Cyanoaromatic ring derivative and pharmaceutical use thereof

Abstract

The present invention relates to a cyanoaromatic ring derivative and pharmaceutical use thereof. The pharmaceutical formulation comprises a therapeutically effective amount of active ingredient M and a pharmaceutically acceptable excipient. The active ingredient M is selected from the compound represented by general formula (I) or a stereoisomer thereof, a tautomer thereof, or a pharmaceutically acceptable salt thereof. The pharmaceutical formulation comprises 1-800 mg of the active ingredient M. The present invention also relates to use of the pharmaceutical formulation for preparing a related drug for treating cancer. B-L-K (I).

Description

A cyano aromatic ring derivative and its pharmaceutical application Technical Field

[0001] This invention belongs to the field of pharmaceutical formulations, specifically relating to a pharmaceutical formulation comprising a therapeutically effective amount of an active ingredient M and a pharmaceutical excipient, wherein the active ingredient M is selected from compounds of general formula (I) or their stereoisomers, tautomers, or pharmaceutically acceptable salts. This invention also relates to the use of the pharmaceutical formulation in the preparation of cancer-related drugs. Background Technology

[0002] Prostate cancer is a type of cancer that is often discovered in its early stages. Its causes are frequently related to genetic factors, a high-fat diet, and endocrine disorders. Generally, the incidence of prostate cancer is higher in developed countries than in developing countries. In 2016, there were 120,000 new cases of prostate cancer in China, and this number is projected to reach 237,000 by 2030, with a market share of $4.8 billion. Early-stage prostate cancer patients can be treated radically and have a longer survival time. However, late-stage patients with metastatic cancer undergo castration combined with anti-androgen therapy, and the disease progresses to castration-resistant prostate cancer. Clinical studies show that most patients with castration-resistant prostate cancer overexpress the androgen receptor (AR). Inhibiting the androgen receptor (AR) signaling pathway has a significant therapeutic effect on patients with hormone-resistant prostate cancer; therefore, inhibiting the androgen receptor (AR) is an effective way to directly block this pathway.

[0003] PROTAC (proteolysis targeting chimera) molecules are bifunctional compounds that can simultaneously bind to target proteins and E3 ubiquitin ligases. These compounds can be recognized by the cell's proteasome, causing the degradation of the target protein and effectively reducing its concentration in the cell. By introducing ligands that can bind to different target proteins into PROTAC molecules, it has become possible to apply PROTAC technology to the treatment of various diseases, and this technology has received widespread attention in recent years.

[0004] Therefore, it is necessary to develop novel PROTAC drugs targeting the androgen receptor (AR) for the treatment of androgen receptor-related tumors. Summary of the Invention

[0005] The present invention aims to provide a pharmaceutical formulation comprising a therapeutically effective amount of an active ingredient M and a pharmaceutical excipient, wherein the active ingredient M is selected from compounds of general formula (I) or their stereoisomers, tautomers, or pharmaceutically acceptable salts, and the pharmaceutical formulation has a dosage strength of 1-800 mg. The present invention also relates to the use of the pharmaceutical formulation in the preparation of drugs for treating cancer-related diseases.

[0006] The pharmaceutical formulation of this invention has the advantages of stable quality, good dissolution, good absorption, low irritation, good safety, high bioavailability, and oral absorption. The pharmaceutical formulation of this invention exhibits good inhibitory efficacy against LNCAP and VCAP subcutaneous tumor models.

[0007] This invention relates to a pharmaceutical formulation comprising a therapeutically effective amount of an active ingredient M and a pharmaceutical excipient. The pharmaceutical formulation may be in unit dosage form.

[0008] This invention relates to a pharmaceutical formulation comprising an active ingredient M and a pharmaceutical excipient, wherein the active ingredient M is selected from compounds of general formula (I) or their stereoisomers, tautomers, or pharmaceutically acceptable salts, BLK (I).

[0009] L is selected from -Ak1-Cy1-Ak2-Cy2-Ak3-Cy3-Ak4-Cy4-Ak5-;

[0010] Ak1, Ak2, Ak3, Ak4, and Ak5 are each independently selected from -(CH2). q -、-(CH2) q -O-, -O-(CH2) q -、-(CH2) q -S-、-S-(CH2) q -、-(CH2) q -NR L -、-NR L -(CH2) q -、-(CH2) q -NR L C(=O)-、-(CH2) q -C(=O)NR L -, -C(=O)-, -C(=O)-(CH2) q -NR L -、-(C≡C) q - or bond, wherein the -CH2- is optionally selected from one or two elements selected from deuterium, halogen, =O, OH, CN, C 1-4 Alkyl or C 3- Substituents of 6-cycloalkyl groups;

[0011] q is selected from 0, 1, 2, or 3;

[0012] R L Each element is independently selected from H, deuterium, and C. 1-4 Alkyl or deuterated C 1-4 alkyl;

[0013] Cy1, Cy2, Cy3, or Cy4 are each independently selected from the key or arbitrarily selected by 1 to 4 Rs. L2 The substituted group is one of the following: 4-7 membered heteromonocyclic group, 4-12 membered heterocyclic group, 5-13 membered heterospirocyclic group, 7-12 membered heterobridged cyclic group, C 3-7 Monocycloalkyl, C 4-7 Monocyclic alkenyl, C 4- 12 cycloalkyl, C 5-13 Spirocycloalkyl, C 5-12 Bridged cycloalkyl, 5-10 membered heteroaryl or C 6-10 Aryl;

[0014] B is selected from

[0015] X1 is selected from N or CR x1 X2 is selected from N or CR x2 X3 is selected from N or CR. x3 X4 is selected from N or CR x4 X5 is selected from N or CR x5 ;

[0016] At most two of X1, X2, X3, X4, and X5 are selected from N;

[0017] Z1 is selected from N or CR z1 Z2 is selected from N or CR. z2 Z3 is selected from N or CR. z3 Z4 is selected from N or CR z4 ;

[0018] At most 3 of Z1, Z2, Z3, and Z4 are selected from N;

[0019] Y1 and Y2 are each independently selected from -CR y1 R y2 -、-(CR y1 R y2 )2-、-(CR y1 R y2 )3-;

[0020] R 1 Selected from H, deuterium, and C 1-4 Alkyl or C 3-6Cycloalkyl groups, wherein the alkyl or cycloalkyl group is optionally composed of 1 to 4 elements selected from deuterium, halogen, OH, NH2, CN, C. 1-4 Alkyl, C 1-4 Substituents of alkoxy groups;

[0021] R x1 R x2 R x3 R x4 R x5 R z1 R z2 R z3 R z4 Each is independently selected from H, deuterium, halogen, OH, NH2, CN, NO2, COOH, CONH2, and NHC. 1-4 Alkyl, N(C) 1-4 Alkyl)2, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, -OC 1-4 Alkyl, -SC 1-4 Alkyl, -C 0-4 Alkylene-C 3-6 Carbocyclic group, -C 0-4 alkylene-4 to 6-membered heterocyclic groups, 5 to 6-membered heteroaryl groups, -OC 3-6 Carbocyclic, -O-4 to 6-membered heterocyclic groups, wherein the alkylene, alkyl, alkenyl, alkynyl, carbocyclic, heterocyclic, or heteroaryl groups are optionally surrounded by 1 to 4 R groups. s Replaced;

[0022] Or R x1 R x2 R x3 R x4 Each is independently selected from -S(=O)2NH2 and -S(=O)2C 1-4 alkyl;

[0023] Or, R x1 With R x2 R x2 With R x3 R x3 With R x4 R x4 With R x5 R 1 With R z3 R 1 With R z1 Direct connection forms C 4-6 A carbocyclic group or a 4- to 7-membered heterocyclic group, wherein the carbocyclic group or heterocyclic group is optionally surrounded by 1 to 4 R groups. s Replaced;

[0024] R2 R 3 R y1 R y2 Each is independently selected from H, deuterium, halogen, OH, NH2, CN, NO2, COOH, CONH2, C 1- 4-alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, -OC 1-4 Alkyl, -SC 1-4 Alkyl groups, wherein the alkyl, alkenyl, or alkynyl groups are optionally composed of 1 to 4 groups selected from deuterium, halogen, OH, NH2, CN, C. 1-4 Alkyl, C 1-4 Substituents of alkoxy groups;

[0025] K is selected from

[0026] This indicates whether the ring in question is aromatic or non-aromatic.

[0027] F6, F7, and F8 are each independently selected from N, C, CH, or CR. k1 Furthermore, F6, F7, and F8 contain at most two N's;

[0028] G is selected from CH, CD, or N;

[0029] E1 is selected from N, CH, or CD;

[0030] E2 is selected from C, N, CH, or CD;

[0031] Q is selected independently from the following: -O-, -S-, -CH2-, -CD2-, -NR. q -、-C(=O)-、-NR q C(=O)-、-C(=O)NR q -;

[0032] Q and G cannot directly form nitrogen-nitrogen bonds or nitrogen-oxygen bonds;

[0033] R q Each is independently selected from H, deuterium, and C. 1-4 Alkyl or deuterated C 1-4 alkyl;

[0034] R k1 Each is independently selected from deuterium, halogen, OH, NH2, CN, COOH, CONH2, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Alkoxy, C 3-6Cycloalkyl, 4- to 6-membered heterocyclic alkyl, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, or heterocyclic alkyl is optionally surrounded by 1 to 4 R... s Replaced;

[0035] p1 is selected from 0, 1, or 2;

[0036] R L2 R s Each is independently selected from deuterium, halogens, OH, CN, ⁻, CF₃, SF₅, NO₂, NH₂, and NHC. 1-4 Alkyl, N(C) 1- 4-alkyl)2, COOH, CONH2, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Alkoxy, -SC 1-4 Alkyl, -C 0-4 Alkylene-C 3-6 cycloalkyl, -C 0-4 Alkylene-4 to 6-membered heterocyclic group, wherein the alkylene, alkylene, alkoxy, alkenyl, alkynyl, or cycloalkyl group is optionally selected from one to four of deuterium, F, Cl, Br, I, OH, CN, C. 1-4 Alkyl, C 1-4 Substituents of alkoxy groups;

[0037] Or R s Each independently selected from -OC 3-6 cycloalkyl;

[0038] In some implementation schemes, B is selected from

[0039] In some implementation schemes, B is selected from

[0040] In some implementation schemes, B is selected from

[0041] In some implementation schemes, R x2 R x3 R x4 Each is independently selected from H, deuterium, F, Cl, Br, I, OH, NH2, CN, NO2, COOH, CONH2, N(CH3)2, NHCH3, CF3, CHF2, CH2F, OCF3, OCH2F, OCD3, CH2OH, methyl, ethyl, vinyl, ethynyl, methoxy, ethoxy, methylthio, cyclopropyl, -CH2-cyclopropyl, -O-cyclopropyl;

[0042] In some implementation schemes, Rx2 R x4 Each is independently selected from -S(=O)2CH3, -O-CH2-propynyl, -O-CH2-cyclopropyl, -O-CH2CH2-OCH3, -O-CH2CH2-O-cyclopropyl,

[0043] In some implementation schemes, R x2 Each is independently selected from H, deuterium, and methyl;

[0044] In some implementation schemes, R x3 Each is independently selected from F, Cl, Br, CN, vinyl, and ethynyl groups;

[0045] In some implementation schemes, R x4 Each is independently selected from F, Cl, Br, CF3, CHF2, CH2F, methoxy, ethoxy, OCF3, OCH2F, OCD3, -O-CH2-cyclopropyl, -O-CH2CH2-OCH3, -O-CH2CH2-O-cyclopropyl, -O-CH2-propynyl;

[0046] In some implementations, L is selected from -Cy1-, -Cy1-Ak2-, -Cy1-CH2-, -Ak1-Cy1-, -Cy1-Cy2-, -Cy1-CH2-Cy2-, -Cy1-Cy2-Cy3-, -Cy1-CH2-Cy2-Cy3-, -Cy1-Cy2-CH2-Cy3-;

[0047] In some implementations, L is selected from -Cy1-Ak2-Cy2- and -Cy1-O-Cy2-.

[0048] In some implementations, L is selected from -Cy1-, -Cy1-CH2-, and -Cy1-O-Cy2-.

[0049] In some embodiments, Cy1, Cy2, and Cy3 are each independently selected from one of the following optionally substituted groups: When substituted, it is replaced by 1 to 4 substituents selected from deuterium, F, CF3, OH, =O, COOH, CN, NH2, hydroxymethyl, methyl, methoxy, and cyclopropyl.

[0050] In some embodiments, Cy1, Cy2, and Cy3 are each independently selected from one of the following optionally substituted groups: When substituted, it is replaced by 1 to 4 substituents selected from deuterium, F, CF3, OH, =O, COOH, CN, NH2, hydroxymethyl, methyl, methoxy, and cyclopropyl.

[0051] In some implementations, L is selected from one of the structural segments shown in Table L-1:

[0052] Table L-1

[0053] In some implementation schemes, K is selected from

[0054] In some implementation schemes, K is selected from

[0055] In some implementation schemes, K is selected from

[0056] In some implementation schemes, K is selected from

[0057] In some implementations, Q is independently selected from bonds, NH, N(CH3), O, S, NHC(=O), C(=O)NH, N(CH3)C(=O), and C(=O)N(CH3);

[0058] In some implementations, Q is independently selected from bonds, NH, and C(=O)NH;

[0059] In some implementations, Q is independently selected from C(=O)NH;

[0060] In some implementation schemes, Selected from

[0061] In some implementation schemes, R k1 Each is independently selected from deuterium, F, Cl, Br, I, OH, NH2, CN, COOH, CONH2, CF3, CHF2, CH2F, OCF3, OCH2F, CH2OH, methyl, ethyl, isopropyl, methoxy, ethoxy, isopropoxy, and cyclopropyl.

[0062] In some implementations, p1 is selected from 0, 1, or 2;

[0063] In some embodiments, the structure of the compound of general formula (I) is selected from one of the structures shown in Table S-1;

[0064] Table S-1

[0065] In some embodiments, the compound of general formula (I) is selected from the following structures:

[0066] The total content of all components in any of the pharmaceutical preparations described in this invention is 100%.

[0067] The pharmaceutical formulation described in any one of the present invention is a solid dosage form. The present invention relates to a pharmaceutical formulation comprising the aforementioned active ingredient M (calculated as free base, hereinafter the same) and a pharmaceutical excipient, wherein the pharmaceutical formulation comprises 1-800 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 5-750 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 5-500 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 5-400 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 5-300 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 5-200 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 10-700 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 10-400 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 10-300 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 10-200 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 10-100 mg of active ingredient M; in some embodiments… The pharmaceutical formulation comprises 50-500 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 50-400 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 50-300 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 50-200 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 50-100 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 100-500 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 100-400 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 100-300 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 100-200 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 20-100 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 20-75 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 50-100 mg of active ingredient M.

[0068] In some embodiments, the amount of active ingredient M in a unit formulation of the pharmaceutical preparation includes, but is not limited to, 1 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 110 mg, 120 mg, 125 mg, 130 mg, etc. mg, 140mg, 150mg, 160mg, 170mg, 175mg, 180mg, 190mg, 200mg, 210mg, 220mg, 225mg, 230mg, 240m g, 250mg, 275mg, 300mg, 350mg, 400mg, 450mg, 500mg, 550mg, 600mg, 650mg, 700mg, 750mg, 800mg.

[0069] In some embodiments, the formulation strengths of the pharmaceutical preparations described in this invention include, but are not limited to, 1-500mg, 2-500mg, 3-500mg, 5-500mg, 6-500mg, 10-500mg, 20-500mg, 25-500mg, 30-500mg, 40-500mg, 50-500mg, 60-500mg, 70-500mg, 75-500mg, 80-500mg, 90-500mg, 100-500mg, 200-500mg, 300-500mg, 400-500mg. 1-400mg, 2-400mg, 3-400mg, 5-400mg, 6-400mg, 10-400mg, 20-400mg, 25-400mg, 30-400mg, 40-400mg, 50-400mg, 60-400mg, 7 0-400mg, 75-400mg, 80-400mg, 90-400mg, 100-400mg, 200-400mg, 300-400mg, 1-300mg, 2-300mg, 5-300mg, 10-300mg, 20-300m g, 25-300mg, 30-300mg, 40-300mg, 50-300mg, 60-300mg, 70-300mg, 75-300mg, 80-300mg, 90-300mg, 100-300mg, 125-300mg, 1 50-300mg, 200-300mg, 250-300mg, 1-200mg, 2-200mg, 5-200mg, 10-200mg, 20-200mg, 25-200mg, 30-200mg, 40-200mg, 50-200m g, 60-200mg, 70-200mg, 75-200mg, 80-200mg, 90-200mg, 100-200mg, 125-200mg, 150-200mg, 1-100mg, 2-100mg, 5-100mg, 10- 100mg, 15-100mg, 20-100mg, 25-100mg, 30-100mg, 40-100mg, 50-100mg, 60-100mg, 70-100mg, 75-100mg, 80-100mg, 90-100mg;In some embodiments, the formulation strengths of the drug include, but are not limited to, 1 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 110 mg, 120 mg, 125 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 175 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 225 mg, 230 mg, 240 mg, 250 mg, 275 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, and 800 mg.

[0070] The pharmaceutical formulation according to any one of the present invention comprises the aforementioned active ingredient M and a pharmaceutical excipient, wherein the content of active ingredient M is 0.1%-90%; in some embodiments, it is 0.2%-90%; in some embodiments, it is 0.3%-90%; in some embodiments, it is 0.5%-90%; in some embodiments, it is 1%-90%; in some embodiments, it is 1%-80%; in some embodiments, it is 1%-70%; in some embodiments, it is 1%-60%; in some embodiments, it is 1%-50%; in some embodiments, it is 1%-40%; in some embodiments, it is 1%-30%; in some embodiments, it is 1%-20%; in some embodiments, it is 1%-10%; in some embodiments, it is 5%-90%; in some embodiments, it is 5%-80%; in some embodiments, it is... The percentage varies depending on the implementation plan: 5%-70%; in some implementation plans: 5%-60%; in some implementation plans: 5%-50%; in some implementation plans: 0.5%-47.5%; in some implementation plans: 5%-40%; in some implementation plans: 5%-35%; in some implementation plans: 5%-30%; in some implementation plans: 5%-20%; in some implementation plans: 5%-10%; in some implementation plans: 10%-90%; in some implementation plans: 10%-80%; in some implementation plans: 10%-70%; in some implementation plans: 10%-60%; in some implementation plans: 10%-50%; in some implementation plans: 10%-40%; in some implementation plans: 10%-30%; in some implementation plans: 10%-20%.

[0071] In some embodiments, the weight ratio of the active ingredient M to the pharmaceutical excipient is 1:0.01 to 1:100, 1:0.5 to 1:10, 1:1.5 to 1:9, or 1:4 to 1:9;

[0072] In some embodiments, the pharmaceutical excipient comprises a filler, optionally further comprising a disintegrant; in some embodiments, the pharmaceutical excipient comprises one or more of a filler and a disintegrant; in some embodiments, the pharmaceutical excipient comprises one or more of a filler, a disintegrant, a binder, a flow aid, a lubricant, and a pH adjuster; in some embodiments, the pharmaceutical excipient comprises one or more of a filler, a binder, a disintegrant, a flow aid, a lubricant, a flavoring agent, an antioxidant, a preservative, a light-blocking agent, and a film-coating premix; in some embodiments, the pharmaceutical excipient... The excipients include fillers, disintegrants, binders, flow aids, and lubricants; in some embodiments, the pharmaceutical excipients further include pH adjusters; in some embodiments, the pharmaceutical excipients include fillers, disintegrants, binders, and lubricants; in some embodiments, the pharmaceutical excipients include fillers and disintegrants; in some embodiments, the pharmaceutical excipients further include one or more of binders, flow aids, and lubricants; in some embodiments, the pharmaceutical excipients further include one or more of binders, flow aids, lubricants, and pH adjusters.

[0073] In some embodiments, in any of the foregoing embodiments, the filler is selected from one or more of microcrystalline cellulose, mannitol, lactose, sucrose, sorbitol, dextran, anhydrous dicalcium phosphate, pregelatinized starch, dicalcium phosphate, and starch; in some embodiments, the filler is selected from microcrystalline cellulose or lactose; in some embodiments, the filler is selected from a mixture of microcrystalline cellulose and lactose.

[0074] In some embodiments, the filler content in the pharmaceutical formulation is 40%-90% in any of the foregoing embodiments; in other embodiments, the content is 40%-85%.

[0075] In some embodiments, in any of the foregoing embodiments, the disintegrant is selected from one or more of sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, crospovidone, crospovidone sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, and dry starch; in some embodiments, the disintegrant is selected from one or more of crospovidone, crospovidone sodium carboxymethyl cellulose, sodium carboxymethyl starch, and dry starch; in some embodiments, the disintegrant is selected from crospovidone.

[0076] In some embodiments, the disintegrant in any of the foregoing embodiments is 0%-10% in the pharmaceutical preparation; in some embodiments, the content is 1%-10%; and in some embodiments, the content is 2%-8%.

[0077] In some embodiments, in any of the foregoing embodiments, the adhesive is selected from one or more of povidone, hydroxypropyl cellulose, hydroxypropyl methylcellulose, and methylcellulose; in some embodiments, the adhesive is selected from povidone; in some embodiments, the adhesive is selected from hydroxypropyl methylcellulose.

[0078] In some embodiments, the adhesive content in the pharmaceutical preparation is 0%-3% in any of the foregoing embodiments; in other embodiments, the content is 1%-3%.

[0079] In some embodiments, in any of the foregoing embodiments, the flow aid is selected from one or more of talc, silica, micronized silica gel, polyethylene glycol, and magnesium dodecyl sulfate; in some embodiments, the flow aid is selected from silica.

[0080] In some embodiments, in any of the foregoing embodiments, the content of the gliding agent in the pharmaceutical preparation is 0%-3%.

[0081] In some embodiments, the lubricant in any of the foregoing embodiments is selected from magnesium stearate, calcium stearate, stearic acid, and sodium stearate fumarate; in some embodiments, the lubricant is sodium stearate fumarate; in some embodiments, the lubricant content is 0%-3%. The pharmaceutical formulation according to any one of the present invention comprises the active ingredient M in any of the foregoing embodiments and a pharmaceutical excipient, wherein the pharmaceutical excipient comprises magnesium stearate, povidone, crospovidone, hydroxypropyl methylcellulose, microcrystalline cellulose, and lactose.

[0082] In some embodiments, in any of the foregoing embodiments, the pH adjuster is selected from fumaric acid; in any of the foregoing embodiments, the content of the pH adjuster in the pharmaceutical preparation is 0%-5%.

[0083] This invention provides a pharmaceutical formulation comprising the active ingredient M from any of the foregoing embodiments and a pharmaceutical excipient, wherein:

[0084] (i) Active ingredient M, preferably compound 11, and optionally...

[0085] (ii) Pharmaceutical excipients include fillers, optionally further comprising one or more of binders, wetting agents, disintegrants, flow aids, lubricants, and pH adjusters.

[0086] Preferably, the filler includes, but is not limited to, one or more of microcrystalline cellulose, mannitol, lactose, sucrose, sorbitol, dextran, pregelatinized starch, dicalcium phosphate, and starch;

[0087] Preferably, the adhesive includes, but is not limited to, one or more of polyvinylpyrrolidone, hydroxypropyl cellulose, hydroxypropyl methylcellulose, and methylcellulose;

[0088] Preferably, the wetting agent includes, but is not limited to, one or more of water and ethanol;

[0089] Preferably, the disintegrant includes, but is not limited to, one or more of sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, crospovidone, crospovidone carboxymethyl cellulose sodium, and calcium carboxymethyl cellulose;

[0090] Preferably, the gliding agent includes, but is not limited to, one or more of talc, silica, polyethylene glycol, and magnesium dodecyl sulfate;

[0091] Preferably, the lubricant includes, but is not limited to, magnesium stearate, calcium stearate, stearic acid, sodium stearate fumarate, and hydrogenated castor oil;

[0092] Preferably, the pH adjuster includes, but is not limited to, fumaric acid, sodium carbonate, sodium hydroxide, sodium bicarbonate, and calcium bicarbonate;

[0093] Optionally, the pharmaceutical formulation further includes one or more of the following: flavoring agents, antioxidants, preservatives, light-blocking agents, and film-coating premixes.

[0094] The present invention provides a pharmaceutical formulation comprising the active ingredient M, a filler, a disintegrant, and a binder as described in any of the foregoing embodiments, optionally further comprising a flow aid and a lubricant, wherein the content of the active ingredient M is 0.5%-47.5%, the content of the filler is 40%-90%, the content of the disintegrant is 1%-10%, the content of the binder is 1%-10%, the content of the lubricant is 0.5%-3%, and the content of the flow aid is 0%-3%.

[0095] This invention provides a pharmaceutical preparation comprising the aforementioned active ingredient M, microcrystalline cellulose, crospovidone, hydroxypropyl methylcellulose, and magnesium stearate;

[0096] This invention provides a pharmaceutical formulation comprising the aforementioned active ingredient M, lactose, crospovidone, povidone, and magnesium stearate;

[0097] The present invention provides a pharmaceutical preparation comprising 5%-35% of active ingredient M, 60%-90% of lactose or microcrystalline cellulose, 1%-10% of crospovidone, 1%-10% of povidone or hydroxypropyl methylcellulose, and 1%-3% of magnesium stearate.

[0098] Optionally, the pharmaceutical preparation described in any of the above embodiments can be prepared as a formulation selected from oral tablets, capsules, granules, and powders.

[0099] The pharmaceutical preparation of the present invention is prepared by one or more of the following processes: direct mixing, wet granulation, dry granulation, fluidized bed granulation, spray drying, and hot melt extrusion.

[0100] Optionally, the pharmaceutical formulation described above is characterized in that the binder can be added in a solution state or in a powder state; the disintegrant can be added internally, externally, or both.

[0101] The present invention also provides an application in the preparation of a medicament for treating cancer (e.g., prostate cancer), wherein the administration method includes: oral administration, once a day, twice a day, three times a day, once a week, once every two weeks, or once a month.

[0102] This invention provides the use of a pharmaceutical preparation in the manufacture of a medicament for cancer (e.g., prostate cancer), wherein the use is achieved by administering a subject a pharmaceutical preparation comprising the aforementioned pharmaceutical preparation, wherein the dosage of the active ingredient M is selected from 1-2000 mg / dose, 1-1000 mg / dose, 1-800 mg / dose, 1-600 mg / dose, 1-400 mg / dose, 1-350 mg / dose, 1-300 mg / dose, 1-5 mg / dose, 5- 10mg / dose, 10-20mg / dose, 20-25mg / dose, 25-50mg / dose, 50-75mg / dose, 75-100mg / dose, 100-125mg / dose, 125-150mg / dose, 150-175mg / dose, 175-200mg / dose, 200-225mg / dose, 225-250mg / dose, 250-275mg / dose, 275-300mg / dose.

[0103] A method for treating a disease in a mammal, the method comprising administering an active ingredient M to a subject at a daily dose of 1-5000 mg / day, said daily dose being a single dose or divided doses, and in some embodiments, the daily dose including but not limited to 10-5000 mg / day, 25-5000 mg / day, 50-5000 mg / day, 100-4500 mg / day, 100-4000 mg / day, 100-3000 mg / day, 50-3000 mg / day, 50-2500 mg / day, 50-2000 mg / day, 50-1500 mg / day, 20-1500 mg / day, 50-1000 mg / day, 100-1000 mg / day, 10 0-800mg / day, 200-800mg / day, 25-400mg / day, 50-400mg / day, 100-400mg / day, 200-400mg / day. In some embodiments, the daily dose includes, but is not limited to, 10mg / day, 20mg / day, 25mg / day, 50mg / day, 100mg / day, 120mg / day, 125mg / day, 150mg / day, 200mg / day, 240mg / day, 400mg / day, 600mg / day, 800mg / day, 900mg / day, 1000mg / day, 1500mg / day, 2000mg / day, 3500mg / day, 4500mg / day, and 4000mg / day.

[0104] Unless otherwise stated, the terms used in this specification and claims have the following meanings.

[0105] The compounds of this invention include their racemic, stereoisomer, tautomer, isotopic compounds, solvates, prodrugs, metabolites, pharmaceutically acceptable salts, or cocrystals.

[0106] "Product specification" refers to the weight of the active pharmaceutical ingredient (M) contained in each vial, tablet, or other unit of preparation.

[0107] "Excipient" or "pharmaceutical excipient" refers to an agent that is not itself a therapeutic agent but is used as a diluent, filler, binder, and / or medium for addition to a pharmaceutical composition to improve its disposal or storage properties or to allow or facilitate the formation of a unit dosage form of the compound or pharmaceutical composition for administration. As is known to those skilled in the art, pharmaceutical excipients can provide a variety of functions and can be described as including, but not limited to, wetting agents, buffers, suspending agents, lubricants, emulsifiers, disintegrants, solubilizers, binders, flow aids, wetting agents, pH adjusters, absorbents, preservatives, surfactants, colorants, flavoring agents, sweeteners, buffers, effervescent agents, plasticizers, dispersants, parenteral solutions, and coatings. The term "excipient" encompasses pharmaceutical carriers.

[0108] "Fillers" refer to pharmaceutical excipients used to improve the properties of materials or increase the weight or volume of pharmaceutical preparations to facilitate accurate dispensing during use and to aid in the shaping of pharmaceutical preparations. These include, but are not limited to, fillers for tablets, capsules, and granules, and inert diluents (such as water or other solvents) for liquid preparations.

[0109] "Adhesive" refers to an excipient that is used to bind the components together when the active pharmaceutical ingredient itself is not sticky or has low stickiness.

[0110] "Disintegrants" are excipients that swell and dissolve when wet, causing the formulation to decompose in the body and release the active ingredient for absorption. These include, but are not limited to, cross-linked polymers (such as crospovidone, crospovidone sodium carboxymethyl cellulose, etc.) and starch, modified starch (such as sodium glycolate starch).

[0111] "Flow aids" are substances used to promote powder flow by reducing interparticle adhesion, including but not limited to silica, anhydrous silica gel, starch, and talc.

[0112] "Lubricant" refers to a substance that prevents components from adhering to and / or clumping together in a machine used to prepare a pharmaceutical formulation, including but not limited to metal stearate salts (such as magnesium stearate), stearic acid, stearate esters, and stearoyl fumarate.

[0113] "Wetting agent" refers to a liquid that can induce viscosity in materials to be granulated or tableted, in order to facilitate granulation or tableting. It is not viscous itself and includes, but is not limited to, water, ethanol or ethanol of different concentrations.

[0114] "pH adjuster" refers to a substance used to adjust the pH of a composition.

[0115] "Solubilizer" refers to a surfactant that can increase the solubility of the active ingredients in a drug.

[0116] The carbon, hydrogen, oxygen, sulfur, nitrogen, or F, Cl, Br, I involved in the groups and compounds described in this invention include their isotopes, and the carbon, hydrogen, oxygen, sulfur, or nitrogen involved in the groups and compounds described in this invention may optionally be further replaced by one or more of their corresponding isotopes, wherein the isotopes of carbon include 12 C 13 C and 14 C, the isotopes of hydrogen include protium (H), deuterium (D, also called heavy hydrogen), and tritium (T, also called superheavy hydrogen), and the isotopes of oxygen include 16 O、 17 O and 18 O, isotopes of sulfur include 32 S, 33 S, 34 S and 36S, nitrogen isotopes include 14 N and 15 N, isotopes of fluorine include 17 F and 19 F, isotopes of chlorine include 35 Cl and 37 Cl, isotopes of bromine include 79 Br and 81 Br. In some embodiments, at the position of the deuterium substituent, the deuterium isotope abundance is greater than the natural deuterium isotope abundance (0.015%), preferably greater than 50%, more preferably greater than 60%, greater than 70%, greater than 80%, greater than 90%, greater than 95%, greater than 96%, greater than 97%, greater than 98%, greater than 99%, greater than 99.5%, or 100%. In some cases, such as when “hydrogen” and “deuterium” appear as parallel terms, or when “hydrogen” is replaced by “deuterium”, the term “hydrogen” represents the isotope abundance of hydrogen. 1 "H", while "deuterium" represents the isotope of hydrogen. 2 H”; or it should be understood that at this position in the compound, hydrogen, existing in its natural abundance at various isotopes at that position, is replaced by deuterium, existing in its abundance at a level greater than that of the natural deuterium isotopes (e.g., deuterium abundance greater than 50%, greater than 60%, greater than 70%, greater than 80%, greater than 90%, greater than 95%, greater than 96%, greater than 97%, greater than 98%, greater than 99%, greater than 99.5%, or 100%).

[0117] "alkyl" refers to a substituted or unsubstituted straight-chain or branched saturated aliphatic hydrocarbon group, including but not limited to alkyl groups with 1 to 20 carbon atoms, alkyl groups with 1 to 8 carbon atoms, alkyl groups with 1 to 6 carbon atoms, and alkyl groups with 1 to 4 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, tert-pentyl, neopentyl, n-hexyl, and their various branched isomers; the alkyl group can be monovalent, divalent, trivalent, or tetravalent.

[0118] "Alkylene" refers to substituted or unsubstituted straight-chain and branched divalent saturated hydrocarbon groups, including -(CH2)d v - (dv is an integer from 1 to 10), alkylene examples include but are not limited to methylene, ethylene, propylene, and butylene.

[0119] "Cycloalkyl" refers to a substituted or unsubstituted saturated carbocyclic hydrocarbon group, typically having 3 to 12 carbon atoms. Cycloalkyl groups can be monocyclic, fused, bridged, or spirocyclic. Non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclobutyl-cyclobutyl, cyclobutylspirobutyl, bicyclo[1.1.1]pentane, bicyclo[2.2.2]octane, adamantane, etc. Cycloalkyl groups can be monovalent, divalent, trivalent, or tetravalent.

[0120] "Heterocyclic alkyl" refers to a substituted or unsubstituted saturated cyclic hydrocarbon group containing heteroatoms, including but not limited to 3 to 12 atoms or 3 to 8 atoms, containing 1 to 3 heteroatoms selected from N, O, S, or Se. The C, N, S, and Se on the ring of the heterocyclic alkyl group can be oxidized to various oxidation states. Heterocyclic alkyl groups can be monocyclic, fused, bridged, or spirocyclic. Heterocyclic alkyl groups can be attached to heteroatoms or carbon atoms. Non-limiting examples include epoxyethyl, aziridinepropyl, oxacyclobutyl, aziridinebutyl, tetrahydrofuranyl, tetrahydro-2H-pyranyl, dioxopentyl, dioxohexyl, pyrrolylalkyl, piperidinyl, imidazoalkyl, oxazolidinyl, oxazinylalkyl, morpholinyl, hexahydropyrimidinyl, piperazineyl, etc. Heterocyclic alkyl groups can be monovalent, divalent, trivalent, or tetravalent.

[0121] "Alkenyl" refers to a substituted or unsubstituted straight-chain and branched unsaturated hydrocarbon group having at least one, typically one, two, or three, carbon-carbon double bonds. The main chain has, but is not limited to, 2 to 10, 2 to 6, or 2 to 4 carbon atoms. Examples of alkenyl groups include, but are not limited to, vinyl, allyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-methyl-1-butenyl, 2-methyl-1-butenyl, 2... -Methyl-3-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 1-methyl-1-pentenyl, 2-methyl-1-pentenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 1-octenyl, 3-octenyl, 1-nonenyl, 3-nonenyl, 1-decenyl, 4-decenyl, 1,3-butadiene, 1,3-pentadiene, 1,4-pentadiene, and 1,4-hexadiene, etc.; the alkenyl group can be monovalent, divalent, trivalent, or tetravalent.

[0122] "Alynyl" refers to a substituted or unsubstituted straight-chain and branched unsaturated hydrocarbon group having at least one, typically one, two, or three, carbon-carbon triple bonds. The main chain comprises 2 to 10 carbon atoms, including but not limited to having 2 to 6 carbon atoms on the main chain, or 2 to 4 carbon atoms on the main chain. Examples of alkynyl groups include, but are not limited to, ethynyl, propynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, and 4-pentynyl. The alkynyl group can be monovalent, divalent, trivalent, or tetravalent.

[0123] "Alkoxy" refers to a substituted or unsubstituted -O-alkyl group. Non-limiting examples include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, and n-hexoxy.

[0124] "Carbocyclic group" or "carbocyclic ring" refers to a substituted or unsubstituted aromatic or non-aromatic ring. The aromatic or non-aromatic ring can be a 3- to 8-membered monocyclic ring, a 4- to 12-membered bicyclic ring, a 10- to 15-membered tricyclic ring, or a 12- to 18-membered quaternary system. The carbocyclic group can be attached to an aromatic or non-aromatic ring, and the ring can be optionally a monocyclic, fused, bridged, or spirocyclic ring. Non-limiting examples include cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, 1-cyclopentyl-1-enyl, 1-cyclopentyl-2-enyl, 1-cyclopentyl-3-enyl, cyclohexyl, 1-cyclohexyl-2-enyl, 1-cyclohexyl-3-enyl, cyclohexenyl, benzene ring, naphthalene ring, etc. "Carbocyclic group" or "carbon ring" can be monovalent, divalent, trivalent or tetravalent.

[0125] "Heterocyclic group" or "heterocyclic" refers to a substituted or unsubstituted aromatic or non-aromatic ring. The aromatic or non-aromatic ring can be a 3- to 8-membered monocyclic ring, a 4- to 12-membered bicyclic ring, a 10- to 15-membered tricyclic ring, or a 12- to 18-membered quaternary system, and contains one or more (including but not limited to 2, 3, 4 or 5) heteroatoms selected from N, O, S or Se. The C, N, S or Se selectively substituted in the ring of the heterocyclic group can be oxidized to various oxidation states. The heterocyclic group can be attached to a heteroatom or a carbon atom, and can be attached to an aromatic ring or a non-aromatic ring. The heterocyclic group is optionally a monocyclic, bridged, fused, or spirocyclic ring. Non-limiting examples include epoxyethyl, aziridinepropyl, oxacyclobutyl, aziridinebutyl, 1,3-dioxopentyl, 1,4-dioxopentyl, 1,3-dioxahexane, aziridineheptyl, pyridinyl, furanyl, thiophene, pyranyl, N-alkylpyrroleyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, piperidinyl, morpholinyl, thiomorpholinyl, 1,3-dithioyl, dihydrofuranyl, dihydropyranyl, dithiapentylcycloyl. Tetrahydrofuranyl, tetrahydropyrrolyl, tetrahydroimidazolyl, tetrahydrothiazolyl, tetrahydropyranyl, benzimidazolyl, benzopyridyl, pyrrolopyridyl, benzodihydrofuranyl, pyrrolyl, pyrazolyl, thiazolyl, oxazolyl, pyrazinyl, indazolyl, benzothiophene, benzofuranyl, benzopyrrolyl, benzimidazolyl, benzothiazolyl, benzooxazolyl, benzopyridyl, benzopyrimidinyl, benzopyrazinyl, piperazinyl, azabicyclo[3.2.1]octyl, azabicyclo[5.2.0]nonyl, oxatricyclo[5.3.1.1]dodecyl, azaadamantyl, oxaspiro[3.3]heptyl, "Heterocyclic group" or "heterocyclic" can be monovalent, divalent, trivalent or tetravalent.

[0126] A "spirocyclic" or "spirocyclic group" refers to a polycyclic group in which substituted or unsubstituted rings share a single atom (called a spiro atom). The number of ring atoms in a spirocyclic system includes, but is not limited to, 5 to 20, 6 to 14, 6 to 12, or 6 to 10. One or more rings may contain zero or more (including but not limited to 1, 2, 3, or 4) double bonds, and optionally, may contain 0 to 5 double bonds selected from N, O, or S (=O). dn Heteroatoms (dn is 0, 1, or 2). Non-limiting embodiments include: "Spirocyclic" or "spirocyclic group" can be monovalent, divalent, trivalent or tetravalent.

[0127] "Circular fused" or "circular fused group" refers to a polycyclic group in which each ring in a system shares a pair of adjacent atoms with other rings in the system. One or more rings may contain zero or more (including but not limited to 1, 2, 3 or 4) double bonds and may be substituted or unsubstituted. Each ring in a circular fused system may contain 0 to 5 heteroatoms or groups containing heteroatoms (including but not limited to those selected from N, S (=O)). dn (Or O, dn is 0, 1, or 2). The number of ring atoms in a cyclic system includes, but is not limited to, 5 to 20, 5 to 14, 5 to 12, and 5 to 10. Non-limiting examples include: "Cyclone" or "cyclone base" can be monovalent, divalent, trivalent, or tetravalent.

[0128] A "bridged ring" or "bridged ring group" refers to a substituted or unsubstituted polycyclic group containing any two atoms that are not directly connected. It may contain zero or more double bonds. Any ring in a bridged ring system may contain zero to five groups selected from heteroatoms or containing heteroatoms (including but not limited to N, S (=O)). dn Or O, where dn is 0, 1, or 2). The number of ring atoms includes, but is not limited to, 5 to 20, 5 to 14, 5 to 12, or 5 to 10. Non-limiting examples include: Cubicane, adamantane. "Bridged ring" or "bridged ring group" can be monovalent, divalent, trivalent, or tetravalent.

[0129] "Carbon spirocyclic", "spirocyclic carbon cyclic", "spirocarbon cyclic", or "carbon spirocyclic" refers to a spirocyclic system composed only of carbon atoms.

[0130] "Carbon fused ring", "fused cyclic carbon cyclic group", "fused carbon cyclic group" or "carbon fused cyclic group" refers to a ring system composed only of carbon atoms.

[0131] "Carbon bridged ring", "bridged ring carbon cyclo group", "bridged carbon cyclo group" or "carbon bridged ring group" refers to a ring system composed only of carbon atoms.

[0132] "Hybrid monocyclic", "monocyclic heterocyclic group" or "hybrid monocyclic group" refers to the "heterocyclic group" or "heterocyclic" in a monocyclic system.

[0133] "Hydrocyclic ring", "hydrocyclic cyclic group", "fused cyclic heterocyclic group" or "fused heterocyclic group" refers to a "fused ring" containing heteroatoms.

[0134] "Heterospirocyclic", "heterospirocyclic group", "spirocyclic heterocyclic group" or "spiroheterocyclic group" refers to a "spirocycle" containing heteroatoms.

[0135] "Hybrid-bridged ring", "hybrid-bridged ring group", "bridged ring heterocyclic group" or "bridged heterocyclic group" refers to a "bridged ring" containing heteroatoms.

[0136] "Aryl" or "aromatic ring" refers to a substituted or unsubstituted aromatic hydrocarbon group having a monocyclic or fused ring, wherein the number of ring atoms in the aromatic ring includes, but is not limited to, 6 to 18, 6 to 12, or 6 to 10 carbon atoms. The aryl ring can be fused to a saturated or unsaturated carbon ring, wherein the ring connected to the parent structure is the aryl ring. Non-limiting embodiments include benzene rings, naphthalene rings, etc. The "aryl" or "aryl ring" can be monovalent, divalent, trivalent, or tetravalent. When it is divalent, trivalent, or tetravalent, the linking site is located on the aryl ring.

[0137] "Heteroaryl" or "heteroary ring" refers to a substituted or unsubstituted aromatic hydrocarbon group containing 1 to 5 heteroatoms or a group containing heteroatoms (including but not limited to N, O, S (=O)). dn OrSe(=O) dn (dn is 0, 1, or 2), the number of ring atoms in the heteroaromatic ring includes, but is not limited to, 5 to 15, 5 to 10, or 5 to 6. The atoms C, N, and S on the ring are optionally oxidized (i.e., C (=O), NO, S (=O)). dm Se (=O) dm (dm is 1, 2), non-limiting embodiments of heteroaryl groups include, but are not limited to, pyridyl, furanyl, thiophenyl, selenophenyl, pyridyl, pyranyl, N-alkylpyrrolithyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazoleyl, benzopyrazolyl, benzimidazolyl, benzopyridyl, pyrrolopyridyl, pyridinoneyl, etc. The heteroaryl ring may be fused to a saturated or unsaturated carbon ring or heterocycle, wherein the ring connected to the parent structure is an aryl ring. Non-limiting embodiments include: The heteroaryl groups mentioned in this article are defined in accordance with this definition. Heteroaryl groups can be monovalent, divalent, trivalent, or tetravalent. When divalent, trivalent, or tetravalent, the linkage site is located on an aromatic ring.

[0138] "Substituted" or "substituted" means substituted by one or more (including but not limited to 2, 3, 4, or 5) substituents, including but not limited to H, F, Cl, Br, I, alkyl, cycloalkyl, alkoxy, haloalkyl, thiol, hydroxyl, nitro, mercapto, amino, cyano, isocyano, aryl, heteroaryl, heterocyclic, bridged cyclic, spirocyclic, fused cyclic, hydroxyalkyl, =O, carbonyl, aldehyde, carboxylic acid, formate, and -(CH2). dn -C(=O)-R da -O-(CH2) dn -C(=O)-R da -(CH2) dn -C(=O)-NR db R dc -(CH2) dnS(=O) dn R da -(CH2) dn -Alkenyl-R da OR dd Or -(CH2) dn -alkynyl-R da (where dn is 0, 1, or 2), arylthio, thiocarbonyl, silyl, or -NR db R dc Groups, wherein R db With R dc Independently selected from H, hydroxyl, amino, carbonyl, alkyl, alkoxy, cycloalkyl, heterocyclic, aryl, heteroaryl, sulfonyl, trifluoromethanesulfonyl, R db With R dc It can form five- or six-membered cycloalkyl or heterocyclic groups, R da With R dd Each group is independently selected from aryl, heteroaryl, alkyl, alkoxy, cycloalkyl, heterocyclic, carbonyl, ester, bridged cyclic, spirocyclic, or fused cyclic groups.

[0139] "Deuterated alkyl" refers to an alkyl group that has been deuterated by one or more (including but not limited to 2, 3, 4, 5 or 6) deuterium atoms. 2 H) substitution, wherein the alkyl group is as defined above, for example, "deuterated C" 1-6 Alkyl", C 1-6 "Deuterated alkyl", etc., specific examples include but are not limited to: CD3, CH2CD3, etc.

[0140] "1 to X substituents selected from..." means substituted by 1, 2, 3...X substituents selected from..., where X is any integer between 1 and 10. For example, "1 to 4 R..." k "Replace" refers to being replaced by 1, 2, 3, or 4 Rs. k Substitution. For example, "substituted by 1 to 5 substituents selected from..." means that the ring is substituted by 1, 2, 3, 4 or 5 substituents selected from... For example, "the heterobridged ring is optionally substituted by 1 to 4 substituents selected from H or F" means that the heterobridged ring is optionally substituted by 1, 2, 3 or 4 substituents selected from H or F.

[0141] The XY-membered rings (where X and Y are integers, and 3 ≤ X < Y, X < Y ≤ 20, selected from any integer between 4 and 20) include rings of the X, X+1, X+2, X+3, X+4…Y-membered elements. These rings include heterocyclic rings, carbocyclic rings, aromatic rings, aryl groups, heteroaryl groups, cycloalkyl groups, heteromonocyclic rings, heterofused rings, heterospirocyclic rings, or heterobridged rings. For example, "4-7-membered heteromonocyclic rings" refers to heteromonocyclic rings of 4, 5, 6, or 7 members, and "5-10-membered heterofused rings" refers to heterofused rings of 5, 6, 7, 8, 9, or 10 members.

[0142] C x-y Carbocyclic rings (including aryl, cycloalkyl, monocyclic, spirocyclic, fused, or bridged carbocyclic rings) include C x C x+1 C x+2 C x+3 C x+4 ….C y A ring of elements (x is an integer, and 3 ≤ x < y, where y is any integer between 4 and 20), for example, "C". 3-6 "Cycloalkyl" refers to C3, C4, C5, or C6 cycloalkyl groups.

[0143] When a functional group has one or more connectable sites, any one or more of these sites can be linked to other functional groups via chemical bonds. When the chemical bond connection is non-directional and a hydrogen atom is present at the connectable site, the number of hydrogen atoms at that site decreases accordingly with the number of bonds being formed, resulting in a functional group with a corresponding valence. For example... This indicates that any connectable site on the piperidinyl group can be linked to other groups via a single chemical bond, including at least... These four connection methods, even if an H atom is drawn on -N-, This also includes For example This indicates that the R group on the piperidinyl group can be located on C or N, and at least includes [missing information]. For example, the general formula segment is: When X is selected from CH2 or NH, it means that the R group on the general formula fragment can be located on C or X. When X is selected from CH2, the general formula fragment can be... When X is selected from NH, the general formula fragment can be:

[0144] Unless otherwise stated, key This indicates that the configuration is not specified; that is, if chiral isomers exist in the chemical structure, the bond... It can be or Or simultaneously include or Two configurations. Using wedge-shaped solid line keys. and wedge-shaped dashed key The absolute configuration of the center of a solid is represented by a straight solid line key. and straight dashed key Represents the relative configuration of the center of a solid.

[0145] When the listed linking groups do not specify their linking direction, the linking direction includes the direction of the reading order from left to right and from right to left. For example, when ALB is selected from -MW-, it includes AMWB and AWMB.

[0146] "Optional" or "optionally" means that the event or environment described below may but does not have to occur, and the description includes the possibility or possibility that the event or environment may or may not occur. For example, "optionally substituted F alkyl" means that the alkyl group may but does not have to be substituted with F, and the description includes the case where the alkyl group is substituted with F and the case where the alkyl group is not substituted with F.

[0147] "Pharmaceutically acceptable salt" or "its pharmaceutically acceptable salt" means that the compound of the present invention retains the bioavailability and properties of a free acid or a free base, and that the free acid is obtained by reacting with a non-toxic inorganic or organic base, and the free base is obtained by reacting with a non-toxic inorganic or organic acid.

[0148] "Prodrug" refers to a compound of the present invention that can be metabolized in vivo and converted into a biologically active compound. The prodrug of the present invention is prepared by modifying the amino or carboxyl groups in the compound of the present invention. This modification can be performed through conventional procedures or removed in vivo to obtain the parent compound. When the prodrug of the present invention is administered to a mammalian individual, the prodrug is cleaved to form free amino or carboxyl groups.

[0149] "Co-crystal" refers to a crystal formed by the bonding of an active pharmaceutical ingredient (API) and a co-crystal form (CCF) through hydrogen bonds or other non-covalent bonds. Both API and CCF are solids at room temperature in their pure states, and a fixed stoichiometric ratio exists between the components. Co-crystal is a multi-component crystal, encompassing both binary co-crystals formed between two neutral solids and multi-component co-crystals formed between a neutral solid and a salt or solvate.

[0150] "Stereoisomers" refer to isomers that are produced by different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers, diastereomers, and conformational isomers.

[0151] "Tautomers" refer to functional group isomers that are produced by the rapid movement of an atom in two positions within a molecule, such as keto-enol isomers and amide-imine alcohol isomers.

[0152] "Animals" refers to mammals, such as humans, companion animals, zoo animals, and livestock, with humans, horses, or dogs being preferred.

[0153] IC 50 "It refers to the concentration of a drug or inhibitor required to inhibit a specified biological process (or a component of that process, such as an enzyme, receptor, or cell) by half."

[0154] Unless otherwise specified, the “content” of a substance in the pharmaceutical preparation of this application refers to the percentage of the weight of the substance in the total weight of the pharmaceutical preparation. Attached Figure Description

[0155] Figure 1 shows the efficacy of the drug in the VCAP subcutaneous tumor model. * represents P<0.05, ** represents P<0.01, and *** represents P<0.001. Differences in tumor volume were analyzed at the last measurement, comparing the drug-treated group with the vehicle-one-way ANOVA Brown-Forsythe and Welch ANOVA tests.

[0156] Figure 2 shows the efficacy of the drug in the LNCAP subcutaneous tumor model. * represents P<0.05, ** represents P<0.01, and *** represents P<0.001. Differences in tumor volume were analyzed at the last measurement, comparing the drug-treated group with vehicle-one-way ANOVA Brown-Forsythe and Welch ANOVA tests. Detailed Implementation

[0157] The following embodiments illustrate the technical solution of the present invention in detail, but the scope of protection of the present invention includes, but is not limited to, these embodiments.

[0158] The compounds used in the reactions described herein were prepared according to organic synthesis techniques known to those skilled in the art, and were derived from commercially available chemicals and / or compounds described in chemical literature. “Commercially available chemicals” are obtained from legitimate commercial sources, and suppliers include: Titan Technology, Energetic Chemicals, Shanghai Demo, Chengdu Kelon Chemical, Shaoyuan Chemical Technology, Nanjing Yaoshi, WuXi AppTec, and Bailingwei Technology, among others.

[0159] The structure of the compound was determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts (δ) were expressed in 10⁻¹⁰ increments. -6 The unit (ppm) is given. NMR measurements were performed using a Bruker Avance III 400 and Bruker Avance 300 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD). The internal standard was tetramethylsilane (TMS).

[0160] MS determination was performed using (Agilent 6120B (ESI) and Agilent 6120B (APCI));

[0161] HPLC determinations were performed using an Agilent 1260DAD high-performance liquid chromatograph (Zorbax SB-C18 100×4.6mm, 3.5μM).

[0162] Thin-layer chromatography silica gel plates used were from Yantai Huanghai HSGF. 254 Or Qingdao GF 254 Silica gel plates: The silica gel plates used in thin-layer chromatography (TLC) have a diameter of 0.15mm-0.20mm, while those used for TLC separation and purification of products have a diameter of 0.4mm-0.5mm.

[0163] Column chromatography typically uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier;

[0164] Synthesis Examples

[0165] Synthesis of intermediate 1:

[0166] Step 1: Preparation of 1-B

[0167] 1-A (2.8 g, 13.58 mmol) (synthetic method referred to Bioorganic & Medicinal Chemistry Letters, 2016, 26, 5877-5882) was dissolved in dichloromethane (50 mL), Boc₂O (5.93 g, 27.17 mmol) was added, and DMAP (3.32 g, 27.18 mmol) was added. The reaction mixture was reacted at room temperature for 16 h. The reaction mixture was washed with 0.5 mol / L hydrochloric acid (50 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (ethyl acetate: petroleum ether (v / v) = 0:1-1:9) to give the racemic form of 1-B (3.4 g, yield: 82%).

[0168] The racemic mixture of 1-B is chirally separated as follows:

[0169] 1. Instrument: SFC Prep 150AP; Column: Daicel IC-H (19mm×250mm).

[0170] 2. Dissolve the sample in methanol and filter it through a 0.45μm filter to prepare a sample solution.

[0171] 3. Preparative chromatographic conditions: a. The mobile phase consists of system A and system B: mobile phase A: CO2; mobile phase B: methanol / isopropanol (v / v) = 1:1; b. Isogradient elution, with mobile phase B containing 20%; c. Flow rate of 40 mL / min.

[0172] Peak times: chiral isomer 1 (compound 1-B): 5.7 min, chiral isomer 2 (compound 2-A): 6.47 min.

[0173] According to the MicroED structure determination of compound 2-B, compound 1-B has an R configuration and compound 2-A has an S configuration.

[0174] LCMS m / z = 307.3[M+1] + .

[0175] Step 2: Preparation of 1-C

[0176] 1-B (1.8 g, 5.88 mmol) was dissolved in acetonitrile (50 mL), and NBS (1.05 g, 5.90 mmol) was added. The reaction mixture was reacted at room temperature for 1 h. The reaction mixture was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (ethyl acetate: petroleum ether (v / v) = 0:1-1:9) to give 1-C (1.6 g, yield: 71%).

[0177] LCMS m / z = 385.3 [M+1] + .

[0178] Step 3: Preparation of 1-D

[0179] 1-C (0.77 g, 2.0 mmol), 1-C′ (1.67 g, 4.0 mmol) (synthetic method see WO2022235945), Pd(dppf)Cl2·DCM (0.16 g, 0.20 mmol), and cesium carbonate (1.30 g, 4.0 mmol) were added to a reaction flask, along with 1,4-dioxane (30 mL) and water (3 mL). The reaction was carried out at 100 °C for 20 h under a nitrogen atmosphere. The reaction system was cooled to room temperature, and 50 mL of water and 50 mL of ethyl acetate were added. The aqueous phase was extracted with 50 mL of ethyl acetate, and the organic phase was washed with 30 mL of saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1:0-9:1) to give 1-D (0.70 g, yield: 59%).

[0180] Step 4: Preparation of 1-E

[0181] 1-D (0.70 g, 1.18 mmol) was dissolved in THF (20 mL), and 10% palladium on carbon (0.63 g) was added. The mixture was reacted at 45 °C for 20 h under a hydrogen balloon atmosphere. The reaction system was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (MeOH:DCM (v / v) = 0:1-5:95) to give 1-E (0.25 g, yield: 51%).

[0182] LCMS m / z = 418.1[M+1] + .

[0183] Step 5: Preparation of Intermediate 1

[0184] 1-E (250 mg, 0.6 mmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (2 mL) was added. The mixture was reacted at room temperature for 3 h. The reaction mixture was concentrated under reduced pressure, and 5 mL of dichloromethane and 1 mL of triethylamine were added. The mixture was then concentrated under reduced pressure to obtain crude intermediate 1 (190 mg).

[0185] LCMS m / z = 318.3[M+1] + .

[0186] Synthesis of intermediate 2:

[0187] Step 1: Preparation of 2-B

[0188] 2-A (1.4 g, 4.57 mmol) was dissolved in acetonitrile (50 mL), and NBS (0.81 g, 4.55 mmol) was added. The reaction mixture was reacted at room temperature for 1 h. The reaction mixture was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (ethyl acetate: petroleum ether (v / v) = 0:1-1:9) to give 2-B (1.6 g, yield: 92%).

[0189] LCMS m / z = 385.3 [M+1] + .

[0190] Compound 2-B was determined to be S-configuration by MicroED.

[0191] Step 2: Preparation of 2-C

[0192] 2-B (1.6 g, 4.16 mmol), 2-B′ (3.46 g, 8.29 mmol), Pd(dppf)Cl2·DCM (0.34 g, 0.42 mmol), and cesium carbonate (2.70 g, 8.3 mmol) were added to a reaction flask, along with 1,4-dioxane (50 mL) and water (5 mL). The reaction was carried out at 100 °C for 20 h under a nitrogen atmosphere. The reaction system was cooled to room temperature, and 50 mL of water and 50 mL of ethyl acetate were added. The aqueous phase was extracted with ethyl acetate (50 mL), and the organic phase was washed with 30 mL of saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1:0-9:1) to give 2-C (1.4 g, yield: 56%).

[0193] Step 3: Preparation of 2-D

[0194] 2-C (1.4 g, 2.35 mmol) was dissolved in THF (50 mL), and 10% palladium on carbon (1.25 g) was added. The mixture was reacted at 45 °C for 20 h under a hydrogen balloon atmosphere. The reaction system was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (MeOH:DCM (v / v) = 0:1-5:95) to give 2-D (0.85 g, yield: 87%).

[0195] LCMS m / z = 418.1[M+1] + .

[0196] Step 4: Preparation of Intermediate 2

[0197] 2-D (620 mg, 1.49 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (3 mL) was added. The mixture was reacted at room temperature for 3 h. The reaction mixture was concentrated under reduced pressure, and 5 mL of dichloromethane and 1 mL of triethylamine were added. The mixture was then concentrated under reduced pressure to give crude intermediate 2 (460 mg).

[0198] LCMS m / z = 318.3[M+1] + .

[0199] Preparation of intermediate 3:

[0200] Step 1: Preparation of 3-A

[0201] Under a nitrogen atmosphere, 1-C (7.00 g, 18.17 mmol) and 70 mL of tetrahydrofuran were added to a reaction flask. A 2.5 mol / L n-butyllithium solution in n-hexane (14.50 mL, 36.25 mmol) was slowly added dropwise at -78 °C. The mixture was stirred at -78 °C for 1.5 h, then the carbon dioxide was replaced three times. The reaction was carried out under a carbon dioxide balloon atmosphere, maintaining the system temperature below -40 °C for 0.5 h. The reaction system was brought back to room temperature, 20 mL of ethyl acetate was added, and the pH was adjusted to 2 with 1 mol / L hydrochloric acid. The mixture was extracted with ethyl acetate (30 mL × 3). The organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 10:1-2:1) to give 3-A (2.4 g, yield: 38%).

[0202] 1H NMR(400MHz,DMSO-d6)δ7.42(d,1H),6.64(d,1H),4.00–3.82(m,3H),3.23–3.09(m,1H),3.08–2.90(m ,1H),2.90–2.77(m,1H),2.76–2.60(m,3H),2.05–1.90(m,1H),1.65–1.50(m,1H),1.49–1.35(m,9H).

[0203] Step 2: Preparation of Intermediate 3

[0204] 3-A (2.4 g, 6.85 mmol), HATU (3.13 g, 8.23 ​​mmol), diisopropylethylamine (2.66 g, 20.58 mmol), and 10 mL of DMF were added to a reaction flask. After stirring at room temperature for 10 min, 3-amino-2,6-piperidinidone hydrochloride (1.35 g, 8.20 mmol) was added, and the mixture was reacted at room temperature for 30 min. The reaction solution was poured into 100 mL of water, filtered, and the filter cake was washed with 50 mL of water. The filter cake was dried under reduced pressure to give intermediate 3 (2.0 g, yield: 63%).

[0205] LCMS m / z = 461.2[M+1] +

[0206] Preparation of intermediate 4:

[0207] Intermediate 4 was obtained from compound 2-B using the same synthetic method as intermediate 3.

[0208] LCMS m / z = 461.2[M+1] +

[0209] Preparation of intermediate 5:

[0210] Step 1: Preparation of 5-B

[0211] 5-A (10 g, 42.02 mmol) was dissolved in 80 mL of DMSO, and 5-A′ (7.41 g, 46.25 mmol) and DIPEA (16.29 g, 126.03 mmol) were added. The reaction mixture was reacted at 50 °C for 3 h. The reaction mixture was cooled to room temperature, and 300 mL of water and 400 mL of ethyl acetate were added. The organic phase was washed with 300 mL of saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give crude 5-B (6.2 g).

[0212] Step 2: Preparation of 5-C

[0213] The crude product 5-B (6.2 g) was added to 150 mL of methanol and 30 mL of water, along with iron powder (9.15 g, 163.4 mmol) and ammonium chloride (8.77 g, 163.96 mmol). The reaction mixture was reacted at 70 °C for 20 h. The reaction system was cooled to room temperature, filtered through a diatomaceous earth filter, and the filtrate was concentrated under reduced pressure. 300 mL of water and 400 mL of ethyl acetate were added to the residue. The organic phase was washed with 300 mL of saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (methanol:dichloromethane (v / v) = 0:1-1:9) to give 5-C (1.9 g, two-step yield from compound 5-A: 13%).

[0214] Step 3: Preparation of 5-D

[0215] 5-C (1.9 g, 5.46 mmol) and TEA (1.66 g, 16.40 mmol) were dissolved in 20 mL of dichloromethane, and chloroacetyl chloride (0.99 g, 8.77 mmol) was added dropwise. The reaction was carried out at room temperature for 20 h. 50 mL of water was added to the reaction solution, and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain crude 5-D (2.1 g).

[0216] Step 4: Preparation of 5-E

[0217] The crude 5-D (0.95 g) was added to 15 mL of acetic acid and reacted at 50 °C for 20 h. The reaction system was cooled to 0 °C, the pH was adjusted to 8 with 25% ammonia, and the mixture was extracted with 150 mL of ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (methanol:dichloromethane (v / v) = 0:1-1:9) to give 5-E (0.85 g, two-step yield from compound 5-C: 85%).

[0218] Step 5: Preparation of 5-F

[0219] 5-E (7.3 g, 17.95 mmol) was dissolved in 80 mL THF, and 20 mL of 1 mol / L potassium tert-butoxide in tetrahydrofuran solution was added. The reaction was carried out at room temperature for 19 h. 30 mL of water and 50 mL of ethyl acetate were added to the reaction system. The organic phase was washed with 300 mL of saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (methanol:dichloromethane (v / v) = 0:1-1:9) to give 5-F (3.6 g, yield: 54%).

[0220] Step 6: Preparation of 5-G

[0221] 5-F (2.0 g, 5.4 mmol), 2,6-bisbenzyloxypyridine-3-boronic acid pinacol ester (3.4 g, 8.15 mmol), Pd(dppf)Cl2·DCM (0.44 g, 0.54 mmol), and cesium carbonate (3.5 g, 10.74 mmol) were added sequentially to a reaction flask, followed by 1,4-dioxane (100 mL) and water (10 mL). The reaction was carried out at 100 °C for 20 h under a nitrogen atmosphere. The reaction system was cooled to room temperature, and 300 mL of water and 300 mL of ethyl acetate were added. The organic phase was washed with 100 mL of saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1:0-9:1) to obtain 5-G (2.6 g, yield: 83%).

[0222] LCMS m / z = 581.4 [M+1] + .

[0223] Step 7: Preparation of 5-H

[0224] 5-G (2.6 g, 4.48 mmol) was dissolved in THF (10 mL) and methanol (30 mL), and 10% palladium on carbon (2.5 g) was added. The reaction was carried out at 40 °C for 20 h under a hydrogen balloon atmosphere. The reaction system was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to obtain crude 5-H (0.98 g).

[0225] LCMS m / z = 403.3[M+1] + .

[0226] Step 8: Preparation of Intermediate 5

[0227] The crude product 5-H (100 mg) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added. The mixture was reacted at room temperature for 3 h. The reaction system was concentrated under reduced pressure, and 5 mL of methanol and 150 mg of sodium bicarbonate solid were added. After stirring for 10 min, the mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain crude intermediate 5 (80 mg).

[0228] Example 1: Preparation of trifluoroacetate of compound 1

[0229] Dissolve 1a (0.23 g, 0.47 mmol) (synthetic method referred to WO2021127443) in 1,2-dichloroethane (10 mL), add the above crude intermediate 2 (0.15 g), add 1 mL of acetic acid, stir at room temperature for 1 h, then add sodium triacetoxyborohydride (0.30 g, 1.42 mmol), and react at room temperature for 16 h. 20 mL of saturated sodium bicarbonate aqueous solution was slowly added to the reaction solution, and the mixture was extracted with 50 mL of dichloromethane. The organic phase was washed with 50 mL of saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (dichloromethane:methanol (v / v) = 10:1). The crude product was then prepared by HPLC (preparation method: the DMSO solution of the crude product was filtered through a 0.45 μm filter membrane to prepare the sample solution. Mobile phase system: water (containing 0.1% TFA) / acetonitrile. Gradient elution method: acetonitrile was used to elute 50% by a 10% gradient (elution time 15 min)). The product was lyophilized to obtain trifluoroacetate of compound 1 (10 mg).

[0230] 1 H NMR(400MHz,CD3OD)δ7.80–7.72(m,2H),7.52(d,1H),7.08–7.00(m,2H),6.91(d,1H) ),6.71(d,1H),6.64–6.60(m,1H),6.55(dd,1H),4.26(s,1H),4.18–4.03(m,2H),4.0 2–3.83(m,6H),3.80–3.60(m,2H),3.43–3.30(m,1H),3.28–3.06(m,4H),3.04–2.57( m,7H),2.33–2.02(m,4H),2.01–1.70(m,3H),1.56–1.40(m,2H),1.36–1.19(m,12H).

[0231] LCMS m / z = 791.4 [M+1] +

[0232] The crude product obtained from this step can also be prepared by neutral HPLC (preparation method: the crude product's DMSO solution is filtered through a 0.45 μm filter membrane to prepare the sample solution. Mobile phase system: 10 mmol / L ammonium bicarbonate aqueous solution / acetonitrile. Gradient elution method: acetonitrile is used to elute 87% from 47% gradient (elution time 28 min)), and lyophilized to obtain compound 1.

[0233] 1H NMR(400MHz, CDCl3)δ7.91(s,1H),7.73–7.64(m,2H),7.45(d,1H),6.95–6.87(m,2H),6.72 (d,1H),6.53–6.44(m,2H),6.40(dd,1H),6.10(d,1H),4.18–4.11(m,1H),4.04(s,1H),3.9 1(s,3H),3.89–3.72(m,3H),3.65–3.54(m,1H),3.14–3.00(m,1H),2.97–2.72(m,7H),2.70 –2.57(m,2H),2.32–2.11(m,5H),1.99–1.82(m,4H),1.81–1.65(m,2H),1.40–1.19(m,14H).

[0234] Compound 1 was chirally resolved to obtain chiral isomer 1 and chiral isomer 2.

[0235] First chiral separation and purification conditions:

[0236] 1. Instrument: SHIMADZU LC-20AP; Column: Chiral Whelk column.

[0237] 2. Dissolve the sample in acetonitrile and filter it through a 0.45μm filter to prepare the sample solution.

[0238] 3. Preparative chromatographic conditions: a. The mobile phase consists of system A and system B: Mobile phase A: n-hexane; Mobile phase B: a mixed solvent of ethanol / acetonitrile containing 0.1% ammonia; b. Isogradient elution, with mobile phase B containing 70%; c. Flow rate is 100 mL / min.

[0239] Second chiral separation and purification conditions:

[0240] 1. Instrument: Waters 150Prep-SFC; Column: Chiral AD column.

[0241] 2. Dissolve the sample in acetonitrile and filter it through a 0.45μm filter to prepare the sample solution.

[0242] 3. Preparative chromatographic conditions: a. The mobile phase consists of system A and B: mobile phase A: CO2; mobile phase B: a mixed solvent of isopropanol / acetonitrile containing 0.1% ammonia; b. Isogradient elution, with mobile phase B containing 70%; c. Flow rate is 100 mL / min.

[0243] Chiral analysis methods:

[0244] 1. Instrument: SHIMAZU LC-20AD; Column: Chiral Whelk column.

[0245] 2. Analytical chromatographic conditions: a. The mobile phase consists of system A and system B: Mobile phase A: n-hexane; Mobile phase B: ethanol and acetonitrile solution containing 0.1% isopropylamine; b. Isogradient elution, with mobile phase B containing 60%; c. Flow rate is 1 mL / min.

[0246] Peak elution times: chiral isomer 1: 2.594 min, chiral isomer 2: 5.557 min.

[0247] Example 3: Preparation of trifluoroacetate of compound 3

[0248] 2 mL of dichloromethane and 1 mL of trifluoroacetic acid were added to intermediate 4 (160 mg, 0.35 mmol), and the reaction was carried out at room temperature for 2 h. The reaction system was concentrated under reduced pressure, and 5 mL of dichloromethane and 1 mL of triethylamine were added. The mixture was then concentrated under reduced pressure to give crude product 1 (120 mg). 1a (0.160 g, 0.33 mmol) was dissolved in 1,2-dichloroethane (10 mL), and crude product 1 (0.120 g) was added. 1 mL of acetic acid was added, and the mixture was stirred at room temperature for 1 h. Then, sodium triacetoxyborohydride (0.21 g, 0.99 mmol) was added, and the mixture was reacted at room temperature for 16 h. 20 mL of saturated sodium bicarbonate aqueous solution was slowly added to the reaction solution, and the mixture was extracted with 50 mL of dichloromethane. The organic phase was washed with 50 mL of saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (dichloromethane:methanol (v / v) = 10:1). The crude product was then prepared by HPLC (preparation method: the DMSO solution of the crude product was filtered through a 0.45 μm filter membrane to prepare the sample solution. Mobile phase system: water (containing 0.1% TFA) / acetonitrile. Gradient elution method: acetonitrile was used to elute 50% by a 10% gradient (elution time 15 min)). The product was lyophilized to obtain trifluoroacetate of compound 3 (45 mg).

[0249] 1H NMR (400MHz, CD3OD) δ7.80–7.72(m,2H),7.60–7.48(m,2H),7.10–7.00(m,2H),6.76(d,1 H),6.65–6.60(m,1H),6.55(dd,1H),4.85–4.72(m,1H),4.30–4.10(m,3H),4.02–3.88(m, 5H),3.82–3.63(m,2H),3.60–3.43(m,1H),3.33–3.10(m,4H),3.07–2.62(m,7H),2.37–2 .06(m,4H),2.02–1.88(m,2H),1.85–1.72(m,1H),1.57–1.42(m,2H),1.35–1.17(m,12H).

[0250] LCMS m / z = 834.6 [M+1] +

[0251] Example 5: Preparation of trifluoroacetate of compound 5

[0252] Step 1: Preparation of 5A trifluoroacetate

[0253] 3-A (650 mg, 1.86 mmol) was dissolved in 2 mL of dichloromethane, and 2 mL of trifluoroacetic acid was added. The mixture was reacted at room temperature for 3 h. The reaction mixture was then concentrated under reduced pressure to obtain crude trifluoroacetate of 5A (700 mg).

[0254] Step 2: Preparation of 5b trifluoroacetate

[0255] Dissolve 5a (3.00 g, 11.75 mmol) in 5 mL of dichloromethane, add 5 mL of trifluoroacetic acid, and react at room temperature for 3 h. Concentrate the reaction system under reduced pressure to obtain crude trifluoroacetate of 5b (3.2 g).

[0256] Step 3: Preparation of 5C

[0257] Add 3.2 g of trifluoroacetate, 1.85 g (11.00 mmol) of crude product 5b, 3.04 g (22.0 mmol) of potassium carbonate, and 15 mL of dimethyl sulfoxide to the reaction flask, and react at 100 °C for 16 h. Cool the reaction system to room temperature, add 100 mL of water and 100 mL of ethyl acetate to the reaction solution, separate the layers, extract the aqueous phase with ethyl acetate (50 mL × 3), combine the organic phases, wash the organic phase with 50 mL of saturated brine, dry with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and purify the crude product by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1:0-1:1) to give 5c (2.40 g, yield: 72%).

[0258] Step 4: Preparation of 5D

[0259] Add 5c (2.40 g, 7.91 mmol), potassium hydroxide (1.78 g, 31.79 mmol), 10 mL ethanol, 10 mL tetrahydrofuran, and 10 mL water to the reaction flask, and stir at room temperature for 16 h. Concentrate the reaction system under reduced pressure, adjust the pH to 2 with 2 mol / L hydrochloric acid, filter, collect the filter cake, and dry the filter cake under reduced pressure to obtain crude product 5d (1.50 g).

[0260] LCMS m / z = 276.3[M+1] +

[0261] Step 5: Preparation of 5e

[0262] Add the above crude product 5d (0.6 g), HATU (0.99 g, 2.60 mmol), diisopropylethylamine (1.41 g, 10.91 mmol), and 10 mL DMF to the reaction flask. After stirring at room temperature for 0.5 h, add trans-4-(3-amino-2,2,4,4-tetramethylcyclobutoxy)-2-methoxybenzyl nitrile hydrochloride (0.66 g, 2.12 mmol) (synthetic method see CN115175901), and stir at room temperature for 2 h. Add 100 mL water and 100 mL ethyl acetate to the reaction solution, separate the layers, extract the aqueous phase with ethyl acetate (50 mL × 3), combine the organic phases, wash the organic phase with 50 mL saturated brine, dry with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and purify the crude product by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1:0-0:1) to give 5e (1.10 g, yield: 98%).

[0263] LCMS m / z = 532.6 [M+1] +

[0264] Step 6: Preparation of 5f

[0265] 5e (1.10 g, 2.07 mmol), Dess-Martin oxidant (1.05 g, 2.48 mmol), and 10 mL of dichloromethane were added to the reaction flask, and the reaction was carried out at room temperature for 4 h. The reaction system was filtered through a diatomaceous earth liner, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (methanol / dichloromethane (v / v) = 0:1-1:9) to give 5f (0.52 g, yield: 47%).

[0266] LCMS m / z = 530.2[M+1] +

[0267] Step 7: Preparation of 5g

[0268] Add 5f (0.25 g, 0.47 mmol), trifluoroacetate of the above crude product 5A (0.18 g), 0.2 mL glacial acetic acid, and 3 mL DMAc to the reaction flask. After reacting at room temperature for 1 h, add sodium triacetoxyborohydride (0.30 g, 1.42 mmol) and react at room temperature for 16 h. Add 20 mL of water and 20 mL of ethyl acetate to the reaction solution, separate the layers, extract the aqueous phase with ethyl acetate (20 mL × 3), combine the organic phases, wash the organic phase with 50 mL of saturated brine, dry with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and purify the crude product by silica gel column chromatography (methanol / dichloromethane (v / v) = 0:1-1:9) to obtain 5 g (0.20 g, yield: 56%).

[0269] Step 8: Preparation of trifluoroacetate of compound 5

[0270] Add 5 g (0.10 g, 0.13 mmol), (S)-3-aminopiperidine-2,6-dione hydrochloride (0.043 g, 0.26 mmol), EDCI (0.037 g, 0.19 mmol), HOBt (0.026 g, 0.19 mmol), N-methylmorpholine (0.065 g, 0.64 mmol), and 4 mL DMF to the reaction flask, and react at room temperature for 16 h. Pass the reaction solution through Pre-HPLC (instrument and preparative column: Waters 2767 preparative HPLC, SunFire@Prep C18, 5 μm, inner diameter × length = 19 mm × 250 mm). Preparation method: Filter the crude DMSO solution through a 0.45 μm filter membrane to prepare the sample solution. Mobile phase system: water (containing 0.1% TFA) / acetonitrile. Gradient elution method: Acetonitrile was used to elute 50% by a 10% gradient (elution time 15 min), and the product was lyophilized to obtain trifluoroacetate of compound 5 (20 mg).

[0271] 1H NMR(400MHz,CD3OD)δ7.74(d,2H),7.57–7.48(m,2H),7.02(d,2H),6.75(d,1H),6.62(d,1 H),6.55(dd,1H),4.82–4.74(m,1H),4.30–4.23(m,1H),4.23–4.17(m,1H),4.13(s,1H),3 .93(s,3H),3.68–3.49(m,2H),3.48–3.30(m,5H),3.28–3.08(m,4H),3.02–2.90(m,1H),2 .88–2.65(m,5H),2.35–2.05(m,5H),1.89–1.82(m,2H),1.82–1.65(m,5H),1.26(d,12H).

[0272] LCMS m / z = 874.4 [M+1] +

[0273] Example 7: Preparation of trifluoroacetate of compound 7

[0274] The trifluoroacetate of compound 7 was obtained from compound 4-A using the synthesis method described in Example 5.

[0275] 1 H NMR (400MHz, CD3OD) δ7.81–7.71(m,2H),7.57–7.48(m,2H),7.13–7.01(m,2H),6.75(d ,1H),6.62(d,1H),6.55(dd,1H),4.80–4.74(m,1H),4.26(s,1H),4.24–4.16(m,1H),4 .13(s,1H),3.93(s,3H),3.67–3.50(m,2H),3.50–3.15(m,9H),3.01–2.91(m,1H),2.9 1–2.63(m,5H),2.35–2.07(m,5H),1.94–1.84(m,2H),1.84–1.67(m,5H),1.26(d,12H).

[0276] LCMS m / z = 437.8 [M / 2+1] +

[0277] The following compounds were obtained by synthesizing according to the methods described in other embodiments:

[0278] Example 11: Preparation of Compound 11

[0279] Compound 11 was prepared by neutral preparation (aqueous solution of ammonium acetate (5 mmol / L) / acetonitrile) and lyophilized, following the synthesis method of Example 5.

[0280] 1 H NMR(400MHz,DMSO-d6)δ10.82(s,1H),7.94(t,1H),7.78–7.69(m,2H),7.64(d,1H),7.47(d,1H) ),7.36(d,1H),7.00–6.90(m,2H),6.73–6.61(m,2H),6.54(dd,1H),4.78–4.65(m,1H),4.27(s, 1H),4.05(d,1H),3.95–3.79(m,6H),3.16–3.06(m,1H),2.97–2.61(m,8H),2.57–2.50(m,1H),2 .26–2.14(m,2H),2.13–1.88(m,4H),1.87–1.72(m,4H),1.66–1.52(m,1H),1.28–1.09(m,14H).

[0281] LCMS m / z = 418.0 [M / 2+1] +

[0282] Example 13: Preparation of Compound 13

[0283] Compound 13 was obtained by neutral preparation (aqueous solution of ammonium acetate (5 mmol / L) / acetonitrile) lyophilization, using compounds 1a and 7A as raw materials, following the synthesis method of Example 11.

[0284] 1H NMR(400MHz,DMSO-d6)δ10.82(s,1H),7.94(t,1H),7.74(d,2H),7.64(d,1H),7.47(d,1H),7 .36(d,1H),6.95(d,2H),6.74–6.61(m,2H),6.54(dd,1H),4.82–4.60(m,1H),4.27(s,1H),4 .05(d,1H),3.95–3.79(m,6H),3.17–3.04(m,1H),2.97–2.87(m,2H),2.87–2.62(m,6H),2.5 7–2.52(m,1H),2.25–1.88(m,6H),1.88–1.70(m,4H),1.67–1.51(m,1H),1.29–1.08(m,14H).

[0285] LCMS m / z = 418.0 [M / 2+1] +

[0286] Example 15: Preparation of Compound 15

[0287] Step 1: Preparation of 15b

[0288] 15a (1.28 g, 3.48 mmol) (synthetic method see Bioorg. Med. Chem. Lett. 2016, 26, 5877-5882), cesium carbonate (2.27 g, 6.97 mmol), palladium acetate (0.16 g, 0.71 mmol), XantPhos (0.20 g, 0.35 mmol), and 15A (0.89 g, 4.91 mmol) were added to a 1,4-dioxane solution (40 mL), and reacted at 105 °C for 3 h under a nitrogen atmosphere. The reaction system was cooled to room temperature, water (100 mL) was added, and the mixture was extracted with ethyl acetate (80 mL × 3). The organic phase was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1:0-7:3) to give 15b (1.30 g, yield: 80%).

[0289] LCMS m / z = 468.4[M+1] +

[0290] Step 2: Preparation of 15C

[0291] 15b (1.30 g, 2.78 mmol) was added to methanol (10 mL), along with 10% palladium on carbon (1.33 g) and ammonium acetate (1.32 g, 17.13 mmol). The mixture was reacted at room temperature for 12 h under a hydrogen balloon atmosphere. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1:0-1:1) to give 15c (0.76 g, yield: 90%).

[0292] LCMS m / z = 304.4[M+1] +

[0293] Step 3: Preparation at 15 days

[0294] 15c (0.76 g, 2.55 mmol), 15B (0.75 g, 7.49 mmol), and DIPEA (0.97 g, 7.50 mmol) were added sequentially to ethanol (50 mL), and the reaction was carried out at 100 °C for 48 h. The reaction system was cooled to room temperature, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (methanol / dichloromethane (v / v) = 0:1-6:94) to obtain 15d (0.70 g, yield: 68%).

[0295] LCMS m / z = 404.2[M+1] +

[0296] Step 4: Preparation of 15e

[0297] 15d (700 mg, 1.74 mmol) and DIPEA (671 mg, 5.19 mmol) were added to THF (20 mL), and triphosgene (565 mg, 1.90 mmol) was slowly added. The reaction was carried out at room temperature for 1 h. Ammonia (5 mL) was added to the reaction system, and the reaction was carried out at 50 °C for 2 h. The reaction system was cooled to room temperature, water (50 mL) was added, and the mixture was extracted with ethyl acetate (50 mL × 2). The organic phase was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (methanol / dichloromethane (v / v) = 0:1-1:9) to obtain 15e (700 mg, yield: 90%).

[0298] Step 5: Preparation of 15f

[0299] 15e (0.70 g, 1.57 mmol) was added to acetonitrile (30 mL), followed by 2 mL of a 40% benzyltrimethylammonium hydroxide methanol solution. The reaction mixture was reacted at 60 °C for 2 h. The reaction system was cooled to room temperature, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (methanol / dichloromethane (v / v) = 0:1-8:92) to obtain 15f (530 mg, yield: 84%).

[0300] LCMS m / z = 401.4[M+1] +

[0301] Step 6: Preparation of Compound 15

[0302] Add 15f (0.15 g, 0.37 mmol), 2 mL trifluoroacetic acid, and 3 mL dichloromethane to the reaction flask and react at room temperature for 2 h. Concentrate the reaction solution under reduced pressure, add triethylamine to adjust the pH to 7, concentrate under reduced pressure, dissolve the residue in 5 mL DMA, and add 1a (0.23 g, 0.47 mmol), 0.5 mL acetic acid, and sodium triacetoxyborohydride (0.24 g, 1.13 mmol) sequentially, reacting at room temperature for 16 h. Add 10 mL methanol to the reaction solution and concentrate under reduced pressure. Pass the reaction solution through Pre-HPLC (instrument and preparative column: Waters 2767 preparative HPLC, SunFire@Prep C18, 5 μm, inner diameter × length = 19 mm × 250 mm). Preparation method: Filter the crude product's DMSO solution through a 0.45 μm filter membrane to prepare the sample solution. Mobile phase system: aqueous solution of ammonium acetate (5 mmol / L) / acetonitrile. Gradient elution method: Acetonitrile was eluted from 70% by a 10% gradient (elution time 12 min), and the compound 15 was obtained by lyophilization (0.20 g, yield: 70%).

[0303] 1 H NMR(400MHz,DMSO-d6)δ10.21(s,1H),7.74(d,2H),7.64(d,1H),7.47(d,1H),6.99–6.91(m,3H ),6.91–6.85(m,1H),6.85–6.77(m,1H),6.64(d,1H),6.54(dd,1H),4.27(s,1H),4.05(d,1H),3 .97–3.81(m,5H),3.80–3.72(m,1H),3.70–3.61(m,2H),2.99–2.73(m,6H),2.73–2.57(m,4H),2 .24–2.13(m,2H),2.13–2.02(m,1H),1.93–1.72(m,5H),1.69–1.54(m,1H),1.29–1.10(m,14H).

[0304] LCMS m / z = 774.6 [M+1] + .

[0305] Example 16: Preparation of trifluoroacetate of compound 16

[0306] Step 1: Preparation of 16b trifluoroacetate

[0307] Dissolve 16a (2.77 g, 11.48 mmol) in 5 mL of dichloromethane, add 5 mL of trifluoroacetic acid, and react at room temperature for 3 h. Concentrate the reaction system under reduced pressure to obtain crude trifluoroacetate of 16b (3.0 g).

[0308] Step 2: Preparation of 16C

[0309] Add 3.0 g of trifluoroacetate, 1.85 g (11.00 mmol) of crude product 16b, 3.04 g (22.00 mmol) of potassium carbonate, and 15 mL of dimethyl sulfoxide to the reaction flask, and react at 100 °C for 16 h. Cool the reaction system to room temperature, add 100 mL of water and 100 mL of ethyl acetate to the reaction solution, separate the layers, extract the aqueous phase with ethyl acetate (50 mL × 3), combine the organic phases, wash the organic phase with 50 mL of saturated brine, dry with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and purify the crude product by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1:0-1:1) to give 16c (2.5 g, yield: 79%).

[0310] Step 3: Preparation of 16 days

[0311] 16c (2.4 g, 8.29 mmol), potassium hydroxide (1.86 g, 33.21 mmol), 10 mL ethanol, 10 mL tetrahydrofuran, and 10 mL water were added to the reaction flask, and the mixture was reacted at room temperature for 16 h. The reaction system was concentrated under reduced pressure, the pH was adjusted to 2 with 2 mol / L hydrochloric acid, filtered, and the filter cake was collected and dried under reduced pressure to obtain crude product 16d (1.80 g).

[0312] LCMS m / z = 262.1[M+1] +

[0313] Step 4: Preparation of 16e

[0314] The crude product 16d (0.68 g), HATU (0.99 g, 2.60 mmol), diisopropylethylamine (1.41 g, 10.91 mmol) and 10 mL DMF were added to the reaction flask. After stirring at room temperature for 0.5 h, trans-4-(3-amino-2,2,4,4-tetramethylcyclobutoxy)-2-methoxybenzyl nitrile hydrochloride (0.57 g, 1.83 mmol) (synthetic method see CN115175901) was added, and the reaction was carried out at room temperature for 2 h. Add 100 mL of water and 100 mL of ethyl acetate to the reaction solution, separate the liquid and extract the aqueous phase with ethyl acetate (50 mL × 3), combine the organic phases, wash the organic phase with 50 mL of saturated brine, dry with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and purify the crude product by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1:0-0:1) to give 16e (1.10 g, two-step yield from compound 16c: 68%).

[0315] LCMS m / z = 518.5[M+1] +

[0316] Step 5: Preparation of 16F

[0317] 16e (1.10 g, 2.13 mmol), Dess-Martin oxidant (1.08 g, 2.55 mmol), and 10 mL of dichloromethane were added to the reaction flask, and the reaction was carried out at room temperature for 4 h. The reaction system was filtered through a diatomaceous earth liner, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (methanol / dichloromethane (v / v) = 0:1-1:9) to give 16f (0.9 g, yield: 82%).

[0318] LCMS m / z = 516.2[M+1] +

[0319] Step 6: Preparation of 16g

[0320] Add 16f (0.25 g, 0.485 mmol), trifluoroacetate of the above crude product 5A (0.18 g), 0.2 mL glacial acetic acid, and 3 mL DMAc to the reaction flask. After reacting at room temperature for 1 h, add sodium triacetoxyborohydride (0.30 g, 1.42 mmol) and react at room temperature for 16 h. Add 20 mL of water and 20 mL of ethyl acetate to the reaction solution, separate the layers, extract the aqueous phase with ethyl acetate (20 mL × 3), combine the organic phases, wash with 50 mL saturated brine, dry with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and purify the crude product by silica gel column chromatography (methanol / dichloromethane (v / v) = 0:1-1:9) to obtain 16 g (0.25 g, yield: 69%).

[0321] LCMS m / z = 750.4 [M+1] +

[0322] Step 7: Preparation of compound 16 trifluoroacetate

[0323] Add 16 g (0.2 g, 0.27 mmol), (S)-3-aminopiperidine-2,6-dione hydrochloride (0.069 g, 0.42 mmol), EDCI (0.13 g, 0.68 mmol), HOBt (0.073 g, 0.54 mmol), N-methylmorpholine (0.17 g, 1.68 mmol), and 4 mL of DMF to the reaction flask, and react at room temperature for 16 h. Pass the reaction solution through Pre-HPLC (instrument and preparative column: Waters 2767 preparative HPLC, SunFire@Prep C18, 5 μm, inner diameter × length = 19 mm × 250 mm). Preparation method: Filter the crude product's DMSO solution through a 0.45 μm filter membrane to prepare the sample solution. Mobile phase system: water (containing 0.1% TFA) / acetonitrile. Gradient elution method: Acetonitrile was used to elute 50% by a 10% gradient (elution time 15 min), and the product was lyophilized to obtain trifluoroacetate of compound 16 (50 mg).

[0324] 1 H NMR (400MHz, CD3OD) δ7.79–7.69(m,2H),7.59–7.47(m,2H),7.07–6.98(m,2H),6.76(d,1H),6.62(d,1H),6 .55(dd,1H),4.79–4.74(m,1H),4.29–4.18(m,2H),4.13(s,1H),3.93(s,3H),3.86–3.72(m,1H),3.67–3.54 (m,2H),3.51–3.40(m,1H),3.40–3.30(m,4H),3.26–3.15(m,1H),3.08–2.94(m,1H),2.89–2.74(m,4H),2. 74–2.65(m,1H),2.47–2.37(m,2H),2.35–2.25(m,1H),2.21–2.05(m,4H),1.87–1.70(m,5H),1.26(d,12H).

[0325] LCMS m / z = 431.0[M / 2+1] +

[0326] The following compounds were obtained by synthesizing according to the methods described in other embodiments:

[0327] Example 18: Preparation of trifluoroacetate of compound 18

[0328] Compound 18 trifluoroacetate was obtained from compounds 16f and 7A using the synthesis method described in Example 16.

[0329] 1 H NMR (400MHz, CD3OD) δ7.77–7.69(m,2H),7.59–7.47(m,2H),7.05–6.98(m,2H),6.76(d,1H),6.66–6.60(m,1 H),6.55(dd,1H),4.79–4.74(m,1H),4.28–4.18(m,2H),4.13(s,1H),3.93(s,3H),3.86–3.72(m,1H),3.67–3 .54(m,2H),3.51–3.40(m,1H),3.40–3.31(m,4H),3.27–3.15(m,1H),3.10–2.94(m,1H),2.89–2.74(m,4H),2 .74–2.63(m,1H),2.48–2.37(m,2H),2.36–2.25(m,1H),2.21–2.05(m,4H),1.88–1.70(m,5H),1.26(d,12H).

[0330] LCMS m / z = 860.4 [M+1] +

[0331] Example 20: Preparation of trifluoroacetate of compound 20

[0332] Step 1: Preparation of 20b

[0333] Trans-4-(3-amino-2,2,4,4-tetramethylcyclobutoxy)-2-methoxybenzyl nitrile hydrochloride (3.93 g, 12.64 mmol) (synthetic method see CN115175901) was dissolved in 30 mL of DMF, and DIPEA (5.84 g, 45.18 mmol), HATU (5.16 g, 13.57 mmol), and 20a (2.0 g, 9.04 mmol) were added. The reaction was carried out at room temperature under a nitrogen atmosphere for 16 h. 50 mL of ethyl acetate and 50 mL of purified water were added to the reaction solution. The aqueous phase was extracted with ethyl acetate (25 mL × 2). The organic phases were combined, washed with 25 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate (v / v) = 1:2) to give 20b (4.0 g, yield: 93%).

[0334] LCMS m / z = 478.3[M+1] +

[0335] Step 2: Preparation of 20C

[0336] 20b (4.0 g, 8.38 mmol) was dissolved in 80 mL of dichloromethane, and Dysmartin oxidant (4.27 g, 10.07 mmol) was added. The mixture was refluxed for 3 h. The reaction mixture was cooled to room temperature, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate (v / v) = 1:2) to give 20c (2.5 g, yield: 63%).

[0337] LCMS m / z = 476.2[M+1] +

[0338] Step 3: Preparation of compound 20 trifluoroacetate

[0339] Add 1-E (0.3 g, 0.72 mmol), 1 mL trifluoroacetic acid, and 3 mL dichloromethane to the reaction flask and react at room temperature for 3 h. Concentrate the reaction solution under reduced pressure, add triethylamine to adjust the pH to 7, concentrate under reduced pressure, add 5 mL N,N-dimethylacetamide to the residue, then add 20C (0.34 g, 0.72 mmol), 0.3 mL acetic acid, and sodium triacetoxyborohydride (0.31 g, 1.46 mmol) sequentially, and react at room temperature for 16 h. Add 15 mL of dichloromethane and 15 mL of saturated sodium bicarbonate aqueous solution to the reaction solution. Extract the aqueous phase with dichloromethane (15 mL × 3). Combine the organic phases, wash with 15 mL of saturated brine, dry with anhydrous sodium sulfate, concentrate under reduced pressure, and purify the crude product using a silica gel column (dichloromethane:methanol (v / v) = 10:1). Perform Pre-HPLC on the obtained crude product (instrument and preparative column: Waters 2767 preparative HPLC, SunFire@Prep C18, 5 μm, inner diameter × length = 19 mm × 250 mm). Preparation method: Filter the crude product's DMSO solution through a 0.45 μm filter membrane to prepare the sample solution. Mobile phase system: water (containing 0.1% TFA) / acetonitrile. Gradient elution method: Elute 50% of the compound 20 with 10% gradient elution (elution time 15 min), and lyophilize to obtain trifluoroacetate of compound 20 (50 mg).

[0340] 1H NMR(400MHz,DMSO-d6)δ10.75(s,1H),7.74(d,2H),7.64(d,1H),7.48(d,1H),6.97(d,2H),6.78 (d,1H),6.66–6.57(m,2H),6.54(dd,1H),4.27(s,1H),4.05(d,1H),3.97–3.85(m,5H),3.81(dd ,1H),3.77–3.68(m,1H),3.02–2.86(m,3H),2.86–2.75(m,2H),2.75–2.55(m,4H),2.50–2.39(m ,2H),2.35–2.22(m,1H),2.18–2.04(m,1H),2.03–1.80(m,5H),1.67–1.43(m,3H),1.19(d,12H).

[0341] LCMS m / z = 389.4 [M / 2+1] +

[0342] Example 21: Preparation of trifluoroacetate of compound 21

[0343] Compound 21 trifluoroacetate was obtained from compounds 20c and 2-D using the synthesis method described in Example 20.

[0344] 1 H NMR(400MHz,DMSO-d6)δ10.76(s,1H),7.79–7.70(m,2H),7.64(d,1H),7.48(d,1H),7.01–6.91 (m,2H),6.78(d,1H),6.67–6.49(m,3H),4.27(s,1H),4.05(d,1H),3.97–3.85(m,5H),3.81(dd, 1H),3.78–3.68(m,1H),3.01–2.86(m,3H),2.86–2.75(m,2H),2.75–2.54(m,4H),2.50–2.39(m, 2H),2.36–2.22(m,1H),2.18–2.04(m,1H),2.03–1.80(m,5H),1.67–1.43(m,3H),1.19(d,12H).

[0345] LCMS m / z = 777.4 [M+1] +

[0346] Example 22: Preparation of trifluoroacetate of compound 22

[0347] Step 1: Preparation of 22a

[0348] Add 3-A (0.5 g, 1.43 mmol), 2 mL trifluoroacetic acid, and 6 mL dichloromethane to a reaction flask and react at room temperature for 3 h. Concentrate the reaction solution under reduced pressure, add 5 mL N,N-dimethylacetamide to the residue, followed by 20c (0.68 g, 1.43 mmol), 0.3 mL acetic acid, and sodium triacetoxyborohydride (0.61 g, 2.88 mmol), and react at room temperature for 16 h. Add 15 mL dichloromethane and 15 mL saturated sodium bicarbonate aqueous solution to the reaction solution, extract the aqueous phase with dichloromethane (15 mL × 3), combine the organic phases, wash with 15 mL saturated brine, dry to anhydrous sodium sulfate, concentrate under reduced pressure, and purify the crude product by silica gel column chromatography (dichloromethane:methanol (v / v) = 10:1) to give 22a (0.4 g, yield: 39%).

[0349] LCMS m / z = 710.3[M+1] +

[0350] Step 2: Preparation of compound 22 trifluoroacetate

[0351] Add 22a (0.2 g, 0.28 mmol), EDCI (0.11 g, 0.57 mmol), HOBt (0.038 g, 0.28 mmol), and N-methylmorpholine (0.14 g, 1.38 mmol) to the reaction flask. Under nitrogen protection, add (S)-3-aminopiperidine-2,6-dione hydrochloride (0.092 g, 0.56 mmol). React at room temperature for 16 h under nitrogen atmosphere. Add 15 mL of dichloromethane and 15 mL of purified water to the reaction solution. Extract the aqueous phase with dichloromethane (15 mL × 3). Combine the organic phases, dry the organic phase with anhydrous sodium sulfate, concentrate under reduced pressure, and pass the crude product through Pre-HPLC (instrument and preparative column: Waters 2767 preparative HPLC, preparative column model: SunFire@Prep C18, 5 μm, inner diameter × length = 19 mm × 250 mm). Preparation method: The crude product was filtered through a 0.45 μm filter membrane to prepare the sample solution. Mobile phase system: water (containing 0.1% TFA) / acetonitrile. Gradient elution method: Acetonitrile was used to elute 50% by a 10% gradient (elution time 15 min), and the solution was lyophilized to obtain compound 22 trifluoroacetate (50 mg).

[0352] 1H NMR(400MHz,DMSO-d6)δ10.81(s,1H),7.93(t,1H),7.74(d,2H),7.64(d,1H),7.48(d,1H), 7.35(d,1H),6.97(d,2H),6.71–6.62(m,2H),6.54(dd,1H),4.78–4.61(m,1H),4.27(s,1H) ,4.05(d,1H),3.98–3.78(m,6H),3.13–3.02(m,1H),3.02–2.91(m,2H),2.87–2.62(m,6H), 2.57–2.41(m,2H),2.36–2.21(m,1H),2.17–1.80(m,6H),1.66–1.44(m,3H),1.19(d,12H).

[0353] LCMS m / z = 820.4 [M+1] +

[0354] The following compounds were obtained by synthesizing according to the methods described in other embodiments:

[0355] Example 24: Preparation of trifluoroacetate of compound 24

[0356] Compound 24 trifluoroacetate was obtained from compounds 20c and 4-A using the synthesis method described in Example 22.

[0357] 1 H NMR(400MHz,DMSO-d6)δ10.81(s,1H),7.98–7.88(m,1H),7.78–7.70(m,2H),7.68–7.61(m,1H) ,7.49(d,1H),7.35(d,1H),6.97(d,2H),6.73–6.61(m,2H),6.54(dd,1H),4.79–4.61(m,1H),4. 27(s,1H),4.05(d,1H),3.98–3.81(m,6H),3.14–3.03(m,1H),3.02–2.90(m,2H),2.87–2.61(m, 6H),2.58–2.42(m,2H),2.36–2.22(m,1H),2.17–1.81(m,6H),1.66–1.43(m,3H),1.19(d,12H).

[0358] LCMS m / z = 820.4 [M+1] +

[0359] Example 26: Preparation of Compound 26

[0360] Step 1: Preparation of 26b

[0361] Add 26a hydrochloride (5.0 g, 29.48 mmol), ethyl 4-fluorobenzoate (4.96 g, 29.50 mmol), potassium carbonate (10.19 g, 73.7 mmol), and 50 mL of dimethyl sulfoxide to the reaction flask, and react at 100 °C for 16 h. Cool the reaction system to room temperature, add 100 mL of water and 100 mL of ethyl acetate to the reaction solution, separate the layers, extract the aqueous phase with ethyl acetate (50 mL × 3), combine the organic phases, wash the organic phase with 50 mL of saturated brine, dry with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and purify the crude product by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1:0-1:1) to give 26b (5.0 g, yield: 60%).

[0362] Step 2: Preparation of 26c

[0363] Add 26b (5.0 g, 17.77 mmol), potassium hydroxide (4 g, 71.43 mmol), 50 mL ethanol, 50 mL tetrahydrofuran, and 50 mL water to the reaction flask, and react at room temperature for 16 h. Concentrate the reaction system under reduced pressure, adjust the pH to 2 with 2 mol / L hydrochloric acid, extract the aqueous phase with ethyl acetate (50 mL × 3), combine the organic phases, wash with 50 mL saturated brine, dry with anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure to obtain crude product 26c (3.0 g).

[0364] LCMS m / z = 254.1[M+1] +

[0365] Step 3: Preparation of 26 days

[0366] Add the above crude product 26c (0.82 g), HATU (2.46 g, 6.47 mmol), diisopropylethylamine (2.09 g, 16.17 mmol), and 10 mL DMF to the reaction flask. After stirring at room temperature for 0.5 h, add trans-4-(3-amino-2,2,4,4-tetramethylcyclobutoxy)-2-methoxybenzyl nitrile hydrochloride (1.0 g, 3.22 mmol) (synthetic method see CN115175901), and react at room temperature for 2 h. Add 100 mL water and 100 mL ethyl acetate to the reaction solution, separate the layers, extract the aqueous phase with ethyl acetate (50 mL × 3), combine the organic phases, wash the organic phase with 50 mL saturated brine, dry with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and purify the crude product by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1:0-0:1) to obtain 26d (1.5 g, yield: 91%).

[0367] LCMS m / z = 510.3[M+1] +

[0368] Step 4: Preparation of 26e

[0369] 26d (0.9 g, 1.77 mmol), Dess-Martin oxidant (0.9 g, 2.12 mmol), and 10 mL of dichloromethane were added to the reaction flask, and the reaction was carried out at room temperature for 4 h. The reaction solution was washed with 10 mL of saturated sodium bicarbonate aqueous solution, the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give crude product 26e (0.9 g).

[0370] Step 5: Preparation of Compound 26

[0371] Add the above crude product 26e (0.25 g), crude intermediate 1 (0.2 g), 0.2 mL glacial acetic acid, and 3 mL DMAc to the reaction flask. After reacting at room temperature for 1 h, add sodium triacetoxyborohydride (0.31 g, 1.46 mmol) and react at room temperature for 16 h. Pass the reaction solution through Pre-HPLC (instrument and preparative column: Waters 2767 preparative HPLC, SunFire@Prep C18, 5 μm, inner diameter × length = 19 mm × 250 mm). Preparation method: Filter the crude product's DMSO solution through a 0.45 μm filter membrane to prepare the sample solution. Mobile phase system: water (containing 10 mmol / L ammonium bicarbonate) / acetonitrile. Gradient elution method: Elute 50% of the product with acetonitrile in a 10% gradient (elution time 15 min), and lyophilize to obtain compound 26 (91 mg, two-step yield from compound 26d: 23%).

[0372] 1H NMR(400MHz,DMSO-d6)δ10.75(s,1H),7.75(d,2H),7.64(d,1H),7.48(d,1H),7.00(d,2H),6.79(d,1 H),6.67–6.58(m,2H),6.54(dd,1H),4.27(s,1H),4.05(d,1H),3.91(s,3H),3.85–3.77(m,1H),3.75 –3.59(m,3H),3.19–3.07(m,2H),3.04–2.89(m,3H),2.80–2.64(m,3H),2.63–2.52(m,4H),2.31–2.2 1(m,1H),2.18–2.05(m,1H),2.04–1.88(m,4H),1.88–1.68(m,3H),1.64–1.51(m,1H),1.18(d,12H).

[0373] LCMS m / z = 809.4 [M+1] +

[0374] Example 28: Preparation of Compound 28

[0375] Compound 28 was prepared from compounds 1-C and 15A by neutral preparation (ammonium bicarbonate aqueous solution (10 mmol / L) / acetonitrile) lyophilization, following the synthesis method of Example 15.

[0376] 1 H NMR(400MHz,DMSO-d6)δ10.33(s,1H),7.74(d,2H),7.64(d,1H),7.47(d,1H),7.00–6.90(m,3H),6 .72(d,1H),6.64(d,1H),6.54(dd,1H),4.27(s,1H),4.05(d,1H),3.91(s,3H),3.89–3.81(m,2H),3 .79–3.71(m,1H),3.60(t,2H),3.04–2.95(m,1H),2.94–2.85(m,2H),2.84–2.57(m,7H),2.19(d,2H ),2.11–2.00(m,1H),1.94–1.85(m,1H),1.85–1.71(m,4H),1.67–1.54(m,1H),1.28–1.10(m,14H).

[0377] LCMS m / z = 792.6 [M+1] +

[0378] Example 29: Preparation of compound 29

[0379] Compound 29 was prepared from compounds 2-B and 15A by neutral preparation (ammonium bicarbonate aqueous solution (10 mmol / L) / acetonitrile) lyophilization, following the synthesis method of Example 15.

[0380] 1 H NMR(400MHz,DMSO-d6)δ10.33(s,1H),7.77–7.70(m,2H),7.64(d,1H),7.47(d,1H),7.00–6.91(m,3H) ,6.76–6.68(m,1H),6.64(d,1H),6.54(dd,1H),4.27(s,1H),4.05(d,1H),3.91(s,3H),3.91–3.81(m, 2H),3.79–3.71(m,1H),3.60(t,2H),3.06–2.94(m,1H),2.94–2.85(m,2H),2.84–2.55(m,7H),2.19(d ,2H),2.11–2.00(m,1H),1.94–1.85(m,1H),1.85–1.71(m,4H),1.67–1.53(m,1H),1.29–1.11(m,14H).

[0381] Example 30: Preparation of compound 30

[0382] Step 1: Preparation of 30b trifluoroacetate

[0383] 30a (2.00 g, 5.06 mmol) (synthetic method according to US2023135173) was dissolved in 20 mL of dichloromethane, and 10 mL of trifluoroacetic acid was added. The reaction was carried out at room temperature for 3 h. The reaction system was concentrated under reduced pressure to obtain crude trifluoroacetate of 30b (3.00 g).

[0384] LCMS m / z = 296.2[M+1] +

[0385] Step 2: Preparation of 30C

[0386] Add 30A (1.43 g, 6.07 mmol) (synthetic method referred to CN115974840), HATU (2.87 g, 7.56 mmol), diisopropylethylamine (1.96 g, 15.18 mmol), and 10 mL of DMF to the reaction flask. After stirring at room temperature for 0.5 h, add the trifluoroacetate salt of the above crude product 30b (3.00 g) and stir at room temperature for 2 h. Add 100 mL of water to the reaction solution, filter, collect the filter cake, and purify the filter cake by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1:0-0:1) to obtain 30c (1.50 g, yield: 58%).

[0387] LCMS m / z = 513.2[M+1] +

[0388] Step 3: Preparation for 30 days

[0389] Add 30C (1.50 g, 2.93 mmol), Dess-Martin oxidant (2.49 g, 5.86 mmol), and 20 mL of dichloromethane to the reaction flask, and react at room temperature for 30 min. Filter the reaction mixture through a diatomaceous earth liner, and concentrate the filtrate under reduced pressure to obtain crude product 30d (2.00 g).

[0390] LCMS m / z = 511.3[M+1] +

[0391] Step 4: Preparation of Compound 30

[0392] Add crude product 30d (0.20 g), crude intermediate 1 (0.15 g), 0.2 mL glacial acetic acid, and 5 mL DMAc to the reaction flask. After reacting at room temperature for 1 h, add sodium triacetoxyborohydride (0.25 g, 1.17 mmol) and react at room temperature for 16 h. Pass the reaction solution through Pre-HPLC (instrument and preparative column: SHIMADZU LC-20AP preparative HPLC, column type C18, 5 μm, inner diameter × length = 19 mm × 250 mm). Preparation method: Filter the DMF solution of the crude product through a 0.45 μm filter membrane to prepare the sample solution. Mobile phase system: water (containing 10 mmol / L ammonium bicarbonate) / acetonitrile. Gradient elution method: Elute 82% acetonitrile with a 52% gradient (elution time 15 min), lyophilize to obtain compound 30 (16.2 mg, two-step yield from compound 30c: 7%).

[0393] 1H NMR(400MHz,DMSO-d6)δ10.76(s,1H),9.13–9.08(m,1H),8.74(dd,1H),8.30(d,1H),7.80–7. 69(m,3H),7.52(d,1H),6.96(d,3H),6.79(d,1H),6.62(d,1H),4.49(s,1H),4.15(d,1H),3.90 –3.76(m,3H),3.76–3.66(m,1H),3.00–2.84(m,3H),2.84–2.64(m,5H),2.64–2.54(m,1H),2. 54–2.45(m,1H),2.25–1.99(m,4H),1.99–1.68(m,6H),1.66–1.51(m,1H),1.35–1.11(m,14H).

[0394] LCMS m / z = 812.4[M+1] +

[0395] The following compounds were obtained by synthesizing according to the methods described in other embodiments:

[0396] Example 36: Preparation of compound 36

[0397] Step 1: Preparation of 36b

[0398] Dissolve 36a (10.00 g, 52.32 mmol) in 100 mL of DMF, add potassium carbonate (14.46 g, 104.64 mmol) and deuterated iodomethane (9.10 g, 62.78 mmol), and react at room temperature for 19 h. Pour the reaction solution into 200 mL of water, extract with 100 mL of ethyl acetate, dry the organic phase with anhydrous sodium sulfate, concentrate under reduced pressure, and purify the crude product by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 10:1-2:1) to obtain crude product 36b (11.5 g).

[0399] Step 2: Preparation of 36c

[0400] The crude product 36b (11.5 g) was dissolved in 100 mL of NMP, and cuprous cyanide (11.30 g, 126.17 mmol) was added. The mixture was reacted at 180 °C for 19 h. The reaction solution was cooled to room temperature, and 10 mL of concentrated ammonia and 200 mL of water were added. The mixture was extracted with 100 mL of ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 10:1-2:1) to give 36c (7.5 g, two-step yield from compound 36a: 93%).

[0401] Step 3: Preparation of 36 days

[0402] 36h (8.37 g, 34.38 mmol) was dissolved in 50 mL of THF, and 60% sodium hydride (1.65 g) was added at 0 °C. After reacting at 0 °C for 30 min, 36c (5.30 g, 34.38 mmol) was added, and the reaction was carried out at room temperature for 19 h. The reaction solution was poured into 100 mL of water and extracted with 100 mL of ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 10:1-2:1) to give 36d (2.5 g, yield: 19%).

[0403] LCMS m / z = 378.3[M+1] +

[0404] Step 4: Preparation of 36e hydrochloride

[0405] 36e (2.5 g, 6.62 mmol) was dissolved in 5 mL of ethyl acetate, and 10 mL of 4 mol / L ethyl hydrochloride solution was added. The mixture was reacted at room temperature for 5 h. The reaction mixture was filtered, and the filter cake was collected and dried under reduced pressure to obtain crude 36e hydrochloride (3.5 g).

[0406] LCMS m / z = 278.2[M+1] +

[0407] Step 5: Preparation of 36F

[0408] Add 30A (1.35 g, 5.74 mmol) (synthetic method referred to CN115974840), HATU (2.73 g, 7.17 mmol), diisopropylethylamine (1.85 g, 14.34 mmol), and 10 mL of DMF to the reaction flask. After stirring at room temperature for 0.5 h, add the hydrochloride salt of the above crude product 36e (2.5 g) and stir at room temperature for 2 h. Add 100 mL of water to the reaction solution, filter, collect the filter cake, and purify the filter cake by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1:0-0:1) to obtain 36f (1.6 g, two-step yield from compound 36d: 68%).

[0409] LCMS m / z = 495.3 [M+1] +

[0410] Step 6: Preparation of 36g

[0411] Add 36f (99 mg, 0.20 mmol), Dess-Martin oxidant (0.17 g, 0.40 mmol), and 3 mL of dichloromethane to the reaction flask, and react at room temperature for 30 min. Add 10 mL of saturated sodium bicarbonate aqueous solution to the reaction system, extract with 30 mL of dichloromethane, dry the organic phase with anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure to obtain 36 g (110 mg) of crude product.

[0412] Step 7: Preparation of Compound 36

[0413] 36 g (0.11 g) of the crude product, crude intermediate 1 (0.12 g), 0.1 mL of glacial acetic acid, and 5 mL of DMAc were added to the reaction flask. After reacting at room temperature for 1 h, sodium triacetoxyborohydride (0.13 g, 0.6 mmol) was added, and the reaction was continued at room temperature for 16 h. The reaction solution was then passed through Pre-HPLC (instrument and preparative column: SHIMADZU LC-20AP preparative HPLC, SunFire@Prep C18, 5 μm, inner diameter × length = 19 mm × 250 mm). Preparation method: The DMSO solution of the crude product was filtered through a 0.45 μm filter membrane to prepare the sample solution. Mobile phase system: water (containing 10 mmol / L NH4HCO3) / acetonitrile. Gradient elution method: Acetonitrile was used to elute 83% from 53% (elution time 15 min), and the solution was lyophilized to obtain compound 36 (46 mg, two-step yield from compound 36f: 29%).

[0414] 1 H NMR(400MHz, CDCl3)δ7.89(s,1H),7.68(d,2H),7.45(d,1H),6.91(d,2H),6.78– 6.65(m,1H),6.54–6.43(m,2H),6.40(dd,1H),6.10(d,1H),4.15(d,1H),4.04(s ,1H),3.93–3.71(m,3H),3.68–3.50(m,1H),3.16–2.99(m,1H),2.99–2.71(m,7H ),2.71–2.55(m,2H),2.35–2.07(m,5H),2.02–1.61(m,6H),1.39–1.17(m,14H).

[0415] LCMS m / z = 794.7 [M+1] +

[0416] The following compounds were obtained by synthesizing according to the methods described in other embodiments:

[0417] Example 40: Preparation of Compound 40

[0418] Compound 40 was obtained from compound 36g and 5A trifluoroacetate as raw materials, following the synthesis method described in Example 11.

[0419] 1 H NMR(400MHz,DMSO-d6)δ10.82(s,1H),7.94(t,1H),7.78–7.68(m,2H),7.64(d,1H),7.47 (d,1H),7.36(d,1H),6.95(d,2H),6.74–6.59(m,2H),6.54(dd,1H),4.79–4.65(m,1H),4. 27(s,1H),4.05(d,1H),3.93–3.76(m,3H),3.17–3.05(m,1H),3.00–2.61(m,8H),2.58–2 .44(m,1H),2.25–1.88(m,6H),1.87–1.70(m,4H),1.67–1.51(m,1H),1.28–1.11(m,14H).

[0420] LCMS m / z = 419.3[M / 2+1] +

[0421] Example 42: Preparation of compound 42

[0422] Compound 42 was obtained from the hydrochloride salt of compound 36e and 5d, using the synthesis method described in Example 36.

[0423] 1H NMR (400MHz, DMSO-d6) δ10.75(s,1H),7.72(d,2H),7.64(d,1H),7.47(d,1H),6.95(d,2H),6.78(d,1H),6. 65–6.57(m,2H),6.56–6.50(m,1H),4.26(s,1H),4.04(d,1H),3.81(dd,1H),3.74–3.62(m,1H),3.29–3.22 (m,2H),3.21–3.12(m,2H),2.99–2.78(m,3H),2.75–2.54(m,4H),2.54–2.45(m,2H),2.43–2.34(m,2H),2. 17–1.90(m,5H),1.89–1.72(m,2H),1.71–1.63(m,2H),1.63–1.50(m,3H),1.50–1.39(m,2H),1.18(d,12H).

[0424] LCMS m / z = 834.7 [M+1] +

[0425] The following compounds were obtained by synthesizing according to the methods described in other embodiments:

[0426] Example 48: Preparation of Compound 48

[0427] Compound 48 was obtained by using compound 48a (synthesis method referred to WO2021231927) and intermediate 1 as raw materials, and by referring to the synthesis method of Example 36.

[0428] 1 H NMR(400MHz,DMSO-d6)δ10.75(s,1H),7.90(d,1H),7.73(d,2H),7.45(d,1H),7.23–7.17(m,1 H),7.03–6.97(m,1H),6.95(d,2H),6.79(d,1H),6.62(d,1H),4.32(s,1H),4.05(d,1H),3.90 –3.77(m,3H),3.76–3.67(m,1H),2.99–2.85(m,3H),2.83–2.55(m,6H),2.54–2.45(m,1H),2. 21–2.14(m,2H),2.14–2.02(m,2H),1.98–1.73(m,6H),1.65–1.53(m,1H),1.26–1.08(m,14H).

[0429] LCMS m / z = 795.3 [M+1] +

[0430] The following compounds were obtained by synthesizing according to the methods described in other embodiments:

[0431] Example 54: Preparation of compound 54

[0432] Compound 54 was obtained from compound 54a hydrochloride (synthesis method referred to WO2021231927) and 5d as raw materials, according to the synthesis method of Example 42.

[0433] 1H NMR(400MHz,DMSO-d6)δ10.74(s,1H),7.90(d,1H),7.72(d,2H),7.45(d,1H),7.20(d,1H),7.04–6.90(m,3H),6.78 (d,1H),6.60(d,1H),4.32(s,1H),4.04(d,1H),3.81(dd,1H),3.74–3.62(m,1H),3.28–3.23(m,2H),3.20–3.15(m, 2H),2.97–2.79(m,3H),2.74–2.56(m,4H),2.55–2.51(m,2H),2.42–2.36(m,2H),2.14–2.03(m,2H),2.00–1.90(m, 3H),1.89–1.81(m,1H),1.81–1.73(m,1H),1.71–1.63(m,2H),1.62–1.50(m,3H),1.50–1.40(m,2H),1.17(d,12H).

[0434] LCMS m / z = 835.4 [M+1] +

[0435] The following compounds were obtained by synthesizing according to the methods described in other embodiments:

[0436] Example 60: Preparation of compound 60, trifluoroacetic acid

[0437] Step 1: Preparation of 60b

[0438] 60a (0.51 g, 2.15 mmol) (synthetic method referred to WO2023066350) was dissolved in 15 mL DMF. Under nitrogen protection, trans-4-(3-amino-2,2,4,4-tetramethylcyclobutoxy)-2-methoxybenzyl nitrile hydrochloride (1.00 g, 3.22 mmol) (synthetic method referred to CN115175901), N,N-diisopropylethylamine (1.39 g, 10.75 mmol) and HATU (0.98 g, 2.58 mmol) were added, and the reaction was carried out at room temperature for 16 h. Add 30 mL of ethyl acetate and 30 mL of water to the reaction solution, separate the liquid and extract the aqueous phase with ethyl acetate (15 mL × 3), combine the organic phases, wash the organic phase with 15 mL of saturated brine, dry with anhydrous sodium sulfate, concentrate under reduced pressure, and purify the crude product by silica gel column chromatography (petroleum ether: ethyl acetate (v / v) = 1:2) to give 60b (0.8 g, yield: 75%).

[0439] LCMS m / z = 494.2[M+1] +

[0440] Step 2: Preparation of 60C

[0441] 60b (0.8 g, 1.62 mmol) was dissolved in 15 mL of dichloromethane, and Dys-Martin oxidant (0.82 g, 1.94 mmol) was added. The reaction mixture was reacted at room temperature for 2 h. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate (v / v) = 1:2) to give 60c (0.6 g, yield: 75%).

[0442] LCMS m / z = 492.4 [M+1] +

[0443] Step 3: Preparation of compound 60 trifluoroacetate

[0444] Compound 60 trifluoroacetate was obtained from compound 60c and intermediate 1 using the synthesis method described in Example 1.

[0445] 1 H NMR(400MHz,DMSO-d6)δ10.78(s,1H),8.23(d,1H),7.86(d,1H),7.64(d,1H),7.42(d,1H),6.88(d,1H),6.80(d,1H ),6.67(d,1H),6.57(dd,1H),4.60–4.48(m,2H),4.40(s,1H),4.14–4.05(m,1H),4.04–3.98(m,1H),3.92(s,3H),3 .89–3.82(m,1H),3.70–3.57(m,2H),3.39–3.30(m,1H),3.19–2.98(m,6H),2.93–2.82(m,1H),2.81–2.62(m,3H),2 .56–2.52(m,1H),2.31–2.21(m,1H),2.20–2.06(m,1H),2.06–1.79(m,4H),1.70–1.56(m,1H),1.34–1.14(m,14H).

[0446] LCMS m / z = 793.8 [M+1] +

[0447] Example 62: Preparation of compound 62 trifluoroacetic acid

[0448] Compound 62 trifluoroacetate was obtained by acidic preparation (water (containing 0.1% TFA) / acetonitrile) freeze-drying, using compounds 60c and 28f as raw materials, following the synthesis method described in Example 15.

[0449] 1 H NMR (400MHz, DMSO-d6) δ10.36(s,1H),8.23(d,1H),7.86(d,1H),7.64(d,1H),7.42(d,1H),7.03(d,1H),6. 96–6.86(m,1H),6.67(d,1H),6.57(dd,1H),4.61–4.47(m,2H),4.40(s,1H),4.14–4.08(m,1H),4.04–3.98 (m,1H),3.91(s,3H),3.68–3.58(m,4H),3.43–3.33(m,1H),3.19–3.00(m,6H),2.95–2.82(m,1H),2.81–2. 61(m,4H),2.31–2.17(m,1H),2.08–1.97(m,1H),1.95–1.80(m,2H),1.72–1.57(m,1H),1.32–1.12(m,14H).

[0450] LCMS m / z = 794.6[M+1]+

[0451] Example 64: Preparation of compound 64 trifluoroacetic acid

[0452] Step 1: Preparation of 64b

[0453] 64a (1.12 g, 4.74 mmol) (synthetic method referred to WO2021249534A1) was dissolved in 30 mL DMF. Under nitrogen protection, trans-4-(3-amino-2,2,4,4-tetramethylcyclobutoxy)-2-methoxybenzyl nitrile hydrochloride (2.21 g, 7.11 mmol) (synthetic method referred to CN115175901), N,N-diisopropylethylamine (3.06 g, 23.70 mmol) and HATU (2.16 g, 5.69 mmol) were added, and the reaction was carried out at room temperature for 16 h. Add 50 mL of ethyl acetate and 50 mL of water to the reaction solution, separate the layers, extract the aqueous phase with ethyl acetate (15 mL × 3), combine the organic phases, wash the organic phase with 15 mL of saturated brine, dry with anhydrous sodium sulfate, concentrate under reduced pressure, and purify the crude product by silica gel column chromatography (petroleum ether: ethyl acetate (v / v) = 1:2) to obtain 64b (1.5 g, yield: 64%).

[0454] LCMS m / z = 493.2[M+1]+

[0455] Step 2: Preparation of 64C

[0456] 64b (1.5 g, 3.04 mmol) was dissolved in 30 mL of dichloromethane, and Dys-Martin oxidant (1.55 g, 3.65 mmol) was added. The reaction mixture was reacted at room temperature for 2 h. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate (v / v) = 1:2) to give 64c (1.4 g, yield: 93%).

[0457] LCMS m / z = 491.2[M+1] +

[0458] Step 3: Preparation of compound 64 trifluoroacetate

[0459] Compound 64 trifluoroacetate was obtained from compound 64c and intermediate 1 using the synthesis method described in Example 1.

[0460] 1 H NMR(400MHz,DMSO-d6)δ10.78(s,1H),8.63(d,1H),8.00(dd,1H),7.68–7.58(m,2H),6.97–6.91(m,1H) ,6.88(d,1H),6.80(d,1H),6.64(d,1H),6.54(dd,1H),4.48–4.37(m,2H),4.26(s,1H),4.13–4.02(m,2H ),3.91(s,3H),3.89–3.82(m,1H),3.68–3.57(m,2H),3.40–3.30(m,1H),3.19–2.81(m,7H),2.81–2.58( m,3H),2.57–2.45(m,1H),2.27–2.07(m,2H),2.05–1.79(m,4H),1.70–1.57(m,1H),1.27–1.10(m,14H).

[0461] LCMS m / z = 396.8 [M / 2+1] +

[0462] The following compounds were obtained by synthesizing according to the methods described in other embodiments:

[0463] Example 72: Preparation of compound 72

[0464] Step 1: Preparation of 72a

[0465] Add 5d (1.00 g, 3.63 mmol), HATU (2.07 g, 5.45 mmol), diisopropylethylamine (1.41 g, 10.89 mmol), and 8 mL of DMF to the reaction flask. Stir at room temperature for 0.5 h, then add the crude product 30b trifluoroacetate (1.0 g) and stir at room temperature for 2 h. Add 100 mL of water to the reaction solution, filter, and purify the filter cake by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1:0-0:1) to obtain 72a (0.97 g, yield: 48%).

[0466] LCMS m / z = 553.4 [M+1] +

[0467] Step 2: Preparation of 72b

[0468] Add 72a (0.97 g, 1.76 mmol), Dess-Martin oxidant (1.49 g, 3.52 mmol), and 20 mL of dichloromethane to the reaction flask, and react at room temperature for 30 min. Filter the reaction mixture through a diatomaceous earth liner, and concentrate the filtrate under reduced pressure to obtain crude product 72b (1.50 g).

[0469] LCMS m / z = 551.3[M+1] +

[0470] Step 3: Preparation of Compound 72

[0471] Add the above crude product 72b (0.10 g), crude intermediate 1 (0.15 g), 0.2 mL glacial acetic acid, and 5 mL DMAc to the reaction flask. After reacting at room temperature for 1 h, add sodium triacetoxyborohydride (0.25 g, 1.17 mmol) and react at room temperature for 16 h. Pass the reaction solution through Pre-HPLC (instrument and preparative column: SHIMADZU LC-20AP preparative HPLC, column model: SunFire@Prep C18, 5 μm, inner diameter × length = 19 mm × 250 mm). Preparation method: Filter the crude product's DMSO solution through a 0.45 μm filter membrane to prepare the sample solution. Mobile phase system: water (containing 10 mmol / L NH4HCO3) / acetonitrile. Gradient elution method: Acetonitrile was eluted from 83% to 53% (elution time 15 min), and lyophilized to give compound 72 (27.0 mg, two-step yield from compound 72a: 27%).

[0472] 1H NMR(400MHz,DMSO-d6)δ10.76(s,1H),9.10(dd,1H),8.74(dd,1H),8.30(d,1H),7.80–7.68(m,3H),7 .54(d,1H),7.01–6.90(m,3H),6.78(d,1H),6.61(d,1H),4.49(s,1H),4.15(d,1H),3.81(dd,1H),3.7 4–3.63(m,1H),3.30–3.24(m,4H),3.22–3.14(m,2H),2.98–2.78(m,3H),2.78–2.46(m,5H),2.44–2. 35(m,2H),2.19–1.72(m,7H),1.72–1.63(m,2H),1.63–1.51(m,2H),1.50–1.41(m,2H),1.26(d,12H).

[0473] LCMS m / z = 426.8 [M / 2+1] +

[0474] Example 78: Preparation of Compound 78

[0475] Compound 78 was prepared by neutral preparation (ammonium bicarbonate aqueous solution (10 mmol / L) / acetonitrile) and lyophilization, using compound 78a (synthetic method referred to WO2021127443) and intermediate 2 as raw materials, following the synthesis method of Example 1.

[0476] 1 H NMR(400MHz,DMSO-d6)δ10.75(s,1H),7.73(d,2H),7.44(d,1H),6.95(d,2H),6.79(d,1 H),6.73(s,2H),6.62(d,1H),4.22(s,1H),4.03(d,1H),3.90–3.76(m,3H),3.76–3.67( m,1H),3.00–2.85(m,3H),2.84–2.64(m,5H),2.64–2.54(m,1H),2.54–2.46(m,1H),2.4 3(s,6H),2.25–2.00(m,4H),1.99–1.70(m,6H),1.67–1.52(m,1H),1.26–1.07(m,14H).

[0477] LCMS m / z = 789.4 [M+1] +

[0478] The following compounds were obtained by synthesizing according to the methods described in other embodiments:

[0479] Example 83: Preparation of Compound 83

[0480] Step 1: Preparation of 83b

[0481] 83a (3.3 g, 8.96 mmol) (synthetic method referred to Tetrahedron Letters, 2018, 59, 2030–2033) was dissolved in 20 mL of methanol, and 10% palladium on carbon (2.38 g) was added. The mixture was purged with hydrogen three times, and the reaction was carried out at 45 °C for 20 h under a hydrogen atmosphere. The reaction system was cooled to room temperature, filtered, and the filtrate was collected. The filtrate was concentrated under reduced pressure to obtain crude product 83b (2.55 g).

[0482] LCMS m / z = 290.2[M+1] +

[0483] Step 2: Preparation of 83C

[0484] The crude product 83b (2.55 g) was dissolved in 20 mL of acetonitrile, and NBS (1.54 g, 8.64 mmol) was added. The mixture was stirred at room temperature for 1 h. The reaction system was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (ethyl acetate: petroleum ether (v / v) = 0:1-1:9) to obtain 83c (3.1 g, yield: 98%).

[0485] Step 3: Preparation of 83d

[0486] 83C (1.00 g, 2.72 mmol), 2,6-bisbenzyloxypyridine-3-borate pinacol ester (1.70 g, 4.08 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (0.22 g, 0.27 mmol), and cesium carbonate (1.77 g, 5.44 mmol) were added to the reaction flask, nitrogen gas was purged three times, 30 mL of 1,4-dioxane and 3 mL of water were added, and the reaction was carried out at 100 °C for 20 h under a nitrogen atmosphere. The reaction system was cooled to room temperature, and 50 mL of water and 50 mL of ethyl acetate were added. The mixture was separated, and the aqueous phase was extracted with 50 mL of ethyl acetate. The organic phases were combined, washed with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1:0-9:1) to give 83d (1.10 g, yield: 70%).

[0487] LCMS m / z = 579.3 [M+1] +

[0488] Step 4: Preparation of 83e

[0489] 83d (1.10 g, 1.90 mmol) was dissolved in 20 mL of tetrahydrofuran, and 10% palladium on carbon (1.0 g) was added. The hydrogen gas was replaced three times, and the reaction was carried out at 45 °C for 20 h under a hydrogen atmosphere. The reaction system was cooled to room temperature, filtered through a diatomaceous earth liner, and the filter cake was washed with 30 mL of tetrahydrofuran. The filtrate was collected, and the filter cake was dried under reduced pressure to obtain 83e (0.70 g, yield: 92%).

[0490] LCMS m / z = 401.2[M+1] +

[0491] Step 5: Preparation of 83f

[0492] Dissolve 200 mg (0.50 mmol) of 83e in 2 mL of dichloromethane, add 2 mL of trifluoroacetic acid, and react at room temperature for 3 h. Concentrate the reaction system under reduced pressure, add 5 mL of dichloromethane and 1 mL of triethylamine, and concentrate under reduced pressure to obtain crude 83f (150 mg).

[0493] LCMS m / z = 301.3[M+1] +

[0494] Step 6: Preparation of Compound 83

[0495] Compound 83 was obtained by using crude products 83f and 1a as raw materials, following the synthesis method of Example 1, and by neutral preparation (ammonium bicarbonate aqueous solution (10 mmol / L) / acetonitrile) freeze-drying.

[0496] 1 H NMR(400MHz,DMSO-d6)δ10.76(s,1H),7.80–7.69(m,3H),7.64(d,1H),7.46(d,1H),7.11–7.04(m,1H ),7.00–6.90(m,2H),6.64(d,1H),6.54(dd,1H),4.65–4.49(m,1H),4.27(s,1H),4.05(d,1H),3.93– 3.79(m,5H),3.68(dd,1H),3.27–3.17(m,1H),2.99–2.86(m,2H),2.85–2.56(m,6H),2.56–2.50(m,1 H),2.25–2.08(m,3H),2.02–1.87(m,3H),1.87–1.70(m,4H),1.66–1.52(m,1H),1.29–1.08(m,14H).

[0497] LCMS m / z = 774.5 [M+1] +

[0498] Example 85: Preparation of Compound 85

[0499] Compound 85 was prepared by neutral preparation (ammonium bicarbonate aqueous solution (10 mmol / L) / acetonitrile) and lyophilization, using compound 36e hydrochloride and compound 60a as raw materials, following the synthesis method of Example 60.

[0500] 1 H NMR(400MHz,DMSO-d6)δ10.76(s,1H),8.24(d,1H),7.82(d,1H),7.64(d,1H),7.36( d,1H),6.79(d,1H),6.69–6.52(m,3H),4.55–4.44(m,2H),4.40(s,1H),4.00(d,1H), 3.81(dd,1H),3.76–3.67(m,1H),3.09–2.84(m,5H),2.79–2.54(m,4H),2.54–2.44( m,1H),2.24–2.00(m,4H),1.99–1.70(m,6H),1.66–1.52(m,1H),1.28–1.05(m,14H).

[0501] LCMS m / z = 796.6 [M+1] +

[0502] The following compounds were obtained by synthesizing according to the methods described in other embodiments:

[0503] Example 89: Preparation of Compound 89

[0504] Compound 89 was prepared by neutral preparation (water (containing 10 mmol / L ammonium bicarbonate) / acetonitrile) lyophilization, using compound 30b trifluoroacetate and compound 60a as raw materials, following the synthesis method of Example 60.

[0505] 1H NMR(400MHz,DMSO-d6)δ10.75(s,1H),9.10(d,1H),8.76(d,1H),8.33–8.24(m,2H),7.83(d,1H),7.79–7 .72(m,1H),7.36(d,1H),7.01(d,1H),6.79(d,1H),6.62(d,1H),4.61(s,1H),4.56–4.44(m,2H),4.12(d, 1H),3.85–3.77(m,1H),3.76–3.69(m,1H),3.10–2.85(m,5H),2.78–2.64(m,3H),2.64–2.45(m,2H),2.23 –2.15(m,2H),2.14–2.01(m,2H),1.99–1.72(m,6H),1.67–1.53(m,1H),1.28(d,12H),1.20–1.07(m,2H).

[0506] LCMS m / z = 814.6 [M+1] +

[0507] Example 93: Preparation of Compound 93

[0508] Compound 93 was obtained from compound 93a (synthetic method referred to in Tetrahedron Letters, 2018, 59, 2030–2033) as a starting material, following the synthetic method described in Example 83.

[0509] 1 H NMR(400MHz,DMSO-d6)δ10.76(s,1H),7.80–7.67(m,3H),7.64(d,1H),7.46(d,1H),7.11–7.03(m, 1H),6.95(d,2H),6.64(d,1H),6.54(dd,1H),4.66–4.49(m,1H),4.27(s,1H),4.05(d,1H),3.93–3. 78(m,5H),3.68(dd,1H),3.29–3.15(m,1H),2.99–2.87(m,2H),2.86–2.56(m,6H),2.56–2.45(m,1H ),2.25–2.06(m,3H),2.02–1.87(m,3H),1.87–1.69(m,4H),1.66–1.52(m,1H),1.29–1.08(m,14H).

[0510] The following compounds were obtained by synthesizing according to the methods described in other embodiments:

[0511] Example 97: Preparation of compound 97 trifluoroacetic acid

[0512] Step 1: Preparation of 97b

[0513] Add 97a (5.0 g, 36.47 mmol), 1-bromo-2-butyne (9.7 g, 72.94 mmol), potassium carbonate (10.08 g, 72.94 mmol), potassium iodide (0.61 g, 3.67 mmol), and 100 mL of acetone to the reaction flask, and react at room temperature for 12 h. Add 200 mL of water to the reaction system, extract with ethyl acetate (100 mL × 3), combine the organic phases, dry to anhydrous sodium sulfate, concentrate under reduced pressure, and purify the crude product by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1:0-1:1) to obtain 97b (5.0 g, yield: 72%).

[0514] The trifluoroacetate of compound 97 was obtained by acidic preparation (water (containing 0.1% TFA) / acetonitrile) freeze-drying, using compounds 97b, 36h and intermediate 2 as raw materials, referring to the synthesis method of Example 36.

[0515] 1 H NMR (400MHz, DMSO-d6) δ10.78(s,1H),7.76(d,2H),7.64(d,1H),7.46(d,1H),6.99(d,2H),6.88(d,1H),6. 80(d,1H),6.69(d,1H),6.60(dd,1H),5.00–4.92(m,2H),4.25(s,1H),4.10–4.03(m,2H),3.95–3.78(m,3H ),3.68–3.57(m,2H),3.41–3.28(m,1H),3.21–2.97(m,4H),2.93–2.77(m,3H),2.77–2.60(m,3H),2.58–2. 48(m,1H),2.20–2.06(m,2H),2.05–1.91(m,2H),1.91–1.76(m,5H),1.71–1.56(m,1H),1.38–1.09(m,14H).

[0516] LCMS m / z = 829.6 [M+1] +

[0517] The following compounds were obtained by synthesizing according to the methods described in other embodiments:

[0518] Example 101: Preparation of trifluoroacetate of compound 101

[0519] Step 1: Preparation of 101b

[0520] 101a (10 g, 72.93 mmol), 2-cyclopropoxyethyl 4-methylbenzenesulfonic acid (22.62 g, 88.25 mmol), and cesium carbonate (35.64 g, 109.38 mmol) were dissolved in 100 mL of DMF and reacted at 90 °C for 16 h. The reaction mixture was cooled to room temperature and added to 300 mL of water. The mixture was extracted with ethyl acetate (200 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate (v / v) = 1:0-1:1) to give 101b (10 g, yield: 62%).

[0521] The trifluoroacetate of compound 101 was obtained by acidic preparation (water (containing 0.1% TFA) / acetonitrile) freeze-drying, using compounds 101b, 36h and intermediate 2 as raw materials, following the synthesis method of Example 36.

[0522] 1 H NMR(400MHz,DMSO-d6)δ10.79(s,1H),7.76(d,2H),7.64(d,1H),7.48(d,1H),6.99(d,2H),6.88(d,1H) ,6.80(d,1H),6.65(d,1H),6.55(dd,1H),4.29–4.20(m,3H),4.13–3.99(m,2H),3.95–3.74(m,5H),3.70 –3.56(m,2H),3.46–3.38(m,1H),3.38–3.28(m,1H),3.24–2.98(m,4H),2.93–2.59(m,6H),2.56–2.46( m,1H),2.21–2.05(m,2H),2.05–1.75(m,4H),1.70–1.55(m,1H),1.37–1.08(m,14H),0.55–0.38(m,4H).

[0523] LCMS m / z = 861.6 [M+1] +

[0524] Example 105: Preparation of compound 105

[0525] Step 1: Preparation of 105b

[0526] 105a (1.6 g, 11.67 mmol), 1-bromo-2-methoxyethane (3.80 g, 15.18 mmol), potassium carbonate (4.84 g, 35.02 mmol), potassium iodide (0.19 g, 1.14 mmol), and 15 mL of DMF were added to the reaction flask, and the reaction was carried out at room temperature for 12 h. 100 mL of water was added to the reaction system, and the mixture was extracted with ethyl acetate (100 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1:0–3:1) to give 105b (1.1 g, yield: 48%).

[0527] LCMS m / z = 196.1 [M+1] +

[0528] Compound 105 was obtained from compounds 105b and 36h using the synthesis method described in Example 36.

[0529] 1 H NMR(400MHz,DMSO-d6)δ10.76(s,1H),7.73(d,2H),7.64(d,1H),7.46(d,1H),6.95(d,2H),6. 79(d,1H),6.70–6.58(m,2H),6.54(dd,1H),4.30–4.21(m,3H),4.04(d,1H),3.90–3.77(m,3H) ,3.76–3.66(m,3H),3.34(s,3H),3.00–2.85(m,3H),2.83–2.64(m,5H),2.64–2.54(m,1H),2. 54–2.46(m,1H),2.23–1.99(m,4H),1.99–1.72(m,6H),1.66–1.52(m,1H),1.27–1.09(m,14H).

[0530] LCMS m / z = 835.3 [M+1] +

[0531] The following compounds were obtained by synthesizing according to the methods described in other embodiments:

[0532] Example 109: Preparation of compound 109

[0533] Step 1: Preparation of 109b

[0534] 109a (5.0 g, 36.47 mmol), (bromomethyl)cyclopropane (6.4 g, 47.41 mmol), potassium carbonate (15.12 g, 109.41 mmol), potassium iodide (0.61 g, 3.67 mmol), and 50 mL of DMF were added to the reaction flask, and the reaction was carried out at room temperature for 12 h. 200 mL of water was added to the reaction system, and the mixture was extracted with ethyl acetate (100 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1:0–3:1) to give 109b (6.2 g, yield: 89%).

[0535] LCMS m / z = 192.1 [M+1] +

[0536] Compound 109 was obtained from compounds 109b and 36h using the synthesis method described in Example 36.

[0537] 1 H NMR(400MHz,DMSO-d6)δ10.76(s,1H),7.73(d,2H),7.63(d,1H),7.46(d,1H),6.95(d,2H),6.79( d,1H),6.66–6.58(m,2H),6.52(dd,1H),4.25(s,1H),4.04(d,1H),3.98(d,2H),3.89–3.77(m,3H ),3.76–3.67(m,1H),3.01–2.84(m,3H),2.83–2.64(m,5H),2.64–2.52(m,2H),2.24–1.98(m,4H) ,1.98–1.71(m,6H),1.66–1.51(m,1H),1.32–1.06(m,15H),0.64–0.57(m,2H),0.40–0.34(m,2H).

[0538] LCMS m / z = 831.5[M+1] +

[0539] Example 113: Preparation of compound 113 trifluoroacetic acid

[0540] Step 1: Preparation of 113b

[0541] 113a (5 g, 24.63 mmol) was dissolved in 100 mL of tetrahydrofuran, cooled to -78 °C under nitrogen protection, and 24.63 mL of a 1 mol / L tetrahydrofuran solution of cyclopropylmagnesium bromide was added dropwise. The mixture was slowly heated to room temperature and reacted for 3 h. 100 mL of saturated ammonium chloride aqueous solution was added to the reaction solution, and the mixture was extracted with ethyl acetate (100 mL × 3). The organic phases were combined, washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate (v / v) = 2:1) to obtain crude product 113b (3.3 g).

[0542] Step 2: Preparation of 113c

[0543] The crude product 113b (4.5 g) was dissolved in 20 mL of dichloromethane, and triethylsilane (3.20 g, 27.52 mmol) and trifluoroacetic acid (12.56 g, 110.16 mmol) were added at 0 °C. The mixture was reacted at 40 °C for 2 h. The reaction solution was cooled to room temperature, and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate (v / v) = 5:1) to give 113c (4.0 g, two-step yield from compound 113a: 52%).

[0544] Step 3: Preparation of 113d

[0545] 113c (4.0 g, 17.46 mmol) was added to 40 mL of DMF. Under nitrogen protection, zinc cyanide (3.08 g, 26.23 mmol) and tetraphenylphosphine palladium (2.02 g, 1.75 mmol) were added, and the reaction was carried out at 110 °C for 2 h. The reaction solution was cooled to room temperature, and 60 mL of water and 60 mL of ethyl acetate were added. The mixture was separated, and the aqueous phase was extracted with ethyl acetate (50 mL × 3). The organic phases were combined, washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate (v / v) = 5:1) to give 113d (2.2 g, yield: 72%).

[0546] LCMS m / z = 176.1 [M+1] +

[0547] The trifluoroacetate of compound 113 was obtained by acidic preparation (water (containing 0.1% TFA) / acetonitrile) freeze-drying using compounds 113d and 36h as raw materials, following the synthesis method of Example 36.

[0548] 1H NMR(400MHz,DMSO-d6)δ10.79(s,1H),7.81–7.65(m,3H),7.50(d,1H),7.04–6.93(m,3H),6.91–6.83 (m,2H),6.80(d,1H),4.25(s,1H),4.12–4.00(m,2H),3.96–3.79(m,3H),3.68–3.55(m,2H),3.41–3.2 7(m,1H),3.19–2.98(m,4H),2.93–2.59(m,8H),2.57–2.46(m,1H),2.21–2.05(m,2H),2.05–1.75(m, 4H),1.70–1.54(m,1H),1.38–1.09(m,14H),1.09–0.96(m,1H),0.57–0.47(m,2H),0.31–0.24(m,2H).

[0549] LCMS m / z = 408.4 [M / 2+1] +

[0550] The following compounds were obtained by synthesizing according to the methods described in other embodiments:

[0551] Example 117: Preparation of Compound 117

[0552] Step 1: Preparation of 117b

[0553] 117a (5.00 g, 25.0 mmol), cyclopropylboronic acid (2.79 g, 32.48 mmol), palladium acetate (0.56 g, 2.49 mmol), tricyclohexylphosphine (0.76 g, 2.71 mmol), potassium phosphate (15.92 g, 75.0 mmol), and 100 mL of toluene were added to the reaction flask, and the reaction was carried out at 100 °C for 3 h under nitrogen protection. The reaction system was cooled to room temperature, filtered through a diatomaceous earth liner, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1:0-9:1) to obtain 117b (3.8 g, yield: 94%).

[0554] Compound 117 was obtained from compounds 117b and 36h using the synthesis method described in Example 36.

[0555] 1H NMR(400MHz,DMSO-d6)δ10.75(s,1H),7.79–7.60(m,3H),7.45(d,1H),6.95(d,2H),6 .79(dd,2H),6.62(d,1H),6.45(d,1H),4.24(s,1H),4.03(d,1H),3.93–3.76(m,3H),3 .76–3.64(m,1H),3.00–2.84(m,3H),2.84–2.55(m,6H),2.55–2.46(m,1H),2.25–2.00 (m,5H),1.99–1.70(m,6H),1.68–1.52(m,1H),1.29–1.05(m,16H),0.86–0.74(m,2H).

[0556] LCMS m / z = 801.4[M+1] +

[0557] Example 119: Preparation of compound 119

[0558] Compound 119 was obtained by using compound 117f as a starting material and following the synthesis method of Example 5, through neutral preparation (10 mmol / L ammonium bicarbonate aqueous solution / acetonitrile) lyophilization.

[0559] 1 H NMR(400MHz,DMSO-d6)δ10.82(s,1H),7.94(t,1H),7.79–7.63(m,3H),7.46(d,1H),7.36(d,1H), 6.95(d,2H),6.79(dd,1H),6.68(d,1H),6.45(d,1H),4.79–4.66(m,1H),4.24(s,1H),4.03(d,1H ),3.95–3.74(m,3H),3.17–3.05(m,1H),2.98–2.62(m,8H),2.58–2.48(m,1H),2.25–1.97(m,6H) ,1.97–1.87(m,1H),1.87–1.69(m,4H),1.68–1.51(m,1H),1.25–1.07(m,16H),0.85–0.75(m,2H).

[0560] LCMS m / z = 844.5[M+1]+

[0561] The following compounds were obtained by synthesizing according to the methods described in other embodiments:

[0562] Example 127: Preparation of Compound 127

[0563] Step 1: Preparation of 127b

[0564] 127a (4.98 g, 18.35 mmol) (synthetic method according to CN112912376) was dissolved in 20 mL of dichloromethane, and 10 mL of trifluoroacetic acid was added. The reaction was carried out at room temperature for 3 h. The reaction solution was concentrated under reduced pressure, dissolved in 50 mL of DMSO, and DIPEA (11.86 g, 91.76 mmol) and tert-butyl 4-fluorobenzoate (3.6 g, 18.35 mmol) were added separately. The reaction was carried out at 100 °C for 19 h. The reaction solution was cooled to room temperature, and 100 mL of water and 100 mL of ethyl acetate were added. The mixture was separated, and the aqueous phase was extracted with ethyl acetate (80 mL × 3). The organic phases were combined, washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate (v / v) = 5:1-1:1) to give 127b (2.75 g, yield: 43%).

[0565] Step 2: Preparation of 127c

[0566] 127b (0.4 g, 1.15 mmol) was dissolved in 2 mL of dichloromethane, and triethylamine (0.58 g, 5.73 mmol) was added. The system was cooled to -50 °C and trifluoromethanesulfonic anhydride (0.39 g, 1.38 mmol) was slowly added dropwise. The mixture was then allowed to return to room temperature and reacted for 3 h. The reaction solution was concentrated under reduced pressure to obtain crude 127c (1.0 g).

[0567] Step 3: Preparation of 127 days

[0568] The crude product 127c (1.0 g) was dissolved in 5 mL of acetonitrile, and DIPEA (0.59 g, 4.6 mmol) and crude intermediate 1 (0.30 g) were added separately. The reaction was carried out at room temperature for 19 h. 10 mL of water and 20 mL of ethyl acetate were added to the reaction solution, and the mixture was separated. The aqueous phase was extracted with ethyl acetate (80 mL × 3), and the organic phases were combined. The organic phase was washed with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate (v / v) = 5:1-2:1) to give 127d (0.45 g, two-step yield from compound 127b: 60%).

[0569] Step 4: Preparation of 127e hydrochloride

[0570] 0.2 g (0.31 mmol) of 127e was dissolved in 2 mL of dichloromethane, and 5 mL of 4 mol / L ethyl acetate hydrochloride solution was added. The mixture was reacted at room temperature for 5 h. The reaction solution was concentrated under reduced pressure to give 0.22 g of crude 127e hydrochloride.

[0571] LCMS m / z = 591.3 [M+1] +

[0572] Step 5: Preparation of Compound 127

[0573] The crude product 127e hydrochloride (0.22 g) was dissolved in 5 mL of DMF, and DIPEA (0.2 g, 1.55 mmol), HATU (0.18 g, 0.47 mmol), and 123A hydrochloride (96 mg, 0.31 mmol) were added separately. The reaction was carried out at room temperature under a nitrogen atmosphere for 16 h. 50 mL of ethyl acetate and 50 mL of purified water were added to the reaction solution. The aqueous phase was extracted with ethyl acetate (25 mL × 2). The organic phases were combined, washed with 25 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate (v / v) = 1:2) to give compound 127 (110 mg, two-step yield from compound 127d: 42%).

[0574] 1 H NMR(400MHz, CDCl3)δ7.88(s,1H),7.67(d,2H),7.45(d,1H),6.96–6.86(m,2H),6.72(d,1H) ,6.53–6.43(m,2H),6.40(dd,1H),6.10(d,1H),4.33–4.20(m,1H),4.18–4.10(m,1H),4.04(s ,1H),3.91(s,3H),3.84–3.72(m,1H),3.71–3.56(m,3H),3.56–3.46(m,1H),3.12–2.71(m,9H ),2.70–2.59(m,2H),2.32–2.07(m,6H),2.03–1.84(m,4H),1.79–1.62(m,4H),1.24(d,12H).

[0575] LCMS m / z = 847.4 [M+1] +

[0576] The following compounds were obtained by synthesizing according to the methods described in other embodiments:

[0577] Formulation Examples

[0578] Formulation Example 1 (100mg / tablet):

[0579] Preparation method: Compound 11 is pulverized and passed through a 60-mesh sieve. It is then mixed thoroughly with microcrystalline cellulose, crospovidone, and hydroxypropyl methylcellulose in a multi-directional motion mixer. Finally, magnesium stearate is added, and mixing continues for 5 minutes. The resulting powder mixture is then compressed into tablets using a high-speed tableting machine.

[0580] Formulation Example 2 (200mg / tablet):

[0581] Preparation method: Compound 11 is pulverized and passed through a 60-mesh sieve. It is then mixed with lactose, crospovidone, and povidone in a multi-directional motion mixer until homogeneous. Finally, magnesium stearate is added, and mixing continues for 5 minutes. The resulting powder mixture is then compressed into tablets using a high-speed tableting machine.

[0582] Biological test cases

[0583] 1. Detection of AR degradation in VCap cells

[0584] VCap is a human prostate cancer cell line purchased from ATCC. Culture conditions: DMEM + 10% FBS + 1% antibiotics, cultured at 37°C in a 5% CO2 incubator. Cells were seeded into 6-well plates at 5 × 10⁶ cells / well. 5 Cells / well. After plating, different concentrations of compounds were added, and the cells were incubated at 37°C and 5% CO2 for 24 hours. After the culture, the cells were collected, and RIPA lysis buffer (beyotime, Cat. P0013B) was added and lysed on ice for 15 minutes. After centrifugation at 12000 rpm and 4°C for 10 minutes, the supernatant protein sample was collected. After protein quantification using the BCA kit (Beyotime, Cat. P0009), the protein was diluted to 0.25 mg / mL, and the expression of AR (CST, Cat. 5153S) and internal control β-actin (CST, Cat. 3700S) was detected using a fully automated Western blot quantitative analyzer (Proteinsimple). The relative peak area of ​​AR when the internal control area was 10000 was calculated using the "Compass for SW" software. The proportion of AR relative to the solvent control group at different drug concentrations was calculated according to Equation (1), where AR treat AR represents the relative peak area of ​​the treatment group. solvent The relative peak area is for the solvent control group. The data processed according to equation (1) were analyzed using GraphPad Prism 8.3.0 software, and a four-parameter nonlinear regression model was employed to calculate DC. 50Value; the degradation rate at a specific concentration is measured as 100-AR%; AR% = AR treat / AR solvent ×100% Formula 1

[0585] DCs that degrade AR proteins in VCaP cells 50 The results are shown in Table 1.

[0586] Table 1. DCs of the compound of the present invention degrading AR protein in VCaP cells 50 value

[0587] Table 2. Degradation rate of AR protein in VCaP cells by the compounds of the present invention at a concentration of 30 nM.

[0588] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good AR degradation activity.

[0589] 2. Rat pharmacokinetic assay

[0590] Experimental objective: This experiment aimed to evaluate the pharmacokinetic characteristics of the test substance in rats by administering a single dose of the test substance intravenously and by gavage to SD rats, measuring the concentration of the test substance in rat plasma, and administering it intravenously and by gavage.

[0591] Experimental animals: Male SD rats, 200-220g, 6 rats / compound. Purchased from Chengdu Dashuo Experimental Animal Co., Ltd.

[0592] Experimental method: On the day of the experiment, 6 SD rats were randomly divided into groups according to their body weight. They were fasted for 12-14 hours before administration but allowed free access to water. They were fed 4 hours after administration.

[0593] Table 3. Dosage information for rat pharmacokinetic tests

[0594] *Dosage is calculated based on free base.

[0595] Sampling: Before and after administration of isoflurane anesthesia, 0.1 mL of blood was collected via the orbital cavity and placed in an EDTAK2 centrifuge tube. The plasma was collected by centrifugation at 5000 rpm and 4°C for 10 min.

[0596] Plasma collection time points for IV&PO groups: 0, 5 min, 15 min, 30 min, 1, 2, 4, 7, 24, 48 h.

[0597] All samples were stored at -60℃ before analysis and testing. Quantitative analysis of the samples was performed using LC-MS / MS.

[0598] Table 4. Pharmacokinetic parameters of the compounds of the present invention in rat plasma

[0599] # The oral solvent is 5% DMSO + 5% Solutol + 30% PEG-400 + 60% (20% SBE-β-CD).

[0600] ## The oral solvent is 5% DMSO + 5% Solutol + 10% PEG400 + 80% (20% SBE-β-CD).

[0601] ### The oral solvent is 5% DMSO + 5% Solutol + 90% (20% SBE-β-CD).

[0602] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good oral absorption in rats.

[0603] 3. Experiment on inhibiting VCaP cell proliferation

[0604] Prostate cancer cells (VCaP) were purchased from ATCC. The cell culture medium was RPMI 1640 + 10% FBS, and the cells were cultured in a 37°C, 5% CO2 incubator. Before the experiment, the cells cultured in normal medium were passaged into phenol red-free RPMI 1640 containing 10% activated charcoal-adsorbed FBS and cultured for 3 days. On day 4, cells were digested with phenol red-free digestion solution (Trypsin LE trypsin). Digestion was stopped with phenol red-free RPMI 1640 medium containing 1% activated charcoal-adsorbed FBS, 0.5% PS, and 0.1 nM R1881. Cells were centrifuged and resuspended, and viable cell counts were performed using a Vi-Cell XR cell counter. The cell suspension was adjusted to an appropriate concentration with phenol red-free RPMI 1640 medium containing 1% activated charcoal-adsorbed FBS, 0.5% PS, and 0.1 nM R1881. 180 μl of cell suspension was added to each well of a 96-well cell culture plate to achieve a cell density of 7500 cells / well. T0 wells were also seeded simultaneously. The next day, R1881 at a final concentration of 0.1 nM and different concentrations of the compound were added, and the plates were incubated for another 7 days. The T0 plates were detected using the CellTiter-Glo (CTG) (Promega, product number: G7572) kit, and the results were recorded as RLU0. After culture, 75 μl of pre-melted and equilibrated CellTiter-Glo solution was added to each well, mixed with a microplate shaker for 2 minutes, and incubated at room temperature for 10 minutes before measuring the fluorescence signal using an Envision 2104 plate reader. Results were processed according to equation (2), and the cell growth rate at each compound concentration was calculated in Excel. GraphPad software was used to calculate the GI concentration of the compound at a proliferation rate of 50%. 50 Values. Among them, RLU... compound For the drug treatment group, RLU readingscontrol This represents the average value of the solvent control group. Growth% = (RLU) compound -RLU0) / (RLU control -RLU0)×100% Equation (2)

[0605] GI that inhibits VCaP cell proliferation 50 The results are shown in Table 5.

[0606] Table 5. GI of the compounds of this invention in inhibiting VCaP cell proliferation 50 value

[0607] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good inhibitory activity against VCaP cell proliferation.

[0608] 4. Caco2 permeability test

[0609] The experiment used monolayers of Caco-2 cells, incubated in triple parallel in 96-well Transwell plates. A transport buffer solution (HBSS, 10 mM HEPES, pH 7.4±0.05) containing either the compound of the present invention (2 μM) or the control compounds digoxin (10 μM), naldolol (2 μM), and metoprolol (2 μM) was added to the dosing well on the apical or basal side. A transport buffer solution containing DMSO was added to the corresponding receiving well. After incubation at 37±1 °C for 2 hours, the cell plate was removed, and appropriate amounts of sample were transferred from both the apical and basal sides to new 96-well plates. Acetonitrile containing an internal standard was then added to precipitate the protein. The samples were analyzed using LC MS / MS to determine the concentrations of the compound of the present invention and the control compounds. The concentration data were used to calculate the apparent permeability coefficients for transport from the apical to the basal side of the monolayer cells, and from the basal side to the apical side, thereby calculating the efflux rate. Leakage of fluorescein was used to evaluate the integrity of the monolayer cells after 2 hours of incubation.

[0610] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good CaCO2 permeability.

[0611] 5. Pharmacokinetic assays in mice

[0612] Experimental objective: This experiment aimed to evaluate the pharmacokinetic characteristics of the test substance in mice by administering a single dose of the test substance intravenously and by gavage to ICR mice, measuring the concentration of the test substance in mouse plasma, and administering the test substance in vivo.

[0613] Experimental animals: 6 male ICR mice per compound. Purchased from Chengdu Dashuo Experimental Animal Co., Ltd.

[0614] Experimental method: On the day of the experiment, 6 SD mice were randomly divided into groups according to their body weight. They were fasted for 12-14 hours before administration but allowed free access to water. They were fed 4 hours after administration.

[0615] Table 6. Dosage information for mouse pharmacokinetic tests

[0616] *Dosage is calculated based on free base.

[0617] Sampling: Before and after administration of isoflurane anesthesia, 0.15 mL of blood was collected via the orbital cavity and placed in an EDTAK2 centrifuge tube. The plasma was collected by centrifugation at 5000 rpm and 4°C for 10 min.

[0618] Plasma collection time points for the PO group: 0, 5 min, 15 min, 30 min, 1, 2, 4, 7, 24, 48 h.

[0619] Plasma collection time points for Group IV: 0, 5 min, 15 min, 30 min, 1, 2, 4, 7, 24, 48 h.

[0620] All samples were stored at -60℃ before analysis and testing. Quantitative analysis of the samples was performed using LC-MS / MS.

[0621] Table 7-1 Pharmacokinetic parameters of the compounds of the present invention in mouse plasma

[0622] Table 7-2 Pharmacokinetic parameters of the compounds of the present invention in mouse plasma

[0623] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good oral absorption in mice.

[0624] 6. Pharmacokinetics of Beagle Dogs

[0625] Experimental animals: Male beagles, weighing approximately 8-10 kg, 6 per compound, purchased from Beijing Mars Biotechnology Co., Ltd.

[0626] Experimental method: On the day of the experiment, 6 beagle dogs were randomly divided into groups according to their weight. They were fasted for 14-18 hours before administration but allowed free access to water. They were fed 4 hours after administration.

[0627] Table 8. Dosing information for beagle pharmacokinetic studies

[0628] Note: Intravenous administration solvent: 5% DMA + 5% Solutol + 90% NS;

[0629] Oral (oral) administration solvent: 5% DMSO + 5% Solutol + 10% PEG400 + 80% (20% SBE-CD);

[0630] *Dosage is calculated based on free base.

[0631] Blood samples (1 ml) were collected via jugular or limb veins before and after drug administration and placed in EDTAK2 centrifuge tubes. Plasma was collected by centrifugation at 5000 rpm and 4°C for 10 min. Blood collection time points for both the intravenous and gavage groups were: 0, 5, 15, 30 min, 1, 2, 4, 6, 8, 10, 12, 24, and 48 h. All samples were stored at -80°C before analysis and quantitative analysis was performed using LC-MS / MS.

[0632] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good oral absorption in dogs.

[0633] 7. Pharmacokinetics in monkeys

[0634] Experimental animals: Male cynomolgus monkeys, 3–5 kg, 3–6 years old, 4 animals / compound. Purchased from Suzhou Xishan Biotechnology Co., Ltd.

[0635] Experimental method: On the day of the experiment, four monkeys were randomly divided into groups according to their body weight. They were fasted for 14-18 hours before administration but allowed free access to water. They were fed 4 hours after administration.

[0636] Table 9. Dosing Information for Monkey Pharmacokinetic Tests

[0637] Note: Intravenous administration solvent: 5% DMA + 5% Solutol + 90% NS;

[0638] Oral (oral) administration solvent: 5% DMSO + 5% Solutol + 10% PEG400 + 80% (20% SBE-CD);

[0639] *Dosage is calculated based on free base.

[0640] Blood samples of 1.0 mL were collected via the jugular vein before and after drug administration and placed in EDTAK2 centrifuge tubes. Plasma was collected by centrifugation at 5000 rpm and 4°C for 10 min. Blood collection time points for both the intravenous and gavage groups were: 0, 5, 15, 30 min, 1, 2, 4, 6, 8, 10, 12, 24, and 48 h. Before analysis, all samples were stored at -60°C and quantitatively analyzed using LC-MS / MS.

[0641] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good oral absorption in monkeys.

[0642] 8. hERG potassium ion channel function test

[0643] Experimental platform: Electrophysiological manual patch-clamp system

[0644] Cell line: Chinese hamster ovary (CHO) cell line stably expressing hERG potassium channels

[0645] Experimental Methods: CHO (Chinese Hamster Ovary) cells stably expressing hERG potassium channels were used to record hERG potassium channel currents at room temperature using whole-cell patch-clamp technique. Glass microelectrodes were fabricated from glass electrode blanks (BF150-86-10, Sutter) using a stretching device. The tip resistance after perfusion with electrode fluid was approximately 2-5 MΩ. The glass microelectrodes were inserted into the amplifier probe to connect to the patch-clamp amplifier. Clamp voltage and data recording were controlled and recorded using pClamp 10 software via computer, with a sampling frequency of 10 kHz and a filtering frequency of 2 kHz. After obtaining whole-cell recordings, the cells were clamped at -80 mV to induce hERG potassium current (Ig). hERG The step voltage was applied from -80 mV to +20 mV for 2 seconds, then repolarized to -50 mV for 1 second before returning to -80 mV. This voltage stimulation was applied every 10 seconds, and the drug administration process began after the hERG potassium current stabilized (at least 1 minute). Each test concentration of the compound was administered for at least 1 minute, and at least 2 cells (n≥2) were tested for each concentration.

[0646] Data processing: Data analysis and processing were performed using pClamp 10, GraphPad Prism 5, and Excel software. The degree of inhibition of hERG potassium current (the peak hERG tail current induced at -50mV) by different compound concentrations was calculated using the following formula:

[0647] Inhibition%=[1–(I / Io)]×100%

[0648] Where Inhibition% represents the percentage of inhibition of hERG potassium current by the compound, and I and Io represent the amplitude of hERG potassium current before and after drug administration, respectively.

[0649] Compound IC 50 The following equation was used to fit and calculate the result using GraphPad Prism 5 software: Y=Bottom+(Top-Bottom) / (1+10^((LogIC)) 50 -X)*HillSlope))

[0650] Where X is the Log value of the detected concentration of the test sample, Y is the inhibition percentage at the corresponding concentration, and Bottom and Top are the minimum and maximum inhibition percentages, respectively.

[0651] Conclusion: The compounds of the present invention, such as those in the examples, do not have a significant inhibitory effect on hERG potassium channel current. Specifically, the trifluoroacetate salts of compounds 1, 2, 3, and 4, and compounds 11, 28, 30, 31, 34, 35, 73, 79, and 118 show an inhibitory activity (IC50) on hERG potassium channel current. 50 The values ​​are all greater than 40 μM.

[0652] 9. Liver microsomal stability test

[0653] This experiment used liver microsomes from five genera—human, monkey, dog, rat, and mouse—as in vitro models to evaluate the metabolic stability of the test substance.

[0654] At 37°C, 1 μM of the test substance was co-incubated with microsomal protein and coenzyme NADPH. The reaction was terminated by adding ice-cold acetonitrile containing an internal standard after certain time intervals (5, 10, 20, 30, 60 min). The concentration of the test substance in the sample was detected by LC-MS / MS. T was calculated using the ln value of the drug residue in the incubation system and the incubation time. 1 / 2 Furthermore, the intrinsic clearance rate (CL) of liver microsomes was calculated. int(mic) and hepatic intrinsic clearance rate CL int(Liver) .

[0655] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good liver microsomal stability.

[0656] 10. CYP450 enzyme inhibition test

[0657] The aim of this study was to evaluate the effects of test substances on the activities of five isoenzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4) of human liver microsomal cytochrome P450 (CYP) using an in vitro assay system. Specific probe substrates for CYP450 isoenzymes were co-incubated with human liver microsomes and different concentrations of the test substances. Reduced nicotinamide adenine dinucleotide phosphate (NADPH) was added to initiate the reaction. After the reaction, the metabolites produced by the specific substrates were quantitatively detected by liquid chromatography-tandem mass spectrometry (LC-MS / MS) after sample processing, and the changes in CYP enzyme activity were measured. The IC50 values ​​were calculated. 50 The value is used to evaluate the inhibitory potential of the test substance against each CYP enzyme subtype.

[0658] Conclusion: The compounds of the present invention, such as the compounds in the examples, do not have a significant inhibitory effect on the five isoenzymes of CYP.

[0659] 11. Human CD34+ hematopoietic stem cell proliferation inhibition experiment

[0660] Human CD34+ Hematopoietic stem cells (TPCS, Cat.hmPB34-P-2CW) are CD34-positive stem cells obtained from human PBMCs through immunomagnetic bead sorting. The culture conditions were: DPBS (Gibco, Cat.14190-144) + StemSpan SFEMII (STEMCELL, Cat.9655) + 1X StemSpan CD34+Expansion Supplement (STEMCELL, Cat.2691). First, 40 nL of DMSO, the positive control Talazoparib, or the test compound were added to each well of a 384-well plate (Corning, Cat.3764). Then, cell suspension was added at a concentration of 400 cells / 40 μL / well, resulting in a final concentration of 0.1% DMSO, 3 μM Talazoparib, or different concentrations of the compound per well. The plates were centrifuged at 1000 rpm for 1 minute at room temperature and then cultured at 37°C in a 5% CO2 incubator for 7 days. After culture, 20 μL of CellTiter-Glo Reagent (Promega, Cat. G7573) was added directly to each well, centrifuged at 1000 rpm for 1 minute at room temperature, and then incubated in the dark for 20 minutes. After incubation, the chemiluminescent signal (CFU) was read and recorded using an Envision multi-mode microplate reader (PerkinElmer, Cat. 2104). The cell proliferation inhibition rate of different concentrations of the compound was calculated according to equation (3), and the CFU was recorded. high control The average signal value of the DMSO group, CFU low control The average signal value of the Talazoparib group, CFU compound The average signal value of the compound group is given. The data processed according to equation (3) were subjected to curve fitting using XLfit or GraphPad Prism software with four parameters to calculate the IC50 concentration of the compound when the inhibition rate was 50%. 50 value. Inhibiton%=(CFU high control -CFU compound ) / (CFU high control -CFU low control ) ×100% (Equation 3)

[0661] Conclusion: The compounds of the present invention, such as those in the examples, have no significant inhibitory effect on the proliferation of CD34+ hematopoietic stem cells. Specifically, the trifluoroacetate of compound 7, compound 11, and compound 31 show no significant inhibitory activity (IC50) on the proliferation of CD34+ hematopoietic stem cells. 50 The values ​​are all greater than 10000nM.

[0662] 12. Study on AR degradation activity in MDA-PCA-2B cells

[0663] Human prostate cancer cells MDA-PCA-2B were cultured in a complete medium consisting of F-12K + 20% FBS + 25 ng / ml cholera toxin + 10 ng / ml mouse epidermal growth factor + 0.005 mM phosphoethanolamine + 100 pg / ml hydrocortisone + 45 nM sodium selenite + 0.005 mg / ml insulin at 37°C in a 5% CO2 incubator. Cells in the exponential growth phase were collected and the cell suspension was adjusted to the appropriate concentration using hormone-deprived medium (containing 10% carbon-adsorbed serum) for plating. Cells were seeded in 12-well plates at 5 × 10⁶ cells / well. 5 Cells / well. After plating, incubate in an incubator for 72 hours, then replace with experimental medium (containing 1% carbon-adsorbed serum), add different concentrations of compounds, and incubate at 37°C in a 5% CO2 incubator for 24 hours. After culture, cells were washed with pre-chilled PBS and then lysed on ice for 15 minutes with complete cell lysis buffer containing a Protease / Phosphatase Inhibitor Cocktail (cell lysis buffer information: CST, Cat. 9803, diluted to 1X before use; Protease / Phosphatase Inhibitor Cocktail (100X) information: Cat. 5872. The Cocktail was then diluted 100-fold with the 1X cell lysis buffer to obtain the complete cell lysis buffer). Cells were scraped into new pre-chilled EP tubes and centrifuged at 13500 rpm, 4°C for 20 minutes. The supernatant protein sample was collected, and protein quantification was performed using a BCA kit (Thermofisher, Cat. 23225). The protein concentration was then diluted to 2 mg / mL. The prepared sample was loaded into a 4-12% precast gel at a loading volume of 5 μL (10 μg), and Androgen Receptor (D6F11) was detected using a conventional Western blot method. The expression levels of Rabbit mAb (CST, Cat. 5153S) and the internal control β-Actin (CST, Cat. 3700S) were measured. Secondary antibodies were HRP-labeled Anti-rabbit IgG Antibody (CST, Cat. 7074V) and Anti-mouse IgG Antibody (CST, Cat. 7076V). The expression levels of AR and the internal control were calculated using the protein expression quantification software "Image Studio". The degradation rate (%Degradation) of the compounds at different concentrations was calculated according to equation (4), where R... compound AVE_R represents the relative expression level of AR in different concentration groups. DMSO The value represents the average relative expression level of AR in the DMSO control group. Then, inhibition curves and DC values ​​were calculated using GraphPad Prism software. 50 %Degradation=(AVE_R DMSO -R compound ) / AVE_R DMSO ×100% Equation (4)

[0664] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good degradation activity against AR proteins in MDA-PCA-2B cells.

[0665] 13. Experiment on inhibiting the proliferation of MDA-PCA-2B cells

[0666] Human prostate cancer cells MDA-PCA-2B were purchased from ATCC. The complete culture medium consisted of F-12K + 20% FBS + 25 ng / ml cholera toxin + 10 ng / ml mouse epidermal growth factor + 0.005 mM phosphoethanolamine + 100 pg / ml hydrocortisone + 45 nM sodium selenite + 0.005 mg / ml human recombinant insulin + 1% PS. Cells were cultured at 37°C in a 5% CO2 incubator. At the start of the experiment, cells were digested with 0.25% trypsin-EDTA (1x), phenol red, and the digestion was terminated with the complete culture medium followed by centrifugation. The cells were resuspended in experimental medium (DMEM / F-12, phenol red-free + 1% carbon-adsorbed FBS + 0.5% PS + 25 ng / ml cholera toxin + 10 ng / ml mouse epidermal growth factor + 0.005 mM phosphoethanolamine + 100 pg / ml hydrocortisone + 45 nM sodium selenite + 0.005 mg / ml human recombinant insulin), and viable cells were counted using a cell counter. The cell suspension was adjusted to an appropriate concentration, and 100 μl was seeded into each well of a 96-well cell culture plate at a cell density of 30,000 cells / well. The cells were incubated overnight. The following day, 100 μL of diluted 2x working solution (containing different concentrations of test samples and exogenous androgen R1881, for a total volume of 200 μL, with a final R1881 concentration of 0.2 nM) was added and incubated for another 7 days. Simultaneously, T0 wells were set up, and the Day 0 cell viability reading was measured using a CellCounting-lite 2.0 (Vazyme, DD1101-03) kit on the compound treatment day, denoted as RLU0. After culture, 100 μL of supernatant was discarded from each well, and 60 μL of pre-melted and equilibrated CellCounting-lite 2.0 solution was added. The mixture was shaken for 2 minutes using a microplate shaker, incubated at room temperature for 30 minutes, and then the luminescence signal value was measured using a BMG multi-mode microplate reader.

[0667] The results were processed according to equation (5). The inhibition rate of each concentration of the compound was calculated in Excel, and the concentration GI of the compound when the inhibition rate was 50% was calculated using GraphPad software. 50Values. Where RLU compound represents the readings of the drug-treated group, and RLU control represents the average value of the solvent-controlled group.

[0668] inhibition%=1-(RLU compound-RLU0) / (RLU control-RLU0)×100% Formula (5)

[0669] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good inhibitory activity against the proliferation of MDA-PCA-2B cells.

[0670] 14. Inhibition of LNCaP AR F877L Cell proliferation experiment

[0671] LNCaP AR prostate cancer cells F877L The cells were constructed using WuXi AppTec, with RPMI 1640 culture medium containing 10% FBS, and cultured in a 37℃, 5% CO2 incubator. Before the experiment, cells cultured in normal medium were passaged into phenol red-free RPMI 1640 culture medium containing 10% activated carbon-adsorbed FBS and cultured for 3 days. On day 4, cells were digested with phenol red-free trypsin LE. Digestion was stopped with phenol red-free RPMI 1640 medium containing 1% activated charcoal-adsorbed FBS, 0.5% PS, and 0.1 nM R1881. Cells were centrifuged, resuspended, and viable cell counts were performed using a Vi-Cell XR cell counter. The cell suspension was adjusted to an appropriate concentration with phenol red-free RPMI 1640 medium containing 1% activated charcoal-adsorbed FBS, 0.5% PS, and 0.1 nM R1881. 180 μl of cell suspension was added to each well of a 96-well cell culture plate to achieve a cell density of 2500 cells / well. T0 wells were also seeded simultaneously. The next day, R1881 at a final concentration of 0.1 nM and different concentrations of the compound were added, and the plates were incubated for another 7 days. The T0 plates were detected using the CellTiter-Glo (CTG) (Promega, product number: G7572) kit and recorded as RLU0. After incubation, 75 μl of pre-melted and equilibrated CellTiter-Glo solution was added to each well, mixed with a microplate shaker for 2 minutes, and incubated at room temperature for 10 minutes before measuring the fluorescence signal value (RLU) using an Envision 2104 plate reader. The results were processed according to equation (6), and the inhibition rate (Inhibition%) of each compound concentration was calculated in Excel. The concentration (GI) of the compound at an inhibition rate of 50% was calculated using GraphPad software. 50 Values. Among them, RLU... compound For the drug treatment group, RLU readings control This represents the average value of the solvent control group. Inhibition% = 1 - (RLU)compound -RLU0) / (RLU control -RLU0)×100% Equation (6)

[0672] Conclusion: The compounds of the present invention, such as the compounds in the examples, are effective against LNCaP AR. F877L It exhibits good inhibitory activity against cell proliferation.

[0673] 15. LNCaP AR F877L Study on AR degradation activity in cells

[0674] LNCaP AR prostate cancer cells F877L The assay was constructed using WuXi AppTec, with cells cultured in RPMI 1640 medium containing 10% FBS and 1% PS at 37°C in a 5% CO2 incubator. Before the experiment, cells cultured in normal medium were passaged into phenol red-free RPMI 1640 medium containing 10% activated charcoal-adsorbed FBS and cultured for 3 days. On day 4, cells were digested with phenol red-free digestion solution (Trypsin LE trypsin), and the cell suspension was adjusted to the appropriate concentration using hormone-deprived medium (containing 1% carbon-adsorbed serum) for plating. Cells were plated in 24-well plates at 50,000 cells / well. After plating, cells were incubated for 24 hours, and then different concentrations of the compound were added, followed by another 24 hours of incubation at 37°C in a 5% CO2 incubator. After culture, cells were digested with phenol red-free trypsin LE, and digestion was stopped with phenol red-free RPMI 1640 medium containing 1% activated charcoal-adsorbed FBS + 0.5% PS. Cells were collected in 1.5 mL centrifuge tubes, centrifuged and resuspended, and washed twice with PBS. 20 μL of pre-chilled (ice-bathed) complete cell lysis buffer containing a Protease / Phosphatase Inhibitor Cocktail was added to each tube. (RIPA information for cell lysis buffer: Sigma, Cat. R0278; complete) TM Mini protease inhibitor mixture information: Roche, Cat. 04693124001; Mix RIPA and cOmplete according to the instructions. TM The Mini protease inhibitor mixture (prepared as a complete cell lysis buffer) was lysed on ice for 30 minutes, followed by centrifugation at 12000×g, 4°C for 10 minutes. The supernatant protein sample was collected, and SDS-PAGE protein loading buffer (5X) was added. The sample was heated at 100°C for 10 minutes. The prepared sample was loaded in 15 μL volumes into a 4-12% precast gel, and Androgen Receptor (D6F11) was detected using a conventional Western blot method. Rabbit mAb (CST, Cat. 5153S) and internal control β-Actin (CST, Cat. 3700S) expression were used as the secondary antibody. Anti-rabbit IgG (H+L) (DyLight) was used as the secondary antibody. TM 800 4X PEG Conjugate)(CST,Cat.5151), Anti-mouse IgG(H+L)(DyLight TM 680 Conjugate)(CST, Cat. 5470). The expression levels of AR and internal control were calculated using the protein expression quantification software "Image Studio". The degradation rate (% Degradation) of the compound at different concentrations was calculated according to Equation (7), where R compound AVE_R represents the relative expression level of AR in different drug concentration groups. DMSO The value represents the average relative expression level of AR in the DMSO control group. Then, inhibition curves and DC values ​​were calculated using GraphPad Prism software. 50 %Degradation=(AVE_R DMSO -R compound ) / AVE_R DMSO ×100% Equation (7)

[0675] Conclusion: The compounds of the present invention, such as the compounds in the examples, are effective against LNCaP AR. F877L AR proteins in cells exhibit good degradation activity.

[0676] 16. Inhibition of LNCaP cell proliferation experiment

[0677] LNCaP prostate cancer cells were purchased from ATCC. The cell culture medium was RPMI 1640 + 10% FBS, and the cells were cultured in a 37°C, 5% CO2 incubator. Before the experiment, the cells cultured in normal medium were passaged into phenol red-free RPMI 1640 containing 10% activated charcoal-adsorbed FBS and cultured for 3 days. On day 4, cells were digested with phenol red-free digestion solution (Trypsin LE trypsin). Digestion was stopped with phenol red-free RPMI 1640 medium containing 1% activated charcoal-adsorbed FBS, 0.5% PS, and 0.1 nM R1881. Cells were centrifuged and resuspended, and viable cell counts were performed using a Vi-Cell XR cell counter. The cell suspension was adjusted to an appropriate concentration with phenol red-free RPMI 1640 medium containing 1% activated charcoal-adsorbed FBS, 0.5% PS, and 0.1 nM R1881. 180 μl of cell suspension was added to each well of a 96-well cell culture plate to achieve a cell density of 5000 cells / well. The next day, R1881 and different concentrations of compounds were added to a final concentration of 0.1 nM, and the plates were incubated for another 7 days. After culture, 75 μl of pre-melted and equilibrated CellTiter-Glo solution was added to each well, mixed with a microplate shaker for 2 minutes, and incubated at room temperature for 10 minutes before fluorescence signal values ​​were measured using an Envision 2104 plate reader. Results were processed according to equation (8), and the cell growth inhibition rate of each compound concentration was calculated in Excel. The IC50 concentration of the compound at a 50% inhibition rate was calculated using GraphPad software. 50 Values. Among them, RLU... compound For the drug treatment group, RLU readings control This represents the average value of the solvent control group. Inhibition rate % = (1 - RLU) compound / RLU control Equation (8) is calculated as follows: (1) × 100%

[0678] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good inhibitory activity against the proliferation of LNCaP cells.

[0679] 17. Study on AR T878A degradation activity in LNCAP cells

[0680] Human prostate cancer cells LNCAP (AR T878A mutant expression) were cultured in complete medium of RPMI 1640 + 10% FBS + 1% penicillin-drug antibiotics at 37°C in a 5% CO2 incubator. Cells in the exponential growth phase were collected and the cell suspension was adjusted to the appropriate concentration using experimental medium (phenol red-free RPMI 1640 + 1% carbon-adsorbed fetal bovine serum + 0.5% PS). Cells were seeded in 12-well plates at 1.5 × 10⁶ cells / well. 5Cells / well. After plating, the compound diluted with experimental culture medium was added, and the cells were incubated in a CO2 incubator for 24 hours. After incubation, cells were collected, and RIPA lysis buffer (cocktail (Beyotime, Cat. P1010), RIPA (Beyotime, Cat. P0013B, RIPA lysis buffer is prepared by mixing cocktail and RIPA at a ratio of 1:100) was added and lysed on ice for 15 minutes. After centrifugation at 12000 rpm and 4°C for 10 minutes, the supernatant protein sample was collected. Protein quantification was performed using a BCA kit (Beyotime, Cat. P0009). The protein was then diluted to 0.4 mg / mL, and the expression of AR (Abcam, Cat. ab133273) and the internal control β-Actin Mouse mAb (CST, Cat. 3700S) was detected using a fully automated Western blot quantitative analyzer (Proteinsimple). The expression level of AR T878A relative to the internal reference was calculated using the fully automated protein expression quantification software "Compass for SW". The data processed according to equation (9) were then analyzed using GraphPad Prism 8.0 software, and a four-parameter nonlinear regression model was used to calculate DC. 50 The values ​​are: Protein% = Protein administration / Protein solvent × 100% (Equation 9)

[0681] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good degradation activity against AR T878A protein in LNCAP cells.

[0682] 18. Plasma stability test

[0683] This experiment used plasma samples from five species—human, monkey, dog, rat, and mouse—to evaluate the stability of the test compound in plasma. Compound 11 or the working solution of the control compound was diluted to 2000 ng / mL plasma samples using blank plasma from humans, dogs, rats, and mice (organic reagent content less than 5%). The plasma samples were incubated at 37°C for 0 min, 6 h, and 24 h, after which acetonitrile containing an internal standard was added to terminate the reaction.

[0684] The concentration of analytes in samples was semi-quantitatively determined using liquid chromatography-tandem mass spectrometry (LC-MS / MS), excluding standard curves and quality control samples. The concentration in the sample was expressed as the ratio of the analyte peak area to the internal standard peak area. Retention times of analytes and internal standards, chromatogram acquisition, and chromatogram integration were performed using Analyst software (Sciex, Framingham, Massachusetts, USA). Plasma stability = 100 * T 6h or 24h / T 0min

[0685] Conclusion: The compounds of the present invention, such as compound 11, exhibit good stability in plasma at 6 h and 24 h.

[0686] 19. Efficacy evaluation of VCAP subcutaneous tumor model

[0687] Experimental reagents: Human prostate cancer cells (VCaP cells) were purchased from ATCC; DMEM medium was purchased from ATCC (catalog number 30-2002); fetal bovine serum was purchased from HyClone (catalog number SV30208.02); penicillin-streptomycin was purchased from Gibco (catalog number 15140-122); 0.5% Trypsin-EDTA was purchased from Hyclone (catalog number SH30042.01); PBS was purchased from Sangon Biotech (catalog number E607008-0500); and Matrigel was purchased from Corning (catalog number 354234).

[0688] Experimental methods: Animal information: CB17 SCID mice, male, 7-9 weeks old, weighing approximately 18-22 grams, Zhejiang Vitonlihua Technology Co., Ltd. The mice were housed in an SPF-grade environment with individual air supply and exhaust for each cage. All animals had free access to standard certified commercial laboratory food and water.

[0689] Cell culture: Human prostate cancer cell line VCaP was cultured in vitro under the following conditions: DMEM containing 10% fetal bovine serum and 1% penicillin-streptomycin, at 37°C and 5% CO2. Once cells adhered and reached 80%–90% confluence, they were passaged every 3 or 4 days in a 37°C, 5% CO2 cell culture incubator. The cell expansion cycle was approximately 30–40 days. When the desired cell count was achieved, the cells were harvested and counted.

[0690] Cell seeding: 0.2 mL of VCAP cell suspension (containing 5 x 10⁻⁶ cells) 6 100 cells (1:1 volume ratio of matrix gel) were subcutaneously inoculated into the axilla of each mouse.

[0691] Grouped administration: When solid tumors can be measured in most animals, with an average tumor volume of 80–100 mm. 3 Following standard operating procedures (SOPs), animals were anesthetized with isoflurane and then castrated, with postoperative care provided. After castration, the tumors were allowed to grow to 100–150 mm. 3 Tumors were randomly assigned to groups based on tumor volume, with the grouping day designated as Day 0.

[0692] Tumor measurement and experimental parameters: Tumor diameter was measured twice weekly using calipers. Tumor volume was calculated using the formula: V = 0.5a × b, where a and b represent the major and minor diameters of the tumor, respectively. Mouse body weight was measured twice weekly. The tumor-suppressive efficacy of the test drug was evaluated using the tumor growth inhibition rate (TGI) (%).

[0693] The formula for calculating tumor growth inhibition rate is: TGI(%) = [1 - (T Vt -T V0 ) / (C Vt -C V0 )]×100%

[0694] T Vt T represents the average tumor volume at the end of a given treatment group. V0 The mean tumor volume at the time of grouping for this treatment group; C Vt C represents the average tumor volume at the end of the Vehicle group. V0 The average tumor volume when the Vehicle group was grouped. T Vt and C Vt Take data from the same day.

[0695] Statistical analysis: Data are expressed as mean and standard error (SEM). Statistical analysis was performed using Graphpad Prism with one-way ANOVA and Dunnett's multiple comparison test. In Figure 1, * represents P < 0.05, ** represents P < 0.01, and *** represents P < 0.001. In the analysis of variance between the drug group and the vehicle group, a P value < 0.05 was considered statistically significant.

[0696] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good inhibitory effects on the VCAP subcutaneous tumor model. Specifically, the results for compound 11 are shown in Figure 1.

[0697] 20. Evaluation of drug efficacy in LNCAP subcutaneous tumor model

[0698] Experimental reagents: Human prostate cancer cells (LNCaP) were purchased from ATCC; RPMI-1640 medium was purchased from Gibco (catalog number 22400-089); fetal bovine serum was purchased from HyClone (catalog number SH30406.05); penicillin-streptomycin was purchased from Gibco (catalog number 15140-122); 0.5% Trypsin-EDTA was purchased from Gibco (catalog number 15400-054); PBS was purchased from Sangon Biotech (catalog number E607008-0500); and Matrigel was purchased from Corning (catalog number 354234).

[0699] Experimental methods: Animal information: CB17 SCID mice, male, 7-9 weeks old, weighing approximately 18-22 grams, Zhejiang Vitonlihua Technology Co., Ltd. The mice were housed in an SPF-grade environment with individual air supply and exhaust for each cage. All animals had free access to standard certified commercial laboratory food and water.

[0700] Cell culture: Human prostate cancer cell line LNCaP was cultured in vitro under the following conditions: RPMI-1640 medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin, at 37°C and 5% CO2. Once cells adhered and reached 80%–90% confluence, they were passaged every 3 or 4 days in a 37°C, 5% CO2 cell culture incubator for approximately 30 days to allow for cell expansion. When the desired cell count was achieved, the cells were harvested and counted.

[0701] Cell seeding: 0.2 mL of LNCAP cell suspension (containing 1 x 10⁻⁶ cells) was added. 7 100 cells (1:1 volume ratio of matrix gel) were subcutaneously inoculated into the axilla of each mouse.

[0702] Grouped administration: When solid tumors can be measured in most animals, with an average tumor volume of 80–120 mm. 3 Following standard operating procedures (SOPs), animals were anesthetized with isoflurane and then castrated, with postoperative care provided. After castration, the tumors were allowed to grow to 100–150 mm. 3 Tumors were randomly assigned to groups based on tumor volume, with the grouping day designated as Day 0.

[0703] Tumor measurement and experimental parameters: Tumor diameter was measured twice weekly using calipers. Tumor volume was calculated using the formula: V = 0.5a × b, where a and b represent the major and minor diameters of the tumor, respectively. Mouse body weight was measured twice weekly. The tumor-suppressive efficacy of the test drug was evaluated using the tumor growth inhibition rate (TGI) (%).

[0704] The formula for calculating tumor growth inhibition rate is: TGI(%) = [1 - (T Vt -T V0) / (C Vt -C V0 )]×100%

[0705] T Vt T represents the average tumor volume at the end of a given treatment group. V0 The mean tumor volume at the time of grouping for this treatment group; C Vt C represents the average tumor volume at the end of the Vehicle group. V0 The average tumor volume when the Vehicle group was grouped. Vt and C Vt Take data from the same day.

[0706] Statistical analysis: Data are expressed as mean and standard error (SEM). Statistical analysis was performed using Graphpad Prism with one-way ANOVA and Dunnett's multiple comparison test. In Figure 2, * represents P < 0.05, ** represents P < 0.01, and *** represents P < 0.001. In the analysis of variance between the drug group and the vehicle group, a P value < 0.05 was considered statistically significant.

[0707] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good inhibitory effects on the LNCAP subcutaneous tumor model. Specifically, the results for compound 11 are shown in Figure 2.

Claims

1. A pharmaceutical formulation comprising an active ingredient M and a pharmaceutical excipient, wherein the active ingredient M is selected from compounds of general formula (I) or their stereoisomers, tautomers, or pharmaceutically acceptable salts. BLK(I); L is selected from -Ak1-Cy1-Ak2-Cy2-Ak3-Cy3-Ak4-Cy4-Ak5-; Ak1, Ak2, Ak3, Ak4, and Ak5 are each independently selected from -(CH2). q -、-(CH2) q -O-, -O-(CH2) q -、-(CH2) q -S-、-S-(CH2) q -、-(CH2) q -NR L -、-NR L -(CH2) q -、-(CH2) q -NR L C(=O)-、-(CH2) q -C(=O)NR L -, -C(=O)-, -C(=O)-(CH2) q -NR L -、-(C≡C) q - or bond, wherein the -CH2- is optionally selected from one or two elements selected from deuterium, halogen, =O, OH, CN, C 1- 4-alkyl or C 3-6 Substituents of cycloalkyl groups; q is selected from 0, 1, 2, or 3; R L Each element is independently selected from H, deuterium, and C. 1-4 Alkyl or deuterated C 1-4 alkyl; Cy1, Cy2, Cy3, or Cy4 are each independently selected from the key or arbitrarily selected by 1 to 4 Rs. L2 The substituted group is one of the following: 4-7 membered heteromonocyclic group, 4-12 membered heterocyclic group, 5-13 membered heterospirocyclic group, 7-12 membered heterobridged cyclic group, C 3-7 Monocycloalkyl, C 4-7 Monocyclic alkenyl, C 4-12 cycloalkyl, C 5-13 Spirocycloalkyl, C 5-12 Bridged cycloalkyl, 5-10 membered heteroaryl or C 6-10 Aryl; B is selected from X1 is selected from N or CR x1 X2 is selected from N or CR x2 X3 is selected from N or CR. x3 X4 is selected from N or CR x4 X5 is selected from N or CR x5 ; At most two of X1, X2, X3, X4, and X5 are selected from N; Z1 is selected from N or CR z1 Z2 is selected from N or CR. z2 Z3 is selected from N or CR. z3 Z4 is selected from N or CR z4 ; At most 3 of Z1, Z2, Z3, and Z4 are selected from N; Y1 and Y2 are each independently selected from -CR y1 R y2 -、-(CR y1 R y2 )2-、-(CR y1 R y2 )3-; R 1 Selected from H, deuterium, C 1-4 Alkyl or C 3-6 Cycloalkyl groups, wherein the alkyl or cycloalkyl group is optionally composed of 1 to 4 elements selected from deuterium, halogen, OH, NH2, CN, C. 1-4 Alkyl, C 1-4 Substituents of alkoxy groups; R x1 R x2 R x3 R x4 R x5 R z1 R z2 R z3 R z4 Each is independently selected from H, deuterium, halogen, OH, NH2, CN, NO2, COOH, CONH2, and NHC. 1-4 Alkyl, N(C) 1-4 Alkyl)2, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, -OC 1-4 Alkyl, -SC 1-4 Alkyl, -C 0-4 Alkylene-C 3-6 carbonyl group, -C 0-4 alkylene-4 to 6-membered heterocyclic groups, 5 to 6-membered heteroaryl groups, -OC 3-6 Carbocyclic, -O-4 to 6-membered heterocyclic groups, wherein the alkylene, alkyl, alkenyl, alkynyl, carbocyclic, heterocyclic, or heteroaryl groups are optionally surrounded by 1 to 4 R groups. s Replaced; Or R x1 R x2 R x3 R x4 Each is independently selected from -S(=O)2NH2 and -S(=O)2C 1-4 alkyl; Or, R x1 With R x2 R x2 With R x3 R x3 With R x4 R x4 With R x5 R 1 With R z3 R 1 With R z1 Direct connection forms C 4-6 A carbocyclic group or a 4- to 7-membered heterocyclic group, wherein the carbocyclic group or heterocyclic group is optionally surrounded by 1 to 4 R groups. s Replaced; R 2 R 3 R y1 R y2 Each is independently selected from H, deuterium, halogen, OH, NH2, CN, NO2, COOH, CONH2, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, -OC 1-4 Alkyl, -SC 1-4 Alkyl groups, wherein the alkyl, alkenyl, or alkynyl groups are optionally composed of 1 to 4 groups selected from deuterium, halogen, OH, NH2, CN, C. 1-4 Alkyl, C 1-4 Substituents of alkoxy groups; K is selected from This indicates whether the ring in question is aromatic or non-aromatic. F6, F7, and F8 are each independently selected from N, C, CH, or CR. k1 Furthermore, F6, F7, and F8 contain at most two N's; G is selected from CH, CD, or N; E1 is selected from N, CH, or CD; E2 is selected from C, N, CH, or CD; Q is selected independently from the following: -O-, -S-, -CH2-, -CD2-, -NR. q -、-C(=O)-、-NR q C(=O)-、-C(=O)NR q -; Q and G cannot directly form nitrogen-nitrogen bonds or nitrogen-oxygen bonds; R q Each is independently selected from H, deuterium, and C. 1-4 Alkyl or deuterated C 1-4 alkyl; R k1 Each is independently selected from deuterium, halogen, OH, NH2, CN, COOH, CONH2, C 1-4 Alkyl, C 2-4 alkenyl, C 2- 4-Alynyl group, C 1-4 Alkoxy, C 3-6 Cycloalkyl, 4- to 6-membered heterocyclic alkyl, wherein the alkyl, alkenyl, alkoxy, cycloalkyl, or heterocyclic alkyl is optionally surrounded by 1 to 4 R... s Replaced; p1 is selected from 0, 1, or 2; R L2 R s Each is independently selected from deuterium, halogens, OH, CN, ⁻, CF₃, SF₅, NO₂, NH₂, and NHC. 1-4 Alkyl, N(C) 1-4 Alkyl group 2, COOH, CONH2, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Alkoxy, -SC 1-4 Alkyl, -C 0- 4-alkylene-C 3-6 cycloalkyl, -C 0-4 Alkylene-4 to 6-membered heterocyclic group, wherein the alkylene, alkylene, alkoxy, alkenyl, alkynyl, or cycloalkyl group is optionally selected from one to four of deuterium, F, Cl, Br, I, OH, CN, C. 1-4 Alkyl, C 1-4 Substituents of alkoxy groups; Or R s Each independently selected from -OC 3-6 cycloalkyl; The pharmaceutical preparation contains 1-800 mg of active ingredient M; Preferably, the content (wt%) of the active ingredient M is 0.1%-99%; The total content of all components in the pharmaceutical preparation is 100%.

2. The pharmaceutical preparation according to claim 1, wherein, The compounds of general formula (I) are selected from the compounds of general formula (II). B is selected from Or B is selected from Preferably, B is selected from R x2 R x3 R x4 Each is independently selected from H, deuterium, F, Cl, Br, I, OH, NH2, CN, NO2, COOH, CONH2, N(CH3)2, NHCH3, CF3, CHF2, CH2F, OCF3, OCH2F, OCD3, CH2OH, methyl, ethyl, vinyl, ethynyl, methoxy, ethoxy, methylthio, cyclopropyl, -CH2-cyclopropyl, -O-cyclopropyl; Or R x2 R x4 Each is independently selected from -S(=O)2CH3, -O-CH2-propynyl, -O-CH2-cyclopropyl, -O-CH2CH2-OCH3, -O-CH2CH2-O-cyclopropyl, Preferably, R x3 Each is independently selected from F, Cl, Br, CN, vinyl, and ethynyl groups; Preferably, R x4 Each is independently selected from F, Cl, Br, CF3, CHF2, CH2F, methoxy, ethoxy, OCF3, OCH2F, OCD3, -O-CH2-cyclopropyl, -O-CH2CH2-OCH3, -O-CH2CH2-O-cyclopropyl, -O-CH2-propynyl; L is selected from -Cy1-, -Cy1-Ak2-, -Cy1-CH2-, -Ak1-Cy1-, -Cy1-Cy2-, -Cy1-CH2-Cy2-, -Cy1-Cy2-Cy3-, -Cy1-CH2-Cy2-Cy3-, -Cy1-Cy2-CH2-Cy3-; Alternatively, L can be selected from -Cy1-Ak2-Cy2- or -Cy1-O-Cy2-. Preferably, L is selected from -Cy1-, -Cy1-CH2-, and -Cy1-O-Cy2-; Cy1, Cy2, and Cy3 are each independently selected from one of the following groups that can be substituted: When substituted, it is replaced by 1 to 4 substituents selected from deuterium, F, CF3, OH, =O, COOH, CN, NH2, hydroxymethyl, methyl, methoxy, and cyclopropyl. Preferably, Cy1, Cy2, and Cy3 are each independently selected from one of the following optionally substituted groups: When substituted, it is replaced by 1 to 4 substituents selected from deuterium, F, CF3, OH, =O, COOH, CN, NH2, hydroxymethyl, methyl, methoxy, and cyclopropyl. Preferably, L is selected from one of the structural segments shown in Table L-1; K is selected from Preferably, K is selected from Q is independently selected from bonds, NH, N(CH3), O, S, NHC(=O), C(=O)NH, N(CH3)C(=O), and C(=O)N(CH3); Preferably, Q is independently selected from bonds, NH, and C(=O)NH; Preferably, Selected from R k1 Each is independently selected from deuterium, F, Cl, Br, I, OH, NH2, CN, COOH, CONH2, CF3, CHF2, CH2F, OCF3, OCH2F, CH2OH, methyl, ethyl, isopropyl, methoxy, ethoxy, isopropoxy, and cyclopropyl. p1 is selected from 0, 1, or 2.

3. The pharmaceutical formulation according to claim 2, wherein the structure of the compound of general formula (I) is selected from one of the structures shown in Table S-1.

4. The pharmaceutical formulation according to claim 1, wherein the compound of general formula (I) is selected from the following structures:

5. The pharmaceutical formulation according to any one of claims 1-4, wherein the pharmaceutical formulation comprises an active ingredient M at a unit dosage.

6. The pharmaceutical preparation according to any one of claims 1-5, wherein the pharmaceutical preparation comprises 5-750 mg, 5-500 mg, 5-300 mg, 10-700 mg, 10-400 mg, 10-300 mg, 50-500 mg, 50-400 mg, 50-300 mg, 100-500 mg, 100-400 mg, or 100-300 mg of active ingredient M, preferably 5 mg, 10 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 50 mg, 55 mg, 60 mg, 65 mg, 100 mg, 120 mg, 150 mg, 200 mg, 250 mg, 300 mg, 400 mg, or 500 mg.

7. The pharmaceutical preparation according to any one of claims 1-6, wherein the content of the active ingredient M is 1%-80%, 1%-50%, 0.5%-47.5%, 1%-40%, 5%-40%, 5%-35%, 5%-30%, 5%-20%, or 10-30%.

8. The pharmaceutical preparation according to claims 1-7, The pharmaceutical excipient includes a filler; The pharmaceutical excipient may optionally further contain a disintegrant; Preferably, the filler is selected from one or more of microcrystalline cellulose, mannitol, lactose, sucrose, sorbitol, dextran, anhydrous dicalcium phosphate, pregelatinized starch, dicalcium phosphate, and starch, and is preferably microcrystalline cellulose or lactose. Preferably, the disintegrant is selected from one or more of sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, crospovidone, crospovidone carboxymethyl cellulose sodium, calcium carboxymethyl cellulose, and dry starch.

9. The pharmaceutical preparation according to claims 1-8, The filler content in the pharmaceutical preparation is 40%-90% or 40%-85%. The disintegrant is present in the pharmaceutical preparation at a concentration of 0%-10% or 2%-8%.

10. The pharmaceutical formulation according to claim 1, wherein the pharmaceutical excipient further comprises one or more of a binder, a flow aid, a lubricant, a wetting agent, and a pH adjuster; Preferably, The adhesive is selected from one or more of copovidone, hydroxypropyl cellulose, hydroxypropyl methylcellulose, and methylcellulose; The gliding agent is selected from one or more of talc, silica, micronized silica gel, polyethylene glycol, and magnesium dodecyl sulfate; The lubricant is selected from one or more of magnesium stearate, calcium stearate, stearic acid, and sodium stearate fumarate; The wetting agent is selected from one or more of water and ethanol; pH adjusters are selected from fumaric acid, sodium carbonate, sodium hydroxide, sodium bicarbonate, and calcium bicarbonate. Preferably, The content of the adhesive in the pharmaceutical preparation is 0%-10%; The content of gliding agents in pharmaceutical preparations is 0%-3%; The content of lubricant in pharmaceutical preparations is 0%-3%; The content of pH adjusters in pharmaceutical preparations is 0%-5%.

11. A pharmaceutical formulation comprising the active ingredient M as described in any one of claims 1-4, a filler, a disintegrant, and a binder, optionally further comprising a flow aid and a lubricant, wherein the content of the active ingredient M is 0.5%-47.5%, the content of the filler is 40%-90%, the content of the disintegrant is 1%-10%, the content of the binder is 1%-10%, the content of the lubricant is 0.5%-3%, and the content of the flow aid is 0%-3%.

12. The pharmaceutical preparation according to claim 8, wherein, The filler is one or a mixture of microcrystalline cellulose or lactose. The disintegrant is crospovidone; The adhesive is one or a mixture of copovidone or hydroxypropyl methylcellulose; The lubricant is magnesium stearate; The gliding agent is silicon dioxide.

13. The pharmaceutical preparation according to any one of claims 8-12, wherein the pharmaceutical excipient further comprises one or more of a flavoring agent, an antioxidant, a preservative, a light-blocking agent, and a film-coating premix.

14. The pharmaceutical preparation according to any one of claims 1-13, wherein the dosage form is an oral tablet, capsule, or granule.

15. Use of the pharmaceutical preparation according to any one of claims 1-14 in the preparation of a medicament for treating cancer (e.g., prostate cancer).

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