] 1) Prepare complete medium and mix well.

[0692] 2) Revive the cells and pass them for two generations or so to select cell lines with good growth conditions.

[0693] 3) Take the cell culture bottle out of the incubator and check the cell name and medium type marked on the bottle.

[0694] 4) Adherent cells: aspirate the medium, wash once with trypsin, discard the waste liquid, and add 3 mL of fresh trypsin to the culture bottle for digestion. When the cells are loose and about to detach from the bottle wall, stop the trypsin digestion by adding 8 mL of complete medium and gently mix. Use a pipette to transfer the cell suspension to a centrifuge tube and centrifuge at 1000 rpm for 5 minutes.

[0695] 5) Discard the supernatant.

[0696] 6) Add an appropriate volume of medium to the centrifuge tube and gently blow to resuspend the cells evenly.

[0697] 7) Use the Vi-Cell XR cell counter to count.

[0698] 8) Adjust the cell suspension to the appropriate concentration.

[0699] 9) Add the cell suspension to the 96-well plate, 100 μL / well. Label the cell name, seeding density, date, etc. detailed information, and place the culture plate in the CO2 incubator overnight.

[0700] 2.3.4 Drug Addition

[0701] After mixing the compounds, add them to the cells, allow them to settle naturally for 10 min, observe, and place them in the incubator for incubation.

[0702] 2.3.5 Plate Reading

[0703] 1) Thaw CytoTox-Glo Buffer at 37°C water bath. Equilibrate lyophilized AAF-Glo substrate to room temperature.

[0704] 2) Add CytoTox-Glo Buffer to AAF-Glo substrate and mix well.

[0705] 3) Remove cell plate from equilibration at room temperature.

[0706] 4) Add 50 μL of mixed CytoTox-Glo reagent to each well, avoid light, shake 10 min, incubate at room temperature for 15 min.

[0707] 5) Transfer 162 μl of Digitonin solution to 25 ml of CytoTox-Glo, mix well, add 50 μL to half of the DMSO control wells for lysis.

[0708] 6) Place the plate into Enspire plate reader, record luminescence readout results.

[0709] 2.3.6 Data Analysis

[0710] Inhibition rate is calculated as follows:

[0711] 1) Inhibition rate (%) = (1 - (RLU compound - RLU blank) / (RLU DMSO - RLU blank)) x 100%.

[0712] 2) Use XLFit to plot the pharmacological inhibition rate curve and calculate IC 50 value. Use 4-parameter model [fit = (A + ((B-A) / (1+((C / x)^D)))).

[0713] 2.3.7 Test Results

[0714]

[0715] Note: - means no activity or not detected

[0716] From the above results, it can be seen that the compound of the present application is a pan-KRAS inhibitor, which has good inhibitory activity against G12C mutant, G12D mutant and G12V mutant.

[0717] All documents referred to in the present application are incorporated herein by reference as if each were individually incorporated by reference. In addition, it is to be understood that the application can be carried out by specifically different embodiments and that embodiments can be practiced without materials details (specific materials and shapes), conditions (especially, explicitly stated conditions), processes, machines, manufacture's equipment, etc. that are of a speculative or hypothetical nature. The application is not limited to the embodiments described herein which are presented as examples but cover every novel, practical and inventive feature and combination of features.

Claims

1. A compound of Formula I, or a pharmaceutically acceptable salt thereof, wherein wherein single or double bond, as valence allows; X is selected from N or CH; R1 is selected from C6-C18 aryl or 5-18 membered heteroaryl, optionally substituted with one or more Ra, independently selected from C1-C4 alkyl, C1-C4 alkoxy, halogen, C2-C4 alkynyl, hydroxyl; R2 is selected from H, fluorine, chlorine, bromine, cyano; R3is selected from a 5-18 membered heterocyclyl group, which can be monocyclic or bicyclic forming a bridged or spirocyclic ring, which heterocyclyl group contains 1-3 heteroatoms which are O, S and N, and which heterocyclyl group is optionally substituted with one or more R b substituted, R b is independently selected from C1-C4alkyl, C1-C4alkoxy, carbonyl; A is each independently O or N; n is each independently 0, 1 or 2; R4 is selected from the group consisting of H, D and C1-C4 alkyl; C is selected from a 5-10 membered heterocyclyl, which can be monocyclic or spiro or bridged bicyclic, which contains 1-3 heteroatoms, which are O, S and N, and which is optionally substituted by one or more R c substituted, R c independently selected from C1-C4 alkyl, C1-4 alkoxy, halogen; L is a divalent linker selected from: wherein F1, F2, F3 and F4 are selected from O, NH or absent; n1, n2, n3 and n4 are selected from 0, 1, 2, 3 or 4; G1 is each independently selected from the group consisting of C3-C10 membered aliphatic ring, C6-C10 membered aromatic ring or 5-10 membered heteroaromatic ring; wherein said aliphatic ring is a monocyclic ring, or a spiro or bridged bicyclic ring; and 0, 1, 2, or 3 R g substituted, said R g selected from the group consisting of C1-4alkyl, C1-C4oxyalkyl, halogen, C3-C10cycloalkyl; and the selected 5-10 membered heteroaromatic ring contains 1-3 heteroatoms selected from O, N, S; E is selected from a protein binding fragment of Formula III: wherein R5 is selected from methyl and H; R6 is selected from methyl and hydroxymethyl; R7 is selected from methyl and H; R8 is selected from -NH, 2. The compound of claim 1, wherein R1 is phenyl, naphthalene ring; Ra is independently selected from C1-C4 alkyl, halogen, hydroxyl; R2 is F.

3. The compound of claim 1, wherein R3 is selected from the group consisting of: and each chiral isomer thereof.

4. The compound of claim 1, wherein The compound is of the structure of Formula I-1: The remaining substituents are as described in claim 1.

5. The compound of claim 1, wherein C is selected from the group consisting of:

6. The compound of claim 1, wherein L is selected from the group consisting of: wherein n1, n2, n3 and n4 are as defined in claim 1.

7. The compound of claim 1, wherein The compound is selected from the group consisting of:

8. A pharmaceutical composition comprising: (1) a compound of any one of claims 1-7, a tautomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof; and (2) and a pharmaceutically acceptable carrier.

9. Use of a compound according to any one of claims 1 to 7, a tautomer, a racemate, an enantiomer, a diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 8, characterized in that, The use is for the manufacture of a medicament for preventing, treating and / or ameliorating a disease mediated by or dependent on KRAS protein.

10. The use according to claim 9, characterized in that, The disease is a tumor.

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