Preparation method of compound containing aza-bridged ring

Abstract

The invention discloses a preparation method of a compound containing an aza-bridged ring. The invention provides a preparation method of a compound shown as a formula I. The preparation method comprises the following steps: (i) reacting a compound shown as a formula II with a Grignard reagent R2MgX in a solvent at-30 DEG C to-5 DEG C to obtain a reaction solution; and (ii) in a solvent, in the presence of an inorganic base, carrying out a hydrolysis reaction on the reaction liquid obtained in the step (i) to obtain the compound shown in the formula I. The method overcomes the defects of complex route and difficult purification in the prior art, and has remarkable advantages in the aspects of reducing the production cost and being suitable for commercialized large-scale production.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of medicine, and particularly relates to a preparation method of a compound containing a nitrogen heterobridging ring. BACKGROUND

[0002] RAS (rat sarcoma) is one of the oncogenes with the highest mutation rate in tumors, and its mutation exists in about 30% of human malignancies. The RAS family includes KRAS, NRAS and HRAS, among which KRAS (kirsten rat sarcoma viral oncogene) is more prone to mutation than the other two RAS subtypes, accounting for about 85%, and is particularly common in solid tumors. KRAS gene mutations exist in 30-40% of colorectal cancers, 90% of pancreatic cancers and 15-20% of lung cancers. After KRAS is activated, it regulates cell proliferation, differentiation and survival functions through downstream signaling pathways such as RAF-MEK-ERK and PI3K-AKT-mTOR. After KRAS gene mutation, the protein is continuously in an activated state, leading to continuous activation of downstream signaling pathways and promoting tumor occurrence. Therefore, KRAS is an important target for tumor treatment efforts, including targeting KRAS protein itself, or its post-translational modification, membrane localization, protein-protein interaction and RAS downstream signaling pathway.

[0003] At present, according to the mode of action of the inhibitor, KRAS inhibitors can be divided into:

[0004] 1) Directly targeting KRAS inhibitors, such as KRAS G12C, KRAS G12D, KRAS G12R and KRAS G12S inhibitors;

[0005] 2) Indirectly acting on KRAS inhibitors, such as SHP2, SOS1 inhibitors and KRAS (on) inhibitors.

[0006] As one of the common KRAS mutations, KRAS G12D drives a highly immunosuppressive tumor microenvironment and exhibits strong oncogenic potential. Therefore, developing KRAS G12D inhibitors and other pan-KRAS inhibitors is a new direction for KRAS-targeted therapy.

[0007] The compound of formula I (the structure is shown below) is an important intermediate for synthesizing KRAS G12D-targeted protein degradation agent molecules.

[0008]

[0009] The synthesis route of the compound of formula I disclosed in patent WO2023 / 225302A1 is as follows:

[0010]

[0011] The route has five steps, and the reagent sec-butyl lithium is used when introducing the chiral side chain in the second step. The sec-butyl lithium needs to be strictly controlled in terms of reaction equipment and anhydrous and oxygen-free environment, and the step produces four diastereoisomers (3a, 3b, 3c and 3d). In addition to column chromatography, the target product 3a can be separated and purified only by SFC chiral separation. The route does not have the developability of scale-up production. Secondly, the route first introduces the benzyl protecting group, then removes the benzyl protecting group, and then introduces the Boc protecting group. The selection of the protecting group is not efficient.

[0012] The synthesis route of the compound of formula I disclosed in patent WO2023 / 030385A1 is as follows:

[0013]

[0014] The route has four steps, two steps of deep cold reaction, and one step of using metal palladium. The yield of the second step of oxidation reaction is low, and the product quality is poor. At the same time, the by-product 3b will also continue to be derived, which will affect the subsequent production process and product quality. The process of introducing chiral methyl in the third step of methyl Grignard reagent produces four diastereoisomers (4aa, 4ab, 4ba and 4bb). Under the reaction condition of-78℃, the selectivity of 4aa and 4ab is 5:1. A large amount of by-product brings great challenges to the purification and product quality of the product, and greatly reduces the developability of scale-up production of the route.

[0015] In view of the comprehensive consideration of reducing cost and commercial production, it is particularly necessary to provide a new method for preparing a KRAS G12D degrader intermediate. SUMMARY

[0016] In view of the above problems, the present application provides a preparation method of a compound containing a nitrogen hetero bridge ring. The method overcomes the defects of complex route and difficult purification in the prior art, and has significant advantages in reducing production cost and being suitable for commercial scale-up production.

[0017] The present application mainly solves the above technical problems through the following technical solutions.

[0018] The present application provides a preparation method of a compound of formula I, which comprises the following steps:

[0019] (i) reacting a compound of formula II and a Grignard reagent R 2 MgX in a solvent, the reaction being carried out at-30℃ to-5℃ to obtain a reaction liquid;

[0020] (ii) hydrolysis of the reaction solution of step (i) in a solvent in the presence of an inorganic base to give a compound of formula I,

[0021]

[0022] wherein R a is an amino protecting group: -COO-C 1-6 alkyl (e.g. a Boc group) or -C 1-6 alkyl-phenyl (e.g. benzyl or PMB), said phenyl being optionally substituted by one or more C 1-6 alkoxy, halo, C 1-6 alkyl;

[0023] X is halo;

[0024] R 2 is C 1-6 alkyl or C 1-6 alkyl substituted by one or more deuterium (D).

[0025] In step (i), the solvent can be an ether solvent, such as tetrahydrofuran and / or dimethyltetrahydrofuran.

[0026] In step (i), the Grignard reagent can be a methyl magnesium halide or a deuterated methyl magnesium halide, such as CH3MgBr.

[0027] In step (i), the volume to mass ratio of the solvent to the compound of formula II can be 5 to 15 L / kg, such as 10 L / kg or 11 L / kg.

[0028] In step (i), the molar ratio of the compound of formula II to Grignard reagent can be 1 : (1 to 3), such as 1 :3, 1 :1.8, 1 :1.7 or 1 :1.2, preferably 1 :1.2.

[0029] In step (i), the Grignard reagent can be used in the form of a Grignard reagent solution in dimethyltetrahydrofuran, such as a Grignard reagent solution in dimethyltetrahydrofuran at 1 to 4 mol / L (e.g. 3 mol / L). The volume ratio of the Grignard reagent solution in dimethyltetrahydrofuran to the tetrahydrofuran can be 1 : (4 to 10), such as 1 :6.

[0030] In step (i), when the Grignard reagent is used in the form of a methyl Grignard reagent solution in dimethyltetrahydrofuran, the volume to mass ratio of the methyl Grignard reagent solution in dimethyltetrahydrofuran to the compound of formula II can be 0.9 to 2.2 L / kg, such as 1.5 L / kg.

[0031] The step (i) can be carried out in two temperature stages, the first temperature stage can be -25°C to -5°C (e.g. -10 to -20°C), and the second temperature stage can be 10 to 40°C (e.g. 20 to 30°C). The skilled person can determine the timing of the transition from the first stage temperature (-10 to -20°C) to the second stage temperature (10 to 40°C) based on the progress of the reaction, for example, based on the progress of the conversion of the compound of formula II, and the skilled person can confirm the appropriate timing, for example, based on the progress of the conversion of the compound of formula II. The reaction reagents can be at the first stage temperature for 1 to 2 hours, and the second temperature stage can be for 5 to 8 hours.

[0032] The step (i) can further comprise the following steps:

[0033] Step a, first at -5 to -25°C for 0.5 to 3 hours, for example, at -10 to -20°C for 1 to 2 hours;

[0034] Step b, and then at 10 to 40°C for 2 to 10 hours, for example, at 20 to 30°C for 5 to 8 hours.

[0035] After the reaction in step (i) is completed, the following post-treatment steps can be included: cooling, quenching the reaction (for example, with water), and concentration (for example, under reduced pressure to a volume a; the volume a can have a mass to volume ratio of (0.5 to 1.5) L / kg (preferably, 1 to 1.5 L / kg) with respect to the compound of formula II).

[0036] Preferably, after step (i) is completed, the solvent in step (i) can be removed.

[0037] In step (ii), the solvent can be a mixture of an alcoholic solvent and water (the mixture of the alcoholic solvent and water can be added in step (ii)); the alcoholic solvent can be ethanol; and the volume ratio of the alcoholic solvent to water can be (1 to 3):(1 to 3), for example, 1:1.

[0038] In step (ii), the mass to volume ratio of the compound of formula II to the solvent can be 0.02 to 0.3 kg / L, for example, 0.1 kg / L.

[0039] In step (ii), the mass to volume ratio of the compound of formula II to the alcoholic solvent can be 0.05 to 0.3 kg / L, for example, 0.2 kg / L.

[0040] In step (ii), the inorganic base can be an alkali metal hydroxide, for example, sodium hydroxide.

[0041] In step (ii), the molar ratio of the compound of formula II to the inorganic base can be 1:(5 to 10), for example, 1:8.

[0042] In step (ii), the temperature of the reaction can be 65-90 °C, for example 75-80 °C.

[0043] In step (ii), the time of the reaction can be 10-24 h, for example 16-20 h.

[0044] In step (ii), after the reaction is completed, the following post-treatment steps can be further included: concentration of the reaction solution (for example, concentration under reduced pressure at 30-55 °C (for example 40-45 °C)), extraction (for example, extraction with water and an organic solvent (for example, ethyl acetate)), separation (preferably further comprising extraction of the aqueous phase with an organic solvent (for example, ethyl acetate)), washing (for example, saturated brine washing), concentration, purification (for example, column chromatography), to obtain the compound of formula I.

[0045] The step (ii) can further comprise the following steps: adding the alcohol solvent and water mixed solvent, the inorganic base, and carrying out the reaction at 65-90 °C.

[0046] In the present application, in step (i) and step (ii), the progress of the reaction is detected by using the conventional monitoring methods in the art (for example, TLC, HPLC or NMR), and the reaction end point is generally when the raw materials of the reaction disappear or no longer react, or the compound of formula I no longer increases.

[0047] In the present application, the reactants of the preparation method of the compound of formula I can be the compound of formula II, the Grignard reagent, the inorganic base and the solvent.

[0048] In the present application, the preparation method of the compound of formula I can comprise the following steps: in an ether solvent, the compound of formula II and the methyl Grignard reagent are reacted at -10 to -20 °C, and the reaction is continued by warming to 20-30 °C.

[0049] In the present application, the C 1-6 The alkyl group can be methyl, ethyl, n-propyl, isopropyl or tert-butyl, for example methyl or tert-butyl.

[0050] In the present application, the R a may be a Boc group.

[0051] In the present application, the R 2 may be methyl.

[0052] In the present application, the preparation method of the compound of formula I can further comprise the preparation method of the compound of formula II, which comprises the following steps: in a solvent, the compound of formula III is subjected to an oxidation reaction in the presence of an oxidizing agent and an organic base to generate the compound of formula II;

[0053]

[0054] Ra as any one of the schemes of the present application.

[0055] In the present application, the solvent in the oxidation reaction can be a halogenated alkane solvent, such as dichloromethane.

[0056] In the present application, the oxidant in the oxidation reaction is preferably dimethylsulfoxonium chloride, and the oxidant is preferably prepared by the following steps: reacting oxalyl chloride and DMSO in a solvent to obtain the oxidant. The prepared oxidant can be directly used in the oxidation reaction.

[0057] In the present application, the solvent in the preparation of the oxidant can be a halogenated alkane solvent, such as dichloromethane.

[0058] In the present application, the mass ratio of the solvent to the oxalyl chloride in the preparation of the oxidant can be (15-25):1, such as 19.3:1.

[0059] In the present application, the preparation of the oxidant can be carried out at -70 to -40℃, such as -60 to -50℃.

[0060] In the present application, the organic base in the oxidation reaction can be an alkylamine (such as N(C 1-6 alkyl)3), such as triethylamine.

[0061] In the present application, the volume-to-mass ratio of the solvent to the compound of formula III in the oxidation reaction can be 10-25 L / kg.

[0062] In the present application, the compound of formula III in the oxidation reaction can be used in the form of a mixture of the compound of formula III and a halogenated alkane solvent (such as added to the reaction solution of the oxidant), and the volume-to-mass ratio of the compound of formula III to the halogenated alkane solvent can be 1-10 L / kg, such as 5 L / kg.

[0063] In the present application, the molar ratio of the compound of formula III to oxalyl chloride in the oxidation reaction can be 1:(1-3), such as 1:2.

[0064] In the present application, the molar ratio of the compound of formula III to DMSO in the oxidation reaction can be 1:(1-4), such as 1:2.5.

[0065] In the present application, the molar ratio of the compound of formula III to the organic base in the oxidation reaction can be 1:(5-10), such as 1:8.

[0066] In the present application, the oxidation reaction can be carried out at -70 to -40℃, such as -60 to -50℃.

[0067] In the present application, the organic base is preferably added during the reaction of the compound of formula III with the oxidizing agent (the organic base is added after the compound of formula III is added), for example, the compound of formula III is added for 1-3 h before the organic base is added, preferably, the compound of formula III is added for 3 h before the organic base is added.

[0068] In the present application, the oxidation reaction can further comprise the following steps:

[0069] Step S1, the DMSO and oxalyl chloride are reacted in a solvent at -70 to -40 °C (for example, the reaction is carried out for 1-3 h), for example, the reaction is carried out at -60 to -50 °C (for example, for 1 h);

[0070] Step S2, the reaction solution of step S1 and the compound of formula III are reacted at -70 to -40 °C (for example, the reaction is carried out for 1-3 h), for example, the reaction is carried out at -60 to -50 °C (for example, for 2 h);

[0071] Step S3, the reaction solution of step S2 and the organic base are reacted at -70 to -40 °C (for example, the reaction is carried out for 0.5-5 h), for example, the reaction is carried out at -60 to -50 °C (for example, for 1-2 h).

[0072] In the present application, the oxidation reaction can comprise the following quenching step: after the reaction is completed, the reaction solution is mixed with an aqueous citric acid solution (for example, 10% aqueous citric acid solution) to quench the reaction.

[0073] In the present application, the post-treatment process of the oxidation reaction can comprise the following steps: after the reaction is completed, quenching, extraction and separation (for example, dichloromethane extraction), washing (for example, 10% salt water washing), drying (for example, sodium sulfate drying), concentration, and purification to obtain the compound of formula II.

[0074] In the present application, the progress of the oxidation reaction is detected by using conventional monitoring methods in the art (for example, TLC, HPLC or NMR), and the reaction endpoint is generally when the compound of formula III disappears or no longer reacts, or when the compound of formula II no longer increases.

[0075] In the present application, the reactants of the method for preparing the compound of formula II can be the compound of formula III, the oxidizing agent, the solvent and the organic base.

[0076] In the present application, the method for preparing the compound of formula II can comprise the following steps: the compound of formula III and the oxidizing agent are reacted in a halogenated alkane at -60 to -50 °C, and the organic base is added for reaction.

[0077] In the present application, the preparation method of the compound of formula I can further comprise a preparation method of a compound of formula III, which comprises the following steps: performing a substitution reaction on a compound of formula IV and X-Cbz in a solvent in the presence of a base, X being halogen, to generate a compound of formula III;

[0078]

[0079] R a as described in any of the schemes of the present application.

[0080] In the present application, in the substitution reaction, the solvent can be a mixed solvent of water and an organic solvent; the organic solvent can be an ester solvent, such as ethyl acetate.

[0081] In the present application, in the substitution reaction, X can be chlorine (X-Cbz is benzyl chloroformate).

[0082] In the present application, in the substitution reaction, the base can be an alkali metal acid salt (such as an alkali metal bicarbonate salt), for example, an alkali metal bicarbonate salt, for example, sodium bicarbonate.

[0083] In the present application, in the substitution reaction, the volume-to-mass ratio of the ester solvent to the compound of formula IV can be 5-15 L / kg, for example, 10 L / kg.

[0084] In the present application, in the substitution reaction, the volume-to-mass ratio of water to the compound of formula IV can be 1-8 L / kg, for example, 3 L / kg.

[0085] In the present application, in the substitution reaction, the molar ratio of the compound of formula IV to the base can be 1:(3-5), for example, 1:4.

[0086] In the present application, in the substitution reaction, the molar ratio of the compound of formula IV to X-Cbz can be 1:(1-2), for example, 1:1.2.

[0087] In the present application, the substitution reaction can comprise the following post-treatment steps: after the reaction is completed, mixing the reaction liquid of the substitution reaction with a saturated aqueous ammonium chloride solution, performing liquid-liquid extraction, washing, drying (for example, sodium sulfate washing), to obtain the compound of formula III.

[0088] In the present application, the progress of the substitution reaction is detected by using a conventional monitoring method in the art (such as TLC, HPLC or NMR), and the reaction end point is generally when the compound of formula IV disappears or no longer reacts, or when the compound of formula III no longer increases.

[0089] In the present application, the reactant materials of the preparation method of the compound of formula III can be the solvent, the compound of formula IV, the base and benzyl chloroformate.

[0090] In the present application, the preparation method of the compound of formula III can comprise the following steps: substitution reaction of the compound of formula IV, a base and benzyl chloroformate in a mixed solvent of water and an ester solvent at 20-25℃.

[0091] The present application also provides a preparation method of the compound of formula II, which comprises the following steps: oxidation reaction of the compound of formula III in the presence of an oxidizing agent and an organic base in a solvent to generate the compound of formula II.

[0092]

[0093] R a as described in any of the schemes of the present application;

[0094] In the preparation method of the compound of formula II, each reaction condition can be as described above; preferably, after the reaction is completed, the reaction solution is quenched by mixing with an aqueous citric acid solution (for example, 10% aqueous citric acid solution).

[0095] The preparation method of the compound of formula II can further comprise the preparation method of the compound of formula III as described in any of the schemes of the present application.

[0096] The present application also provides a preparation method of the compound of formula X, which comprises the following steps:

[0097]

[0098] Step 1: substitution reaction of the compound of formula I and the compound of formula a in the presence of an inorganic base in an organic solvent to generate the compound of formula b; R a and R 2 as described in any of the schemes of the present application;

[0099] In the step 1, the organic solvent can be a conventional organic solvent in the art, for example, a cyclic ether solvent, and for example, tetrahydrofuran;

[0100] In the step 1, the inorganic base can be a conventional base in the art, for example, an alkali metal hydride, and for example, sodium hydride;

[0101] In the step 1, the step 1 can be carried out at room temperature, for example, at 25℃.

[0102] Step 2: ring and reaction of the compound of formula b in the presence of an organic base and a coupling reagent in an organic solvent to generate the compound of formula c;

[0103] In the step 2, the organic solvent can be a conventional organic solvent in the art, for example, a halogenated alkane solvent, and for example, dichloromethane;

[0104] In the step 2, the coupling reagent can be a coupling reagent conventional in the art, for example, BOPCl;

[0105] In the step 2, the organic base can be a base conventional in the art, for example, an alkylamine, for example, N,N-diisopropylethylamine;

[0106] In the step 2, the reaction can be carried out at room temperature, for example, at 25°C.

[0107] Step 3: the compound of formula c is subjected to an oxidation reaction to generate a compound of formula d in an organic solvent under the action of an oxidizing reagent;

[0108] In the step 3, the organic solvent can be a halogenated alkane solvent, for example, dichloromethane.

[0109] In the step 3, the oxidizing reagent can be a peroxy acid oxidizing reagent, for example, m-CPBA.

[0110] In the step 3, the reaction can be carried out at room temperature.

[0111] Step 4: the compound of formula d and the compound of formula e are subjected to a substitution reaction to generate the compound of formula b in an organic solvent under the action of a base.

[0112] In the step 4, the organic solvent can be an ether solvent, for example, tetrahydrofuran.

[0113] In the step 4, the reaction can be carried out at -10°C to 10°C, for example, at 0°C.

[0114] In the step 4, the base can be a base conventional in the art, for example, an alkali metal hydride, for example, sodium hydride.

[0115] Step 5: the compound of formula f and the compound of formula g are subjected to a coupling reaction to generate the compound of formula h in a solvent in the presence of an inorganic base and a catalyst.

[0116] wherein, R 3 is R 3-1 is C 1-6 alkyl or C 3-6 cycloalkyl (for example, trifluoromethyl), the C 1-6 alkyl or C 3-6 cycloalkyl being optionally substituted by one or more deuterium, halogen, C 1-6 alkyl, R 3-2 is OH, amino, O-hydroxyl protecting group (preferably, -C 1-6 alkyl-O-C 1-6 alkyl, for example, -MOM) or NH-amino protecting group (preferably, -CO-O-C 1-6alkyl, for example, -Boc group);

[0117] In the step 5, the solvent can be a mixture of an organic solvent and water, and the organic solvent can be a conventional organic solvent in the art, for example, a cyclic ether solvent, and for example, 1,4-dioxane;

[0118] In the step 5, the base can be a conventional base in the art, for example, an alkali metal carbonate, and for example, cesium carbonate or potassium phosphate;

[0119] In the step 5, the catalyst can be a metal catalyst, for example, a palladium catalyst and a ligand, and for example, Pd(dppf)Cl2;

[0120] In the step 5, the reaction can be carried out at 80-120°C, for example, at 100°C;

[0121] Step 6: in an organic reagent, the compound of formula h is deprotected to generate the compound of formula i;

[0122] In the step 6, the organic reagent can be an organic acid reagent or an organic acid and a halogenated alkyl reagent, and preferably a carboxylic acid reagent, for example, acetic acid;

[0123] In the step 6, the reaction can be carried out at 40-80°C, for example, at 60°C or 50°C;

[0124] Step 7: in an organic solvent, the compound of formula i and the compound of formula j undergo a reductive amination reaction under the action of a reducing agent to generate the compound of formula k;

[0125] In the step 7, the organic solvent can be a conventional organic solvent in the art, for example, a halogenated alkyl solvent, and for example, dichloromethane;

[0126] In the step 7, an organic acid can be further included, and the acid can be a conventional acid in the art, for example, an organic acid, and for example, acetic acid;

[0127] In the step 7, the catalyst can be a titanate coupling agent, for example, tetraisopropyl titanate;

[0128] In the step 7, the following step can be further included, that is, a reducing reagent is added, and the reducing reagent can be NaBH(OAc)3;

[0129] Step 8: in an organic solvent, the compound of formula k is deprotected to generate the compound of formula X;

[0130] In the step 8, the organic solvent can be a conventional organic solvent in the art, for example, a cyclic ether solvent, and for example, 1,4-dioxane;

[0131] The deprotection in step 8 can be carried out in the presence of an acidic reagent, such as, for example, a mineral acid, and, for example, hydrochloric acid.

[0132] R 4 is R 3-1 R 3-2 is -OH or amino. Preferably, the process for preparing a compound of formula X further comprises the process for preparing a compound of formula I according to any of the aspects of the present application.

[0133] Preferably, the compound of formula X is

[0134] R 2 , R 3-1 , R 3-2a according to any of the aspects of the present application.

[0135] Preferably, the compound of formula X is

[0136] Explanation of terms:

[0137] The term "C 1-6 alkyl" can be methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl or t-butyl, for example methyl.

[0138] The term "halogen" can be fluorine, chlorine or bromine, for example chlorine.

[0139] The term "C 1-6 alkoxy" can be -O-C 1-6 alkyl, C 1-6 alkyl is defined as above.

[0140] The term "C 3-6 cycloalkyl" can be cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

[0141] Without deviating from the general knowledge in the art, the above-mentioned preferred conditions can be combined in any way, resulting in preferred embodiments of the present application.

[0142] The reagents and starting materials used in the present application are commercially available.

[0143] The positive progress effect of the present application is that:

[0144] 1. The preparation process is simple and suitable for scale-up production.

[0145] 2. The starting materials are readily available, the side reactions are few and the yield is high, which can significantly reduce the production cost. DETAILED DESCRIPTION

[0146] The application will be further described in the following by way of examples without limiting the application to the examples described. The experimental methods in the following examples, unless otherwise specified, are carried out according to the conventional methods and conditions, or according to the instructions of the commercial products.

[0147] Example 1

[0148]

[0149] Step one: Preparation of compound of formula III

[0150]

[0151] The compound of formula IV (24.6 kg, 1.0 eq), ethyl acetate (250.0 L, 10.0 v / w) were added into the reaction kettle, after stirring, water (75.0 L, 3.0 v / w) and sodium bicarbonate (19.3 kg, 4.0 eq) were added, the reaction kettle was cooled to 0-10 °C, then benzyl chloroformate (11.7 kg, 1.2 eq) was added dropwise, and the temperature was raised to 20-25 °C and stirred for 16-20 hours. After the reaction was completed, the material in the reaction kettle was added to saturated aqueous ammonium chloride solution (125.0 L), stirred for 30 minutes, then separated, the lower aqueous phase was extracted with ethyl acetate, then combined with the upper organic phase, after combination, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the compound of formula III (23.7 kg), purity 88.9%, yield 100%.

[0152] 1 H NMR (400 MHz, D6-DMSO): δ 7.29-7.39 (m, 5H), 5.04 (dd, J = 12.8 Hz, J = 16.4 Hz, 2H), 4.76-4.79 (m, 1H), 4.25 (s, 1H), 4.11 (s, 1H), 3.94-4.02 (m, 1H), 3.75-3.78 (m, 1H), 3.42-3.53 (m, 2H), 3.31-3.35 (m, 1H), 1.82-1.96 (m, 2H), 1.67-1.70 (m, 1H), 1.53-1.58 (m, 1H), 1.37 (s, 9H).

[0153] Step two: Preparation of compound of formula II

[0154]

[0155] Oxalyl chloride (14.5 kg, 2.0 eq), dichloromethane (280.0 kg, 10.0 v / w) were charged into a reaction kettle, the reaction kettle was cooled to -60 ~ -50 °C, DMSO (11.2 kg, 2.5 eq) was slowly added dropwise into the reaction kettle, and the temperature of the reaction kettle was controlled at -60 ~ -50 °C, after stirring for 1 hour, the compound of formula III (21.2 kg, 1.0 eq) / DCM (108.0 L, 5.0 v / w) mixture was added dropwise into the reaction kettle, and the temperature of the reaction kettle was controlled at -60 ~ -50 °C, after stirring for 2 hours, triethylamine (46.2 kg, 8.0 eq) was slowly added dropwise into the reaction kettle, and the temperature of the reaction kettle was controlled at -60 ~ -50 °C, and stirred for 1 ~ 2 hours. After the reaction was completed, the material in the reaction kettle was added to 10% citric acid aqueous solution (322.0 L, 15.0 v / w), stirred for 30 minutes, then separated, the upper aqueous phase was extracted with dichloromethane, then combined with the lower organic phase, after combination, the organic phase was washed with 10% salt water, dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was column chromatographed and concentrated to give the compound of formula II (19.7 kg), purity 92.6%, yield 93%.

[0156] 1 H NMR (400 MHz, D6-DMSO): δ 9.42 (d, J = 3.2 Hz, 1H), 7.31-7.37 (m, 5H), 5.11 (s, 2H), 4.22 (s, 1H), 4.02-4.07 (m, 1H), 3.73 (s, 1H), 3.57 (d, J = 11.6 Hz, 1H), 3.16-3.19 (m, 1H), 1.85-1.99 (m, 2H), 1.70-1.78 (m, 2H), 1.41 (s, 9H).

[0157] LCMS: (ESI, m / z): [M+H] + = 375.0

[0158] Step three: preparation of the compound of formula I

[0159]

[0160] Stage one: the compound of formula II (19.70 kg, 1.0 eq), tetrahydrofuran (180.0 kg, 10.0 v / w) were added into the reaction kettle, the reaction kettle was cooled to -10~ -20 °C, 3M CH3MgBr dimethyl tetrahydrofuran solution (30.0 L, 1.7 eq) was slowly added into the reaction kettle, and the temperature of the reaction kettle was controlled at -10~ -20 °C, after stirring for 1~2 hours, the reaction kettle was slowly warmed to 20~30 °C, and stirred for 5~8 hours. After the reaction was completed, the reaction kettle was cooled to 0~10 °C, water (28 L, 1.4 v / w) was slowly added into the reaction kettle for quenching, and the reaction solution was concentrated to 20~30 L under reduced pressure at 40~45 °C.

[0161] Stage two: ethanol (100 L, 5 v / w), water (100 L, 5 v / w) were further added into the reaction kettle, sodium hydroxide (17.0 kg, 8.0 eq) was added into the reaction kettle in batches, the reaction kettle was warmed to 75~80 °C and stirred for 16~20 hours. After the reaction was completed, the reaction solution was concentrated to 60~100 L under reduced pressure at 40~45 °C, water (160.0 kg, 8 v / w), ethyl acetate (200 L, 10 v / w) were added into the reaction kettle, after stirring for 30 minutes, the liquid was separated, the lower aqueous phase was extracted with ethyl acetate (100 L x 5 times), then the upper organic phase was combined, after the combination, the organic phase was washed with saturated brine, and then concentrated to obtain the crude product, the crude product was subjected to column chromatography, and then concentrated to obtain the compound of formula I (8 kg), the chemical purity was 99%, the chiral purity was 99%, and the yield was 60%. 1 H NMR (400 MHz, D6-DMSO): δ 4.55 (s, 1H), 3.93-3.95 (m, 1H), 3.77-3.84 (m, 1H), 2.70-2.73 (m, 1H), 2.59-2.62 (m, 1H), 2.39-2.42 (m, 1H), 1.68-1.80 (m, 3H), 1.56-1.60 (m, 1H), 1.40 (s, 9H), 1.03 (d, J = 6.0 Hz, 3H). LCMS: (ESI, m / z): [M+H] + = 257.0.

[0162] Screening of starting temperature in step three of example 1-1

[0163] The reaction operation was the same as the preparation of the compound of formula I in step 3 of example 1, the temperature in the first stage was shown in table 1, and the other operations were the same as step three of example 1, and the reaction results were shown in table 1.

[0164] Table 1

[0165] temperature ratio of hplc peak areas of diastereoisomers to target configuration ，70～-60℃ 14：86 -60～-50℃ 10：90 -50～-40℃ 8：92 -30～-20℃ 5：95 ，20～-10℃ 2：98

[0166] The HPLC analysis method used in step three was shown in table 2:

[0167] Table 2

[0168]

[0169] Example 1-2 Step two of preparing the compound of formula I

[0170] The reaction was operated as the preparation of the compound of formula II in Example 1, Step two. After the reaction was completed, HPLC monitoring was performed, and the results are shown in Table 3. When the quenching reagent was an aqueous ammonium chloride solution, the product had a diastereoisomer, and the separation yield was 70%. It was found that the diastereoisomer could not be racemized. When the quenching reagent was changed to a 10% aqueous citric acid solution, the diastereoisomer was avoided, and the kilogram yield was increased to 93%.

[0171] Table 3

[0172] quenching solution reaction yield aqueous ammonium chloride solution 70％ 10% aqueous citric acid solution 93％

[0173] The HPLC analysis method used in Step two is shown in Table 4:

[0174] Table 4

[0175]

[0176] Example 1-3 Stepwise reaction

[0177]

[0178] Step one:

[0179] The compound of formula II (50 g, 1.0 eq) and tetrahydrofuran (0.5 L, 10.0 v / w) were added to a reaction kettle, which was cooled to -60 to -50°C. A 3M CH3MgBr solution in dimethyltetrahydrofuran (0.53 L, 1.2 eq) was slowly added dropwise to the reaction kettle, and the temperature of the reaction kettle was controlled at -60 to -50°C, and stirred for 2 hours. After the reaction was completed, saturated aqueous ammonium chloride solution was added to the reaction kettle and stirred for 30 minutes, and the upper organic phase was concentrated and purified by column chromatography to obtain the compound of formula M (21.0 g), with a product purity of 84.3%, a diastereoisomer content of 8.1%, and a weight yield of 40.4%.

[0180] Step two:

[0181] The compound of formula M (13 g), 10% Pd / C (10% w / w), MeOH (130 mL, 10 v / w) were added into the reaction flask, and hydrogenation deprotection was carried out under H2 at 20-25 °C. After 16 hours, TLC monitoring found that the reaction was incomplete, and the palladium carbon was easily poisoned by unknown impurities in the system. The reaction solution was filtered, and 10% Pd / C (10% w / w) was added to continue the conversion. After the reaction solution was filtered, concentrated and purified by column chromatography, 6.8 g of the product was finally obtained, with a purity of 91.7% and a weight yield of 80%.

[0182] Example 1-4 Stepwise reaction

[0183]

[0184] Step one:

[0185] The compound of formula II (150 g, 1.0 eq), tetrahydrofuran (1.5 L, 10.0 v / w) were added into the reaction kettle, and the reaction kettle was cooled to

[0186] -60 to -50 °C, 3M CH3MgBr dimethyl tetrahydrofuran solution (1.6 L, 1.2 eq) was slowly added into the reaction kettle, and the temperature of the reaction kettle was controlled at -60 to -50 °C, and stirred for 2 hours. After the reaction was completed, saturated aqueous ammonium chloride solution was added into the reaction kettle and stirred for 30 minutes, and the upper organic phase was concentrated and purified by column chromatography to obtain the compound of formula M (101 g), with a product purity of 86.5% and a weight yield of 65%.

[0187] Step two:

[0188] The compound of formula M (25 g), 10% Pd / C (15% w / w), MeOH (20 v / w), HCOOK (20 eq) were added into the reaction flask, and deprotection reaction was carried out at 20-25 °C. After stirring for 2 hours, TLC monitoring found that the reaction was incomplete, and the palladium carbon was poisoned by unknown impurities in the system. The reaction solution was filtered, and 10% Pd / C (10% w / w) and HCOOK (20 eq) were added to continue the conversion. After the reaction solution was filtered, concentrated and purified by column chromatography, 13.6 g of the product was obtained, with a purity of 94.1% and a weight yield of 83%.

[0189] Example 2

[0190] The following KRAS G12D degraders can be prepared using the processes disclosed below

[0191]

[0192] Starting materials for which no preparation method is given can be obtained using existing approaches in the art, or according to the methods described in PCT / CN2025 / 089926.

[0193] Preparation route as follows:

[0194] 1. Preparation of compound 5

[0195]

[0196] Step 1: Preparation of compound 4

[0197] Compound 3 (2.64 kg, 0.95 eq), DIPEA (1.64 kg, 1.5 eq), 1,4-dioxane (20.0 L) were added into the reaction kettle, and stirred for half an hour at 25-30 °C. The reaction kettle was cooled to 10-15 °C, and compound 2 (2.13 kg, 1.0 eq) was added dropwise into the reaction kettle. After the dropwise addition was completed, the reaction kettle was warmed to 25-30 °C and stirred for 16 hours. After the reaction was completed, the reaction liquid was added into ice water (10.0 L), stirred for 1-2 hours, centrifuged and filtered, 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%.

[0198] 1 H NMR (400 MHz, CDCl3): δ 7.12-7.46 (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 = 6.8 Hz, 3H).

[0199] Step 2: Preparation of compound 5

[0200] Lithium aluminum hydride (140.5 g, 0.55 eq, flaky), tetrahydrofuran (23.5 L) were added into the reaction kettle, and the reaction kettle was cooled to 0-5 °C and stirred until the solid was completely dissolved. The temperature of the reaction kettle was controlled at 0-5 °C, and a tetrahydrofuran (10.5 L) solution of compound 4 (3.35 kg, 1.0 eq) was added dropwise into the reaction kettle. After the dropwise addition was completed, stirring was continued for half an hour. After the reaction was completed, the reaction kettle was cooled to -10 °C, and water (140.5 mL), 15% NaOH solution (140.5 mL), water (421.5 mL) were added dropwise into the reaction kettle in sequence, and then N,N-dimethylformamide (400 mL) was added and stirring was continued for 15 minutes. After filtration, the filtrate was washed with water, saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a white solid crude product. The crude product was recrystallized with 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 obtain 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%.

[0201] 1 H NMR (400 MHz, CDC13): δ 7.12-7.44 (m, 15H), 6.01 (s, 1H), 3.55-3.59 (m, 1H), 3.38 (t, J = 10.4 Hz, 1H), 2.41-3.02 (m, 8H), 1.61-2.12 (m, 3H), 0.72 (d, J = 6.8 Hz, 3H).

[0202] 2. Preparation of intermediate 27:

[0203] Preparation of (5S,5aS,6S,9R)-2-chloro-1-fluoro-5-methyl-12-((R)-2-methyl-3-(4- triphenylphosphoniumylpiperazin-1-yl)propoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[1,8-ab]heptene-14-carboxylic acid tert-butyl ester

[0204] Step 1: Preparation of 3-benzyl 8-(tert-butyl) (1S,2S,5R)-2-(hydroxymethyl)-3,8-diazabicyclo[3.2.1]octane-3,8-dicarboxylate

[0205]

[0206] tert-Butyl (1S,2S,5R)-2-(hydroxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (1.80 g, 3.72 mmol) and sodium bicarbonate (1.87 g, 22.3 mmol) were dissolved in ethyl acetate (20 mL) and water (7 mL), benzyl chloroformate (1.52 g, 8.92 mmol) was added, and the reaction was allowed to proceed at room temperature for 18 hours. After the reaction was completed, the reaction solution was poured into saturated aqueous ammonium chloride solution (30 mL) and extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column chromatography (PE:EtOAc = 10:1 ~ 2:1) to obtain 3-benzyl 8-(tert-butyl) (1S,2S,5R)-2-(hydroxymethyl)-3,8-diazabicyclo[3.2.1]octane-3,8-dicarboxylate.

[0207] LC-MS: (ESI, m / z): [M-Boc+H] + = 277.1.

[0208] Step 2: Preparation of 3-benzyl 8-(tert-butyl) (1S,2S,5R)-2-formyl-3,8-diazabicyclo[3.2.1]octane-3,8-dicarboxylate

[0209]

[0210] DMSO (3.89 g, 49.75 mmol) was added dropwise to a solution of oxalyl chloride (1.37 g, 11.72 mmol) in dichloromethane (20 mL) at -78 °C under nitrogen protection. After stirring for 15 min at the temperature, a solution of 3-benzyl 8-(tert-butyl) (1S,2S,5R)-2-(hydroxymethyl)-3,8-diazabicyclo[3.2.1]octane-3,8-dicarboxylate (2.5 g, 6.63 mmol) in DCM (20 mL) was added. After 1 h of reaction at -78 °C, triethylamine (5.37 g, 5.31 mmol) was added, and the reaction was kept at -78 °C for 2 h. After the reaction was completed, the reaction solution was poured into saturated aqueous ammonium chloride solution (100 mL) and extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column chromatography (PE:EtOAc = 10:1 ~ 2:1) to give 3-benzyl 8-(tert-butyl) (1S,2S,5R)-2-formyl-3,8-diazabicyclo[3.2.1]octane-3,8-dicarboxylate.

[0211] LC-MS: (ESI, m / z): [M+H] + = 375.2.

[0212] Step 3: Preparation of 3-benzyl 8-(tert-butyl) (1S,2S,5R)-2-((S)-1-hydroxyethyl)-3,8- diazabicyclo[3.2.1]octane-3,8-dicarboxylate

[0213]

[0214] A solution of 3-benzyl 8-(tert-butyl) (1S,2S,5R)-2-formyl-3,8-diazabicyclo[3.2.1]octane-3,8-dicarboxylate (2.03 g, 5.43 mmol) in tetrahydrofuran (20 mL) was cooled to -78 °C, and CH3MgBr (2.17 mL, 6.52 mmol, 3.0 M / THF) was added dropwise. After the addition was completed, the mixture was stirred at -78 °C for 3 h. After the reaction was completed, saturated aqueous ammonium chloride solution (30 mL) was added and extracted with ethyl acetate (30 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product, which was purified by silica gel column chromatography (PE / EA = 2 / 1) to give 3-benzyl 8-(tert-butyl) (1S,2S,5R)-2-((S)-1-hydroxyethyl)-3,8-diazabicyclo[3.2.1]octane-3,8-dicarboxylate.

[0215] LC-MS: (ESI, m / z): [M-56+H] + = 335.2.

[0216] Step 4: Preparation of tert-butyl (1S,2S,5R)-2-((S)-1-hydroxyethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

[0217]

[0218] tert-Butyl 3-benzyl 8-(tert-butyl) (1S,2S,5R)-2-((S)-1-hydroxyethyl)-3,8-diazabicyclo[3.2.1]octane-3,8-dicarboxylate (1.83 g, 4.70 mmol) was dissolved in methanol (20 mL), then Pd / C (0.2 g) was added, and the reaction mixture was replaced with hydrogen for three times, and continued to react for 16 hours at 25 °C. After the reaction was completed, the reaction mixture was filtered, and the crude product was concentrated under reduced pressure. The organic phase was concentrated under reduced pressure to obtain tert-butyl (1S,2S,5R)-2-((S)-1-hydroxyethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate, which was used directly in the next step without further purification.

[0219] LC-MS: (ESI, m / z): [M+H] + = 257.2.

[0220] Step 5: Preparation of tert-butyl (1S,2S,5R)-2-((S)-1-((7-chloro-8-fluoro-2- (methylthio)-4-oxo-3,4-dihydropyrido[4,3-d]pyrimidin-5-yl)oxy)ethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

[0221]

[0222] (1S,2S,5R)-2-((S)-1-hydroxyethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (1.07 g, 4.17 mmol) was dissolved in tetrahydrofuran (20 mL), cooled to 0 °C, NaH (0.5 g, 12.5 mmol, 60%) was added, the mixture was stirred at 0 °C for 30 min, then 5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (1.28 g, 4.58 mmol) was added, the mixture was stirred at 25 °C for 3 h. After the reaction was completed, water (10 mL) was added to the reaction solution, then the tetrahydrofuran in the reaction solution was distilled off under reduced pressure. Then the mixture was adjusted to pH about 8 with dilute hydrochloric acid (1 M) until precipitate appeared, the precipitate was filtered and dried to obtain the crude product (1S,2S,5R)-2-((S)-1-((7-chloro-8-fluoro-2-(methylthio)-4-oxo-3,4-dihydropyrido[4,3-d]pyrimidin-5-yl)oxy)ethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester, which was used in the next step without further purification.

[0223] LC-MS: (ESI, m / z): [M+H] + = 500.1.

[0224] Step 6: Preparation of (5S,5aS,6S,9R)-2-chloro-1-fluoro-5-methyl-12-(methylthio)- 5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[1,8-ab]heptene-14-carboxylic acid tert-butyl ester

[0225]

[0226] (1S,2S,5R)-2-((S)-1-((7-chloro-8-fluoro-2-(methylthio)-4-oxo-3,4- dihydropyridine[4,3-d]pyrimidin-5-yl)oxy)ethyl)-3,8-diazabicyclo[3.2.1]octane-8- carboxylic acid tert-butyl ester (1.43 g, 2.87 mmol) and DIEA (0.37 g, 28.7 mmol) were dissolved in DCM (20 mL), then BOPCl (2.19 g, 8.6 mmol) was added to the mixture. The mixture was reacted at 25 °C for 4 h. After the reaction was completed, water (50 mL) was added to the reaction solution, and dichloromethane was extracted (50 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column chromatography (PE:EA = 2:1) to obtain (5S,5aS,6S,9R)-2-chloro-1-fluoro-5-methyl-12-(methylthio)-5a,6,7,8,9,10- hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[1,8-ab]heptene-14- carboxylic acid tert-butyl ester.

[0227] LC-MS: (ESI, m / z): [M+H] + = 482.1.

[0228] Step 7: Preparation of (5S,5aS,6S,9R)-2-chloro-1-fluoro-5-methyl-12-(methylsulfinyl)- 5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[1,8- ab]heptene-14-carboxylic acid tert-butyl ester

[0229]

[0230] (5S,5aS,6S,9R)-2-chloro-l-fluoro-5-methyl-12-(methylthio)-5a,6,7,8,9,10- hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[1,8-ab]heptene- 14-carboxylic acid tert-butyl ester (0.87 g, 1.8 mmol) was dissolved in dichloromethane (10 mL), m-CPBA (0.47 g, 2.7 mmol) was added, and the reaction was allowed to proceed at room temperature for 2 hours. After the reaction was completed, the reaction solution was poured into saturated aqueous sodium thiosulfate (20 mL), washed with saturated aqueous sodium bicarbonate (10 mL), and then extracted with dichloromethane (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give crude (5S,5aS,6S,9R)-2-chloro-l-fluoro-5-methyl-12-(methylsulfinyl)-5a,6,7,8,9,10- hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[1,8-ab]heptene-14- carboxylic acid tert-butyl ester, which was used directly in the next step without purification.

[0231] LC-MS: (ESI, m / z): [M+H] + = 498.1.

[0232] Step 8: Preparation of (5S,5aS,6S,9R)-2-chloro-l-fluoro-5-methyl-12-((R)-2-methyl-3- (4-tritylpiperazin-l-yl)propoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14- pentaaza-6,9-methanonaphtho[1,8-ab]heptene-14-carboxylic acid tert-butyl ester

[0233]

[0234] (R)-2-methyl-3-(4-tritylpiperazin-l-yl)propan-l-ol (0.8 g, 2.0 mmol) was dissolved in tetrahydrofuran (20 mL), cooled to 0 °C, NaH (132 mg, 3.3 mmol) was added slowly, reacted at 25 °C for 30 min, then (5S,5aS,6S,9R)-tert-butyl 2-chloro-l-fluoro-5-methyl-12-((R)-2-methyl-3-(4- tritylpiperazin-l-yl)propoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14- pentaaza-6,9-methanonaphtho[l,8-ab]heptene-14-carboxylate (0.83 g, 1.67 mmol) was added, the mixture was reacted at 0 °C for 1 h. After the reaction was completed, saturated aqueous ammonium chloride solution (10 mL) was added, extracted with ethyl acetate (20 mL x 3). The combined organic phase was washed with saturated NaCl solution, dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column chromatography (DCM:MeOH = 15:1) to obtain (5S,5aS,6S,9R)-tert-butyl 2-chloro-l-fluoro-5-methyl-12-((R)-2-methyl-3-(4- tritylpiperazin-l-yl)propoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14- pentaaza-6,9-methanonaphtho[l,8-ab]heptene-14-carboxylate.

[0235] LC-MS: (ESI, m / z): [M+H] + = 834.3.

[0236] 3. Preparation of compound 237:

[0237] 1-(5-(9-((4-((R)-3-(((5S,5aS,6S,9R)-2-(3-chloro-5-hydroxy-2- (trifluoromethyl)phenyl)-l-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14- pentaaza-6,9-methanonaphtho[l,8-ab]heptan-12-yl)oxy)-2-methylpropyl)piperazin-l-yl) methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidine- 2,4(lH,3H)-dione

[0238] Step 1: Preparation of 2-(3-bromo-5-chloro-4-(trifluoromethyl)phenyl)-4,4,5,5- tetramethyl-l,3,2-dioxaborolane

[0239]

[0240] To a solution of 1-bromo-3-chloro-2-(trifluoromethyl)benzene (3.0 g, 11.56 mmol) in tetrahydrofuran (30 mL) was added 4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2.96 g, 23.13 mmol), (1,5-cyclooctadiene)(methoxy)iridium(I) dimer (766 mg, 1.16 mmol) and 4-tert-butyl-2-(4-tert-butyl-2-pyridyl)pyridine (372 mg, 1.39 mmol). The mixture was degassed and replaced with nitrogen three times, heated to 60 °C and stirred for 3 hours. The reaction was diluted with water (30 mL) and extracted with ethyl acetate (40 mL x 3), the combined organic phase was dried over sodium sulfate and concentrated in vacuo to give 2-(3-bromo-5-chloro-4-(trifluoromethyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane, which was used in the next step without further purification.

[0241] Step 2: Preparation of 3-bromo-5-chloro-4-(trifluoromethyl)phenol

[0242]

[0243] To a solution of 2-(3-bromo-5-chloro-4-(trifluoromethyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (4.45 g, crude) in tetrahydrofuran (30 mL) and water (15 mL) was added AcOH (27.7 g, 0.46 mol) and H2O2 (7.86 g, 0.23 mol), the mixture was stirred at 10 °C for 1 hour. After the reaction was completed, the mixture was diluted with water (30 mL) and extracted with ethyl acetate (30 mL x 3), the combined organic phase was dried over sodium sulfate and then concentrated in vacuo. The concentrate was purified by silica gel column chromatography (PE:EA = 5:1) to give 3-bromo-5-chloro-4-(trifluoromethyl)phenol.

[0244] 1 H NMR (400 MHz, DMSO-d6) δ 11.25 (s, 1H), 7.22 (d, J = 2.0 Hz, 1H), 7.05 (d, J = 1.9 Hz, 1H).

[0245] Step 3: Preparation of 1-bromo-3-chloro-5-(methoxymethoxy)-2- (trifluoromethyl)benzene

[0246]

[0247] To a solution of 3-bromo-5-chloro-4-(trifluoromethyl)phenol (1.4 g, 5.08 mmol) in DCM (20 mL) was added DIEA (1.97 g, 15.54 mmol) and MOMBr (1.27 g, 10.17 mmol) at 0 °C. The mixture was stirred at room temperature for 2 h under nitrogen. After completion of the reaction, the mixture was diluted with water (30 mL) and extracted with dichloromethane (30 mL x 3). The combined organic phase was dried over sodium sulfate and concentrated in vacuo to give 1-bromo-3-chloro-5-(methoxymethoxy)-2-(trifluoromethyl)benzene. The crude product was used in the next step without further purification.

[0248] Step 4: Preparation of 2-(3-chloro-5-(methoxymethoxy)-2- (trifluoromethyl)phenyl)-4,4,5,5-tetramethyl-l,3,2-dioxaborolane

[0249]

[0250] To a solution of 1-bromo-3-chloro-5-(methoxymethoxy)-2-(trifluoromethyl)benzene (1.4 g, 4.38 mmol) and bis(pinacolato)diboron (2.23 g, 8.76 mmol) in 1,4-dioxane (20 mL) was added Pd(dppf)Cl2(321 mg, 0.44 mmol) and potassium acetate (1.29 g, 13.14 mmol). The mixture was stirred at 100 °C for 5 h under nitrogen. The reaction mixture was filtered and the filtrate was concentrated to give the crude product. The concentrate was purified by silica gel column chromatography (PE:EA = 30: 1) to give 2-(3-chloro-5-(methoxymethoxy)-2-(trifluoromethyl)phenyl)-4,4,5,5-tetramethyl-l,3,2-dioxaborolane.

[0251] 1 H NMR (400 MHz, CDC13) δ 7.18 (d, J = 2.3 Hz, 1H), 6.97 (d, J = 2.2 Hz, 1H), 5.19 (s, 2H), 3.46 (s, 3H), 1.36 (s, 12H).

[0252] Step 5: Preparation of (5S,5aS,6S,9R)-2-(3-chloro-5-(methoxymethoxy)-2- (trifluoromethyl)phenyl)-l-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[l,8-ab]heptene-14-carboxylic acid tert-butyl ester 2 Preparation of (5S,5aS,6S,9R)-2-(3-chloro-5-(methoxymethoxy)-2- (trifluoromethyl)phenyl)-l-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[l,8-ab]heptene-14-carboxylic acid tert-butyl ester

[0253]

[0254] To a mixture solution of (5S,5aS,6S,9R)-tert-butyl 2-chloro-l-fluoro-5-methyl- 12-((R)-2-methyl-3-(4-tritylpiperazin-l-yl)propoxy)-5a,6,7,8,9,10-hexahydro-5H- 4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[l,8-ab]heptene-14-carboxylate (400 mg, 0.48 mmol) and 2-(3-chloro-5-(methoxymethoxy)-2- (trifluoromethyl)phenyl)-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (264 mg, 0.72 mmol) in 1,4-dioxane (5 mL) and H2O (0.8 mL) was added Pd(dppf)Cl2(35 mg, 0.0048 mmol) and potassium phosphate (204 mg, 0.96 mmol). The mixture was reacted at 100 °C for 8 h under nitrogen protection. Water (5 mL) was added to the reaction solution, which was then extracted with ethyl acetate (5 mL x 3), the organic phases were combined and dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (DCM:MeOH = 10:1) to obtain (5S,5aS,6S,9R)-tert-butyl 2-(3-chloro-5- (methoxymethoxy)-2-(trifluoromethyl)phenyl)-l-fluoro-5-methyl-5a,6,7,8,9,10- hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[l,8-ab]heptene-14- carboxylate (400 mg, 0.48 mmol) as a white solid. LC-MS: (ESI, m / z): [M + H] = 796.3. 2 -((R)-2-methyl-3-(4-tritylpiperazin-l-yl)propoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxa- 3,10a,11,13,14-pentaaza-6,9-methanonaphtho[l,8-ab]heptene-14-carboxylate.

[0255] LC-MS: (ESI, m / z): [M + H] = 796.3. + = 796.3.

[0256] Step 6: Preparation of (5S,5aS,6S,9R)-tert-butyl 2-(3-chloro-5- (methoxymethoxy)-2-(trifluoromethyl)phenyl)-l-fluoro-5-methyl-5a,6,7,8,9,10- hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[l,8-ab]heptene-14- carboxylate 2 -((R)-2-methyl-3-(4-tritylpiperazin-l-yl)propoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxa- 3,10a,11,13,14-pentaaza-6,9-methanonaphtho[l,8-ab]heptene-14-carboxylate.

[0257]

[0258] Step 6: Preparation of (5S,5aS,6S,9R)-tert-butyl 2-(3-chloro-5- (methoxymethoxy)-2-(trifluoromethyl)phenyl)-l-fluoro-5-methyl-5a,6,7,8,9,10- hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[l,8-ab]heptene-14- carboxylate 2-((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]hepten-14-carboxylic acid tert-butyl ester (220 mg, 0.21 mmol) was dissolved in glacial acetic acid (3 mL) and reacted at 60 °C for 2 hours. After the reaction was completed, the crude product was concentrated and purified by silica gel column chromatography (DCM:MeOH(NH3) = 10:1) to obtain (5S,5aS,6S,9R)-2-(3-chloro-5-(methoxymethoxy)-2-(trifluoromethyl)phenyl)-1-fluoro-5-methyl-λ 2 -((R)-2-methyl-3-(piperazin-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]hepten-14-carboxylic acid tert-butyl ester.

[0259] LC-MS:(ESI,m / z):[M+H] + =796.3.

[0260] Step 7: Preparation of (5S,5aS,6S,9R)-2-(3-chloro-5-(methoxymethoxy)-2-(trifluoromethyl)phenyl)-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-5-methyl-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

[0261]

[0262] (5S,5aS,6S,9R)-2-(3-chloro-5-(methoxymethoxy)-2-(trifluoromethyl)phenyl)-1-fluoro-5-methyl-λ 2To a solution of tert-butyl ((R)-2-methyl-3-(piperazin-l-yl)propoxy)-5a,6,7,8,9,10- hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[l,8-ab]heptene-14- carboxylate (140 mg, 0.176 mmol) in DCM / AcOH (2 mL / 0.2 mL) was added 3-(3-(2,4- dioxotetrahydropyrimidin-l(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecan-9- formaldehyde (90 mg, 0.21 mmol) and stirred at 25 °C for 1 h. The mixture was added with NaBH(OAc)3(93 mg, 0.44 mmol) under ice-bath and stirred for 2 h. The mixture was concentrated, and the concentrate was purified by silica gel column chromatography (DCM:MeOH = 100:1 ~ 10:1) to give tert-butyl (5S,5aS,6S,9R)-2-(3-chloro-5-(methoxymethoxy)-2- (trifluoromethyl)phenyl)-12-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-l(2H)-yl)-4- methoxybenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazin-l-yl)-2-methylpropoxy)-l- fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9- methanonaphtho[l,8-ab]heptene-14-carboxylate.

[0263] LC-MS: (ESI, m / z): [(M-Boc) / 2 + H] + = 554.2.

[0264] Step 8: Preparation of l-(5-(9-((4-((R)-3-(((5S,5aS,6S,9R)-2-(3-chloro-5-hydroxy-2- (trifluoromethyl)phenyl)-l-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14- pentaaza-6,9-methanonaphtho[l,8-ab]heptan-l2-yl)oxy)-2-methylpropyl)piperazin-l-yl)methyl)- 3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidine-2,4(lH,3H)-dione

[0265]

[0266] To a solution of (5S,5aS,6S,9R)-2-(3-chloro-5-(methoxymethoxy)-2- (trifluoromethyl)phenyl)-12-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)- yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazin-1-yl)-2- methylpropoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14- pentaaza-6,9-methanonaphtho[1,8-ab]heptene-14-carboxylic acid tert-butyl ester (150 mg, 0.124 mmol) in 1,4-dioxane (1 mL) was added HCl / 1,4-dioxane (6 N, 1.0 mL) and the reaction was stirred at room temperature for 1 h. The reaction was concentrated under reduced pressure and the concentrate was adjusted to pH > 7 with aqueous NaHCO3solution and extracted with EtOAc (30 mL x 3). The organic phase was washed with saturated aqueous NaCl solution, dried over anhydrous Na2SO4and concentrated. The crude product was purified by high performance liquid chromatography to give 1-(5-(9-((4-((R)-3-(((5S,5aS,6S,9R)-2-(3-chloro-5-hydroxy-2- (trifluoromethyl)phenyl)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14- pentaaza-6,9-methanonaphtho[1,8-ab]heptan-12-yl)oxy)-2-methylpropyl)piperazin-1-yl)methyl)- 3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione.

[0267] LC-MS: (ESI, m / z): [M / 2 + H] + = 532.6.

[0268] 1H NMR (400 MHz, DMSO-d6) δ 10.32 (s, 1H), 8.15 (s, 1H), 7.36 (dd, J = 8.4, 2.1 Hz, 1H), 7.32 (d, J = 2.1 Hz, 1H), 7.18 (d, J = 1.9 Hz, 1H), 7.15 (d, J = 8.6 Hz, 1H), 6.76 (d, J = 65.0 Hz, 1H), 5.13 (d, J = 11.1 Hz, 1H), 4.64 - 4.55 (m, 1H), 4.51 - 4.43 (m, 1H), 4.07 (d, J = 8.4 Hz, 2H), 3.84 (s, 3H), 3.79 - 3.71 (m, 2H), 3.66 - 3.56 (m, 5H), 3.13 (d, J = 12.5 Hz, 2H), 2.68 (t, J = 6.6 Hz, 2H), 2.58 - 2.53 (m, 2H), 2.48 - 2.26 (m, 8H), 2.25 - 2.10 (m, 4H), 1.92 - 1.84 (m, 1H), 1.78 - 1.59 (m, 5H), 1.58 - 1.36 (m, 8H), 1.36 - 1.22 (m, 2H), 1.17 - 0.91 (m, 7H).

[0269] Test Example 1: KRAS-G12D degradation test

[0270] The degradation effect of the compound on KRAS-G12D was investigated on KRAS G12D mutant human lung cancer cell A-427 (ATCC), KRAS G12D mutant human stomach cancer cell AGS, KRAS G12D mutant human metastatic pancreatic cancer cell AsPc-1, KRAS G12D mutant human pancreatic adenocarcinoma cell Panc04.03, KRAS G12D mutant human pancreatic adenocarcinoma cell PK59, KRAS G12D mutant human colon cancer cell GP2D. The specific method is: 0.95 mL of cells were plated in each well of a 24-well cell culture plate to make the cell density 5 x 105cells / mL. 5Cells were plated at 10,000 cells / well in 24-well plates in 1 mL of DMEM medium containing 10% FBS. The cells were incubated overnight at 37°C in a 5% CO2 incubator. Then 50 μL of diluted compound solution was added to the corresponding wells, so that the final concentration of the compound was in the range of 0.03-3000 nM. The final concentration of DMSO was 0.25%. After 24 hours of incubation at 37°C in a 5% CO2 incubator, the medium was removed from the 24-well cell culture plate, which was washed twice with 1 x PBS (Kabi). The A-427 cells adhering to the bottom of the cell culture plate were lysed by adding 120 μL of RIPA (QIAGEN) lysis buffer supplemented with 1 mM phenylmethylsulfonyl fluoride, protease inhibitor cocktail (QIAGEN), and phosphatase inhibitor cocktail (QIAGEN). After 30 minutes of incubation on ice, the protein lysate in each well was transferred to a 1.5 mL centrifuge tube and centrifuged at 15000 g for 20 minutes at 4°C. The cell lysate supernatant was stored at -80°C.

[0271] The prepared cell lysate supernatant sample was taken out from the -80°C refrigerator after thawing, and the total protein concentration in the cell lysate was determined using the BCA protein quantification kit (Tiangen). Then the total protein concentration of the sample was adjusted to 0.5 μg / μL with PBS and 5×SDS-PAGE protein loading buffer (Bi Yun Tian), and 100°C water bath for 15 minutes. Then placed in ice bath for 5 minutes, 14000g, 4°C centrifugation for 1 minute, mixed as the sample for protein immunoblotting test. Add sample to the loading well of 10% precast gel (Kai Ji), the loading amount is 10 μL (total protein is 5 μg), after adding enough Tris-MOPS-SDS electrophoresis liquid (Adam), 120V constant voltage, 55 minutes for electrophoresis. After electrophoresis, 250mA constant current, 55 minutes, transfer the protein on the gel to the PVDF membrane. After the transfer, the PVDF membrane was placed in 1×Quick Block blocking solution (Bi Yun Tian) and incubated at room temperature for 30 minutes. After blocking, the PVDF membrane was diluted with 5% BSA as diluent at a ratio of 1:1000 for KRAS antibody (Abeam) and 1:2000 for β-actin antibody (Abeam), 4°C incubation overnight, then washed the membrane with 1×TBST buffer (2.4g Tris, 8.8g NaCl, 1.5mL Tween 20, adjust pH to 7.4, constant volume to 1L) for 10 minutes / time, a total of 3 times. Then incubate with 5% BSA diluted secondary antibody (Abeam) at room temperature for 2 hours, then wash the membrane with 1×TBST buffer for 10 minutes / time, a total of 3 times. Finally, incubate with Clarity Western ECL Substrate (BIO-RAD) for 5 minutes for color development, and take pictures of the protein map by ChemiScope 6200 Touch chemiluminescence imaging system (Qin Xiang). The protein map was analyzed by gray value analysis software (Qin Xiang). The gray correction value of each sample was calculated using the formula: gray correction value = (target protein gray value / corresponding internal standard gray value) ×10 3 . The degradation rate was calculated by comparing the gray correction value of the control group. Then the log concentration-inhibition rate was analyzed by GraphPad Prism 8 analysis software to obtain the DC 50 and D max values of the compounds (as shown in Table 5).

[0272] Table 5

[0273]

[0274] Test Example 2: 3D cell proliferation inhibition test

[0275] The inhibitory effect of the compound on 3D cell proliferation was investigated on KRAS G12D mutant human lung cancer cells A427, KRAS G12D mutant human gastric cancer cells AGS, KRAS G12D mutant human metastatic pancreatic cancer cells AsPc-1, KRAS G12D mutant human pancreatic cancer cells Panc 04.03, KRAS G12D mutant human pancreatic cancer cells HPAC, KRAS G12D mutant human pancreatic adenocarcinoma cells PK59, and KRAS G12D mutant human colon cancer cells GP2D. The specific method is as follows:

[0276] 10 μL of a gradient dilution solution of the compound was added to the corresponding well of a 96-well low adsorption microplate (PerkinElmer), so that the final concentration of the test compound in the reaction system was in the range of 10000 nM to 0.026 nM. The final concentration of DMSO was 0.2%. Then 90 μL of cells were added to the corresponding well, so that the cell density was 5×10 2 μL / well. In addition to the test compound test well, DMSO control wells and medium control wells were set up at the same time, the DMSO control wells contained DMSO and cells respectively, and the medium wells contained only medium. After adding the sample, the 96-well low adsorption microplate was placed in a 5% carbon dioxide incubator and incubated at 37°C for 7 days. After 7 days, the microplate was taken out, 100 μL of CTG reagent (Promega) was added to each well, and after incubation at room temperature for 60 minutes, the reading was performed on the enzyme label instrument EnVision using the chemiluminescence program. The inhibition rate of the compound at each concentration was calculated by the cell proliferation inhibition percentage calculation formula: inhibition rate % = (DMSO control group average value - single concentration reading value of the compound) / (DMSO control group average value - medium control group average value) x 100, and then the IC 50 value of the compound was obtained by non-linear curve fitting of the logarithmic concentration-inhibition rate using GraphPad Prism 8 (as shown in Table 6).

[0277] Table 6

[0278]

[0279] In Table 6, “-” indicates that it is not detected

[0280] Test Example 3: Mouse pharmacokinetic test

[0281] (1) Experimental reagents

[0282] The test compound in this test is from the compound of the specific embodiment of the present application.

[0283] (2) Experimental animals

[0284] ICR mice, male, 6, source: Shanghai Shengkang Biological Technology Co., Ltd.

[0285] (3) Drug formulation and administration

[0286] ICR mice were administered by single intravenous (IV) injection: the appropriate amount of compound was weighed, 1% volume of DMSO was added and vortexed and ultrasonically dissolved, then 10% volume of Solutol was added and vortexed, and finally 89% volume of 0.25% glucose solution was added and vortexed to obtain a clear solution. Three mice were administered by tail vein injection after fasting overnight, and the administration dose was 1 mg / mL.

[0287] ICR mice were administered by single oral (PO) administration: the appropriate amount of compound was weighed, 2% volume of DMSO was added and vortexed and ultrasonically dissolved, then 10% volume of Solutol was added and vortexed, and finally 88% volume of 0.25% glucose solution was added and vortexed to obtain a clear solution. Three mice were administered orally after fasting overnight, and the administration dose was 30 mg / mL.

[0288] (4) Sample collection

[0289] The sampling time points were: 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after administration. At the above set time points, blood was collected from the submandibular vein, and 30-40 μL of whole blood was collected for each sample, anticoagulated with sodium heparin, and placed on wet ice after collection, and centrifuged to separate plasma (centrifugation conditions: 8000 rpm / min, 10 min, 2-8°C) within 1 hour. The plasma samples were stored in a -80°C refrigerator before analysis.

[0290] Sample analysis pretreatment: 10 μL of plasma sample was taken, 200 μL of precipitant containing internal standard (methanol: acetonitrile = 1:1) was added, vortexed for 1 minute, and centrifuged at 4000 rpm / min for 15 minutes. 100 μL of supernatant was transferred to a 96-well plate, and 100 μL of diluent (water:methanol:formic acid = 80:20:0.1) was added, vortexed for 1 minute, and 15 μL of sample was injected into LC-MS / MS for analysis.

[0291] Control 1: 4-(4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)pyrido[4,3-d]pyrimidin-7-yl)-5-ethynyl-6-fluoronaphthalen-2-ol

[0292] Prepared according to the method of Example 252 in WO2021041671A1.

[0293] MS (ESI) m / z: [M+H] + = 601.1.

[0294] 1H NMR (400 MHz, DMSO-d6) δ 10.14 (s, 1H), 9.03 (s, 1H), 7.97 (dd, J = 9.2 Hz, 6.0 Hz, 1H), 7.49 - 7.43 (m, 1H), 7.39 (d, J = 2.8 Hz, 1H), 7.17 (d, J = 2.4 Hz, 1H), 5.36 - 5.19 (m, 1H), 4.46 (d, J = 12.2 Hz, 1H), 4.31 (d, J = 11.9 Hz, 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).

[0295] Reference Compound 2: 1-(5-(9-((4-((1-(((4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-7-(8- ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-chlorophenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0296] Reference Compound 2 was prepared according to the method described in Example 6 of WO2024083258A1.

[0297] LC-MS: (ESI, m / z): [M+H]+= 1027.2.

[0298] 1 H NMR (400 MHz, CD3OD) δ 8.99 (s, 1H), 7.85 (dd, J = 9.2, 5.6 Hz, 1H), 7.63 (d, J = 8.2 Hz, 1H), 7.53 (d, J = 1.6 Hz, 1H), 7.41 (dd, J = 8.2, 2.0 Hz, 1H), 7.36 - 7.28 (m, 2H), 7.20 (d, J = 2.4 Hz, 1H), 4.85 - 4.33 (m, 5H), 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).

Claims

1. A method for preparing a compound of formula I, characterized in that, It includes the following steps: (i) In a solvent, the compound of formula II and the Grignard reagent R 2 MgX is reacted at -30°C to -5°C to obtain a reaction solution; (ii) In a solvent, in the presence of an inorganic base, the reaction solution from step (i) undergoes a hydrolysis reaction to yield compound I. Among them, R a Amino protecting group: -COO-C 1-6 Alkyl or -C 1-6 alkyl-phenyl, wherein the phenyl group is optionally oxidized by one or more C atoms. 1-6 Alkoxy, halogen, C 1-6 Alkyl substitution; X is a halogen; R 2 C 1-6 Alkyl groups or C groups substituted with one or more deuterium groups 1-6 alkyl.

2. The method for preparing the compound of formula I as described in claim 1, characterized in that, It satisfies at least one of the following conditions: (1) In step (i), the solvent is an ether solvent, such as tetrahydrofuran and / or dimethyltetrahydrofuran; (2) In step (i), the Grignard reagent is methyl magnesium halide or deuterated methyl magnesium halide, for example CH3MgBr; (3) In step (i), the volume-to-mass ratio of the solvent to the compound of formula II is 5 to 15 L / kg, for example 10 L / kg or 11 L / kg; (4) In step (i), the molar ratio of the compound of formula II to the Grignard reagent is 1:(1 to 3), for example 1:3, 1:1.8, 1:1.7 or 1:1.2, preferably 1:1.2; (5) In step (i), the Grignard reagent is used in the form of a Grignard reagent dimethyltetrahydrofuran solution, for example, a 1-4 mol / L methyl Grignard reagent dimethyltetrahydrofuran solution, wherein the volume ratio of the methyl Grignard reagent dimethyltetrahydrofuran solution to the tetrahydrofuran is 1:(4-10), for example, 1:

6. (6) In step (i), when the Grignard reagent is used in the form of a methyl Grignard reagent dimethyltetrahydrofuran solution, the volume-to-mass ratio of the methyl Grignard reagent dimethyltetrahydrofuran solution to the compound of formula II is 0.9 to 2.2 L / kg, for example 1.5 L / kg; (7) In step (i), the reaction is carried out at two temperature stages, the first temperature stage being -25 to -5°C, for example 10 to -20°C, and the second temperature stage being 10 to 40°C, for example 20 to 30°C. (8) Step (i) preferably also includes the following steps: Step a: First, react at -5 to -25°C for 0.5 to 3 hours, for example, at -10 to -20°C for 1 to 2 hours; Step b, and then react at 10-40°C for 2-10 hours, for example, at 20-30°C for 5-8 hours; (9) In step (i), after the reaction is completed, the following post-processing steps may also be included: cooling, quenching the reaction, and concentration; (10) The solvent is a mixture of an alcohol solvent and water, wherein the alcohol solvent may be ethanol; the volume ratio of the alcohol solvent to water may be (1-3):(1-3), for example 1:1; (11) In step (ii), the mass-to-volume ratio of the compound of formula II to the solvent may be 0.02 to 0.3 kg / L, for example 0.1 kg / L; (12) In step (ii), the inorganic base is an alkali metal hydroxide, such as sodium hydroxide; (13) In step (ii), the molar ratio of the compound of formula II to the inorganic base is 1:(5 to 10), for example 1:8; (14) In step (ii), the temperature of the reaction is 65–90°C, for example 75–80°C; (15) In step (ii), the reaction time is 10 to 24 hours, for example 16 to 20 hours; (16) In step (ii), after the reaction is completed, the following post-processing steps may be included: concentrating the reaction solution, extracting, separating, washing, concentrating, purifying, to obtain the compound of formula I; (17) Step (ii) preferably also includes the following steps: adding the alcohol solvent and water mixture, the inorganic base, and reacting at 65-90°C; (18) The reaction materials for the preparation method of the compound of formula I are the compound of formula II, the Grignard reagent, the inorganic base and the solvent; (19) The preparation method of the compound of formula I preferably includes the following steps: in an ether solvent, at -10 to -20°C, the compound of formula II and the methyl Grignard reagent are reacted, and the temperature is raised to 20 to 30°C to continue the reaction; (20) The R a It is a Boc group; (21) The R 2 It is methyl; (22) Remove the solvent described in step (i) after step (i) is completed.

3. The method for preparing the compound of formula I as described in claim 1, characterized in that, It also includes the following steps: in a solvent, the compound of formula III undergoes an oxidation reaction in the presence of an oxidizing agent and an organic base to produce the compound of formula II; R a As described in claim 1.

4. The method for preparing the compound of formula I as described in claim 3, characterized in that, It satisfies at least one of the following conditions: (1) The solvent is a haloalkane solvent, such as dichloromethane; (2) The oxidant in the oxidation reaction is a dimethyl chlorosulfonium salt. Preferably, the oxidant is prepared by the following steps: DMSO and oxalyl chloride react in a solvent to obtain the oxidant. The oxidant can be directly used in the oxidation reaction after preparation. Preferably, the solvent is a haloalkane solvent, such as dichloromethane. More preferably, the mass ratio of the solvent to the oxalyl chloride is (15-25):1, for example, 19.

3. The oxidant can be prepared at -70 to -40°C, for example, -60 to -50°C. (3) The organic base is an alkylamine, such as triethylamine; (4) The volume-to-mass ratio of the solvent to the compound of formula III is 10-25 L / kg; (5) The compound of formula III is used in the form of a mixture of the compound of formula III and a haloalkane solvent, wherein the volume-to-mass ratio of the compound of formula III to the haloalkane solvent is 1 to 10 L / kg, for example 5 L / kg; (6) The molar ratio of the compound of formula III to oxaloyl chloride is 1:(1-3), for example 1:2; (7) The molar ratio of the compound of formula III to DMSO is 1:(1-4), for example 1:2.5; (8) The molar ratio of the compound of formula III to the organic base is 1:(5-10), for example 1:8; (9) The oxidation reaction is carried out at -70 to -40°C, for example at -60 to -50°C; (10) The organic base is preferably added during the reaction of the compound of formula III with the oxidant, preferably 1 to 3 hours after the addition of the compound of formula III; (11) The oxidation reaction preferably includes the following steps: Step S1: In a solvent, DMSO and oxalyl chloride react at -70 to -40°C, for example at -60 to -50°C. Step S2: The reaction solution from step S1 reacts with the compound of formula III at -70 to -40°C, for example, at -60 to -50°C. Step S3: The reaction solution from step S2 reacts with an organic base at -70 to -40°C, for example, at -60 to -50°C. (12) The oxidation reaction may include the following quenching step: after the reaction is completed, the reaction solution is mixed with a citric acid aqueous solution to quench the reaction, for example, a 10% citric acid aqueous solution is mixed to quench the reaction. (13) The post-processing of the oxidation reaction may include the following steps: after the reaction is completed, quench, extract and separate, wash, dry, concentrate, and purify to obtain compound II; (14) The reaction materials for the preparation method of the compound of formula II are the compound of formula III, the oxidant, the solvent and the organic base; (15) The preferred method for preparing the compound of formula II includes the following steps: reacting the compound of formula III with an oxidant in a haloalkane at -60 to -50°C, and then adding an organic base to react.

5. The method for preparing the compound of formula I as described in claim 3, characterized in that, The preparation method of the compound of formula I further includes the following steps: in a solvent, in the presence of a base, the compound of formula IV and X-Cbz undergo a substitution reaction, where X is a halogen, to generate the compound of formula III. R a As described in claim 3.

6. The method for preparing the compound of formula I as described in claim 5, characterized in that, It satisfies at least one of the following conditions: (1) The solvent is a mixture of water and an organic solvent; the organic solvent may be an ester solvent, such as ethyl acetate; (2) X is chlorine; (3) The base is an alkali metal acid salt, such as an alkali metal bicarbonate, or sodium bicarbonate; (4) The volume-to-mass ratio of the ester solvent to the compound of formula IV is preferably 5 to 15 L / kg, for example 10 L / kg; (5) The volume-to-mass ratio of the water to the compound of formula IV is preferably 1 to 8 L / kg, for example 3 L / kg; (6) The molar ratio of the compound of formula IV to the base is 1:(3-5), for example 1:4; (7) The molar ratio of the compound of formula IV and X-Cbz is 1:(1~2), for example 1:1.2; (8) The preferred post-processing steps of the substitution reaction are as follows: after the reaction is completed, the reaction solution of the substitution reaction is mixed with a saturated ammonium chloride aqueous solution, extracted and separated, washed and dried to obtain compound III; (9) The reactants in the preparation method of the compound of formula III are the solvent, the compound of formula IV, the base and benzyl chloroformate; (10) The method for preparing the compound of formula III preferably includes the following steps: in a mixed solvent of water and ester solvent, at 20-25°C, the compound of formula IV, the base and benzyl chloroformate undergo a substitution reaction.

7. A method for preparing a compound of formula II, characterized in that, It includes the following steps: In a solvent, compound III undergoes an oxidation reaction in the presence of an oxidizing agent and an organic base to produce compound II; The oxidation reaction was quenched using an aqueous solution of citric acid.

8. The method for preparing the compound of formula II as described in claim 7, characterized in that, In the preparation method of the compound of formula II, the organic base is an alkylamine, such as triethylamine; Preferably, the conditions and operation of the oxidation reaction are as described in claim 3 or 4; the method for preparing the compound of formula II also preferably includes the method for preparing the compound of formula III as described in claim 5 or 6.

9. A method for preparing a compound of formula X, characterized in that, It includes the following steps: Compound I is prepared by any one of the preparation methods described in claims 1-6, and compound X is prepared by reacting compound I with the following reaction. Step 1: In an organic solvent, under the action of an inorganic base, compound I and compound a undergo a substitution reaction to produce compound b; R a and R 2 As described in claim 1 or 2; Step 2: In an organic solvent, in the presence of an organic base and a coupling agent, compound b undergoes a cyclization reaction to generate compound c. Step 3: In an organic solvent, under the action of an oxidizing agent, compound c undergoes an oxidation reaction to produce compound d; Step 4: In an organic solvent, under the action of a base, compound d and compound e undergo a substitution reaction to produce compound b. Step 5: In a solvent, in the presence of an inorganic base and a catalyst, compound f and compound g undergo a coupling reaction to generate compound h. Among them, R 3 for R 3-1 C 1-6 Alkyl or C 3-6 cycloalkyl, the C 1-6 Alkyl or C 3-6 The cycloalkyl group is optionally reacted with one or more deuterium, halogen, or C. 1-6 Alkyl substitution, R 3-2 It is protected by OH, amino, O-hydroxy or NH-amino groups. Step 6: In an organic reagent, compound h is deprotected to generate compound i; Step 7: In an organic solvent, under the action of a reducing agent, compounds i and j undergo a reductive amination reaction to produce compound k; Step 8: In an organic solvent, compound k is deprotected to form compound X; R 4 for R 3-2a It is -OH or amino.

10. The method for preparing compound X as described in claim 9, characterized in that, Compound X is R 2 R 3-1 R 3-2a As described in claim 9; The preferred compound X is

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