Preparation method and intermediate of nitrogen-containing heterocyclic compound

Abstract

The invention provides a preparation method of a nitrogen-containing heterocyclic compound and an intermediate of the nitrogen-containing heterocyclic compound. Specifically provided is a preparation method of a compound 5, which comprises the following steps: in an organic solvent, in the presence of a reducing agent, carrying out a reduction reaction as shown in the specification on a compound 4 to obtain the compound 5, r1 and R2 are independently hydrogen or deuterium, R3 is independently C1-6 alkyl, the organic solvent is an ether solvent, and the reducing agent is hydroaluminate or deuterated hydroaluminate. The preparation method provided by the invention is simple and convenient to operate, high in yield, good in selectivity and good in industrialization prospect.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology. Specifically, this invention relates to a method for preparing a nitrogen-containing heterocyclic compound and its intermediates. Background Technology

[0002] RAS (rat sarcoma) is one of the most frequently mutated oncogenes in tumors, with mutations present in approximately 30% of human malignancies. The RAS family includes KRAS, NRAS, and HRAS, with KRAS (kirsten rat sarcomaviral oncogene) being more prone to mutation than the other two RAS subtypes, accounting for approximately 85% of mutations, and is particularly common in solid tumors.

[0003] Because KRAS binds to GTP or GDP with ultra-high affinity (picomolar concentration), and because the RAS protein surface is smooth and lacks ideal small molecule binding sites, developing competitive inhibitors that act directly on the RAS protein is considered extremely challenging.

[0004] In recent years, with the continuous deepening of KRAS research, the development of KRAS inhibitors has finally made significant progress.

[0005] KRAS G12D is the most common mutation in colorectal and pancreatic cancer.

[0006] On December 10, 2021, Mirati Pharmaceuticals reported the first non-covalent, potent, and highly selective KRAS-G12D inhibitor, MRTX1133. MRTX1133 inhibits both activated and inactivated KRAS-G12D mutant cells but does not inhibit wild-type tumor cells, exhibiting a specificity of over 1000-fold. It demonstrated dose-dependent inhibition in in vivo transplanted tumor models of pancreatic and colorectal cancer.

[0007] Therefore, the current degradation of KRAS G12D-targeting proteins does not meet clinical needs.

[0008] (S)-2-methyl-3-(piperazin-1-yl)prop-1-ol intermediates are important intermediates in the synthesis of KRAS G12D targeting protein degraders. Currently, the synthetic route and method for related intermediates reported in patent WO2011127567A1 are as follows:

[0009]

[0010] This route starts with methyl (S)-(-)-3-bromoisobutyrate (compound 2), and obtains target compound 3 and racemic byproduct 3a through a substitution reaction with N-protected piperazine. Post-processing requires column chromatography purification, with a separation yield of only 34%. Moreover, methyl (S)-(-)-3-bromoisobutyrate is only a reagent-grade commercial product, not a bulk chemical, and is not readily available on the market. Therefore, this route is not feasible for large-scale production.

[0011] The synthetic route for similar compounds reported in patent WO2016005577A1 is as follows:

[0012]

[0013] This route starts with R-(-)-3-bromo-2-methyl-1-propanol (compound 2), and obtains the target compound 3 through a substitution reaction with N-protected piperazine. However, both R-(-)-3-bromo-2-methyl-1-propanol and (S)-(+)-3-bromo-2-methyl-1-propanol are only reagent-grade commodities, not bulk chemicals, and are not readily available on the market. This poses a huge challenge to the continuous supply of the target product and greatly reduces the feasibility of scaling up the production of this route.

[0014] Given the combined considerations of cost reduction and commercial production, it is particularly necessary to provide a new method for preparing KRAS G12D degradation agent intermediates in order to reduce preparation costs and increase yield. Summary of the Invention

[0015] To address the aforementioned problems, this invention provides a method for preparing nitrogen-containing heterocyclic compounds and its intermediates. This method overcomes the drawbacks of complex routes and low yields in existing technologies, and has significant advantages in terms of suitability for commercial-scale production.

[0016] The present invention mainly solves the above-mentioned technical problems through the following technical solution. The present invention provides a method for preparing compound 5, which includes the following steps: in an organic solvent, in the presence of a reducing agent, compound 4 is subjected to the following reduction reaction to obtain compound 5;

[0017]

[0018] in,

[0019] R 1 and R 2 Independently hydrogen or deuterium;

[0020] R 3 Independently for C 1-6 alkyl;

[0021] The organic solvent is an ether solvent;

[0022] The reducing agent is aluminate or deuterated aluminate.

[0023] In the reduction reaction, when the reducing agent used is aluminate hydride, R 1 and R 2 It can be hydrogen; when the reducing agent used is deuterated aluminate hydride, R 1 and R 2 It can be deuterium.

[0024] In the reduction reaction, the organic solvent may be a cyclic ether solvent, such as tetrahydrofuran.

[0025] In the reduction reaction, the mass-to-volume ratio of compound 4 to the organic solvent can be (50-150) g / L, for example, 98.5 g / L.

[0026] In the reduction reaction, the reducing agent may be lithium aluminum hydride, sodium aluminum hydride, or potassium aluminum hydride, such as lithium aluminum hydride.

[0027] In the reduction reaction, the reducing agent may be lithium deuterated aluminum hydride, sodium deuterated aluminum hydride, or potassium deuterated aluminum hydride, such as lithium aluminum deuterated hydride (LiAlD4).

[0028] In the reduction reaction, the molar ratio of compound 4 to the reducing agent can be 1:(0.1~1), for example 1:0.55.

[0029] In the reduction reaction, the reaction temperature can be 0 to 20°C, for example, 0 to 5°C.

[0030] The reduction reaction preferably includes the following steps: adding water and an inorganic base.

[0031] In the reduction reaction, the inorganic base can be an alkali metal hydroxide, such as NaOH.

[0032] In the reduction reaction, the inorganic base can be used in the form of an aqueous solution of an inorganic base, such as a 15 wt% NaOH aqueous solution.

[0033] In the reduction reaction, the mass-to-volume ratio of the reducing agent to the water can be (0.1-1) g / mL, for example, 1 / 5 g / mL.

[0034] In the reduction reaction, the water and inorganic base may be added slowly, for example, dropwise.

[0035] In the reduction reaction, the water can be added multiple times, in 2-5 portions. For example, some water can be added first, followed by an aqueous solution of an inorganic base, and then water can be added again (when added in three portions, the volume ratio can be 1:1:3).

[0036] In the reduction reaction, the water and inorganic base may be added at 0 to -15°C, for example at -10°C.

[0037] The reduction reaction preferably further includes the following step: adding a purification solvent, wherein the purification solvent is N,N-dimethylformamide (DMF).

[0038] The mass-to-volume ratio of the reducing agent to the purified solvent can be (0.1-1) g / mL, for example, 0.35 g / mL.

[0039] In the reduction reaction, the progress of the reaction can be monitored using conventional monitoring methods in the art (e.g., TLC, HPLC, or NMR), and the reaction endpoint is generally defined as the disappearance of compound 4 or the cessation of the reaction. The reaction time can be 0.1 to 2 hours, for example, 0.5 hours.

[0040] The preparation method may further include the following post-processing steps: after the reduction reaction is completed, an aqueous solution of water and inorganic base is added at 0 to -15°C, followed by the addition of a purification solvent, filtration, washing (e.g., washing the filter cake with ethyl acetate, and washing the filtrate with water and saturated brine), drying (e.g., drying with anhydrous sodium sulfate), and concentration to obtain compound 5.

[0041] The post-processing step may further include a recrystallization step, wherein the solvent for recrystallization may be an ether solvent (e.g., methyl tert-butyl ether) or an alkane solvent (e.g., n-heptane). The recrystallization step may include the following steps: crystallization, filtration, and drying to obtain compound 5.

[0042] The reduction reaction may include the following steps: reacting the mixture of the reducing agent and the ether solvent with the mixture of compound 4 and the ether solvent at 0-5°C.

[0043] The materials for the reduction reaction may be compound 4, lithium aluminum hydride (LAH), and the ether solvents mentioned above.

[0044] The R 1 and R 2 It can be hydrogen independently.

[0045] The R 3 It can be methyl.

[0046] In the reduction reaction, R 1 and R 2 It can be hydrogen, R 3 The solvent is methyl, the organic solvent is a cyclic ether solvent, and the reducing agent is lithium aluminum hydride.

[0047] The preparation method of compound 5 may further include the preparation method of compound 4, which includes the following steps: in an organic solvent, in the presence of an organic base, compound 2 and compound 3 undergo a substitution reaction as shown below to obtain compound 4;

[0048]

[0049] R 3 As described in any embodiment of the present invention.

[0050] In the substitution reaction, the organic solvent may be a cyclic ether solvent (e.g., a six-membered cyclic ether solvent), such as 1,4-dioxane.

[0051] In the substitution reaction, the mass-to-volume ratio of compound 2 to the organic solvent can be (50-150) g / L, for example, 106.5 g / L.

[0052] In the substitution reaction, the molar ratio of compound 2 to compound 3 can be 1:(0.5 to 1.5), for example 1:0.95.

[0053] In the substitution reaction, the organic base may be a tertiary amine compound (e.g., N(C)). 1-6 Alkyl groups, such as N,N-diisopropylethylamine (DIPEA).

[0054] In the substitution reaction, the molar ratio of compound 2 to the organic base can be 1:(0.5-4), for example 1:1.5.

[0055] In the substitution reaction, the reaction temperature can be 10–35°C, for example 25–30°C.

[0056] In the substitution reaction, the progress of the reaction can be monitored using conventional monitoring methods in the art (e.g., TLC, HPLC, or NMR), and the reaction endpoint is generally defined as the disappearance of compound 3 or the cessation of the reaction. The reaction time can be 10–30 hours, for example, 16 hours.

[0057] The preparation method may further include the following post-processing steps: after the substitution reaction is completed, the mixture is filtered (e.g., placed in ice water for cooling and stirring, then filtered), the filter cake is washed (e.g., washed with water), and dried to obtain compound 4.

[0058] The substitution reaction may include the following steps: mixing the compound 3, the organic base and the organic solvent (e.g., mixing at 25-30°C), mixing the mixture with the compound 2 (e.g., mixing at 10-15°C), and reacting at 10-35°C.

[0059] In one embodiment, the materials for the substitution reaction may be compound 3, N,N-diisopropylethylamine, compound 2, and the organic solvent.

[0060] In the substitution reaction, R 3 The solvent may be methyl, the organic solvent may be a cyclic ether solvent, and the base may be N,N-diisopropylethylamine.

[0061] The method for preparing compound 5 may further include a method for preparing compound 2, which includes the following steps: in an organic solvent, in the presence of an organic base, compound 1 and a sulfonating agent undergo a sulfonation reaction as shown below to obtain compound 2;

[0062]

[0063] R 3 As described in any embodiment of the present invention.

[0064] In the sulfonation reaction, the organic solvent is a conventional organic solvent for such reactions in the art, preferably a haloalkane solvent, such as dichloromethane.

[0065] In the sulfonation reaction, the mass-to-volume ratio of compound 1 to the organic solvent can be (50-150) g / L, for example, 100 g / L.

[0066] In the sulfonation reaction, the sulfonating agent is a conventional sulfonating agent for such reactions in the art, preferably trifluoromethanesulfonyl chloride or trifluoromethanesulfonic anhydride, such as trifluoromethanesulfonic anhydride.

[0067] In the sulfonation reaction, the molar ratio of compound 1 to the sulfonating agent can be 1:(0.5-2), for example 1:1.1.

[0068] In the sulfonation reaction, the organic base is a conventional organic base for such reactions in the art, preferably a pyridine organic base, such as 2,6-dimethylpyridine.

[0069] In the sulfonation reaction, the molar ratio of compound 1 to the organic base can be 1:(0.5-2), for example 1:1.1.

[0070] In the sulfonation reaction, the reaction temperature is the conventional reaction temperature for such reactions in the art, preferably 10 to 35°C, for example 25 to 30°C.

[0071] In the sulfonation reaction, the progress of the reaction can be detected using conventional monitoring methods in the art (e.g., TLC, HPLC, or NMR), and the reaction endpoint is generally defined as the disappearance of compound 1 or the cessation of the reaction. The reaction time can be 0.5 to 5 hours, for example, 1 hour.

[0072] The preparation method may further include the following post-processing steps: after the sulfonation reaction is completed, quenching (e.g., quenching by adding ice water), separation (e.g., separation after stirring), washing of the organic phase (e.g., washing successively with hydrochloric acid aqueous solution and saturated saline solution), drying (e.g., drying with anhydrous sodium sulfate), filtration, and concentration to obtain compound 2.

[0073] The sulfonation reaction may include the following steps: mixing the compound 1 and the organic solvent, adding an organic base (e.g., adding at 0-5°C), adding a sulfonating agent (e.g., adding at 0-5°C), and reacting at 10-35°C.

[0074] The materials for the sulfonation reaction can be compounds 1, 2,6-dimethylpyridine, trifluoromethanesulfonic anhydride, and the organic solvents mentioned above.

[0075] In the sulfonation reaction, R 3 The organic solvent can be methyl, and the organic base is a haloalkane solvent. The organic base is 2,6-dimethylpyridine.

[0076] The compound 3 can be prepared by the following steps: in an organic solvent, compound 3a and Trt-X undergo a substitution reaction as shown below to obtain compound 3, where X is a halogen;

[0077]

[0078] In the substitution reaction, the organic solvent can be a conventional organic solvent for such reactions in the art, preferably a mixed solvent of aromatic hydrocarbons and alcohols, such as a mixed solvent of toluene and methanol. The volume ratio of the aromatic hydrocarbons to the alcohols can be (1-5):1, for example, 5:2.

[0079] In the substitution reaction, the mass-to-volume ratio of compound 3a to the organic solvent can be (150-300) g / L, for example (217-218) g / L.

[0080] In the substitution reaction, X can be chlorine (Trt-X is triphenylchloromethane).

[0081] In the substitution reaction, the molar ratio of compound 3a to Trt-X can be (1 to 10):1, for example, 5:1.

[0082] In the substitution reaction, the reaction temperature is the conventional reaction temperature for such reactions in the art, preferably 10–35°C, for example 20–25°C.

[0083] In the substitution reaction, the progress of the reaction can be detected using conventional monitoring methods in the art (e.g., TLC, HPLC, or NMR), and the reaction endpoint is generally defined as the disappearance of the triphenylchloromethane or the cessation of the reaction. The reaction time can be 10–30 hours, for example, 16 hours.

[0084] The preparation method may further include the following post-processing steps: after the substitution reaction is completed, the mixture is separated (e.g., separated after adding water), the organic phase is washed (e.g., washed with water), an aqueous solution of an organic acid (e.g., an aqueous solution of succinic acid) is added, the mixture is filtered, washed (e.g., the filter cake is washed with methyl tert-butyl ether), the mixture is added to an aqueous solution of an inorganic base (e.g., the filter cake is added to an aqueous solution of sodium hydroxide), filtered, washed (e.g., washed with water), and dried to obtain compound 3.

[0085] The substitution reaction preferably includes the following steps: mixing compound 3a and the organic solvent with Trt-X at 0–5°C, and reacting at 10–35°C.

[0086] In one embodiment, the reactants in the substitution reaction may be compound 3a, triphenylchloromethane, and the organic solvent. This invention provides a compound 4;

[0087]

[0088] R 3 As described in any embodiment of the present invention.

[0089] Preferably, compound 4 is

[0090] This invention provides a compound 5;

[0091]

[0092] R 1 and R 2 As described in any embodiment of the present invention, R is preferred. 1 and R 2 They are not both H.

[0093] The present invention provides a method for preparing compound 4, which includes the following steps: in an organic solvent, in the presence of a base, compound 2 and compound 3 undergo a substitution reaction as shown below to obtain compound 4;

[0094]

[0095] R 3 As described in any embodiment of the present invention;

[0096] The organic solvent is a cyclic ether solvent;

[0097] The organic base is a tertiary amine compound;

[0098] In the preparation method described above, the reaction conditions and operations can be as described in any embodiment of the present invention.

[0099] The method for preparing compound 4 preferably further includes the method for preparing compound 2 or 3 as described in any embodiment of the present invention.

[0100] This invention provides a method for preparing compound L, comprising the following steps:

[0101]

[0102] Step 1: In an organic solvent and in the presence of an inorganic base, compound E and compound 5 undergo a substitution reaction to obtain compound F;

[0103] In step 1, the organic solvent may be an ether solvent, such as tetrahydrofuran;

[0104] In step 1, the inorganic base can be an alkali metal hydride, such as NaH;

[0105] In step 1, the reaction can be carried out at -10 to 10°C, for example at 0°C;

[0106] Step 2: In a solvent, in the presence of a catalyst and an inorganic base, compound F and compound G undergo a coupling reaction to obtain compound H;

[0107] In step 2, the solvent can be a mixed solution of ether solvent and water, such as a mixed solution of 1,4-dioxane and water;

[0108] In step 2, the inorganic base can be an alkali metal carbonate, such as cesium carbonate;

[0109] In step 2, the catalyst may be a divalent palladium and a phosphine ligand, such as cataCXium A Pd G3;

[0110] Step 2 can be performed in a protective atmosphere, and the protective atmosphere can be nitrogen.

[0111] In step 2, the reaction can be carried out at 70 to 100°C, for example at 85°C;

[0112] Step 3: In an organic reagent, compound H undergoes a deprotection reaction to obtain compound I;

[0113] In step 3, the organic reagent may be an organic acid reagent, preferably a carboxylic acid reagent, such as acetic acid;

[0114] In step 3, the reaction can be carried out at 40 to 80°C, for example at 60°C;

[0115] Step 4: In an organic solvent and in the presence of a catalyst, compound I and compound J undergo a coupling reaction to obtain compound K;

[0116] In step 4, the organic solvent may be a halogenated hydrocarbon solvent, such as dichloromethane;

[0117] Step 4 may also include an organic acid, which is a carboxylic acid reagent, such as acetic acid;

[0118] In step 4, the catalyst may be a titanate coupling agent, such as tetraisopropyl titanate.

[0119] Step 4 may also include the following step: adding a reducing agent, which may be NaBH(OAc)3;

[0120] In step 4, the reaction can be carried out at room temperature, for example at 25°C;

[0121] Step 5: In an organic solvent and in the presence of an auxiliary agent, compound K undergoes a deprotection reaction to yield compound L;

[0122] In step 5, the organic solvent may be an amide solvent, such as N,N-dimethylformamide;

[0123] In step 5, the reaction can be carried out at room temperature, for example at 25°C;

[0124] In step 5, the auxiliary agent may be an alkali metal fluoride salt, such as cesium fluoride;

[0125] R a It is an amino protecting agent, preferably -CO-OC 1-6 Alkyl groups, such as Boc groups;

[0126] R b It is an alkynyl protecting agent, preferably -Si(C 1-6 Alkyl group 3, such as TIPS group;

[0127] R 1 and R 2 As described in any embodiment of the present invention (e.g., H).

[0128] The method for preparing compound L preferably further comprises the method for preparing compound 5 as described in any embodiment of the present invention.

[0129] This invention provides a method for preparing compound G7, comprising the following steps:

[0130]

[0131] *The C marked is a chiral C, which can be in the S configuration, R configuration, or a mixture thereof;

[0132] Step 1: In an organic solvent and in the presence of an inorganic base, compound A1 and compound 5 undergo a substitution reaction to obtain compound B2;

[0133] R a It is an amino protecting agent, preferably -CO-OC 1-6 Alkyl groups, such as Boc groups;

[0134] R C Halogens (e.g., F), C 1-6 Alkyl or deuterated C 1-6 Alkyl groups (e.g., -CD3);

[0135] R 1 and R 2 As described in any embodiment of the present invention (e.g., H);

[0136] In step 1, the organic solvent may be an ether solvent, such as tetrahydrofuran;

[0137] In step 1, the inorganic base can be an alkali metal carbonate, such as Cs2CO3;

[0138] In step 1, the reaction can be carried out at 80 to 120°C, for example at 90°C;

[0139] Step 2: In a solvent, in the presence of a catalyst and an inorganic base, compound B2 and compound C3 undergo a coupling reaction to obtain compound D4;

[0140] R b It is an alkynyl protecting agent, preferably -Si(C 1-6 Alkyl group 3, such as TIPS group;

[0141] In step 2, the solvent can be a mixed solution of ether solvent and water, such as a mixed solution of 1,4-dioxane and water;

[0142] In step 2, the inorganic base can be an alkali metal carbonate, such as cesium carbonate;

[0143] In step 2, the catalyst may be a divalent palladium and a phosphine ligand, such as cataCXium A Pd G3;

[0144] Step 2 can be performed in a protective atmosphere, and the protective atmosphere can be nitrogen.

[0145] In step 2, the reaction can be carried out at 70 to 100°C, for example at 85°C;

[0146] Step 3: In an organic reagent, compound D4 undergoes a deprotection reaction to obtain compound E5;

[0147] In step 3, the organic reagent may be an organic acid reagent, preferably a carboxylic acid reagent, such as acetic acid;

[0148] In step 3, the reaction can be carried out at 40 to 80°C, for example at 60 or 50°C;

[0149] Step 4: In an organic solvent and in the presence of a catalyst, compound E5 and compound J undergo a coupling reaction to obtain compound F6;

[0150] In step 4, the organic solvent may be a halogenated hydrocarbon solvent, such as dichloromethane;

[0151] Step 4 may further include an organic acid, wherein the organic acid is a carboxylic acid reagent, such as acetic acid;

[0152] In step 4, the catalyst may be a titanate coupling agent, such as tetraisopropyl titanate.

[0153] Step 4 may further include the following step: adding a reducing agent, which may be NaBH(OAc)3;

[0154] In step 4, the reaction can be carried out at room temperature, for example at 25°C;

[0155] Step 5: In an organic solvent and in the presence of an auxiliary agent, compound F6 undergoes a deprotection reaction to obtain compound G7;

[0156] In step 5, the organic solvent may be an amide solvent, such as N,N-dimethylformamide;

[0157] In step 5, the reaction can be carried out at room temperature, for example at 25°C;

[0158] In step 5, the auxiliary agent may be an alkali metal fluoride salt, such as cesium fluoride;

[0159] The method for preparing compound G7 preferably further comprises the method for preparing compound 5 as described in any embodiment of the present invention.

[0160] Preferably, compound L is

[0161] Preferably, compound G7 is The *C is a chiral C, which can be in S configuration, R configuration or a mixture thereof.

[0162] Terminology Explanation:

[0163] Term "C" 1-6 "alkyl" can be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl, such as methyl.

[0164] The term "halogen" can be fluorine, chlorine, or bromine, such as chlorine.

[0165] Based on common knowledge in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0166] The reagents and raw materials used in this invention are all commercially available.

[0167] The significant advantages of this invention are: it provides a KRAS G12D degradation agent intermediate and its preparation method. This preparation process is simple, yields high output, and significantly reduces costs. Detailed Implementation

[0168] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0169] Example 1

[0170]

[0171] Step 1: Preparation of Compound 3

[0172] Piperazine (13.7 kg, 5.0 eq), toluene (45.0 L), and methanol (9.0 L) were added to a reaction vessel. The reaction vessel was cooled to 0–5 °C. A solution of triphenylchloromethane (8.9 kg, 1.0 eq) dissolved in toluene (9.0 L) was added dropwise. After the addition was complete, the reaction vessel was heated to 20–25 °C and stirred for 16 hours. After the reaction was complete, water (9.0 L) was added to the reaction vessel, and the mixture was stirred for 30 minutes. The liquid was separated, and the upper organic phase was washed with water (9.0 L). A solution of succinic acid (4.1 kg) dissolved in water (22.5 L) was added dropwise to the organic phase. After the addition was complete, the reaction vessel was stirred for 2 hours. The mixture was then centrifuged and filtered. The filter cake was washed with methyl tert-butyl ether (5.0 L) and water (5.0 L). The filter cake was added in batches to a sodium hydroxide solution (3.6 kg) dissolved in water (45.0 L). After the addition was complete, the reaction vessel was stirred for 1 hour. The mixture was then centrifuged and filtered. The filter cake was washed with water and dried at 55 °C for 72 hours to obtain compound 3 (8.6 kg) with a purity of 96.1% and a yield of 82%.

[0173] 1 H NMR (400MHz, CDCl3): δ7.13-7.47(m,15H), 2.88-3.13(m,4H), 1.58-2.36(m,4H).

[0174] Step 2: Preparation of Compound 2

[0175] Compound 1 (1.0 kg, 1.0 eq) and dichloromethane (10.0 L) were added to a reaction vessel. The reaction vessel was cooled to 0–5 °C, and 2,6-dimethylpyridine (1.0 kg, 1.1 eq) was added. The reaction vessel temperature was maintained at 0–5 °C, and trifluoromethanesulfonic anhydride (2.6 kg, 1.1 eq) was added dropwise. After the addition was complete, the reaction vessel temperature was raised to 25–30 °C and stirred for 1 hour. After the reaction was complete, the reaction solution was slowly added to ice water (5.0 L) to quench the reaction. After stirring for 30 minutes, the mixture was separated. The organic phase was washed with 6% hydrochloric acid aqueous solution and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 2 (2.13 kg) with a 100% yield, which was directly used in the next step.

[0176] 1 H NMR (400MHz, CDCl3): δ4.68-4.70 (m, 1H), 4.58 (dd, J = 5.2Hz, J = 9.6Hz, 1H), 3.75 (s, 3H), 2.96-2.98 (m, 1H), 1.30 (d, J = 7.2Hz, 3H).

[0177] Step 3: Preparation of Compound 4

[0178] Compound 3 (2.64 kg, 0.95 eq), DIPEA (1.64 kg, 1.5 eq), and 1,4-dioxane (20.0 L) were added to a reaction vessel, and the mixture was stirred at 25–30 °C for half an hour. The reaction vessel was then cooled to 10–15 °C, and compound 2 (2.13 kg, 1.0 eq) was added dropwise. After the addition was complete, the reaction vessel was heated to 25–30 °C and stirred for 16 hours. After the reaction was complete, the reaction solution was added to ice water (10.0 L), stirred for 1–2 hours, filtered by centrifugation, and the filter cake was washed with water and dried at 55 °C for 72 hours to obtain compound 4 (2.3 kg), with a chemical purity of 98.7%, a chiral purity of 99.6%, and a yield of 63%.

[0179] 1H NMR (400MHz, CDCl3): δ7.12-7.46 (m, 15H), 3.61 (s, 3H), 2.63-2.68 (m, 8H), 2.32-2.34 (m, 1H), 1.58-1.60 (m, 2H), 1.10 (d, J = 6.8Hz, 3H).

[0180] Step 4: Preparation of Compound 5

[0181] Lithium aluminum hydride (140.5 g, 0.55 eq, flakes) and tetrahydrofuran (23.5 L) were added to a reaction vessel. The reaction vessel was cooled to 0–5 °C and stirred until the solid was completely dissolved. Maintaining the reaction vessel temperature at 0–5 °C, a tetrahydrofuran solution (10.5 L) of compound 4 (3.35 kg, 1.0 eq) was added dropwise to the reaction vessel. After the addition was complete, stirring was continued for half an hour. After the reaction was complete, the reaction vessel was cooled to -10 °C. Water (140.5 mL), 15% NaOH solution (140.5 mL), and water (421.5 mL) were added dropwise to the reaction vessel in sequence. Then, N,N-dimethylformamide (400 mL) was added and stirring was continued for 15 minutes. The mixture was filtered, and the filter cake was washed with ethyl acetate (6.0 L). The filtrate was washed with water and saturated brine, dried over anhydrous sodium sulfate, and then filtered and concentrated to obtain a white, crude solid. The crude product was recrystallized from methyl tert-butyl ether / n-heptane (1:1, 3 v / w), filtered, and the filter cake was dried at 55 °C for 24 hours to give a white solid compound 5 (1.94 kg) with a chemical purity of 98.6%, a chiral purity of >99.9%, and a yield of 62%.

[0182] 1 H NMR (400MHz, CDCl3): δ7.12-7.44(m,15H),6.01(s,1H),3.55-3.59(m,1H),3.38 (t,J=10.4Hz,1H),2.41-3.02(m,8H),1.61-2.12(m,3H),0.72(d,J=6.8Hz,3H).

[0183] Example 1-1

[0184] Compound 3 (2.48 g, 0.95 eq), DIPEA (1.54 g, 1.5 eq), and N,N-dimethylformamide (20.0 mL) were added to a reaction flask, and the mixture was stirred at 25–30 °C for half an hour. The reaction flask was then cooled to 10–15 °C, and compound 2 (2 g, 1.0 eq) was added dropwise. After the addition was complete, the reaction flask was heated to 25–30 °C and stirred for 16 hours. The reaction was monitored by TLC (PE / EA = 2:1). The starting material disappeared, and a small amount of product was formed.

[0185] Examples 1-2

[0186] Compound 3 (2.48 g, 0.95 eq), DIPEA (1.54 g, 1.5 eq), and dichloromethane (20.0 mL) were added to a reaction flask, and the mixture was stirred at 25–30 °C for half an hour. The reaction flask was then cooled to 10–15 °C, and compound 2 (2 g, 1.0 eq) was added dropwise. After the addition was complete, the reaction flask was heated to 25–30 °C and stirred for 16 hours. The reaction was monitored by TLC (PE / EA = 2:1). The starting material disappeared, and the target product and impurities were formed. The reaction solution was washed with water and concentrated to obtain an oily compound with a purity of less than 80%.

[0187] Examples 1-3

[0188] Compound 3 (5.29 g, 0.95 eq), 2,6-lutidine (2.73 g, 1.5 eq), and 1,4-dioxane (40.0 mL) were added to a reaction flask, and the mixture was stirred at 25–30 °C for half an hour. The reaction flask was then cooled to 10–15 °C, and compound 2 (4.3 g, 1.0 eq) was added dropwise. After the addition was complete, the reaction flask was heated to 25–30 °C and stirred for 16 hours. The reaction was monitored by TLC (PE / EA = 2:1). A large amount of starting material remained, and a small amount of product was formed.

[0189] Examples 1-4

[0190] Compound 4 (5 g, 1.0 eq), methanol (8 mL), and tetrahydrofuran (16.0 mL) were added to a reaction flask, and the mixture was stirred at 25–30 °C for half an hour. The reaction flask was then cooled to 0–5 °C, and NaBH4 (0.88 g, 2.0 eq) was added in portions. After the addition was complete, the reaction flask was heated to 25–30 °C and stirred for 16 hours. The reaction was monitored by TLC (PE / EA = 5:1). A large amount of starting material remained, and a small amount of product was formed.

[0191] Examples 1-5

[0192] Compound 4 (5 g, 1.0 eq), lithium chloride (1 g, 2.0 eq), methanol (8 mL), and tetrahydrofuran (16.0 mL) were added to a reaction flask, and the mixture was stirred at 25–30 °C for half an hour. The reaction flask was then cooled to 0–5 °C, and NaBH4 (0.88 g, 2.0 eq) was added in portions. After the addition was complete, the reaction flask was heated to 25–30 °C and stirred for 16 hours. The reaction was monitored by TLC (PE / EA = 5:1), and the starting material disappeared. The reaction was quenched with saturated ammonium chloride aqueous solution, extracted with ethyl acetate, and concentrated. The concentrate was purified by column chromatography to obtain compound 5 (3 g) with a chemical purity of 97.2% and a chiral purity of 99.0%. The chiral purity of the product was slightly lower than that of the starting material, compound 4 (99.6%).

[0193] Examples 1-6

[0194] Compound 4 (5 g, 1.0 eq) and tetrahydrofuran (25.0 mL) were added to a reaction flask, and the mixture was stirred at 25–30 °C for half an hour. The reaction flask was then cooled to 0–5 °C, and LiBH4 (0.56 g, 2.0 eq) was added in portions. After the addition was complete, the reaction flask was heated to 25–30 °C and stirred for 16 hours. The reaction was monitored by TLC (PE / EA = 5:1). A large amount of starting material remained, and a small amount of product was formed.

[0195] Examples 1-7

[0196] Compound 4 (5 g, 1.0 eq), boron trifluoride diethyl ether (3.3 g, 2.0 eq), and tetrahydrofuran (25.0 mL) were added to a reaction flask, and the mixture was stirred at 25–30 °C for half an hour. The reaction flask was then cooled to 0–5 °C, and NaBH4 (0.88 g, 2.0 eq) was added in portions. After the addition was complete, the reaction flask was heated to 25–30 °C and stirred for 16 hours. The reaction was monitored by TLC (PE / EA = 5:1). The starting material remained, and byproducts of the deprotection of the starting material's Trt protecting group were also detected.

[0197] Example 2

[0198] The following KRAS G12D degrading agent can be prepared using the disclosed process.

[0199]

[0200] The *C is a chiral C, which can be in S configuration, R configuration or a mixture thereof.

[0201] Raw materials for which no preparation method is given may be obtained by means of existing methods in the art, or by means of methods described in PCT / CN2025 / 089926.

[0202] Preparation of compound 117: 1-(5-(9-((4-((R)-3-(((5aS,6S,9R)-2-(8-ethynyl-7-fluoro-3-hydroxynaphthyl-1-yl)-1-fluoro-5a,6,7,8,9,10-hexahydro-5H-4-oxo-3,10a,11,13,14-pentaza-6,9-methylenenaphtho[1,8-ab]hepten-12-yl)oxo)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0203] Step 1: Preparation of (1S,2S,5R)-2-(((7-chloro-8-fluoro-2-(methylthio)-4-oxo-3,4-dihydropyrido[4,3-d]pyrimidin-5-yl)oxy)methyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester,

[0204]

[0205] (1S,2S,5R)-2-(hydroxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (1.99 g, 8.2 mmol) was dissolved in tetrahydrofuran (25 mL), cooled to 0 °C, and NaH (0.98 g, 24.6 mmol) was added. The mixture was reacted at 25 °C for 30 minutes. After cooling to 0 °C, 5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (2.30 g, 8.2 mmol) was added, and the mixture was reacted at 25 °C for 3 hours. After the reaction was complete, water (30 mL) was added to the reaction solution to quench the reaction, and then tetrahydrofuran was removed from the reaction solution by vacuum distillation. The mixture was then adjusted to pH 6 with dilute hydrochloric acid (1.0M) until a precipitate appeared. The mixture was filtered and dried to obtain (1S,2S,5R)-2-(((7-chloro-8-fluoro-2-(methylthio)-4-oxo-3,4-dihydropyridino[4,3-d]pyrimidin-5-yl)oxy)methyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester, which can be used directly in the next step without further purification.

[0206] LC-MS:(ESI,m / z):[M+H] + =486.1.

[0207] Step 2: Preparation of (5aS,6S,9R)-2-chloro-1-fluoro-12-(methylthio)-5a,6,7,8,9,10-hexahydro-5H-4-oxo-3,10a,11,13,14-pentazaza-6,9-methylenenaphtho[1,8-ab]hepten-14-carboxylic acid tert-butyl ester.

[0208]

[0209] (1S,2S,5R)-2-(((7-chloro-8-fluoro-2-(methylthio)-4-oxo-3,4-dihydropyrido[4,3-d]pyrimidin-5-yl)oxy)methyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (3.10 g, 6.37 mmol) and DIEA (5.34 g, 52.80 mmol) were dissolved in DCM (120 mL), and then T3P (12.59 g, 19.8 mmol, 50% EtOAc solution) was added to the mixture. The mixture was reacted at 25 °C for 2 h. After the reaction was complete, water (100 mL) was added, and the mixture was extracted with dichloromethane (50 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated. The crude product was subjected to silica gel column chromatography (PE:EA = 20:1 to 2:1) to obtain (5aS,6S,9R)-2-chloro-1-fluoro-12-(methylthio)-5a,6,7,8,9,10-hexahydro-5H-4-oxo-3,10a,11,13,14-pentaza-6,9-methylenenaphtho[1,8-ab]heptene-14-carboxylic acid tert-butyl ester.

[0210] LC-MS:(ESI,m / z):[M+H] + =468.1.

[0211] Step 3: Preparation of (5aS,6S,9R)-2-chloro-1-fluoro-12-(methylsulfinyl)-5a,6,7,8,9,10-hexahydro-5H-4-oxo-3,10a,11,13,14-pentazaza-6,9-methylenenaphtho[1,8-ab]heptene-14-carboxylic acid tert-butyl ester.

[0212]

[0213] (5aS,6S,9R)-2-chloro-1-fluoro-12-(methylthio)-5a,6,7,8,9,10-hexahydro-5H-4-oxo-3,10a,11,13,14-pentaza-6,9-methylenenaphtho[1,8-ab]hepten-14-carboxylic acid tert-butyl ester (1.60 g, 3.40 mmol) was dissolved in dichloromethane (25 mL), and m-CPBA (1.47 g, 6.85 mmol, 80% wt) was added. The reaction was carried out at room temperature for 2 hours. After the reaction was completed, the reaction solution was poured into a saturated sodium thiosulfate aqueous solution (50 mL), washed with a saturated sodium bicarbonate aqueous solution (30 mL), and then extracted with dichloromethane (50 mL x 3). Combine the organic phases, dry with anhydrous sodium sulfate and filter. Concentrate the filtrate to obtain crude (5aS,6S,9R)-2-chloro-1-fluoro-12-(methylsulfinyl)-5a,6,7,8,9,10-hexahydro-5H-4-oxo-3,10a,11,13,14-pentazaza-6,9-methylenenaphtho[1,8-ab]heptene-14-carboxylic acid tert-butyl ester. The crude product can be used directly in the next step without purification.

[0214] LC-MS:(ESI,m / z):[M+H] + =484.1.

[0215] Step 4: Preparation of (5aS,6S,9R)-2-chloro-1-fluoro-12-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxo-3,10a,11,13,14-pentazaza-6,9-methylenenaphtho[1,8-ab]heptene-14-carboxylic acid tert-butyl ester.

[0216]

[0217] (R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)prop-1-ol (405 mg, 1.01 mmol) was dissolved in tetrahydrofuran (5 mL), cooled to 0 °C, and NaH (58 mg, 1.45 mmol) was slowly added. The mixture was reacted at 25 °C for 30 minutes. Then, (5aS,6S,9R)-2-chloro-1-fluoro-12-(methylsulfinyl)-5a,6,7,8,9,10-hexahydro-5H-4-oxo-3,10a,11,13,14-pentaza-6,9-methylenenaphthalo[1,8-ab]hepten-14-carboxylic acid tert-butyl ester (350 mg, 0.72 mmol) was added, and the mixture was reacted at 0 °C for 2 hours. After the reaction was complete, saturated ammonium chloride aqueous solution (30 mL) was added, and the mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed with saturated NaCl solution, dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH = 15:1) to obtain (5aS,6S,9R)-2-chloro-1-fluoro-12-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxo-3,10a,11,13,14-pentaza-6,9-methylenenaphtho[1,8-ab]heptene-14-carboxylic acid tert-butyl ester.

[0218] LC-MS:(ESI,m / z):[M+H] + =820.3.

[0219] Step 5: Preparation of (5aS,6S,9R)-1-fluoro-2-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-12-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxo-3,10a,11,13,14-pentaza-6,9-methylenenaphtho[1,8-ab]heptene-14-carboxylic acid tert-butyl ester.

[0220]

[0221] Add cataCXium A Pd to a mixed solution of (5aS,6S,9R)-2-chloro-1-fluoro-12-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxo-3,10a,11,13,14-pentaza-6,9-methylenenaphtho[1,8-ab]hepten-14-carboxylic acid tert-butyl ester (200 mg, 0.24 mmol) and 6-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-5-((triisopropylsilyl)ethynyl)naphth-2-ol (228 mg, 0.49 mmol) in 1,4-dioxane (2 mL) and H2O (0.4 mL). G3 (18 mg, 0.024 mmol) and cesium carbonate (159 mg, 0.49 mmol). The mixture was reacted at 85 °C for 8 hours under nitrogen protection. Water (20 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (DCM:MeOH = 10:1) to give (5aS,6S,9R)-1-fluoro-2-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-12-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxo-3,10a,11,13,14-pentaza-6,9-methylenenaphtho[1,8-ab]heptene-14-carboxylic acid tert-butyl ester.

[0222] LC-MS:(ESI,m / z):[M+H] + =1126.4.

[0223] Step 6: Preparation of (5aS,6S,9R)-1-fluoro-2-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-12-((R)-2-methyl-3-(piperazin-1-yl)propoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxo-3,10a,11,13,14-pentaza-6,9-methylenenaphtho[1,8-ab]heptene-14-carboxylic acid tert-butyl ester.

[0224]

[0225] (5aS,6S,9R)-1-fluoro-2-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-12-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxo-3,10a,11,13,14-pentaza-6,9-methylenenaphtho[1,8-ab]hepten-14-carboxylic acid tert-butyl ester (180 mg, 0.16 mmol) was dissolved in glacial acetic acid (3 mL) and reacted at 60 °C for 4 hours. After the reaction was complete, the reaction solution was directly concentrated, and the crude product was subjected to silica gel column chromatography (DCM:MeOH(NH3)=15:1) to obtain (5aS,6S,9R)-1-fluoro-2-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-12-((R)-2-methyl-3-(piperazin-1-yl)propoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxo-3,10a,11,13,14-pentaza-6,9-methylenenaphtho[1,8-ab]heptene-14-carboxylic acid tert-butyl ester.

[0226] LC-MS:(ESI,m / z):[M+H] + =884.3.

[0227] Step 7: Preparation of (5aS,6S,9R)-12-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-1-fluoro-2-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5a,6,7,8,9,10-hexahydro-5H-4-oxo-3,10a,11,13,14-pentaza-6,9-methylenenaphtho[1,8-ab]hepten-14-carboxylic acid tert-butyl ester,

[0228]

[0229] (5aS,6S,9R)-1-fluoro-2-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-12-((R)-2-methyl-3-(piperazin-1-yl)propoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxo-3,10a,11,13,14-pentaza-6,9-methylenenaphtho[1,8-ab]hepten-14-carboxylic acid tert-butyl ester To a stirred solution of DCM / AcOH (2.5 mL / 0.1 mL, 120 mg, 0.14 mmol), 3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-carboxaldehyde (70 mg, 0.16 mmol) and tetraisopropyl titanate (193 mg, 0.68 mmol) were added and the mixture was stirred at 25 °C for 1 hour. The mixture was then further stirred with NaBH(OAc)3 (58 mg, 0.27 mmol) in an ice bath for 2 hours. The mixture was concentrated and purified by silica gel column chromatography (DCM:MeOH = 100:1 to 10:1) to give (5aS,6S,9R)-12-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-1-fluoro-2-(7-fluoro-3-hydroxy-8-(((triisopropylsilyl)ethynyl)naphth-1-yl)-5a,6,7,8,9,10-hexahydro-5H-4-oxo-3,10a,11,13,14-pentaza-6,9-methylenenaphtho[1,8-ab]hepten-14-carboxylic acid tert-butyl ester.

[0230] LC-MS:(ESI,m / z):[M / 2+H] + =648.4.

[0231] Step 8: Preparation of 1-(5-(9-((4-((R)-3-(((5aS,6S,9R)-2-(8-ethynyl-7-fluoro-3-hydroxynaphthyl-1-yl)-1-fluoro-5a,6,7,8,9,10-hexahydro-5H-4-oxo-3,10a,11,13,14-pentaza-6,9-methylenenaphtho[1,8-ab]hepten-12-yl)oxo)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione,

[0232]

[0233] To (5aS,6S,9R)-12-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-1-fluoro-2-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphthalene- Cesium fluoride (32 mg, 0.211 mmol) was added to a DMF (1 mL) solution of 1-yl)-5a,6,7,8,9,10-hexahydro-5H-4-oxo-3,10a,11,13,14-pentaza-6,9-methylenenaphtho[1,8-ab]hepten-14-carboxylic acid tert-butyl ester (80 mg, 0.07 mmol) and stirred at room temperature for 30 minutes. Water (3 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (5 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was dissolved in 1 mL of 1,4-dioxane and HCl / 1,4-dioxane (6N, 1 mL) was added, and the mixture was reacted at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure, and the pH was adjusted to >7 with aqueous NaHCO3 solution and extracted with EtOAc (30 mL x 3). The organic phase was washed with saturated sodium chloride aqueous solution, dried over anhydrous Na2SO4, and concentrated. The crude product was purified by high performance liquid chromatography to obtain 1-(5-(9-((4-((R)-3-(((5aS,6S,9R)-2-(8-ethynyl-7-fluoro-3-hydroxynaphthyl-1-yl)-1-fluoro-5a,6,7,8,9,10-hexahydro-5H-4-oxo-3,10a,11,13,14-pentaza-6,9-methylenenaphtho[1,8-ab]hepten-12-yl)oxo)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione.

[0234] LC-MS:(ESI,m / z):[M+H] + =1039.4.

[0235] 1HNMR(400MHz,DMSO-d6)10.33(s,1H),10.16(s,1H),8.01-7.90(m,1H),7.51-7.41(m ,1H),7.40-7.34(m,2H),7.32(d,J=2.0Hz,1H),7.23-7.03(m,2H),4.91-4.76(m,1H) ,4.57-4.30(m,3H),4.16-3.99(m,3H),3.84(s,3H),3.68-3.48(m,8H),3.12-3.04(m ,1H),2.73-2.61(m,7H),2.47-1.97(m,11H),1.84-1.19(m,12H),1.10-0.94(m,6H).

[0236] Preparation of compound 125:

[0237] 1-(5-(9-((4-((2R)-3-((7-(8-ethynyl-7-fluoro-3-hydroxynaphthyl-1-yl)-8-fluoro-4-(1-(methyl-d3)-5λ) 3 Preparation of -3,8-diazabicyclo[3.2.1]octane-3-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidin-2,4(1H,3H)-dione;

[0238] Step 1: 3-(7-chloro-8-fluoro-2-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-(methyl-d3)-5λ 3 Preparation of tert-butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid,

[0239]

[0240] At room temperature, (R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propanol (538 mg, 1.34 mmol) and 22-P2 (400 mg, 0.90 mmol) were dissolved in tetrahydrofuran (10 mL) and added to a sealed reactor. Cs2CO3 (586 mg, 1.8 mmol) was added, and the reaction was carried out at 90 °C for 16 hours. After the reaction was completed, an aqueous solution (20 mL) was added, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, washed with saturated NaCl solution, dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated. The crude product was subjected to silica gel column chromatography (DCM:MeOH = 20:1) to give 3-(7-chloro-8-fluoro-2-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-(methyl-d3)-5λ 3 -3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester.

[0241] LC-MS:(ESI,m / z):[M-Trt+H] + =567.2.

[0242] Step 2: 3-(8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphth-1-yl)-2-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-(methyl-d3)-5λ 3 Preparation of 3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester

[0243]

[0244] 3-(7-chloro-8-fluoro-2-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-(methyl-d3)-5λ 3CataCXium APd G3 (53 mg, 0.073 mmol) and cesium carbonate (712 mg, 2.18 mmol) were added to a mixed solution of tert-butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid (590 mg, 0.73 mmol) and ((6-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-5-((triisopropylsilyl)ethynyl)naphth-2-yl)oxy)triisopropylsilane (546 mg, 0.87 mmol) in 1,4-dioxane (5 mL) and H2O (1 mL). The mixture was reacted at 85 °C for 16 h under nitrogen protection, water (20 mL) was added, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by column chromatography (DCM:MeOH = 20:1) to give 3-(8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphth-1-yl)-2-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-(methyl-d3)-5λ 3 -3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester.

[0245] LC-MS:(ESI,m / z):[M-Trt+H] + =1029.6.

[0246] Step 3: 3-(8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphth-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-(methyl-d3)-5λ 3 Preparation of 3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester

[0247]

[0248] 3-(8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphth-1-yl)-2-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-(methyl-d3)-5λ 33,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (650 mg, 0.51 mmol) was dissolved in DCM (3 mL), and then glacial acetic acid (3 mL) was added. The mixture was refluxed at 50 °C for 1 hour. After the reaction was completed, the reaction solution was directly concentrated, and the crude product was purified by silica gel column chromatography (DCM:MeOH = 15:1) to obtain 3-(8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthyl-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-(methyl-d3)-5λ 3 -3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester.

[0249] LC-MS:(ESI,m / z):[M / 2+H] + =515.5.

[0250] Step 4: 3-(2-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphth-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methyl-d3)-5λ 3 Preparation of 3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester

[0251]

[0252] 3-(8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphth-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-(methyl-d3)-5λ 3To a stirred solution of tert-butyl octane-8-carboxylate (150 mg, 0.15 mmol) in DCM / AcOH (2 mL / 0.2 mL), 3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-carboxaldehyde (75 mg, 0.17 mmol) and Ti(O-iPr)4 (207 mg, 0.73 mmol) were added, and the mixture was stirred at 25 °C for 2 hours. The mixture was then further stirred in an ice bath with NaBH(OAc)3 (93 mg, 0.44 mmol) for 1 hour. The mixture was concentrated and purified by silica gel column chromatography (DCM:MeOH = 100:1 to 10:1) to give 3-(2-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphth-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methyl-d3)-5λ 3 -3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester.

[0253] LC-MS:(ESI,m / z):[M / 2+H] + =721.1.

[0254] Step 5: 1-(5-(9-((4-((2R)-3-((7-(8-ethynyl-7-fluoro-3-hydroxynaphthyl-1-yl)-8-fluoro-4-(1-(methyl-d3)-5λ) 3 Preparation of -3,8-diazabicyclo[3.2.1]octane-3-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidin-2,4(1H,3H)-dione

[0255]

[0256] To 3-(2-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphth-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methyl-d3)-5λ3 Cesium fluoride (237 mg, 1.56 mmol) was added to a 2 mL solution of tert-butyl octane-8-carboxylic acid (150 mg, 0.104 mmol) in DMF, and the mixture was stirred at room temperature for 4 hours. Water (20 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. HCl / 1,4-dioxane (6 N, 2.0 mL) was added to a stirred solution of 1,4-dioxane (2.0 mL) of the concentrate, and the mixture was reacted at room temperature for 0.5 hours. The mixture was adjusted to pH > 7 with aqueous Na₂CO₃ solution and extracted with EtOAc (20 mL x 3). The organic phase was washed with brine, dried over anhydrous Na₂SO₄, and concentrated. The crude product was purified by high performance liquid chromatography to obtain 1-(5-(9-((4-((2R)-3-((7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-4-(1-(methyl-d3)-5λ 3 -3,8-diazabicyclo[3.2.1]octane-3-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidin-2,4(1H,3H)-dione.

[0257] LC-MS:(ESI,m / z):[M+H] + =1028.4.

[0258] 1 HNMR (400MHz, DMSO-d6) δ10.33(s,1H),9.05(d,J=2.4Hz,1H),7.97(dd,J=9.2,6.0Hz,1H),7.46(t,J=9.0Hz,1H ),7.41-7.33(m,2H),7.31(d,J=1.6Hz,1H),7.22-7.09(m,2H),4.49-4.39(m,2H),4.37-4.28(m,1H),4.20-4.0 0(m,2H),3.93(d,J=2.2Hz,1H),3.84(s,3H),3.63-3.55(m,6H),3.38-3.34(m,2H),2.68(t,J=6.4Hz,2H),2.48 -2.11(m,12H),2.09-2.02(m,2H),1.86-1.64(m,5H),1.56-1.36(m,6H),1.34-1.24(m,2H),1.11-0.91(m,7H).

[0259] Preparation of compound 222:

[0260] 1-(5-(9-((4-((2R)-3-((7-(8-ethynyl-7-fluoro-3-hydroxynaphthyl-1-yl)-8-fluoro-4-(1-fluoro-5λ) 3 -3,8-diazabicyclo[3.2.1]octane-3-yl)pyridin[4,3-d]pyrimidin-2-yl)oxy)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidin-2,4(1H,3H)-dione

[0261] Step 1: 3-(7-chloro-8-fluoro-2-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)pyridin[4,3-d]pyrimidin-4-yl)-1-fluoro-5λ 3 Preparation of 3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester

[0262]

[0263] At room temperature, (R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propanol (450 mg, 1.13 mmol) and intermediate 26-P1 (270 mg, 0.6 mmol) were dissolved in tetrahydrofuran (10 mL) and added to a sealed reactor. Cs₂CO₃ (443 mg, 1.36 mmol) was added, and the reaction was carried out at 90 °C for 16 hours. After the reaction was complete, an aqueous solution (20 mL) was added, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (DCM:MeOH = 50:1) to give 3-(7-chloro-8-fluoro-2-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)pyridin[4,3-d]pyrimidin-4-yl)-1-fluoro-5λ 3 -3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester.

[0264] LC-MS (ESI, m / z): [M+Na] + =832.3.

[0265] Step 2: 1-Fluoro-3-(8-Fluoro-7-(7-Fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphth-1-yl)-2-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)pyridin[4,3-d]pyrimidin-4-yl)-5λ 3 Preparation of 3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester

[0266]

[0267] 3-(7-chloro-8-fluoro-2-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)pyridin[4,3-d]pyrimidin-4-yl)-1-fluoro-5λ 3 Cesium carbonate (419 mg, 1.29 mmol) and cataXium A Pd G3 (63 mg, 0.086 mmol) were added to a mixed solution of 3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (350 mg, 0.43 mmol) and ((6-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-5-((triisopropylsilyl)ethynyl)naphth-2-yl)oxy)triisopropylsilane (325 mg, 0.52 mmol) in 1,4-dioxane (20 mL) and H2O (4 mL), and reacted at 90 °C for 3 hours under N2 protection. The reaction solution was filtered, the filtrate was concentrated, and the concentrate was purified by silica gel column chromatography (DCM:MeOH = 100:1) to obtain 1-fluoro-3-(8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphth-1-yl)-2-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)pyridin[4,3-d]pyrimidin-4-yl)-5λ 3 -3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester.

[0268] LC-MS:(ESI,m / z):[M-Trt+H] + =1030.5.

[0269] Step 3: 1-Fluoro-3-(8-Fluoro-7-(7-Fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphth-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyridin[4,3-d]pyrimidin-4-yl)-5λ 3 Preparation of 3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester

[0270]

[0271] 1-fluoro-3-(8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphth-1-yl)-2-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)pyridin[4,3-d]pyrimidin-4-yl)-5λ 3-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (300 mg, 0.236 mmol) was dissolved in acetic acid (10 mL) and stirred at 50 °C for 2 hours. After the reaction was complete, the mixture was concentrated and purified by silica gel column chromatography (DCM:MeOH = 10:1) to give 1-fluoro-3-(8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthyl-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyridin[4,3-d]pyrimidin-4-yl)-5λ 3 -3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester.

[0272] LC-MS:(ESI,m / z):[M / 2+H] + =515.9.

[0273] Step 4: 3-(2-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthyl-1-yl)pyridin[4,3-d]pyrimidin-4-yl)-1-fluoro-5λ 3 Preparation of 3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester

[0274]

[0275] 1-Fluoro-3-(8-Fluoro-7-(7-Fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphth-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyridin[4,3-d]pyrimidin-4-yl)-5λ 3A solution of tert-butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid (200 mg, 0.194 mmol) and 3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-carboxaldehyde (98 mg, 0.23 mmol) in DCM / AcOH (10 mL / 2 mL) was added to tetraisopropoxytitanium (1 g, 3.52 mmol), and the mixture was stirred at 30 °C for 1 hour. The mixture was then added to NaBH(OAc)3 (165 mg, 0.78 mmol) in an ice bath and stirred for 2 hours. The mixture was concentrated and purified by silica gel column chromatography (DCM:MeOH = 100:1 to 50:1) to obtain 3-(2-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthyl-1-yl)pyridin[4,3-d]pyrimidin-4-yl)-1-fluoro-5λ 3 -3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester.

[0276] LC-MS:(ESI,m / z):[M / 2+H] + =722.0.

[0277] Step 5: 1-(5-(9-((4-((2R)-3-((7-(8-ethynyl-7-fluoro-3-hydroxynaphthyl-1-yl)-8-fluoro-4-(1-fluoro-5λ) 3 Preparation of 3,8-diazabicyclo[3.2.1]octane-3-yl)pyridin[4,3-d]pyrimidin-2-yl)oxy)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidin-2,4(1H,3H)-dione

[0278]

[0279] To 3-(2-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphth-1-yl)pyridin[4,3-d]pyrimidin-4-yl)-1-fluoro-5λ 3Cesium fluoride (342 mg, 2.25 mmol) was added to a stirred solution of tert-butyl octane-8-carboxylic acid (180 mg, 0.125 mmol) in DMF (3 mL), and the mixture was stirred at room temperature for 2 hours. Water (50 mL) was added to the reaction mixture, followed by extraction with ethyl acetate (30 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was dissolved in 1,4-dioxane (10 mL), and HCl / 1,4-dioxane (4.0 M, 5 mL) was added dropwise at room temperature, followed by stirring for 30 minutes at room temperature. The mixture was adjusted to pH > 7 with aqueous Na₂CO₃ solution, followed by extraction with EtOAc (30 mL x 3). The organic phase was washed with brine, dried over anhydrous Na₂SO₄, and concentrated. The crude product was purified by high performance liquid chromatography to obtain 1-(5-(9-((4-((2R)-3-((7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-4-(1-fluoro-5λ) 3 -3,8-diazabicyclo[3.2.1]octane-3-yl)pyridin[4,3-d]pyrimidin-2-yl)oxy)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidin-2,4(1H,3H)-dione.

[0280] LC-MS: (ESI, m / z): [M / 2+H]+=515.5.

[0281] 1H NMR (400MHz, DMSO-d6) δ10.34(s,1H),10.18(s,1H),9.08(d,J=4.4Hz,1H),7.99-7.96(m,1H), 7.48-7.44(m,1H),7.39-7.32(m,3H),7.19-7.14(m,2H),4.89-4.72(m,1H),4.52-4.47(m,1H), 4.33-4.16(m,1H),4.10-4.05(m,1H),3.96(d,J=1.2Hz,1H),3.84(s,3H),3.67-3.59(m,8H),2. 70-2.66(m,2H),2.33-2.13(m,10H),2.05-1.99(m,4H),1.77-1.29(m,12H),1.07-0.97(m,7H).

[0282] Preparation of compound 228:

[0283] 1-(5-(9-((4-((2R)-3-((7-(8-ethynyl-7-fluoro-3-hydroxynaphthyl-1-yl)-8-fluoro-4-(1-fluoro-5λ)3 -3,8-diazabicyclo[3.2.1]octane-3-yl)pyridin[4,3-d]pyrimidin-2-yl)oxy)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidin-2,4(1H,3H)-dione

[0284] Step 1: 3-(7-chloro-8-fluoro-2-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)pyridin[4,3-d]pyrimidin-4-yl)-1-fluoro-5λ 3 Preparation of 3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester

[0285]

[0286] At room temperature, (R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propanol (403 mg, 1.01 mmol) and intermediate 26-P2 (300 mg, 0.67 mmol) were dissolved in tetrahydrofuran (10 mL) and added to a sealed reactor. Cs2CO3 (456 mg, 1.4 mmol) was added, and the reaction was carried out at 90 °C for 16 hours. After the reaction was completed, an aqueous solution (20 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated NaCl solution, dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (DCM:MeOH = 20:1) to obtain 3-(7-chloro-8-fluoro-2-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)pyridin[4,3-d]pyrimidin-4-yl)-1-fluoro-5λ 3 -3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester.

[0287] LC-MS:(ESI,m / z):[M-Trt+H] + =568.2.

[0288] Step 2: 1-Fluoro-3-(8-Fluoro-7-(7-Fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphth-1-yl)-2-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)pyridin[4,3-d]pyrimidin-4-yl)-5λ 3 Preparation of 3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester

[0289]

[0290] 3-(7-chloro-8-fluoro-2-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)pyridin[4,3-d]pyrimidin-4-yl)-1-fluoro-5λ 3 A mixture of tert-butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid (456 mg, 0.56 mmol) and ((6-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-5-((triisopropylsilyl)ethynyl)naphth-2-yl)oxy)triisopropylsilane (421 mg, 0.67 mmol) in 1,4-dioxane (3 mL) and H2O (0.6 mL) was added to CataCXium A Pd G3 (41 mg, 0.056 mmol) and cesium carbonate (549 mg, 1.69 mmol). The mixture was reacted at 85 °C for 16 hours under nitrogen protection, water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by column chromatography (DCM:MeOH = 20:1) to give 1-fluoro-3-(8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphth-1-yl)-2-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)pyridin[4,3-d]pyrimidin-4-yl)-5λ 3 -3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester.

[0291] LC-MS:(ESI,m / z):[(M-Trt)+H] + =516.2.

[0292] Step 3: 1-Fluoro-3-(8-Fluoro-7-(7-Fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphth-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyridin[4,3-d]pyrimidin-4-yl)-5λ 3 Preparation of 3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester

[0293]

[0294] 1-fluoro-3-(8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphth-1-yl)-2-((R)-2-methyl-3-(4-triphenylmethylpiperazin-1-yl)propoxy)pyridin[4,3-d]pyrimidin-4-yl)-5λ 3-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (440 mg, 0.34 mmol) was dissolved in DCM (2 mL) and glacial acetic acid (2 mL) was added. The mixture was refluxed at 50 °C for 1 hour. After the reaction was completed, the crude product was concentrated and purified by silica gel column chromatography (DCM:MeOH = 10:1) to obtain 1-fluoro-3-(8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthyl-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyridin[4,3-d]pyrimidin-4-yl)-5λ 3 -3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester.

[0295] LC-MS:(ESI,m / z):[M / 2+H] + =515.9.

[0296] Step 4: 3-(2-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphthyl-1-yl)pyridin[4,3-d]pyrimidin-4-yl)-1-fluoro-5λ 3 Preparation of 3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester

[0297]

[0298] 1-Fluoro-3-(8-Fluoro-7-(7-Fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphth-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyridin[4,3-d]pyrimidin-4-yl)-5λ 3To a solution of tert-butyl octane-8-carboxylate (150 mg, 0.15 mmol) in DCM / AcOH (2 mL / 0.2 mL), 3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-carboxaldehyde (75 mg, 0.17 mmol) and Ti(O-iPr)4 (207 mg, 0.73 mmol) were added, and the mixture was stirred at 25 °C for 2 hours. The mixture was then stirred for 1 hour with NaBH(OAc)3 (93 mg, 0.44 mmol) added in an ice bath. The mixture was concentrated and purified by silica gel column chromatography (DCM:MeOH = 100:1 to 10:1) to give 3-(2-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphth-1-yl)pyridin[4,3-d]pyrimidin-4-yl)-1-fluoro-5λ 3 -3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester.

[0299] LC-MS:(ESI,m / z):[M / 2+H] + =721.6.

[0300] Step 5: 1-(5-(9-((4-((2R)-3-((7-(8-ethynyl-7-fluoro-3-hydroxynaphthyl-1-yl)-8-fluoro-4-(1-fluoro-5λ) 3 Preparation of 3,8-diazabicyclo[3.2.1]octane-3-yl)pyridin[4,3-d]pyrimidin-2-yl)oxy)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidin-2,4(1H,3H)-dione

[0301]

[0302] To 3-(2-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-8-((triisopropylsilyl)ethynyl)-3-((triisopropylsilyl)oxy)naphth-1-yl)pyridin[4,3-d]pyrimidin-4-yl)-1-fluoro-5λ 3Cesium fluoride (246 mg, 1.62 mmol) was added to a stirred solution of tert-butyl octane-8-carboxylic acid (156 mg, 0.11 mmol) in DMF (2 mL), and the mixture was stirred at room temperature for 4 hours. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. HCl / 1,4-dioxane (6N, 2.0 mL) was added to a stirred solution of 1,4-dioxane (2.0 mL) of the concentrate, and the mixture was reacted at room temperature for 0.5 hours. Na₂CO₃ aqueous solution was added to the reaction mixture to adjust the pH to >7, and the mixture was extracted with EtOAc (10 mL x 3). The organic phase was washed with brine, dried over anhydrous Na₂SO₄, and concentrated. The crude product was purified by high performance liquid chromatography to obtain 1-(5-(9-((4-((2R)-3-((7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-4-(1-fluoro-5λ) 3 -3,8-diazabicyclo[3.2.1]octane-3-yl)pyridin[4,3-d]pyrimidin-2-yl)oxy)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidin-2,4(1H,3H)-dione.

[0303] LC-MS:(ESI,m / z):[M+H] + =1029.3.

[0304] 1 H NMR (400MHz, DMSO-d6) δ10.33(s,1H),9.08(d,J=3.1Hz,1H),7.98(dd,J=9.1,6.0Hz,1H),7.46(t,J=9.0Hz,1H) ,7.41-7.34(m,2H),7.32(s,1H),7.20-7.13(m,2H),4.90-4.69(m,1H),4.52-4.43(m,1H),4.36-4.16(m,1H),4. 14-4.15(m,1H),3.96(d,J=5.0Hz,1H),3.84(s,3H),3.73-7.64(m,2H),3.62-3.51(m,6H),2.68(t,J=6.3Hz,2H ),2.48-2.11(m,12H),2.10-1.83(m,5H),1.80-1.65(m,3H),1.54-1.38(m,6H),1.29(s,2H),1.11-0.95(m,7H).

[0305] Test Example 1: KRAS-G12D Degradation Test

[0306] The degradation effect of compounds on KRAS-G12D was investigated in human lung cancer cells A-427(ATCC) with KRAS G12D mutation, human gastric cancer cells AGS with KRAS G12D mutation, human metastatic pancreatic cancer cells AsPc-1 with KRAS G12D mutation, human pancreatic adenocarcinoma cells Panc04.03 with KRAS G12D mutation, human pancreatic adenocarcinoma cells PK59 with KRAS G12D mutation, and human colon cancer cells GP2D with KRAS G12D mutation. Specifically, 0.95 mL of cells were seeded into each well of a 24-well cell culture plate to achieve a cell density of 5 × 10⁻⁶ cells / well. 5 Cells / well; cell culture plates were incubated overnight at 37°C in a 5% CO2 incubator. Then, 50 μL of diluted compound solution was added to the corresponding wells containing cells, bringing the final compound concentration to the range of 0.03–3000 nM. The final concentration of DMSO was 0.25%. After adding the compound, the cell culture plates were incubated at 37°C in a 5% CO2 incubator for 24 hours. After removing the culture medium from the 24-well cell culture plates, the cells were washed twice with 1×PBS (Kaige). For A-427 cells adhering to the bottom of the cell culture plates, 120 μL of RIPA (Strong) lysis buffer (Beyotime) was added, followed by lysis with 1 mM benzyl sulfonyl fluoride, a mixture of protease inhibitors (Beyotime), and a mixture of protease phosphatase inhibitors (Beyotime). After standing on ice for 30 minutes, the protein lysis buffer from each well was transferred to a 1.5 mL centrifuge tube and centrifuged at 15000 g, 4°C for 20 minutes. The cell lysis supernatant was stored at -80°C for later use.

[0307] After thawing the prepared cell lysis supernatant samples from a -80°C freezer, the total protein concentration in the cell lysis solution was determined using a BCA protein quantification kit (Tiangen). The total protein concentration was then adjusted to 0.5 μg / μL using PBS and 5×SDS-PAGE protein loading buffer (Beyotime), and incubated at 100°C for 15 minutes. Afterward, the samples were placed on ice for 5 minutes, centrifuged at 14000g and 4°C for 1 minute, and mixed thoroughly as the loading sample for Western blotting. 10 μL (5 μg total protein) was loaded into the wells of a 10% precast gel (Kage), and sufficient Tris-MOPS-SDS electrophoresis buffer (Adamas) was added. Electrophoresis was performed at a constant voltage of 120V for 55 minutes. After electrophoresis, the proteins on the gel were transferred to a PVDF membrane at a constant current of 250mA for 55 minutes. After transfer, the PVDF membrane was incubated in 1×Quick Block blocking buffer (Beyotime) at room temperature for 30 minutes. Following blocking, the PVDF membrane was incubated overnight at 4°C with KRAS antibody (Abcam) diluted 1:1000 and β-actin antibody (Abcam) diluted 1:2000 using 5% BSA as a dilution buffer. The membrane was then washed 10 minutes at a time for a total of 3 times with 1×TBST buffer (2.4g Tris, 8.8g NaCl, 1.5mL Tween 20, pH adjusted to 7.4, volume brought to 1L). The membrane was then incubated with 5% BSA-diluted secondary antibody (Abcam) at room temperature for 2 hours, followed by washing 1×TBST buffer for 10 minutes at a time for a total of 3 times. Finally, the membrane was incubated with Clarity Western ECL Substrate (BIO-RAD) for 5 minutes for chemiluminescence development. Color development and protein mapping were performed using a ChemiScope 6200Touch chemiluminescence imaging system (Qingxiang). Protein profiles were analyzed for grayscale values ​​using chemiluminescence analysis software (Qinxiang). The formula used was: Grayscale correction value = (Target protein grayscale value / Corresponding internal reference grayscale value) × 10 3 The grayscale correction value for each sample was calculated. This value was then compared with the grayscale correction value of the control group to calculate the degradation rate. Finally, the DC of the compound was obtained by nonlinear curve fitting using GraphPad Prism 8 analysis software with logarithmic concentration-inhibition rate. 50 and D max Values ​​(Table 1).

[0308] Table 1

[0309]

[0310] Test Example 2: 3D Cell Proliferation Inhibition Assay

[0311] The inhibitory effect of the compound on the proliferation of 3D cells was investigated in human lung cancer cells A427 (KRAS G12D mutation), human gastric cancer cells AGS (KRAS G12D mutation), human metastatic pancreatic cancer cells AsPc-1 (KRAS G12D mutation), human pancreatic cancer cells Panc 04.03 (KRAS G12D mutation), human pancreatic cancer cells HPAC (KRAS G12D mutation), human pancreatic adenocarcinoma cells PK59 (KRAS G12D mutation), and human colon cancer cells GP2D (KRAS G12D mutation). The specific methods were as follows:

[0312] 10 μL of a serially diluted compound solution was added to the corresponding well of a 96-well PerkinElmer microplate to achieve a final concentration of the test compound ranging from 10000 nM to 0.026 nM. The final concentration of DMSO was 0.2%. Then, 90 μL of cells were seeded into each well to achieve a cell density of 5 × 10⁻⁶ cells / well. 2 Cells / well. In addition to the test compound wells, DMSO control wells and culture medium control wells were also set up. The DMSO control wells contained both DMSO and cells, while the culture medium wells contained only culture medium. After adding the samples, the 96-well low-adsorption microplate was placed in a 5% CO2 incubator and incubated at 37°C for 7 days. After 7 days, the microplate was removed, and 100 μL of CTG reagent (Promega) was added to each well. After incubation at room temperature for 60 minutes, the readings were taken using a chemiluminescence immunoassay program on an EnVision microplate reader. The cell proliferation inhibition percentage was calculated using the formula: Inhibition rate % = (DMSO control group average - compound single concentration reading) / (DMSO control group average - culture medium control group average) × 100. The inhibition rate of the compound at each concentration was calculated, and then the IC50 of the compound was obtained by nonlinear curve fitting using GraphPad Prism 8 with logarithmic concentration-inhibition rate. 50 Values ​​(Table 2).

[0313] Table 2

[0314]

[0315]

[0316] In Table 6, "-" indicates that no detection was performed.

[0317] Test Example 3: Mouse Pharmacokinetic Study

[0318] (1) Experimental reagents

[0319] The compounds used in this experiment are derived from compounds in specific embodiments of this invention.

[0320] (2) Laboratory animals

[0321] Six male ICR mice were obtained from Shanghai Shengchang Biotechnology Co., Ltd.

[0322] (3) Drug preparation and administration

[0323] ICR mice were administered a single intravenous (IV) injection: An appropriate amount of the compound was weighed, first dissolved by vortexing and sonication with 1% DMSO, then vortexed with 10% Solutol, and finally vortexed with 89% 0.25% glucose solution to obtain a clear solution. Three mice were fasted overnight and then administered the compound via tail vein injection at a dose of 1 mg / mL.

[0324] ICR mice were administered a single oral (PO) dose: An appropriate amount of the compound was weighed, first dissolved by vortexing and sonication with 2% DMSO, then vortexed with 10% Solutol, and finally vortexed with 88% 0.25% glucose solution to obtain a clear solution. Three mice were fasted overnight before being administered the drug orally at a dose of 30 mg / mL.

[0325] (4) Sample collection

[0326] Sampling time points were: 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after drug administration. Blood was collected via the submandibular vein at the above-defined time points, with 30-40 μL of whole blood collected per sample. Heparin sodium was used for anticoagulation. After collection, the samples were placed on wet ice and centrifuged within 1 hour to separate the plasma (centrifugation conditions: 8000 rpm, 10 minutes, 2-8℃). Plasma samples were stored at -80℃ before analysis.

[0327] Sample pretreatment: Take 10 μL of plasma sample, add 200 μL of precipitant containing internal standard (methanol:acetonitrile = 1:1), vortex for 1 minute, and centrifuge at 4000 rpm for 15 minutes. Transfer 100 μL of supernatant to a 96-well plate, add 100 μL of diluent (water:methanol:formic acid = 80:20:0.1), vortex for 1 minute, and inject 15 μL into LC-MS / MS for analysis.

[0328] Control group 1: 4-(4-(3,8-diazabicyclo[3.2.1]octane-3-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)pyrido[4,3-d]pyrimidin-7-yl)-5-ethynyl-6-fluoronaphthyl-2-ol

[0329] Prepared according to the method of Example 252 in WO2021041671A1.

[0330] MS(ESI)m / z:[M+H] + =601.1.

[0331] 1H NMR (400MHz, DMSO-d6) δ10.14(s,1H),9.03(s,1H),7.97(dd,J=9.2Hz,6.0Hz,1H),7.49-7.43( m,1H),7.39(d,J=2.8Hz,1H),7.17(d,J=2.4Hz,1H),5.36-5.19(m,1H),4.46(d,J=12.2Hz,1H), 4.31(d,J=11.9Hz,1H),4.14-3.98(m,2H),3.92(s,1H),3.67-3.51(m,4H),3.14-2.99(m,3H), 2.87-2.78(m,1H),2.71-2.54(m,1H),2.15-1.96(m,3H),1.89-1.74(m,3H),1.69-1.60(m,4H).

[0332] Control group 2: 1-(5-(9-((4-((1-(((4-(3,8-diazabicyclo[3.2.1]octane-3-yl)-7-(8-ethynyl-7-fluoro-3-hydroxynaphthyl-1-yl)-8-fluoropyridino[4,3-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-chlorophenyl)dihydropyrimidin-2,4(1H,3H)-dione

[0333] Prepared according to the method of Example 6 in WO2024083258A1.

[0334] LC-MS: (ESI, m / z): [M+H]+=1027.2.

[0335] 1 H NMR (400MHz, CD3OD) δ8.99(s,1H),7.85(dd,J=9.2,5.6Hz,1H),7.63(d,J=8.2Hz,1H),7.53(d,J= 1.6Hz,1H),7.41(dd,J=8.2,2.0Hz,1H),7.36-7.28(m,2H),7.20(d,J=2.4Hz,1H),4.85-4.33(m,5 H),3.82-3.64(m,8H),3.45-3.34(m,3H),2.88-2.82(m,2H),2.57-2.36(m,8H),2.19-2.11(m,2H ),1.91-1.73(m,6H),1.61-1.29(m,8H),1.21-1.04(m,4H),0.75-0.66(m,2H),0.54-0.45(m,2H).

[0336] The test results are shown in Table 3. The degrading agent provided by this invention has good pharmacokinetic properties.

[0337] Table 3

[0338]

Claims

1. A method for preparing compound 5, characterized in that, It includes the following steps: in an organic solvent, in the presence of a reducing agent, compound 4 is subjected to the reduction reaction shown below to obtain compound 5; in, R 1 and R 2 Independently hydrogen or deuterium; R 3 Independently for C 1-6 alkyl; The organic solvent is an ether solvent; The reducing agent is aluminate or deuterated aluminate.

2. The preparation method according to claim 1, characterized in that, It meets one or more of the following conditions: (1) The organic solvent is a cyclic ether solvent, such as tetrahydrofuran; (2) The mass-to-volume ratio of compound 4 to the organic solvent is (50-150) g / L, for example 98.5 g / L; (3) The C 1-6 The alkyl group is methyl, ethyl, n-propyl, isopropyl, or tert-butyl, for example, methyl; (4) The molar ratio of compound 4 to the reducing agent is 1:(0.1~1), for example 1:0.55; (5) The reaction temperature is 0 to 20°C, for example, 0 to 5°C; (6) The reduction reaction preferably includes the following steps: adding water and an inorganic base; the mass-to-volume ratio of the reducing agent to the water can be (0.1-1) g / mL, for example 1 / 5 g / mL; the water and the inorganic base are preferably added slowly, for example dropwise; the water is further preferably added in multiple portions, in 2-5 portions, for example, adding a portion of water first, then adding an aqueous solution of the inorganic base, and then adding water; the water and the inorganic base can be added at 0 to -15°C, for example at -10°C; (7) The inorganic base may be an alkali metal hydroxide, such as NaOH, and the inorganic base may be used in the form of an aqueous solution of inorganic base, such as a 15 wt% aqueous solution of NaOH. (8) The reduction reaction preferably further includes the following steps: adding a purification solvent, wherein the purification solvent is N,N-dimethylformamide; the mass-volume ratio of the reducing agent to the purification solvent may be (0.1-1) g / mL, for example 0.35 g / mL; (9) The preparation method preferably includes the following post-processing steps: after the reduction reaction is completed, an aqueous solution of water and inorganic base is added at 0 to -15°C, followed by the addition of a purification solvent, filtration, washing, drying, and concentration to obtain compound 5. (10) The post-processing step preferably further includes a recrystallization step, wherein the recrystallization solvent is an ether solvent or an alkane solvent; the recrystallization step includes the following steps: crystallization, filtration, and drying to obtain compound 5; (11) The reduction reaction preferably includes the following steps: reacting the mixture of the reducing agent and the ether solvent with the mixture of compound 4 and the ether solvent at 0-5°C; (12) The materials for the reduction reaction are compound 4, lithium aluminum hydride and the ether solvents mentioned above; (13) The reducing agent is lithium aluminum hydride, sodium aluminum hydride or potassium aluminum hydride, for example lithium aluminum hydride; Alternatively, the reducing agent may be lithium deuterated aluminum hydride, sodium deuterated aluminum hydride, or potassium deuterated aluminum hydride, such as lithium aluminum deuterated hydride. (14)R 1 and R 2 It is hydrogen; (15)R 3 It is a methyl group.

3. The preparation method according to claim 1, characterized in that, It also includes the following steps: in an organic solvent, in the presence of an organic base, compound 2 and compound 3 undergo a substitution reaction as shown below to obtain compound 4; R 3 As described in claim 1.

4. The preparation method according to claim 3, characterized in that, It meets one or more of the following conditions: (1) The organic solvent is an ether solvent, preferably a six-membered ring ether solvent, such as 1,4-dioxane; (2) The mass-to-volume ratio of compound 2 to the organic solvent is (50-150) g / L, for example, 106.5 g / L; (3) The molar ratio of compound 2 to compound 3 is 1:(0.5 to 1.5), for example 1:0.95; (4) The organic base is a tertiary amine compound, preferably N(C) 1-6 Alkyl group 3, for example, N,N-diisopropylethylamine; (5) The molar ratio of compound 2 to the organic base is 1:(0.5-4), for example 1:1.5; (6) The reaction temperature is 10–35°C, for example 25–30°C; (7) The preparation method preferably further includes the following post-processing steps: after the substitution reaction is completed, the mixture is filtered, the filter cake is washed, and dried to obtain the compound 4. (8) The substitution reaction preferably includes the following steps: mixing the compound 3, the organic base and the organic solvent, mixing the mixture with the compound 2, and reacting at 10-35°C; (9) The materials for the substitution reaction are compound 3, N,N-diisopropylethylamine, compound 2 and the organic solvent.

5. The preparation method according to claim 3, characterized in that, It also includes the following steps: in an organic solvent, in the presence of an organic base, compound 1 and a sulfonating agent undergo a sulfonation reaction as shown below to obtain compound 2; R 3 As described in claim 3.

6. The preparation method according to claim 5, characterized in that, It meets one or more of the following conditions: (1) The organic solvent is a haloalkane solvent, such as dichloromethane; (2) The mass-to-volume ratio of compound 1 to organic solvent is (50-150) g / L, for example, 100 g / L; (3) The sulfonating agent is trifluoromethanesulfonyl chloride or trifluoromethanesulfonic anhydride, for example, trifluoromethanesulfonic anhydride; (4) The molar ratio of compound 1 to the sulfonating agent is 1:(0.5-2), for example 1:1.1; (5) The organic base is a pyridine organic base, such as 2,6-dimethylpyridine; (6) The molar ratio of compound 1 to the organic base is 1:(0.5-2), for example 1:1.1; (7) The reaction temperature is 10-35℃, for example 25-30℃; (8) The preparation method preferably further includes the following post-processing steps: after the sulfonation reaction is completed, the reaction is quenched, separated, washed with organic phase, dried, filtered, and concentrated to obtain compound 2; (9) The sulfonation reaction preferably includes the following steps: mixing the compound 1 and the organic solvent, adding an organic base, adding a sulfonation reagent, and reacting at 10-35°C; (10) The materials for the sulfonation reaction are compound 1, 2,6-dimethylpyridine, trifluoromethanesulfonic anhydride and the organic solvent.

7. The preparation method according to claim 3, characterized in that, It also includes the following steps: in an organic solvent, compound 3a and Trt-X undergo a substitution reaction as shown below to obtain compound 3, where X is a halogen; Preferably, the preparation method satisfies one or more of the following conditions: (1) The organic solvent is a mixed solvent of aromatic hydrocarbons and alcohols, such as a mixed solvent of toluene and methanol; the volume ratio of the aromatic hydrocarbons and alcohols can be (1-5):1, for example 5:2; (2) The mass-to-volume ratio of the compound 3a to the organic solvent is (150-300) g / L, for example (217-218) g / L; (3) The Trt-X is triphenylchloromethane; (4) The molar ratio of compound 3a to Trt-X is (1 to 10):1, for example 5:1; (5) The reaction temperature is 10-35℃, for example 20-25℃; (6) The preparation method preferably further includes the following post-processing steps: after the substitution reaction is completed, the liquid is separated, the organic phase is washed, an aqueous solution of organic acid is added, filtered, washed, an aqueous solution of inorganic base is added, filtered, washed, and dried to obtain compound 3. (7) The substitution reaction preferably includes the following steps: mixing the mixture of compound 3a and the organic solvent with Trt-X at 0-5°C and reacting at 10-35°C; (8) The materials for the substitution reaction are compound 3a, triphenylchloromethane and the organic solvent.

8. A compound 4 or 5; R 1 R 2 and R 3 As described in claim 1 or 2, and R 1 and R 2 Not both H; Preferably, compound 4 is 9. A method for preparing compound 4, characterized in that, It includes the following steps: In an organic solvent, in the presence of a base, compounds 2 and 3 undergo a substitution reaction as shown below to give compound 4; R 3 As described in claim 3 or 4; The organic solvent is a cyclic ether solvent; The organic base is a tertiary amine compound; In the preparation method described above, the reaction conditions and operations may be further as described in claim 3 or 4, and the preparation method of compound 4 preferably further includes the preparation method described in any one of claims 5-7.

10. A method for preparing compound L or compound G7, characterized in that, It includes the following steps: Step (1): Compound 5 is prepared using the preparation method of compound 5 according to any one of claims 1-7; Step (2): Compound L or compound G7 is prepared by using compound 5 and compound E according to the preparation method of Scheme 1 or Scheme 2 below; Option 1 includes the following steps: Step 1: In an organic solvent and in the presence of an inorganic base, compound E and compound 5 undergo a substitution reaction to obtain compound F; R a It is an amino protecting agent, preferably -CO-OC 1-6 Alkyl groups, such as Boc groups; R 1 and R 2 As described in claim 1 or 2; Step 2: In a solvent, in the presence of a catalyst and an inorganic base, compound F and compound G undergo a coupling reaction to yield compound H; R b It is an alkynyl protecting agent, preferably -Si(C 1-6 Alkyl group 3, such as TIPS group; Step 3: In an organic reagent, compound H undergoes a deprotection reaction to obtain compound I; Step 4: In an organic solvent and in the presence of a catalyst, compound I and compound J undergo a coupling reaction to obtain compound K; Step 5: In an organic solvent and in the presence of an auxiliary agent, compound K undergoes a deprotection reaction to yield compound L; Option 2 includes the following steps: *The C marked is a chiral C, which can be in the S configuration, R configuration, or a mixture thereof; Step 1: In an organic solvent and in the presence of an inorganic base, compound A1 and compound 5 undergo a substitution reaction to obtain compound B2; R a It is an amino protecting agent, preferably -CO-OC 1-6 Alkyl groups, such as Boc groups; R C Halogen, C 1-6 Alkyl or deuterated C 1-6 alkyl; R 1 and R 2 As described in claim 1 or 2; Step 2: In a solvent, in the presence of a catalyst and an inorganic base, compound B2 and compound C3 undergo a coupling reaction to obtain compound D4; R b It is an alkynyl protecting agent, preferably -Si(C 1-6 Alkyl group 3, such as TIPS group; Step 3: In an organic reagent, compound D4 undergoes a deprotection reaction to obtain compound E5; Step 4: In an organic solvent and in the presence of a catalyst, compound E5 and compound J undergo a coupling reaction to obtain compound F6; Step 5: In an organic solvent and in the presence of an auxiliary agent, compound F6 undergoes a deprotection reaction to obtain compound G7.

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