PDE4B inhibitors and uses thereof

Abstract

The invention discloses a compound shown as a formula (I), a stereoisomer or pharmaceutically acceptable salt of the compound, or a pharmaceutical composition containing the stereoisomer or the pharmaceutically acceptable salt, and application of the compound serving as a PDE4B inhibitor to preparation of drugs for treating related diseases. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and in particular relates to a small molecule compound with PDE4B selective inhibitory activity, a stereoisomer or a pharmaceutically acceptable salt thereof, and use thereof in preparing medicines for treating related diseases. Background Art

[0002] PDE4 inhibitors produce antidepressant effects in humans and animals by enhancing cAMP signals in the brain. PDE4 inhibitors also play an important role in the treatment of other central nervous system diseases, including Alzheimer's disease, Parkinson's disease, schizophrenia, stroke, Huntington's disease and other diseases. In addition, the research and development of PDE4 inhibitors has also made significant progress in the treatment of respiratory diseases such as asthma and chronic obstructive pulmonary disease. The research and development principle of this type of drug comes from the role of PDE4 in inhibiting the functions of a series of inflammatory cells and resident cells, which is believed to be related to the pathogenesis of such diseases. A large number of clinical studies have shown that cyclic adenosine monophosphate (c AMP) can block the proliferation and chemotaxis of inflammatory cells, inhibit the release of inflammatory and cytotoxic mediators in the lungs, and PDE4 is particularly abundant in immune cells, inflammatory cells and smooth muscle cells.

[0003] PDE4 inhibitors mainly inhibit the hydrolysis of PDE4, increase the level of cAMP in the body, inhibit the release of inflammatory factors, and promote the production of anti-inflammatory mediators to exert anti-inflammatory effects. Roflumilast is used clinically to treat COPD, and has significant anti-inflammatory effects. It can inhibit the release of inflammatory mediators such as TNF-α, interleukins, and chemokines by monocytes, macrophages, and T cells. However, this type of inhibitor generally has serious side effects such as nausea and vomiting, which limits the clinical application of PDE4 inhibitors. A large number of studies have shown that in the human body, subtype B of phosphodiesterase 4 (PDE4B) is related to inflammatory response and participates in the release of various inflammatory mediators in the body, while subtype D is closely related to the production of side effects such as nausea and vomiting. This provides a new idea for the discovery of PDE4 inhibitors with low side effects, that is, the design of PDE4B inhibitors may reduce the impact of side effects and promote further clinical application.

[0004] Phosphodiesterase 4 is highly selective for c AMP and has four subtypes, PDE4A, 4B, 4C and 4D, with at least 25 splice variants. The protein sequences of the catalytic domains of the four subtypes of PDE4 are highly homologous, and inhibitors acting on the catalytic domain will not produce subtype selectivity, while most of the reported classic PDE4 inhibitors act on the catalytic domain. In recent years, new PDE4 inhibitor action modes have been reported, in which the inhibitors act on both the catalytic domain and the regulatory sequence, allowing the regulatory sequence to stabilize the closed conformation of the protein, prevent cAMP from entering, and exert an inhibitory effect. Studies have found that there are amino acid differences between PDE4B and 4D in this regulatory sequence, and inhibitors designed based on this difference can produce subtype selectivity. Therefore, based on the difference in two amino acids in PDE4B Leu674 / PDE4D Gln594 on the downstream regulatory sequence CR3 (Conserved Region 3), it is expected to achieve selectivity for subtype B and reduce the side effects of inhibitors while maintaining activity. A selective PDE4B inhibitor with good activity, high safety and few side effects has been discovered, which has good clinical development prospects and can be used to treat COPD or cancer or other proliferative diseases or conditions. Summary of the invention

[0005] The present invention provides a small molecule compound with PDE4B inhibitory activity, and its stereoisomer or pharmaceutically acceptable salt, which has high activity, low toxic side effects, excellent pharmacokinetic characteristics and bioavailability.

[0006] The present invention provides a compound represented by formula (I) or (II), a stereoisomer or a pharmaceutically acceptable salt thereof,

[0007]

[0008] in,

[0009] Cy2 is selected from Or a 3-6 membered cycloalkyl; In some embodiments, Cy2 is selected from Cyclopropyl; In some embodiments, Cy2 is selected from In some embodiments, Cy2 is selected from In some embodiments, Cy2 is selected from In some embodiments, Cy2 is selected from In some embodiments, Cy2 is selected from or In some embodiments, Cy2 is selected from

[0010] Cy1 is selected from or Indicates the site where Cy1 is linked to Cy2: In some embodiments, Cy1 is selected from In some embodiments, Cy1 is selected from

[0011] Or Cy1-Cy2 together form In some embodiments, Cy1-Cy2 together form or

[0012] R2 is selected from C 2-6 Alkynyl, 3-5 membered cycloalkyl or C 1-4 Alkoxy; in some embodiments, R2 is selected from cyclopropyl, ethynyl, propynyl or ethoxy; in some embodiments, R2 is selected from cyclopropyl, ethynyl, propynyl; in some embodiments, R2 is selected from cyclopropyl;

[0013] q is selected from 1 or 2; in some embodiments, q is selected from 1; in some embodiments, q is selected from 2;

[0014] R 12 is selected from 4-10 membered heterocycloalkylene, 3-10 membered cycloalkylene, 6-10 membered arylene, 5-10 membered heteroarylene, hydroxy-C 1-6 Alkyl, C 1-4 Alkyl -CN, -CRaRb=NOC 1-4 Alkyl, -C(NRaRb)=N-CN, -C(C 1-4 alkyl)=N-CN、-C(NRaRb)=CRa-NO2、-C(CH3)2-CH2-OC(O)NH2、-C(CH3)2-CH2-NH-C(O)O(C 1-4 Alkyl), -C(CH3)2-CH2-OC(O)NHCH3, -CH(CH3)-CH2-OC(O)NH2, -C(CH3)2-CH(CH3)-OC(O)NH2, -CH(CH3)-CH2-NH-C(O)O(C 1-4 alkyl), -CH(CH3)-CH2-OC(O)NHCH3, -C(O)-Ra or -C(O)-(4-6 membered heterocycloalkyl), wherein the heterocycloalkylene, cycloalkylene, arylene, heteroarylene may be further substituted with 1-3 halogens, CN, =O, OH, -SF5, C 1-4 Alkyl, C 1-4 Alkyl-CN, hydroxyl C 1-4 Alkyl, C 1-4 Haloalkyl, -(CH2) t-OC(O)NH2, -CH(CH3)-OC(O)NH2, -(CH2) t -NRa-C(O)NH2, -(CH2) t -NRa-C(=NH)NH2, -(CH2) t -NRa-C(O)-O-(C 1-4 Alkyl), -(CH2) t -NRa-C(C 1-4 alkyl)=N-CN, -(CH2) t -NRa-C(NRaRb)=N-CN、-(CH2) t -NRa-C(NRaRb)=CRa-NO2, -(CH2) t -C(NRaRb)=N-CN、-(CH2) t -C(C 1-4 alkyl)=N-CN, -(CH2) t -C(C 1-4 alkyl)=NO-(C 1-4 alkyl), =NOC 1-4 Alkyl, 5-6 membered heteroaryl, -(CH2) t -OC 1-4 Alkyl, -(CH2) t -OC(O)NHCH3, -(CH2) t -OC 1-2 Haloalkyl, -(CH2) t -OC 3-6 Cycloalkyl, -(CH2) t -OC 1-4 Alkyl-OC 3-6 Cycloalkyl, -(CH2) t -COOH, -(CH2) t -NRaRb, -(CH2) t -C(O)NRaRb, -(CH2) t -NRa-C(O)CH3 group substitution;

[0015] In some embodiments, R 12 Selected from 3-10 membered cycloalkylene, 6-10 membered arylene, hydroxy-C 1-6 Alkyl, C 1-4 Alkyl-CN, -C(CH3)2-CH2-OC(O)NH2, -C(CH3)2-CH2-NH-C(O)O(C 1-4 alkyl), -C(CH3)2-CH(CH3)-OC(O)NH2, wherein the cycloalkylene and arylene groups are optionally further substituted with 1-3 halogens, C1-4 Alkyl-CN, hydroxyl C 1-4 Alkyl, -(CH2) t -OC(O)NH2, -CH(CH3)-OC(O)NH2, -(CH2) t -NRa-C(O)NH2, -(CH2) t -COOH, -(CH2) t -C(O)NRaRb group substitution;

[0016] In some embodiments, R 12 Selected from 3-10 membered cycloalkylene, 6-10 membered arylene, hydroxy-C 1-6 Alkyl, C 1-4 Alkyl-CN, -C(CH3)2-CH2-OC(O)NH2, -C(CH3)2-CH2-NH-C(O)O(C 1-4 alkyl), -C(CH3)2-CH(CH3)-OC(O)NH2, wherein the cycloalkylene and arylene groups are optionally further substituted with 1-3 halogens, C 1-4 Alkyl-CN, hydroxyl C 1-4 Alkyl, -(CH2) t -OC(O)NH2, -CH(CH3)-OC(O)NH2, -(CH2) t -COOH group substitution;

[0017] In some embodiments, R 12 Selected from 3-10 membered cycloalkylene, hydroxy-C 1-6 Alkyl, C 1-4 Alkyl-CN, -C(CH3)2-CH2-OC(O)NH2, -C(CH3)2-CH2-NH-C(O)O(C 1-4 alkyl), -C(CH3)2-CH(CH3)-OC(O)NH2, wherein the cycloalkylene group is optionally further substituted by 1-3 C 1-4 Alkyl-CN, hydroxyl C 1-4 Alkyl, -(CH2) t -OC(O)NH2, -CH(CH3)-OC(O)NH2; in some embodiments, R 12 Selected from 4-5 membered cycloalkyl, hydroxy-C 4-6 Alkyl, C 3-4 Alkyl-CN, -C(CH3)2-CH2-OC(O)NH2, -C(CH3)2-CH2-NH-C(O)O(C 1-2 alkyl), -C(CH3)2-CH(CH3)-OC(O)NH2, wherein the cycloalkylene group is optionally further substituted by 1-3 C1-2 Alkyl-CN, hydroxyl C 1-2 Alkyl, -CH2-OC(O)NH2, -CH(CH3)-OC(O)NH2 group substitution;

[0018] In some embodiments, R 12 Selected from:

[0019]

[0020] Ra and Rb are each independently selected from H, halogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy or 4-6 membered heterocycloalkyl containing 1-2 heteroatoms selected from N, O; in some embodiments, Ra, Rb are each independently selected from H, F, Cl, methyl, ethyl, C 1-2 Haloalkyl; in some embodiments, Ra, Rb are each independently selected from H, F, Cl, methyl, ethyl, -CH2F, -CHF2, -CF3, -CH2CH2F, -CH2CHF2, -CH2CF3, -CHFCH2F, -CHFCHF2, -CHFCF3, -CF2CH2F, -CF2CHF2, -CF2CF3;

[0021] Alternatively, Ra, Rb and the atoms to which they are attached together form C 3-6 Cycloalkyl;

[0022] t is selected from 0, 1, 2 or 3;

[0023] In some embodiments, Selected from:

[0024]

[0025]

[0026] In some embodiments, Selected from:

[0027] The compounds of formula (I) and (II) of the present invention are not the following structures:

[0028]

[0029]

[0030] Specifically, the first technical solution of the present invention provides a compound represented by formula (I) or (II), a stereoisomer or a pharmaceutically acceptable salt thereof,

[0031]

[0032] in,

[0033] Cy2 is selected from 3-6 membered cycloalkyl; in some embodiments, Cy2 is selected from

[0034] Cy1 is selected from or Indicates the site where Cy1 and Cy2 are linked;

[0035] Or Cy1-Cy2 together form

[0036] R2 is selected from C 2-6 Alkynyl, 3-5 membered cycloalkyl or C 1-4 Alkoxy;

[0037] q is selected from 1 or 2;

[0038] R 12 is selected from 4-10 membered heterocycloalkylene, 3-10 membered cycloalkylene, 6-10 membered arylene, 5-10 membered heteroarylene, hydroxy-C 1-6 Alkyl, C 1-4 Alkyl -CN, -CRaRb=NOC 1-4 Alkyl, -C(NRaRb)=N-CN, -C(C 1-4 alkyl)=N-CN、-C(NRaRb)=CRa-NO2、-C(CH3)2-CH2-OC(O)NH2、-C(CH3)2-CH2-NH-C(O)O(C 1-4 Alkyl), -C(CH3)2-CH2-OC(O)NHCH3, -CH(CH3)-CH2-OC(O)NH2, -C(CH3)2-CH(CH3)-OC(O)NH2, -CH(CH3)-CH2-NH-C(O)O(C 1-4 alkyl), -CH(CH3)-CH2-OC(O)NHCH3, -C(O)-Ra or -C(O)-(4-6 membered heterocycloalkyl), wherein the heterocycloalkylene, cycloalkylene, arylene, heteroarylene may be further substituted with 1-3 halogens, CN, =O, OH, -SF5, C 1-4 Alkyl, C 1-4 Alkyl-CN, hydroxyl C 1-4 Alkyl, C 1-4 Haloalkyl, -(CH2) t -OC(O)NH2, -CH(CH3)-OC(O)NH2, -(CH2)t -NRa-C(O)NH2, -(CH2) t -NRa-C(=NH)NH2, -(CH2) t -NRa-C(O)-O-(C 1-4 Alkyl), -(CH2) t -NRa-C(C 1-4 alkyl)=N-CN, -(CH2) t -NRa-C(NRaRb)=N-CN、-(CH2) t -NRa-C(NRaRb)=CRa-NO2, -(CH2) t -C(NRaRb)=N-CN、-(CH2) t -C(C 1-4 alkyl)=N-CN, -(CH2) t -C(C 1-4 alkyl)=NO-(C 1-4 alkyl), =NOC 1-4 Alkyl, 5-6 membered heteroaryl, -(CH2) t -OC 1-4 Alkyl, -(CH2) t -OC(O)NHCH3, -(CH2) t -OC 1-2 Haloalkyl, -(CH2) t -OC 3-6 Cycloalkyl, -(CH2) t -OC 1-4 Alkyl-OC 3-6 Cycloalkyl, -(CH2) t -COOH, -(CH2) t -NRaRb, -(CH2) t -C(O)NRaRb, -(CH2) t -NRa-C(O)CH3 group substitution;

[0039] Ra and Rb are each independently selected from H, halogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy or 4-6 membered heterocycloalkyl containing 1-2 heteroatoms selected from N and O;

[0040] Alternatively, Ra, Rb and the atoms to which they are attached together form C 3-6 Cycloalkyl;

[0041] t is selected from 0, 1, 2 or 3;

[0042] The condition is that the compound of formula (I) or (II) is not the following structure:

[0043]

[0044] Or the compound of formula (I) or (II) is not the following structure: Or the compound of formula (I) or (II) is not the following structure:

[0045]

[0046] The second technical solution of the present invention is a compound described in formula (I) or (II), a stereoisomer or a pharmaceutically acceptable salt thereof, wherein R 12 Selected from 3-10 membered cycloalkylene, 6-10 membered arylene, hydroxy-C 1-6 Alkyl, C 1-4 Alkyl-CN, -C(CH3)2-CH2-OC(O)NH2, -C(CH3)2-CH2-NH-C(O)O(C 1-4 alkyl), -C(CH3)2-CH(CH3)-OC(O)NH2, wherein the cycloalkylene and arylene groups are optionally further substituted with 1-3 halogens, C 1-4 Alkyl-CN, hydroxyl C 1-4 Alkyl, -(CH2) t -OC(O)NH2, -CH(CH3)-OC(O)NH2, -(CH2) t -NRa-C(O)NH2, -(CH2) t -COOH, -(CH2) t -C(O)NRaRb group substitution;

[0047] In some embodiments, R 12 Selected from 3-10 membered cycloalkylene, 6-10 membered arylene, hydroxy-C 1-6 Alkyl, C 1-4 Alkyl-CN, -C(CH3)2-CH2-OC(O)NH2, -C(CH3)2-CH2-NH-C(O)O(C 1-4 alkyl), -C(CH3)2-CH(CH3)-OC(O)NH2, wherein the cycloalkylene and arylene groups are optionally further substituted with 1-3 halogens, C 1-4 Alkyl-CN, hydroxyl C 1-4 Alkyl, -(CH2) t -OC(O)NH2, -CH(CH3)-OC(O)NH2, -(CH2) t -COOH group substitution;

[0048] The definitions of other groups are consistent with those of any technical solution.

[0049] The third technical solution of the present invention is a compound of formula (I) or (II), a stereoisomer or a pharmaceutically acceptable salt thereof, wherein Selected from:

[0050]

[0051] R2 is selected from cyclopropyl, ethynyl, propynyl or ethoxy;

[0052] The definitions of other groups are consistent with those of any technical solution.

[0053] The fourth technical solution of the present invention is a compound described in formula (I) or (II), a stereoisomer or a pharmaceutically acceptable salt thereof, wherein:

[0054] Cy1 is selected from Indicates the site where Cy1 and Cy2 are linked;

[0055] Cy2 is selected from

[0056] R2 is selected from cyclopropyl;

[0057] Selected from:

[0058] Provided that the compound of formula (I) is not

[0059] The compound of the present invention is selected from one of the structures in Table A, its stereoisomers or pharmaceutically acceptable salts,

[0060] Table A:

[0061]

[0062]

[0063]

[0064]

[0065] The present invention also provides a composition or pharmaceutical preparation, which contains the compound described in any of the above schemes, its stereoisomer or pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier and / or excipient. The pharmaceutical composition can be in the form of a unit preparation (unit preparation is also referred to as "preparation specification").

[0066] Furthermore, the composition or pharmaceutical preparation of the present invention contains 1-1500 mg of the compound described in any one of the aforementioned schemes, its stereoisomer or pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier and / or excipient.

[0067] The present invention also provides the use of the compound described in any one of the above schemes, its stereoisomer or pharmaceutically acceptable salt in the preparation of a drug for treating / preventing a PDE4B-mediated disease. Further, the PDE4B-mediated disease is cancer, COPD, idiopathic pulmonary fibrosis or interstitial lung disease.

[0068] "Effective amount" or "therapeutically effective amount" as used herein refers to administering a sufficient amount of a compound disclosed herein that will alleviate one or more symptoms of the disease or condition being treated to some extent. In some embodiments, the result is a reduction and / or alleviation of the signs, symptoms or causes of a disease, or any other desired change in a biological system. For example, an "effective amount" for therapeutic use is the amount of a composition comprising a compound disclosed herein required to provide a clinically significant reduction in disease symptoms. Examples of therapeutically effective amounts include, but are not limited to, 1-1500 mg, 1-1400 mg, 1-1300 mg, 1-1200 mg, 1-1000 mg, 1-900 mg, 1-800 mg, 1-700 mg, 1-600 mg, 1-500 mg, 1-400 mg, 1-300 mg, 1-250 mg, 1-200 mg, 1-150 mg, 1-125 mg, 1-100 mg, 1-80 mg, 1-60 mg, 1-50 mg, 1-40 mg, 1-25 mg, 1- 20mg, 5-1500mg, 5-1000mg, 5-900mg, 5-800mg, 5-700mg, 5-600mg, 5-500mg, 5-400mg, 5-300mg, 5-250mg, 5-200mg, 5 -150mg, 5-125mg, 5-100mg, 5-90mg, 5-70mg, 5-80mg, 5-60mg, 5-50mg, 5-40mg, 5-30mg, 5-25mg, 5-20mg, 10-1500mg, 10-1000mg, 10-900mg, 10-800mg, 10-700mg, 10-600mg, 10-500mg, 10-450mg, 10-400mg, 10-300mg, 10-250mg, 10-20 0mg, 10-150mg, 10-125mg, 10-100mg, 10-90mg, 10-80mg, 10-70mg, 10-60mg, 10-50mg, 10-40mg, 10-30mg, 10-20mg; 2 0-1500mg, 20-1000mg, 20-900mg, 20-800mg, 20-700mg, 20-600mg, 20-500mg, 20-400mg, 20-350mg, 20-300mg, 20-25 0mg, 20-200mg, 20-150mg, 20-125mg, 20-100mg, 20-90mg, 20-80mg, 20-70mg, 20-60mg, 20-50mg, 20-40mg, 20-30mg;50-1500mg, 50-1000mg, 50-900mg, 50-800mg, 50-700mg, 50-600mg, 50-500mg, 50-400mg, 50-300mg, 50-250mg, 50-200mg, 50-150mg, 50-125mg, 5 0-100mg; 100-1500mg, 100-1000mg, 100-900mg, 100-800mg, 100-700mg, 100-600mg, 100-500mg, 100-400mg, 100-300mg, 100-250mg, 100-200mg;

[0069] In some embodiments, the pharmaceutical composition or formulation of the present invention contains the above-mentioned therapeutically effective amount of the compound of the present invention or its stereoisomer, solvate, or pharmaceutically acceptable salt;

[0070] The present invention relates to a pharmaceutical composition or pharmaceutical preparation, which comprises a therapeutically effective amount of the compound of the present invention or its stereoisomer or pharmaceutically acceptable salt and a carrier and / or excipient. The pharmaceutical composition can be in the form of a unit preparation (the amount of the main drug in the unit preparation is also referred to as "preparation specification"). In some embodiments, the pharmaceutical composition includes but is not limited to 1mg, 1.25mg, 2.5mg, 5mg, 10mg, 12.5mg, 15mg, 20mg, 25mg, 30mg, 35mg, 40mg, 45mg, 50mg, 55mg, 60mg, 65mg, 70mg, 75mg, 80mg, 85mg, 90mg, 95mg, 100mg, 110mg, 120mg, 125mg, 130mg, 140mg, 150mg, 160mg, 170mg, 180mg, 190mg, 200mg, 210mg, 220mg, 230mg , 240mg, 250mg, 275mg, 300mg, 325mg, 350mg, 375mg, 400mg, 425mg, 450mg, 475mg, 500mg, 525mg, 550mg, 575mg, 600mg, 625mg, 650mg, 675mg, 700mg, 725mg, 750mg, 775mg, 800mg, 850mg, 900mg, 950mg, 1000mg, 1100mg, 1200mg, 1300mg, 1400mg, 1500mg of a compound of the present invention or a stereoisomer or a pharmaceutically acceptable salt thereof.

[0071] The present invention relates to a kit, which may include a composition in a single-dose or multi-dose form, wherein the kit contains a compound of the present invention or a stereoisomer or a pharmaceutically acceptable salt thereof, and the amount of the compound of the present invention or a stereoisomer or a pharmaceutically acceptable salt thereof is the same as that in the above-mentioned pharmaceutical composition.

[0072] The amount of the compound according to the invention or its stereoisomer or pharmaceutically acceptable salt in the present invention is in each case calculated as the free base.

[0073] "Preparation specifications" refers to the weight of the main drug contained in each vial, tablet or other unit preparation.

[0074] Synthetic route

[0075] Patent documents such as WO2013026797A1 introduce the preparation method of PDE4B inhibitors. Those skilled in the art can combine the documents and known organic synthesis techniques to prepare the compounds of the present invention, and the starting materials are commercially available chemicals and (or) compounds described in chemical literature. "Commercially available chemicals" are obtained from regular commercial sources, and suppliers include: Titan Technology, Anage Chemical, Shanghai Demo, Chengdu Kelon Chemical, Shaoyuan Chemical Technology, Nanjing Yaoshi, WuXi AppTec, and Bailingwei Technology.

[0076] Specific and similar reactants can be selectively identified through indexes of known chemical substances prepared by the Chemical Abstracts Service of the American Chemical Society, which are available in most public and university libraries and online. Chemicals that are known but not commercially available in the catalog are optionally prepared by custom chemical synthesis plants, many of which provide custom synthesis services.

[0077] the term

[0078] Unless otherwise specified in the present invention, the terms of the present invention have the following meanings:

[0079] "Halogen" herein refers to F, Cl, Br, I, or isotopes thereof.

[0080] "Halo" or "halogen substituted" means that a hydrogen atom is replaced by one or more halogens selected from F, Cl, Br, I, or isotopes thereof, and the upper limit of the number of halogen substituents is equal to the sum of the number of hydrogen atoms that can be replaced by the substituted group. Unless otherwise specified, the number of halogen substituents is any integer between 1 and the upper limit. When the number of halogen substituents is greater than 1, they may be substituted by the same or different halogens.

[0081] "Deuterated" or "deuterated substance" refers to the situation where hydrogen atoms on groups such as alkyl, cycloalkyl, alkylene, aryl, heteroaryl, thiol, heterocycloalkyl, alkenyl, alkynyl, etc. are replaced by at least one isotope deuterium, and the upper limit of the number of deuterations is equal to the sum of the number of replaceable hydrogen atoms in the substituted group. Unless otherwise specified, the number of deuterations is any integer between 1 and the upper limit, preferably 1-20 deuterium atoms are replaced, more preferably 1-10 deuterium atoms are replaced, more preferably 1-6 deuterium atoms are replaced, and further preferably 1-3 deuterium atoms are replaced.

[0082] "Alkyl" refers to a monovalent straight or branched saturated aliphatic hydrocarbon group, and unless otherwise specified, is an alkyl group of 1 to 20 carbon atoms, preferably an alkyl group of 1 to 8 carbon atoms, more preferably an alkyl group of 1 to 6 carbon atoms, further preferably an alkyl group of 1 to 4 carbon atoms, and further preferably an alkyl group of 1-2 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, neobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl and various branched isomers thereof.

[0083] "Alkylene" refers to a divalent straight chain or branched chain saturated alkyl group. Examples of alkylene groups include, but are not limited to, methylene, ethylene, propylene, butylene, and the like.

[0084] "Cycloalkyl" refers to a monovalent non-aromatic, partially unsaturated or fully saturated, substituted or unsubstituted carbocyclic hydrocarbon group, which, unless otherwise specified, usually has 3 to 12 carbon atoms, preferably 3 to 10 carbon atoms, more preferably 3 to 6 carbon atoms, and further preferably 3 to 4 carbon atoms. Non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, Or cycloheptyl, etc.

[0085] "Cycloalkylene" refers to a divalent radical of "cycloalkyl", and non-limiting examples include cyclopropylene, cyclobutylene, and the like.

[0086] "Heterocycle" or "heterocyclic group" refers to a substituted or unsubstituted, saturated or unsaturated aromatic or non-aromatic ring, which contains 1 to 3 heteroatoms selected from N, O or S when not specifically defined, including monocyclic heterocycles, bicyclic bridged heterocycles, bicyclic heterocycles and bicyclic spiro heterocycles, etc., and is 3 to 12-membered heterocycles, more preferably 4-12-membered heterocycles, more preferably 4-10-membered heterocycles, and further preferably 4-7-membered heterocycles when not specifically defined. Its definition includes heterocycloalkyl and heteroaryl. N and S in the heterocyclic ring can be oxidized to various oxidation states. The heterocyclic group may be attached to a heteroatom or a carbon atom, and non-limiting examples include oxirane, aziridine, oxetanyl, azetidinyl, 1,3-dioxolanyl, 1,4-dioxolanyl, 1,3-dioxhexacyclyl, azepanyl, pyridinyl, furanyl, thienyl, pyranyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyrazolyl, pyridazinyl, imidazolyl, piperidinyl, piperidinyl, morpholinyl, thiomorpholinyl, 1,3-dithianyl, dithio ... ]octanyl, azabicyclo[5.2.0]nonanyl, oxatricyclo[5.3.1.1]dodecyl, azaadamantyl and oxaspiro[3.3]heptanyl,

[0087]

[0088] wait.

[0089] "Heterocyclylene" is a divalent group corresponding to "heterocyclyl", and non-limiting examples include imidazolyl, piperidinyl, aziridinyl, and the like.

[0090] "Carbocycle" or "carbocyclic group" refers to substituted or unsubstituted, saturated or unsaturated, aromatic or non-aromatic carbocyclic groups, including monocyclic carbocycles, bicyclic bridged rings, bicyclic cyclic rings and bicyclic spirocycles, etc., and unless otherwise specified, has 3 to 12 carbon atoms, preferably 3-10 carbon atoms, and more preferably 3-6 carbon atoms. Its definition includes cycloalkyl and aryl. In non-limiting examples, monocyclic carbocycles include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or phenyl, etc., the bicyclic bridge ring includes etc., double ring and ring include etc., bicyclic spiro ring includes wait.

[0091] "Aryl" refers to a carbon ring having aromatic properties. Non-limiting examples include phenyl, naphthyl, and the like.

[0092] "Alkynyl" refers to a linear or branched monovalent unsaturated hydrocarbon group containing one or more carbon-carbon triple bonds. Unless otherwise specified, the alkynyl group contains 2-6 carbon atoms, preferably 2-4 carbon atoms. Non-limiting examples include ethynyl, propynyl, propargyl, etc.

[0093] "Alkenyl" refers to a linear or branched monovalent unsaturated hydrocarbon group containing one or more carbon-carbon double bonds. Unless otherwise specified, the alkynyl group contains 2-6 carbon atoms, preferably 2-4 carbon atoms. Non-limiting examples are ethenyl, propenyl, allyl, 2-butenyl, 1-butenyl, etc.

[0094] "Alkoxy" or "alkyloxy" refers to -O-alkyl, and unless otherwise specified, -OC 1-8 Alkyl, preferably -OC 1-6 Alkyl, more preferably -OC 1-4 Alkyl, more preferably -OC 1-2 Non-limiting examples include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, n-hexyloxy, cyclopropyloxy, cyclobutyloxy, and the like.

[0095] "Haloalkoxy" refers to -O-haloalkyl, and unless otherwise specified, is -O-haloC 1-8 Alkyl, preferably -O-halogenated C 1-6 Alkyl, more preferably -O-halogenated C 1-4 Alkyl, more preferably -O-halogenated C 1-2 Non-limiting examples include monofluoromethoxy, difluoromethoxy, trifluoromethoxy, difluoroethyloxy, and the like.

[0096] “C 1-4 "Alkyl acyl" refers to C 1-4 Alkyl-C(O)-. Non-limiting examples include formyl, acetyl, propionyl.

[0097] “C 1-4 "Alkylsulfonyl" refers to C 1-4 Alkyl-S(O)2-. Non-limiting examples include methylsulfonyl, ethylsulfonyl, propanesulfonyl.

[0098] "Heteroaromatic ring" or "heteroaryl" refers to a heterocyclic ring having aromatic properties. Non-limiting examples include pyrazolyl, pyrimidinyl, thiazolyl, pyridinyl, furanyl, and the like.

[0099] "Heterocycloalkyl" refers to a non-aromatic, partially unsaturated or fully saturated heterocycle, which generally has 4 to 12 ring members, preferably 4 to 10 ring members, more preferably 4 to 7 ring members, and further preferably 5 or 6 ring members. In addition to carbon atoms, heterocycloalkyl also contains 1-3 heteroatoms selected from N, S, O, Si, P as ring members. Non-limiting examples include azetidinyl, morpholinyl, piperazinyl, piperidinyl, tetrahydropyranyl, oxetanyl, etc.

[0100] "Alkylamino" or "alkylamino" refers to an amino group substituted with a single or double alkyl group, also written as -N-(alkyl)2 or -NH-alkyl, the latter also written as monoalkylamino. Non-limiting examples include dimethylamino, monomethylamino, diethylamino, monoethylamino, and the like.

[0101] "Optional" or "optionally" means that the event or circumstance described later may but need not occur, and the description includes situations where the event or circumstance occurs or does not occur. For example, "alkyl optionally substituted with F" means that alkyl may but need not be substituted with F, and the description includes situations where alkyl is substituted with F and situations where alkyl is not substituted with F.

[0102] "Pharmaceutically acceptable salt" refers to a salt of the compound of the present invention which retains the biological effectiveness and properties of the free acid or free base, and the free acid is obtained by reacting with a non-toxic inorganic base or organic base, or the free base is obtained by reacting with a non-toxic inorganic acid or organic acid.

[0103] A "pharmaceutical composition" refers to a mixture of one or more compounds described herein, or stereoisomers, solvates, pharmaceutically acceptable salts or cocrystals thereof, with other ingredients, wherein the other ingredients include physiologically / pharmaceutically acceptable carriers and / or excipients.

[0104] "Carrier" refers to a system that does not cause significant irritation to the organism and does not eliminate the biological activity and properties of the administered compound, and can change the way the drug enters the human body and its distribution in the body, control the release rate of the drug and deliver the drug to the targeted organ. Non-limiting examples include microcapsules and microspheres, nanoparticles, liposomes, etc.

[0105] "Excipient" refers to a substance that is not a therapeutic agent in itself but is used as a diluent, adjuvant, binder and / or vehicle that is added to a pharmaceutical composition to improve its handling or storage properties or to allow or facilitate the formation of a compound or pharmaceutical composition into a unit dosage form for administration. As known to those skilled in the art, pharmaceutical excipients can serve a variety of functions and can be described as wetting agents, buffers, suspending agents, lubricants, emulsifiers, disintegrants, absorbents, preservatives, surfactants, colorants, flavoring agents and sweeteners. Examples of pharmaceutical excipients include, but are not limited to: (1) sugars such as lactose, glucose and sucrose; (2) starches such as corn starch and potato starch; (3) cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, cellulose acetate, hydroxypropyl methylcellulose, hydroxypropyl cellulose, microcrystalline cellulose and cross-linked carboxymethylcellulose (e.g., cross-linked sodium carboxymethylcellulose); (4) tragacanth powder; (5) malt; (6) gelatin; (7) talc; (8) excipients such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerol, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) pH buffer solutions; (21) polyesters, polycarbonates and / or polyanhydrides; and (22) other nontoxic compatible substances used in pharmaceutical preparations.

[0106] "Isomers" include "stereoisomers" and "tautomers". "Stereoisomers" refer to isomers produced by the same order of connection of atoms or atomic groups in molecules, but different spatial arrangements. Stereoisomers include cis-trans isomers and optical isomers. "Tautomers" refer to the transformation of one functional group to another functional group that can be converted into each other through a reversible chemical reaction called tautomerization, usually caused by the concomitant migration of hydrogen atoms and π bonds (double bonds or triple bonds). For example, the following pairs of compounds: aldehyde / keto-enol, imine-enamine.

[0107] The compounds of the present invention also include their tautomers. For example, when the present invention describes the left-hand compound in which the pyrimidine ring is substituted with OH, the right-hand tautomer compound is also included.

[0108] DETAILED DESCRIPTION

[0109] The content of the present invention will be described in detail below through examples. If no specific conditions are specified in the examples, the experimental method according to conventional conditions is carried out. The examples are given to better illustrate the content of the present invention, but it should not be understood that the content of the present invention is limited to the examples. Those skilled in the art can make non-essential improvements and adjustments to the implementation scheme according to the above invention content, which still belongs to the protection scope of the present invention.

[0110] Dess-Martin reagent; 1,1,1-triacetoxy-1,1-dihydro-1,2-benzidoyl-3-(1H)-one

[0111] DIPEA: N,N-diisopropylethylamine

[0112] NMP; N-methylpyrrolidone

[0113] TBDMSCl: tert-butyldimethylsilyl chloride

[0114] DMAP: 4-dimethylaminopyridine

[0115] HATU: 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate

[0116] T3P: Propylphosphonic acid tricyclic anhydride

[0117] Embodiment 1:

[0118]

[0119] Step 1: Add 5-chloro-2-iodopyrimidine (578 mg, 2.41 mmol), 1,1-bis(diphenylphosphine)dichloropalladium iron (176 mg, 0.24 mmol) and sodium carbonate (766 mg, 7.32 mmol) to compound 1A (0.89 g, 2.65 mmol) (WO2020 / 86533, 2020, A1) dissolved in 1,4-dioxane / water (10 ml / 2 ml) under nitrogen, and react in an oil bath at 90 ° C for 18 hours. After the reaction is completed, cool to room temperature, filter with diatomaceous earth, wash with ethyl acetate, concentrate the filtrate and directly separate and purify the target compound 1B (0.44 g, 52%) by column chromatography (eluent ratio: EA / PE = 0% to 10%).

[0120] LCMS (ESI): m / z = 266.1 [M + H-56]

[0121] Step 2: Compound 1B (200 mg, 0.62 mmol), cyclopropylboronic acid (80 mg, 0.93 mmol), palladium acetate (14 mg, 0.062 mmol), tricyclohexylphosphine (36 mg, 0.12 mmol), potassium phosphate (394 mg, 1.86 mmol), and finally toluene (5 mL) and water (1 mL) were added, and nitrogen was replaced three times. After heating to 100 ° C for 3 h, it was passed through a short silica gel column, washed with ethyl acetate, concentrated, and column chromatography (petroleum ether / ethyl acetate = 10 / 1) was performed to obtain compound 1C (200 mg, 98%).

[0122] LC-MS(ESI):m / z=272.2[M+H-56] +

[0123] Step 3: Dissolve compound 1C (200 mg, 0.63 mmol) in 1,4-dioxane (2 ml) with hydrochloric acid, stir at room temperature for 1 h, and directly spin dry to obtain hydrochloride compound 1D (170 mg).

[0124] LCMS (ESI): m / z = 228.2 [M+H] +

[0125] Step 4: Compound 1E (1 g, 3.48 mmol) (WO2013 / 026797, 2013, A1) was dissolved in dichloromethane (10 mL), trichloroacetyl isocyanate (983 mg, 5.23 mmol) was added under ice bath, and the reaction was continued for 1 h, and then concentrated to obtain 1F (1.5 g, 91%), which was directly used in the next step. LC-MS (ESI): m / z = 475.1 [M+H] +

[0126] Step 5: Dissolve compound 1F (1.5 g, 3.16 mmol) in tetrahydrofuran (20 mL), add water (5 mL) and potassium carbonate (1.31 g, 9.48 mmol), react at room temperature for 2 hours, concentrate, add dichloromethane for extraction, dry, concentrate, and perform column chromatography (dichloromethane / methanol = 10 / 1) to obtain compound 1G (1 g, 96%).

[0127] LC-MS (ESI): m / z = 331.1 [M+H] +

[0128] Step 6: Dissolve compound 1G (200 mg, 0.6 mmol) in 1,4-dioxane (10.0 mL), add compound 1D (137 mg, 0.6 mmol) and diisopropylethylamine (232 mg, 1.8 mmol), heat back to 100 ° C for 3 h, cool to room temperature, concentrate, and column chromatography (dichloromethane / methanol = 10 / 1) to obtain compound 1 (300 mg, 96%).

[0129] LC-MS (ESI): m / z = 522.2 [M+H] +

[0130] Compound 1 splitting: Compound 1 (300 mg) was used for splitting, and after separation, compound 1-1 (retention time: 0.976 min, 135.2 mg, ee% = 100%) and compound 1-2 (retention time: 1.501 min, 108.9 mg, ee% = 99.94%) were obtained.

[0131] Separation conditions: Instrument: Waters 150Prep-SFC E; Column: Chiralcel Cellulose-2 column; Mobile phase: A for CO2; B for 0.1% NH3·H2O in IPA and ACN; Gradient: B 70%; Flow rate: 100 mL / min; Back pressure: 100 bar; Column temperature: 25°C; Wavelength: 220 nm; Cycle: 7.0 min;

[0132] Compound 1-1:

[0133] 1 H NMR (400MHz, DMSO-d6) δ = 8.51 (s, 2H), 7.45 (s, 1H), 6.72 (s, 1H), 6.46 (s, 2H) ,4.48–4.27(m,2H),3.88(d,1H),3.74(d,1H),3.64(s,2H),3.44–3.34(m,1H) ,3.20–3.08(m,3H),2.99-2.81(m,3H),2.73(d,1H),2.43–2.26(m,2H),2.20( s,2H),1.94-1.89(m,1H),1.80(s,2H),1.06–0.98(m,2H),0.86–0.78(m,2H).

[0134] Compound 1-2:

[0135] 1H NMR (400 MHz, DMSO-d6) δ = 8.51 (s, 2H), 7.46 (s, 1H), 6.72 (s, 1H), 6.46 (s, 2H), 4.51-4.28 (m, 2H), 3.88 (d, 1H), 3.74 (d, 1H), 3.65 (d, 2H), 3.46-3.36 (m, 1H), 3.24-3.04 (m, 3H), 3.02-2.81 (m, 3H), 2.74 (d, 1H), 2.42-2.25 (m, 2H), 2.20 (s, 2H), 1.96-1.89 (m, 1H), 1.80 (s, 2H), 1.06-0.98 (m, 2H), 0.89-0.77 (m, 2H). Example 2:

[0136]

[0137] Step 1: Dissolve compound 2A (1 g, 5.75 mmol) in acetone (10 ml), add cesium carbonate (5.62 g, 17.24 mmol) and iodoethane (1.79 g, 11.5 mmol), stir at room temperature for 3 h, add water to quench the reaction after the reaction is completed, extract with ethyl acetate, collect and combine the organic phases, dry over anhydrous sodium sulfate, concentrate, and quickly separate and purify (petroleum ether / ethyl acetate = 10 / 1) to obtain compound 2B (1.11 g, 95%).

[0138] LC-MS (ESI): m / z = 203.1 [M+H] + .

[0139] According to the synthesis method of Example 1, compound 2 (300 mg, 95%) was obtained.

[0140] Compound 2 resolution: Compound 2 (300 mg) was used for resolution, and after separation, compound 2-1 (retention time: 0.891 min, 87.7 mg, ee% = 100%) and compound 2-2 (retention time: 1.273 min, 98.4 mg, ee% = 99.73%) were obtained.

[0141] Separation conditions: instrument: Waters 150Prep-SFC E; column: Chiralcel Cellulose-2 column; mobile phase: A for CO2; B for 0.1% NH3·H2O in IPA and ACN; gradient: B 70%; flow rate: 100 mL / min; back pressure: 100 bar; column temperature: 25°C; wavelength: 220 nm; cycle: 7.0 min.

[0142] Compound 2-1:

[0143] 1 H NMR (400MHz, DMSO-d6)δ=8.48(s,2H),7.45(s,1H),6.62(s,1H),6.46(s,2H),4.48–4.26(m,2H),4.18(q,2H),3.90(t,1H),3.76–3.72(m,1H), 3.65–3.63(m,2H),3.46–3.35(m,1H),3.21–3.08(m,3H),2.99–2.81(m ,3H),2.73(d,1H),2.31(d,2H),2.20(s,2H),1.80(s,2H),1.35(t,3H).

[0144] LC-MS (ESI): m / z = 526.2 [M+H] + .

[0145] Compound 2-2:

[0146] 1 H NMR (400MHz, DMSO-d6)δ=8.48(s,2H),7.45(s,1H),6.62(s,1H),6.46(s,2H),4.40(d,2H),4.18(q,2H),3.87(s,1H),3.76– 3.63(m,3H),3.38(s,1H),3.25–3.03(m,3H),3.01–2.79(m,3H),2.74(d,1H),2.42–2.11(m,4H),1.80(s,2H),1.35(t,3H).

[0147] LC-MS (ESI): m / z = 526.2 [M+H] + .

[0148] Embodiment 3:

[0149]

[0150] Step 1: Add compound 3A (20.0 g, 103 mmol), tert-butyl 3-(hydroxymethyl)piperazine-1-carboxylate (24.6 g, 113 mmol) and N,N-diisopropylethylamine (26.6 g, 206 mmol) to a 500 mL round-bottom flask, then add N-methylpyrrolidone (150 mL), heat to 130 ° C and react overnight. Cool to room temperature, pour the reaction solution into water (600 mL), extract with ethyl acetate (100 mL x 5), combine the organic phases and wash three times with water (150 mL), dry the organic phase with anhydrous sodium sulfate, filter and concentrate, and the crude product is separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 100 / 0 to 50 / 50) to obtain the target compound 3B (21.3 g, 52.9%).

[0151] LC-MS (ESI): m / z = 390.1 [M+H] +

[0152] Step 2: Compound 3B (21.3 g, 54.6 mmol) and potassium tert-butoxide (12.2 g, 109 mmol) were dissolved in tert-butanol (150 mL), heated to 80°C and stirred for 3 h. After the reaction was completed, the temperature was cooled to room temperature, water (400 mL) was added to quench the reaction, and ethyl acetate (100 mL x 5) was used for extraction. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the reaction solution was concentrated under reduced pressure and purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 100 / 0 to 70 / 30) to obtain the target compound 3C (19.4 g, 96.0%).

[0153] LC-MS (ESI): m / z = 370.1 [M+H] + .

[0154] Step 3: Compound 3C (5 g, 13.55 mmol) was dissolved in dry toluene (50 ml), and diboronic acid pinacol ester (0.88 g, 27.1 mmol), [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane complex (1.98 g, 2.71 mmol) and sodium acetate (2.22 g, 27.1 mmol) were added, and nitrogen was replaced three times, and the mixture was heated to 100°C and stirred for 4 h. After the reaction was completed, the mixture was cooled to room temperature, and water (50 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (50 mL x 3), and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the reaction solution was concentrated under reduced pressure and separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 100 / 0 to 70 / 30) to obtain the target compound 3D (4.8 g, 85%).

[0155] LC-MS (ESI): m / z = 418.2 [M+H]+ .

[0156] Step 4: Compound 3D (1.2 g, 2.87 mmol) was dissolved in acetone (20 ml), and hydrogen peroxide (0.24 ml, 8.61 mmol) and sodium bicarbonate (1.69 g, 20.09 mmol) were added. The mixture was stirred at room temperature overnight and tested by TLC. After the reaction was completed, water and dichloromethane were added to dilute the reaction system. The aqueous phase was extracted with dichloromethane, and the organic phases were collected and combined, dried over anhydrous sodium sulfate, concentrated, and quickly separated and purified (petroleum ether / ethyl acetate = 5 / 1) to obtain compound 3E (0.8 g, 91%).

[0157] LC-MS (ESI): m / z = 308.2 [M+H] + .

[0158] Step 5: Compound 3E (0.8 g, 2.61 mmol) was dissolved in acetone (10 ml), and cesium carbonate (2.55 g, 7.83 mmol) and iodoethane (0.82 g, 5.22 mmol) were added. The mixture was stirred at room temperature for 3 h. After the reaction was completed, water was added to quench the reaction. The mixture was extracted with ethyl acetate. The organic phases were collected and combined, dried over anhydrous sodium sulfate, concentrated, and quickly separated and purified (petroleum ether / ethyl acetate = 10 / 1) to obtain compound 3F (0.8 g, 92%).

[0159] LC-MS (ESI): m / z = 336.2 [M+H] + .

[0160] Step 6: Compound 3F (0.8 g) was separated by SFC to obtain two isomers, compound 3G-1 (373 mg, retention time 5.86 min) and compound 3G-2 (340 mg, retention time 7.27 min)

[0161] Preparative chromatographic separation conditions: 1. Instrument: Waters 150AP-SFC; 2. Chromatographic column: AD; 3. Mobile phase system: A for CO2; B for MEOH; 4. Gradient: B 28%; 5. Flow rate: 40 mL / min

[0162] Using 3G-1 (90 mg, 0.26 mmol) as raw material, according to the synthesis method of Example 1, compound 3-1 (80 mg, 63%) was obtained;

[0163] Referring to the above method, 3G-2 was used as the raw material to obtain the target compound 3-2 (85 mg, 67%).

[0164] Compound 3-1:

[0165] 1H NMR(400MHz, DMSO-d6)δ=7.50(d,1H),7.43(s,1H),6.78(d,1H),4.88–4.60(m,3H) ,4.38-4.30(m,2H),4.03–3.90(m,3H),3.71(s,2H),3.49–3.37(m,1H),3.27–3.12 (m,2H),3.05–2.82(m,3H),2.75–2.67(m,1H),2.64–2.57(m,1H),2.40–2.27(m,2H ),2.22–2.16(m,2H),1.82–1.75(m,2H),1.27(t,3H).LC-MS(ESI):m / z=487.2[M+H] +

[0166] Compound 3-2:

[0167] 1 H NMR (400MHz, DMSO-d6) δ = 7.49 (d, 1H), 7.43 (s, 1H), 6.78 (d, 1H), 4.83 (t, 1H), 4.74 (d,2H),4.39-4.30(m,2H),4.06–3.91(m,3H),3.72(d,2H),3.51–3.36(m,1H),3.25 –3.13(m,2H),3.06–2.83(m,3H),2.72(t,1H),2.63–2.58(m,1H),2.40–2.27(m,2H ),2.22–2.17(m,2H),1.82–1.75(m,2H),1.27(t,3H).LC-MS(ESI):m / z=487.2[M+H] +

[0168] Embodiment 4:

[0169]

[0170] Using 1G (100 mg, 0.3 mmol) and 3H-1 as raw materials, according to the synthesis method of Example 1, compound 4-1 (104 mg, 65%) was obtained; using 1G (100 mg, 0.3 mmol) and 3H-2 as raw materials, according to the synthesis method of Example 1, the target compound 4-2 (108 mg, 68%) was obtained.

[0171] Compound 4-1:

[0172] 1H NMR (400MHz, DMSO-d6) δ = 7.68 (s, 1H), 7.49 (d, 1H), 6.78 (d, 1H), 6.50 (s, 2H), 4.75 (t, 2H), 4.50–4.20 (m, 4H), 4.03–3.90 (m, 3H), 3.50–3. 37(m,1H),3.24–3.11(m,2H),3.04–2.79(m,3H),2.76–2.68(m,1H),2 .64–2.59(m,1H),2.41–2.22(m,4H),1.87-1.79(m,2H),1.28(t,3H).

[0173] LC-MS (ESI): m / z = 530.2 [M+H] +

[0174] Compound 4-2:

[0175] 1 H NMR(400MHz, DMSO-d6)δ=7.69(s,1H),7.49(d,1H),6.78(d,1H),6.50(s,2H),4.75(t,2H),4.49–4.24(m,4H),4.04–3.91(m,3H),3. 50–3.39(m,1H),3.27–3.12(m,2H),3.05–2.82(m,3H),2.71(t,1H),2.61(t,1H),2.41–2.19(m,4H),1.89–1.73(m,2H),1.28(t,3H).

[0176] LC-MS (ESI): m / z = 530.2 [M+H] +

[0177] Embodiment 5:

[0178]

[0179] Step 1: At room temperature, add compound 5A (5.0 g, 31.3 mmol), tert-butyl 3-(hydroxymethyl)piperazine-1-carboxylate (6.8 g, 31.3 mmol), potassium hydroxide (5.3 g, 93.9 mmol) and N,N-dimethylformamide (100 mL) in sequence. Heat to 100°C and react for 15 h. After the reaction is completed, filter the reaction solution, extract the filtrate with ethyl acetate (500 mL), wash the organic phase with water (100 mL×3), wash with saturated brine (100 mL), dry the organic phase with anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure to obtain 5B (10 g, 95%).

[0180] LC-MS(ESI):m / z=281.1[M-tBu+H] +

[0181] Step 2: At room temperature and under hydrogen atmosphere, compound 5B (10 g, 29.76 mmol), palladium carbon (2 g) and ethyl acetate (300 mL) were added to a 1 L round-bottom flask and reacted at room temperature for 12 hours. After the reaction, the reaction solution was filtered and the filtrate was concentrated under reduced pressure to obtain 5C (8.0 g, 88%).

[0182] LC-MS (ESI): m / z = 307.1 [M+H] +

[0183] Step 3: At room temperature, tert-butyl nitrite (3.5 g, 33.83 mmol), cuprous iodide (5.2 g, 27.06 mmol) and acetonitrile (30 mL) were added to a 100 mL round-bottom flask in sequence, heated to 65°C, and a solution of 5C (6.9 g, 22.55 mmol) in acetonitrile (40 mL) was added dropwise. After the addition was complete, the mixture was reacted at 65°C for 6 h. After the reaction was completed, the mixture was cooled to room temperature, the reaction solution was concentrated under reduced pressure, and then purified by silica gel column chromatography (eluent EA / PE (v / v) = 0-20%) to obtain 5D (1.5 g, 16%).

[0184] LC-MS (ESI): m / z = 418.4 [M+H] +

[0185] Step 4: Compound 5D (1.5 g) was separated by SFC to obtain two isomers, compound 5D-1 (700 mg, retention time 5.86 min, 47%) and compound 5D-2 (700 mg, retention time 7.27 min, 47%).

[0186] Preparative chromatographic separation conditions: 1. Instrument: Waters 150AP-SFC; 2. Chromatographic column: AD; 3. Mobile phase system: A for CO2; B for MEOH; 4. Gradient: B 28%; 5. Flow rate: 40 mL / min

[0187] LC-MS (ESI): m / z = 418.0 [M+H] +

[0188] Step 5: At room temperature and under nitrogen atmosphere, compound 5D-1 (700 mg, 1.68 mmol), propyne (1M / L, 33.6 mL, 33.6 mmol), bistriphenylphosphine palladium dichloride (119.3 mg, 0.17 mmol), cuprous iodide (64.6 mg, 0.34 mmol) and tetrahydrofuran (10 mL) were added to a 50 mL round-bottom flask, and triethylamine (509.1 mg, 5.04 mmol) was added, and the mixture was reacted at this temperature for 12 h. After the reaction was completed, water (5 mL) was added, and the mixture was extracted twice with ethyl acetate (10 mL), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the reaction solution was concentrated under reduced pressure and purified by silica gel column chromatography (eluent EA / PE (v / v) = 0-30%) to obtain the target compound 5E-1 (500 mg, 90%).

[0189] LC-MS (ESI): m / z = 330.2 [M+H] +

[0190] Referring to the above operation, compound 5D-2 (700 mg, 1.68 mmol) was used as the raw material to obtain the target compound 5E-2 (500 mg, 90%).

[0191] LC-MS (ESI): m / z = 330.2 [M+H] +

[0192] Step 6: At room temperature, compound 5E-1 (500 mg, 1.52 mmol), dichloromethane (4 mL) and trifluoroacetic acid (1 mL) were added to a 25 mL round-bottom flask and stirred for 2 h at this temperature. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain the target compound 5F-1 (325 mg, 93%)

[0193] LC-MS (ESI): m / z = 230.2 [M+H] +

[0194] Referring to the above operation, compound 5E-2 (500 mg, 1.52 mmol) was used as the raw material to obtain the target compound 5F-2 (330 mg, 95%)

[0195] LC-MS (ESI): m / z = 230.2 [M+H] +

[0196] Using 1G (100 mg, 0.3 mmol) and 5F-1 as raw materials, according to the synthesis method of Example 1, compound 5-1 (101 mg, 64%) was obtained; using 1G (100 mg, 0.3 mmol) and 5F-2 as raw materials, according to the synthesis method of Example 1, the target compound 5-2 (103 mg, 66%) was obtained.

[0197] Compound 5-1:

[0198] 1 H NMR(400MHz, DMSO-d6)δ=7.78(d,1H),7.68(s,1H),6.97(d,1H),6.49(s,2H),4.75(d,2H),4.46–4.36(m,4H),4.00–3.95(m,1H),3.49–3. 36(m,2H),3.26–3.16(m,1H),3.05–2.92(m,2H),2.91–2.86(m,1H),2 .79–2.72(m,2H),2.41–2.20(m,4H),2.00(s,3H),1.89–1.77(m,2H).

[0199] LC-MS (ESI): m / z = 524.2 [M+H] +

[0200] Compound 5-2:

[0201] 1 H NMR (400MHz, DMSO-d6)δ=7.78(d,1H),7.68(s,1H),6.97(d,1H),6.49(s,2H),4.74(d,2H),4.47–4.31(m,4H),4.00–3.95(m,1H),3.51–3. 35(m,2H),3.24–3.17(m,1H),3.04–2.93(m,2H),2.92–2.83(m,1H),2 .80–2.72(m,2H),2.41–2.25(m,4H),2.00(s,3H),1.87–1.79(m,2H).

[0202] LC-MS (ESI): m / z = 524.2 [M+H] +

[0203] Embodiment 6:

[0204]

[0205] Step 1: Dissolve compound 6A (5.0 g, 21.81 mmol) in acetonitrile (50 mL), then add pyridine (5.18 g, 65.42 mmol) and di-tert-butyl dicarbonate (7.14 g, 32.71 mmol), stir at room temperature for 4 hours, add ammonia water (10 mL) dropwise, and react at room temperature for 4 hours. After the reaction is complete, concentrate, filter the solid, and wash with acetonitrile to obtain compound 6B (4.5 g, 90%).

[0206] LC-MS(ESI):m / z=173.1[M+H-56] +

[0207] Step 2: Dissolve compound 6B (4.5 g, 19.71 mmol) in dichloromethane (40 mL), add trifluoroacetic acid (10 mL), and stir at room temperature for 1 hour. The reaction solution is concentrated to obtain the crude trifluoroacetate salt of compound 6C, which is directly used for the next step without purification.

[0208] LC-MS (ESI): m / z = 129.1 [M+H] +

[0209] Step 3: The crude trifluoroacetate of compound 6C in the previous step was dissolved in N,N-dimethylformamide (50 mL), and N,N-diisopropylethylamine (6.12 g, 29.57 mmol) and 2,4-dichloro-6,7-dihydrothieno[3,2-d]pyrimidine (12.74 g, 6.87 mmol) were added, and the mixture was heated to 80°C for 16 h. The reaction solution was cooled to room temperature, diluted with ethyl acetate and washed with water, and the organic phase was dried, filtered, concentrated, and column chromatography (dichloromethane / methanol (v / v) = 91 / 9) was performed to obtain compound 6D (720 mg, 12%).

[0210] LC-MS (ESI): m / z = 299.2 [M+H] +

[0211] Step 4: Dissolve the compound 6D in dichloromethane (30 mL), add triethylamine (490 mg, 4.82 mmol) under ice bath, stir for 5 minutes, then add trifluoroacetic anhydride (760 mg, 3.62 mmol) dropwise, stir for 2 hours under ice bath. The reaction solution is quenched with water, extracted with dichloromethane, and the organic phase is dried, filtered, concentrated, and column chromatography (dichloromethane / methanol (v / v) = 91 / 9) is performed to obtain compound 6E (440 mg, 65%).

[0212] LC-MS (ESI): m / z = 281.2 [M+H] +

[0213] Step 5: Compound 6E (500 mg, 1.78 mmol), (S)-(-)-1,1'-bi-2-naphthol (51.0 mg, 0.18 mmol), tetraisopropyloxytitanium (25.0 mg, 0.088 mmol) and water (32.0 mg, 1.78 mmol) were dissolved in dichloromethane (20 mL), nitrogen was replaced three times, and tert-butyl peroxide (256 mg, 1.99 mmol, 70% in water) was added after stirring at room temperature for one hour, and stirred overnight. The solution was quenched with saturated sodium thiosulfate aqueous solution, the aqueous phase was extracted with dichloromethane, dried and concentrated, and column chromatography (dichloromethane / methanol (v / v) = 91 / 9) was performed to obtain compound 6F (520 mg, 98%). LC-MS (ESI): m / z = 297.1 [M+H] +

[0214] Step 6: Using 6F (100 mg, 0.3 mmol) and 5F-1 as raw materials, according to the synthesis method of Example 1, compound 6-1 (85 mg, 58%) was obtained;

[0215] Using 6F (100 mg, 0.3 mmol) and 5F-2 as raw materials and following the synthesis method of Example 1, the target compound 6-2 (100 mg, 68%) was obtained.

[0216] Compound 6-1:

[0217] 1 H NMR (400MHz, DMSO-d6) δ = 8.21 (s, 1H), 7.78 (d, 1H), 6.98 (d, 1H), 4.68 (s, 2H), 4. 46–4.39(m,2H),4.00–3.95(m,1H),3.53–3.37(m,2H),3.33–3.23(s,11H),3.24 –3.15(m,1H),3.09–2.94(m,2H),2.93–2.88(m,1H),2.85–2.69(m,2H),2.46–2. 30(m,2H),2.27–2.21(m,2H),2.00(s,3H),1.96–1.89(m,1H),1.88–1.79(m,1H).

[0218] LC-MS (ESI): m / z = 490.2 [M+H] +

[0219] Compound 6-2:

[0220] 1H NMR (400MHz, DMSO-d6) δ = 8.22 (s, 1H), 7.78 (d, 1H), 6.98 (d, 1H), 4.67 (s, 2H), 4.49–4 .35(m,2H),4.00–3.95(m,1H),3.51–3.39(m,2H),3.34–3.30(m,2H),3.25–3.17(m,1 H),3.09–2.94(m,2H),2.93–2.88(m,1H),2.84–2.73(m,2H),2.47–2.39(m,2H),2.37 –3.32(m,2H),2.29–2.18(m,2H),2.00(s,3H),1.97–1.89(m,1H),1.88–1.79(m,1H).

[0221] LC-MS (ESI): m / z = 490.2 [M+H] +

[0222] Embodiment 7:

[0223]

[0224] Step 1: Dissolve compound 7A (10.0 g, 46.03 mmol) in acetonitrile (200 mL), then add pyridine (10.92 g, 138.09 mmol) and di-tert-butyl dicarbonate (15.07 g, 69.05 mmol), stir at room temperature for 4 hours, add ammonia water (20 mL) dropwise, and react at room temperature for 4 hours. After the reaction is complete, concentrate, filter the solid, and wash with acetonitrile to obtain compound 7B (9.9 g, 99%).

[0225] LC-MS(ESI):m / z=161.1[M+H-56] + .

[0226] Step 2: Compound 7B (9.9 g, 45.77 mmol) was dissolved in dichloromethane (50 mL), trifluoroacetic acid (20 mL) was added, and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated to obtain compound 7C, which was directly used for the next step without purification.

[0227] LC-MS (ESI): m / z = 117.1 [M+H] + .

[0228] Step 3: Dissolve the compound 7C from the previous step in N,N-dimethylformamide (50.0 mL), add N,N-diisopropylethylamine (29.58 g, 228.85 mmol) and 2,4-dichloro-6,7-dihydrothieno[3,2-d]pyrimidine (14.22 g, 26.86 mmol), and heat to 80°C for 16 h. The reaction solution was cooled to room temperature, diluted with ethyl acetate, and washed with water. The organic phase was dried, filtered, concentrated, and column chromatography (dichloromethane / methanol = 10 / 1) was performed to obtain compound 7D (740 mg, 6%).

[0229] LC-MS (ESI): m / z = 287.5 [M+H] + .

[0230] Step 4: Compound 7D (740 mg, 2.58 mmol) was dissolved in dichloromethane (50.0 mL), triethylamine (780 mg, 7.74 mmol) was added under ice bath, and trifluoroacetic anhydride (810 mg, 3.87 mmol) was added dropwise after stirring for five minutes, and stirred under ice bath for 2 hours. The reaction solution was quenched with water, extracted with dichloromethane, and the organic phase was dried, filtered, concentrated, and column chromatography (dichloromethane / methanol = 10 / 1) was performed to obtain compound 7E (590 mg, 85%).

[0231] LC-MS (ESI): m / z = 269.1 [M+H] + .

[0232] Step 5: Compound 7E (590 mg, 2.20 mmol), (S)-(-)-1,1'-bi-2-naphthol (63 mg, 0.22 mmol), tetraisopropyloxytitanium (31 mg, 0.11 mmol) and water (40 mg, 2.20 mmol) were dissolved in dichloromethane (40 mL), nitrogen was replaced three times, and tert-butyl peroxide (340 mg, 2.64 mmol, 70% in water) was added after stirring at room temperature for one hour, and stirred overnight. The solution was quenched with saturated sodium thiosulfate aqueous solution, the aqueous phase was extracted with dichloromethane, dried and concentrated, and column chromatography (dichloromethane / methanol = 10 / 1) was performed to obtain compound 7F (400 mg, 64%).

[0233] LC-MS (ESI): m / z = 285.0 [M+H] + .

[0234] Step 6: Using 7F (100 mg, 0.35 mmol) and 5F-1 as raw materials, according to the synthesis method of Example 1, compound 7-1 (80 mg, 55%) was obtained;

[0235] Using 7F (100 mg, 0.35 mmol) and 5F-2 as raw materials and following the synthesis method of Example 1, the target compound 7-2 (85 mg, 58%) was obtained.

[0236] Compound 7-1:

[0237] 1 H NMR(400MHz, DMSO-d6)δ=7.79(d,1H),7.14(s,1H),6.99(d,1H),4.67(s,1H),4.51–4.38(m,2H),4.02–3.97(m,1H),3.51–3.40(m,2H), 3.28(s,2H),3.26–3.20(m,1H),3.10(t,1H),3.03–2.95(m,1H),2.94–2.87(m,1H),2.82(t,2H),2.01(s,3H),1.53(s,3H),1.52(s,3H).

[0238] LC-MS (ESI): m / z = 478.2 [M+H] +

[0239] Compound 7-2:

[0240] 1 H NMR(400MHz, DMSO-d6)δ=7.79(d,1H),7.14(s,1H),6.99(d,1H),4.66(s,1H),4.48(d,1H),4.43–4.39(m,1H),4.02–3.97(m,1H),3.50–3.40(m,2 H),3.28(t,2H),3.26–3.22(m,1H),3.10(t,1H),3.04–2.95(m,1H),2.93 –2.87(m,1H),2.87–2.78(m,2H),2.01(s,3H),1.53(s,3H),1.52(s,3H).

[0241] LC-MS (ESI): m / z = 478.2 [M+H] +

[0242] Embodiment 8:

[0243]

[0244] Step 1: Dissolve compound 2B (3.00 g, 14.78 mmol, the synthesis method refers to patent WO2012030633A1) in 1,4-dioxane (40 mL), then add potassium carbonate (6.13 g, 44.33 mmol), 8B (9.55 g, 29.55 mmol, containing 25% regioisomers, the synthesis method refers to patent WO2023086575A1) and water (10 mL) in sequence, replace nitrogen three times and add 1,1-bis(diphenylphosphine)diferronichloridepalladium (1.56 g, 2.22 mmol). After the addition was completed, the reaction was heated to 85° C. and stirred for 16 h. After the reaction solution naturally returned to room temperature, water (100 mL) was added to dilute the reaction and extracted three times with ethyl acetate (50 mL). The combined organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo to obtain a crude product. The crude product was purified by column chromatography (eluent: petroleum ether / ethyl acetate = 5 / 1) to obtain 1.30 g of a crude product (containing 25% regioisomers), which was further prepared by HPLC to obtain the target compound 8C (600 mg, yield: 12.71%).

[0245] HPLC analysis method: Instrument: SHIMADZU LC-2020AD, Column: C18 Column; Mobile phase: A: 0.1% TFA in H2O, B: ACN; Gradient: 55-95% B in A; Flow rate: 1.2 mL / min Column temperature: 45°C Wavelength: 210 nm & 254 nm.

[0246] HPLC preparation method: Instrument: SHIMADZU LC-20AP, Column: XB-SiOH Column; Mobile phase: A: Hexane, B: EtOH; Gradient: 35-95% B gradient elution; Flow rate: 45 mL / min, Column temperature: 25°C; Wavelength: 220 nm; Cycle time: 6.0 min Sample preparation: Sample concentration 50 mg / ml, acetonitrile solution injection: 2.0 ml each time. After separation, the fraction was dried by rotary evaporator at a bath temperature of 35°C to obtain compound 8C. LC-MS (ESI): m / z=320.3 [M+H] + .

[0247] Step 2: Compound 8C (600 mg, 1.88 mmol) was dissolved in dichloromethane (10 mL), and trifluoroacetic acid (2 mL) was added under ice bath. After reacting at room temperature for half an hour, the reaction solution was concentrated under reduced pressure to obtain crude compound 8D (700 mg), which was used directly in the next step.

[0248] Step 3: 8E (200 mg, 0.73 mmol, the synthesis method refers to patent WO2019115775A1) was dissolved in 1,4-dioxane (20 mL), and 8D (400 mg, 1.11 mmol) and N,N-diisopropylethylamine (280 mg, 2.18 mmol) were added in sequence. After the addition was completed, the reaction liquid was heated to 85 ° C under nitrogen protection and stirred for 16 hours. After the reaction liquid naturally returned to room temperature, water (50 mL) was added to dilute the reaction, and ethyl acetate (40 mL) was used for extraction three times. The organic phases were combined and dried over anhydrous sodium sulfate and concentrated in vacuo to obtain a crude product. The crude product was purified by column chromatography (eluent: DCM: MeOH = 100: 1 to 10: 1) to obtain the target compound 8F (310 mg, 0.67 mmol, yield: 93.20%).

[0249] LC-MS (ESI): m / z = 459.3 [M+H] +

[0250] Step 4: Dissolve compound 8F (300 mg, 0.65 mmol) in dichloromethane (20 mL) under nitrogen protection, slowly add trichloroacetyl isocyanate (150 mg, 0.80 mmol) dropwise under ice bath stirring. After the addition is complete, stir the reaction solution at 0°C for 1 h. Monitor the disappearance of the starting material by TLC spot plate (developing solvent: DCM / MeOH=10:1), and then concentrate the reaction solution under reduced pressure to obtain crude product 8G (480 mg), which is directly used in the next step.

[0251] Step 5: Dissolve the crude compound 8G (480 mg, 0.66 mmol) in methanol (10 mL), then add potassium carbonate (275 mg, 1.99 mmol) at room temperature, and stir at room temperature for 1 h under nitrogen protection after the addition is complete. After the reaction is complete, the reaction solution is concentrated under reduced pressure to obtain a crude product, which is purified by column chromatography (eluent: DCM: MeOH = 100: 1 to 9: 1) and further purified by reverse phase column (eluent: water: acetonitrile = 90: 1 to 1: 90) to obtain compound 8 (220 mg, 65.99%).

[0252] LC-MS (ESI): m / z = 502.3 [M+H] +

[0253] Step 6: Compound 8 (220 mg, 0.43 mmol) was subjected to chiral separation to obtain compound 8-1 (164.0 mg) and compound 8-2 (30.5 mg). Analysis method: Instrument: SHIMADZU LC-30AD, Column: Chiral AS Column; Mobile phase: A: CO2, B: 0.05% DEA in ETOH; Gradient: 5-40% B in A; Flow rate: 3 mL / min Column temperature: 35°C Wavelength: 220 nm.

[0254] Preparation method: Instrument: Waters 150Prep-SFC, Column: Chiralcel AS Column; Mobile phase: A: CO2, B: 0.1% NH3·H2O in MEOH and ACN; Gradient: 40% B gradient elution flow rate: 100mL / min, column temperature: 25°C Wavelength: 220nm Cycle time: 4.2min Sample preparation: Sample concentration 5mg / ml, ethanol solution injection: 1ml each time. After separation, the fractions were dried by rotary evaporator at a bath temperature of 35°C to obtain compound 8-1 and compound 8-2.

[0255] 8-1:

[0256] SFC retention time: 1.717 min

[0257] 1 H NMR(400MHz, DMSO-d6)δ8.45(s,2H),7.28–7.21(m,1H),6.58–6.44(m,3H),4.50–4.46(m,2H),4.26(s,2H),4.19–4.14(m,2H), 4.03–3.94(m,2H),3.43–3.35(m,1H),3.27–3.20(m,1H),2.96–2.81(m,4H),1.86(m,2H),1.44–1.43(m,6H),1.36–1.32(m,3H).

[0258] LC-MS (ESI): m / z = 502.6 [M+H] +

[0259] 8-2:

[0260] SFC retention time: 2.131 min

[0261] 1 H NMR(400MHz,DMSO-d6)δ8.45(s,2H),7.28–7.21(m,1H),6.57–6.50(m,3H),4.50–4.46(m,2H),4.26(s,2H),4.19–4.14(m,2H), 4.03–3.94(m,2H),3.43–3.35(m,1H),3.26–3.20(m,1H),2.96–2.81(m,4H),1.86(m,2H),1.44–1.43(m,6H),1.36–1.32(m,3H).

[0262] LC-MS (ESI): m / z = 502.6 [M+H]+

[0263] Embodiment 9:

[0264]

[0265] Step 1: Using compound 9A (WO2013 / 50508, 2013, A1) as a raw material, according to the synthesis method of Example 1, compound 9B was obtained.

[0266] Step 2: Dissolve compound 9B (4.15 g, mmol) in dichloromethane (50 ml), add hydrogen peroxide (20 ml), stir at room temperature for 5 h, and after the reaction is completed, extract with dichloromethane, combine the organic phases, dry over anhydrous sodium sulfate, filter and concentrate, and separate and purify by preparative liquid phase (DCM / MeOH=0-15%) to obtain the target compound 9C.

[0267] LC-MS (ESI): m / z = 302.1 [M+H] +

[0268] Split conditions:

[0269] Instrument: Waters 150SFC; Column: Chiralpak IC Column (250×30mm, ID 30mm, 10umparticle size); Mobile phase: A for CO2; B for 0.1% NH3·H2O in IPA and ACN; Gradient: B45%; Flow rate: 100mL / min; Back pressure: 100bar; Column temperature: 25℃; Wavelength: 220nm; Cycle: 2.8min;

[0270] Steps 3, 4 and 5: Compound 9C was used as a raw material and the synthesis method of Example 1 was used to obtain compound 9E. Compound 9-1 (74 mg, 54%) was obtained using 9E (100 mg, 0.29 mmol) and 5F-1 as raw materials and the synthesis method of Example 1; target compound 9-2 (80 mg, 58%) was obtained using 9E (100 mg, 0.29 mmol) and 5F-2 as raw materials and the synthesis method of Example 1.

[0271] Compound 9-1:

[0272] 1H NMR(400MHz, DMSO-d6)δ=7.78(d,1H),6.97(d,1H),6.76(s,1H),6.52(s, 2H),4.78–4.63(m,2H),4.51–4.26(m,4H),4.03–3.93(m,1H),3.38(t,1H) ,3.10–3.06(m,1H),3.00–2.90(m,2H),2.77–2.67(m,3H),2.62–2.58(m,1 H),2.36–2.25(m,5H),2.00(s,3H),1.97–1.94(m,1H),1.88–1.79(m,2H).

[0273] LC-MS (ESI): m / z = 438.2 [M+H] +

[0274] Compound 9-2:

[0275] 1 H NMR (400MHz, DMSO-d6)δ=7.78(s,1H),6.97(s,1H),6.76(s,1H),6.52(s,2H),4.70(t,2H),4.43(t,3H),4.31(s,1H),4.03–3.91(m,1H),3.37(d, 1H),3.10–3.06(m,1H),3.00–2.90(m,2H),2.82–2.63(m,3H),2.62–2.58 (m,1H),2.37–2.20(m,5H),2.00(s,3H),1.95(d,1H),1.89–1.80(m,2H).

[0276] LC-MS (ESI): m / z = 438.2 [M+H] +

[0277] Embodiment 10:

[0278]

[0279] Using 9E (50 mg, 0.14 mmol) and 3H-1 as raw materials, according to the synthesis method of Example 1, compound 10-1 (40 mg, 53%) was obtained; using 9E (50 mg, 0.14 mmol) and 3H-2 as raw materials, according to the synthesis method of Example 1, the target compound 10-2 (36 mg, 47%) was obtained.

[0280] Compound 10-1:

[0281] 1H NMR(400MHz, DMSO-d6)δ=7.49(d,1H),6.77(d,1H),6.73(s,1H),6.52(s,2H),4.70(t,2H),4.39(d,2H),4.31(d,2H),4.03–3.91(m,3H),3. 16(t,1H),3.10–3.06(m,1H),3.00–2.87(m,2H),2.70–2.54(m,4H),2 .41–2.21(m,5H),1.97–1.94(m,1H),1.90–1.77(m,2H),1.27(t,3H).

[0282] LC-MS (ESI): m / z = 544.2 [M+H] +

[0283] Compound 10-2:

[0284] 1 H NMR(400MHz, DMSO-d6)δ=7.49(d,1H),6.77(d,1H),6.73(s,1H),6.52(s,2H),4.70(t,2H),4.44–4.26(m,4H),4.03–3.91(m,3H),3.18( d,1H),2.99–2.96(m,1H),3.01–2.87(m,2H),2.72–2.54(m,4H),2.39–2.21(m,5H),1.97–1.93(m,1H),1.88–1.77(m,2H),1.27(t,3H).

[0285] LC-MS (ESI): m / z = 544.2 [M+H] +

[0286] Embodiment 11:

[0287]

[0288] Step 1: Compound 11A (1.0 g, 3.0 mmol) (synthesis method reference US2022106311), cyclopropylboric acid (356.0 mg, 4.0 mmol), palladium acetate (25.0 mg, 0.15 mmol), tricyclohexylphosphine (84.2 mg, 0.3 mmol), potassium phosphate (2.3 g, 10.5 mmol), and finally toluene (14 mL) and water (0.7 mL) were added, and nitrogen was replaced three times. After heating to 100 ° C for 3 h, it was passed through a short silica gel column, washed with ethyl acetate, concentrated, and column chromatography (petroleum ether / ethyl acetate = 10 / 1) to obtain compound 11B (820.0 mg, 91%).

[0289] LC-MS (ESI): m / z = 302.1 [M+H] + .

[0290] Step 2: Compound 11B (820.0 mg, 2.7 mmol) was dissolved in dichloromethane (20 mL), and trifluoroacetic acid (5.0 mL) was added. After reacting at room temperature for half an hour, the mixture was concentrated to obtain compound 11C (1.0 g, 99%), which was used directly in the next step.

[0291] LC-MS (ESI): m / z = 202.1 [M+H] + .

[0292] Step 3: Weigh compound 6C (2.10 g, 16.38 mmol) and compound 9A (3.62 g, 16.38 mmol) in a 100 mL single-mouth bottle, dissolve in DMF (40 mL), add triethylamine (6.63 g, 65.52 mmol), stir at 80 degrees for 16 h after the addition is complete, detect the reaction is complete by TLC spot plate (petroleum ether: ethyl acetate = 1:1), add water (50 mL), stir for 5 min, extract with ethyl acetate (30 mL), combine the organic phases, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The crude product is purified by column chromatography (eluent: petroleum ether: ethyl acetate = 2:1) to obtain the target compound 11D (1.5 g, yield: 29.2%).

[0293] LCMS m / z=313.1[M+1] + .

[0294] Step 4: Weigh compound 11D (1.5 g, 4.80 mmol) and dissolve it in dichloromethane (20 mL). Add triethylamine (970 mg, 9.60 mmol) under ice bath. Stir for five minutes and then add trifluoroacetic anhydride (1.51 g, 7.20 mmol) dropwise. After the addition, stir in ice bath for 2 hours. The reaction solution was quenched with water (30 mL), extracted with dichloromethane (30 mL), and the organic phase was dried, filtered, concentrated, and column chromatography (dichloromethane / methanol (v / v) = 91 / 9) was performed to obtain compound 11E (1.2 g, yield: 84.8%).

[0295] LCMS m / z=295.1[M+1] +

[0296] Step 5: Weigh compound 11E (1.20 g, 4.07 mmol) into a 100 mL single-mouth bottle, dissolve in dichloromethane (20 mL), add hydrogen peroxide (2.31 g, 20.39 mmol), and stir at room temperature for 12 h after the addition is complete. TLC spot plate (petroleum ether: ethyl acetate = 2:1) detected the formation of the product, added water (50 mL), stirred for 5 min, extracted with dichloromethane (30 mL), dried the organic phase over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by column chromatography (eluent: dichloromethane: methanol = 20:1) to obtain the target compound 11F (0.50 g, yield: 39.5%).

[0297] LCMS m / z=311.1[M+1] +

[0298] Step 6: Compound 11F (500 mg, 1.61 mmol) and dioxane (20 mL) were added to a 100 mL single-mouth bottle to dissolve, and compound 11C (310 mg, 2.40 mmol) and N,N-diisopropylethylamine (1.30 g, 6.46 mmol) were added. After the addition was completed, the system was protected by nitrogen and stirred at 85 ° C for 16 h. The reaction was detected to be complete by TLC spot plate (dichloromethane: methanol = 10:1). Water (50 mL) was added and stirred for 5 min. Dichloromethane (30 mL) was extracted to separate the organic phase, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by column chromatography (eluent: dichloromethane: methanol = 10:1) to obtain the target compound racemate 11G, which was separated by SFC to obtain compound 11-1 (200 mg) and compound 11-2 (210 mg).

[0299] SFC preparation method: instrument: SHIMADZU LC-30AD sf, column: Chiral OX column; mobile phase: A for CO2; B for 0.05% DEA in methanol and acetonitrile; gradient: 50% B gradient elution; flow rate: 3mL / min, column temperature: 35°C; wavelength: 220nm; cycle time: 8.6min; sample preparation: sample concentration 10mg / ml, acetonitrile solution injection: 1.5ml each time. After separation, the fraction was dried by rotary evaporator at a bath temperature of 35°C to obtain compound 11-1 (200mg) and compound 11-2 (210mg).

[0300] Compound 11-1: retention time: 0.975 min; 1H NMR (400MHz, CDCl3) δ8.54(s,2H),7.22(s,1H),7.17(s,1H),4.38(s,2H),3.88-3.95(m,2H),3.36(s,2H),3.05-3.10(m,1H),2.8 7-2.94(m,1H),2.56-2.73(m,4H),2.37-2.44(m,2H),2.23-2.34(m,3H),1.82-2.00(m,4H),1.01-1.05(m,2H),0.81-0.86(m,2H).

[0301] LC-MS (ESI): m / z = 476.2 [M+H] +

[0302] Compound 11-2: retention time: 1.469 min; 1 H NMR (400MHz, CDCl3) δ8.54(s,2H),7.22(s,1H),7.14(s,1H),4.38(s,2H),3.89-3.95(m,2H),3.36(s,2H),3.06-3.12(m,1H),2.8 7-2.94(m,1H),2.58-2.74(m,4H),2.37-2.44(m,2H),2.23-2.34(m,3H),1.82-2.00(m,4H),1.00-1.05(m,2H),0.81-0.86(m,2H).

[0303] LC-MS (ESI): m / z = 476.2 [M+H] +

[0304] Embodiment 12:

[0305]

[0306] Using compound N-Boc-3-amino-3-methylbutyric acid as raw material, compound 12 (800 mg) was synthesized by referring to the synthesis method of Example 11, and then chiral separation was performed to obtain compound 12-1 (260.7 mg) and compound 12-2 (233.6 mg).

[0307] Analytical method: Instrument: SHIMADZU LC-30AD, Column: Chiral AD Column; Mobile phase: A: CO2, B: 0.05% DEA in ETOH; Gradient: 40% B in A; Flow rate: 3mL / min Column temperature: 35℃ Wavelength: 220nm.

[0308] Preparation method: Instrument: Waters 150Prep-SFC, Column: Chiralcel AD Column; Mobile phase: A: CO2, B: 0.1% NH3·H2O in MeOH and ACN; Gradient: 70% B Gradient elution Flow rate: 100mL / min, Column temperature: 25°C Wavelength: 220nm Cycle time: 4.2min Sample preparation: Sample concentration 5mg / ml, ethanol solution injection: 1ml each time. After separation, the fractions were dried by rotary evaporator at a bath temperature of 35°C to obtain compounds 12-1 and 12-2.

[0309] Compound 12-1: SFC retention time: 1.360 min. 1 H NMR(400MHz, CDCl3)δ8.43(s,2H),7.21-7.19(m,1H),5.95(s,1H),4.46(s,2H),4 .02-4.01(m,2H),3.63-3.59(m,1H),3.21-3.16(m,1H),2.97-2.83(m,3H),2.80-2 .77(m,2H),2.62-2.50(m,2H),2.10-2.04(m,1H),1.89-1.83(m,1H),1.66(s,3H) ,1.64(s,3H),1.11-1.06(m,2H),0.80-0.76(m,2H).LC-MS(ESI):m / z=464.5[M+H] +

[0310] Compound 12-2: SFC retention time: 2.721 min. 1 H NMR(400MHz, CDCl3)δ8.43(s,2H),7.21-7.19(m,1H),5.95(s,1H),4.46(s,2H),4 .02-4.01(m,2H),3.63-3.59(m,1H),3.21-3.16(m,1H),2.97-2.83(m,3H),2.80-2 .77(m,2H),2.62-2.47(m,2H),2.10-2.04(m,1H),1.89-1.83(m,1H),1.66(s,3H) ,1.64(s,3H),1.11-1.06(m,2H),0.80-0.76(m,2H).LC-MS(ESI):m / z=464.6[M+H] +

[0311] Embodiment 13:

[0312]

[0313] Using compound 3-amino-3-methyl-2-butanol as raw material, compound 13 (400 mg) was synthesized according to the synthesis method of Example 11, and then chiral separation was performed to obtain compounds 13-1 (22.8 mg), 13-2 (28.9 mg), 13-3 (13.6 mg) and 13-4 (30.3 mg).

[0314] Analytical method: Instrument: SHIMADZU LC-30AD, Column: Chiral AS Column; Mobile phase: A: CO2, B: 0.05% DEA in ETOH; Gradient: 35-80% B in A; Flow rate: 3mL / min Column temperature: 35℃ Wavelength: 220nm.

[0315] Preparation method: Instrument: Waters 150Prep-SFC, Column: Chiralcel AS Column; Mobile phase: A: CO2, B: 0.1% NH3·H2O in MeOH and ACN; Gradient: 55% B gradient elution flow rate: 100mL / min, column temperature: 25°C Wavelength: 220nm Cycle time: 6.0min Sample preparation: Sample concentration 5mg / ml, ethanol solution injection: 1ml each time. After separation, the fractions were dried by rotary evaporator at a bath temperature of 35°C to obtain compounds 13-1, 13-2, 13-3 and 13-4.

[0316] Compound 13-1: SFC retention time: 1.037 min. 1 H NMR(400MHz, CDCl3)δ8.42(s,2H),7.19-7.17(m,1H),6.01(s,1H),5.32(s,1H),4.51-4.4 0(m,2H),4.08-4.05(m,1H),3.97-3.92(m,2H),3.22-3.18(m,1H),2.90-2.84(m,2H),2.7 8(s,2H),2.59-2.53(m,2H),2.10-2.09(m,1H),1.89-1.82(m,1H),1.54(s,3H),1.42(s,3 H),1.20-1.19(m,3H),1.10-1.05(m,2H),0.79-0.75(m,2H).LC-MS(ESI):m / z=469.5[M+H] + .

[0317] Compound 13-2: SFC retention time: 1.281 min. 1H NMR(400MHz, CDCl3)δ8.42(s,2H),7.19-7.17(m,1H),6.09(s,1H),4.50-4.41(m, 2H),4.13(s,1H),3.90-3.85(m,2H),3.22-3.18(m,1H),2.94-2.79(m,4H),2.56-2 .51(m,2H),2.09-2.06(m,1H),1.88-1.82(m,1H),1.56(s,3H),1.41(s,3H),1.21 -1.19(m,3H),1.10-1.05(m,2H),0.79-0.75(m,2H).LC-MS(ESI):m / z=469.5[M+H] + .

[0318] Compound 13-3: SFC retention time: 1.491 min. 1 H NMR(400MHz, CDCl3)δ8.42(s,2H),7.19-7.17(m,1H),6.09(s,1H),4.50-4.41(m, 2H),4.12(s,1H),3.90-3.85(m,2H),3.22-3.18(m,1H),2.94-2.79(m,4H),2.56-2 .51(m,2H),2.09-2.07(m,1H),1.88-1.82(m,1H),1.56(s,3H),1.41(s,3H),1.21 -1.19(m,3H),1.10-1.05(m,2H),0.79-0.75(m,2H).LC-MS(ESI):m / z=469.5[M+H] + .

[0319] Compound 13-4: SFC retention time: 1.829 min. 1H NMR(400MHz, CDCl3)δ8.42(s,2H),7.19-7.17(m,1H),6.01(s,1H),5.32(s,1H),4.51-4.4 0(m,2H),4.08-4.05(m,1H),3.97-3.92(m,2H),3.22-3.18(m,1H),2.93-2.84(m,2H),2.7 8(s,2H),2.59-2.53(m,2H),2.10-2.09(m,1H),1.89-1.82(m,1H),1.54(s,3H),1.42(s,3 H),1.20-1.19(m,3H),1.09-1.07(m,2H),0.78-0.77(m,2H).LC-MS(ESI):m / z=469.5[M+H] + .

[0320] Embodiment 14:

[0321]

[0322] Using compound 2-methyl-2-amino-tert-butoxycarbonylpropylamine as raw material, compound 14 (130 mg) was synthesized referring to the synthesis method of Example 11, and then chiral separation was performed to obtain compound 14-1 (11.6 mg) and compound 14-2 (14.4 mg).

[0323] Analytical method: Instrument: SHIMADZU LC-30AD, Column: Chiral IG Column; Mobile phase: A: CO2, B: 0.05% DEA in ETOH; Gradient: 60% B in A; Flow rate: 3 mL / min Column temperature: 35°C Wavelength: 220 nm.

[0324] Preparation method: Instrument: Waters 150Prep-SFC, Column: Chiralcel IG Column; Mobile phase: A: CO2, B: 0.1% NH3·H2O in i-PrOH and ACN; Gradient: 70% B Gradient elution Flow rate: 100mL / min, Column temperature: 25°C Wavelength: 220nm Cycle time: 4.2min Sample preparation: Sample concentration 5mg / ml, ethanol solution injection: 1ml each time. After separation, the fractions were dried by rotary evaporator at a bath temperature of 35°C to obtain compound 14-1 and compound 14-2.

[0325] Compound 14-1: SFC retention time: 0.977 min. 1H NMR(400MHz, CDCl3)δ8.43(s,2H),7.20-7.19(m,1H),5.81(s,1H),5.34(s,1H),4.4 8(s,2H),4.04-4.02(m,2H),3.67-3.63(m,5H),3.22-3.17(m,1H),2.90-2.84(m,2H ),2.78(s,2H),2.60-2.53(m,2H),2.09-2.06(m,1H),1.89-1.82(m,1H),1.49(s,3H ),1.47(s,3H),1.10-1.06(m,2H),0.79-0.75(m,2H).LC-MS(ESI):m / z=512.6[M+H] + .

[0326] Compound 14-2: SFC retention time: 2.145 min. 1 H NMR(400MHz, CDCl3)δ8.43(s,2H),7.20-7.19(m,1H),5.81(s,1H),5.34(s,1H),4.4 8(s,2H),4.04-4.02(m,2H),3.67-3.63(m,5H),3.22-3.17(m,1H),2.90-2.84(m,2H ),2.78(s,2H),2.64-2.49(m,2H),2.09-2.06(m,1H),1.89-1.82(m,1H),1.49(s,3H ),1.47(s,3H),1.10-1.05(m,2H),0.79-0.75(m,2H).LC-MS(ESI):m / z=512.3[M+H] + .

[0327] Embodiment 15:

[0328]

[0329] Step 1: Dissolve compound 15A (4.0 g, 20.0 mmol) in 1,4-dioxane (60 mL), add bis(triphenylphosphine)palladium dichloride (1.4 g, 2.0 mmol), potassium carbonate (5.6 g, 40.0 mmol), N-Boc-1,2,5,6-tetrahydropyridine-4-boronic acid pinacol ester (7.2 g, 24.0 mmol) and water (6.0 mL), replace nitrogen three times, heat to 90 ° C to react for 14 h, pass through a short silica gel column, elute with ethyl acetate, concentrate, and column chromatography (dichloromethane / methanol = 20 / 1) to obtain compound 15B (6.0 g, 99%).

[0330] LC-MS (ESI): m / z = 300.2 [M+H] + .

[0331] Step 2: Compound 15B (6.0 g, 20.0 mmol) was dissolved in 1,4-dioxane (10 mL) and a solution of hydrogen chloride in dioxane (4.0 M, 20.0 mL). After reacting at room temperature for 1 h, the mixture was concentrated to obtain compound 15C (4.0 g, 99%).

[0332] LC-MS (ESI): m / z = 200.2 [M+H] + .

[0333] Step 3: Compound 15D (660.0 g, 3.0 mmol) was dissolved in 1,4-dioxane (15 mL), and Pd2dba3 (140.0 mg, 0.15 mmol), Xantphos (170.0 mg, 0.3 mmol), ethyl 2-(4-amino-2-fluorophenyl)acetate (710.0 mg, 3.6 mmol) and sodium carbonate (790.0 mg, 7.45 mmol) were added, and nitrogen was replaced three times. After heating to 90 ° C for 14 h, the mixture was passed through a short silica gel column, washed with ethyl acetate, concentrated, and column chromatography (dichloromethane / methanol = 15 / 1) was performed to obtain compound 15E (430.0 mg, 38%).

[0334] LC-MS (ESI): m / z = 382.1 [M+H] + .

[0335] Step 4: Compound 15E (430.0 g, 1.1 mmol) was dissolved in dichloromethane (10 mL), and m-CBPA (490.0 mg, 2.8 mmol) was added. After reacting at room temperature for 14 h, the mixture was concentrated and subjected to column chromatography (dichloromethane / methanol = 10 / 1) to give compound 15F (390.0 mg, 83%).

[0336] LC-MS (ESI): m / z = 414.0 [M+H] + .

[0337] Step 5: Compound 15F (185.0 g, 0.45 mmol) was dissolved in 1,4-dioxane (10 mL), and 15C (11.0 mg, 0.54 mmol) and DIPEA (230.0 mg, 1.8 mmol) were added. The mixture was heated to 90 °C for 14 h, concentrated, and subjected to column chromatography (dichloromethane / methanol = 10 / 1) to give compound 15G (210.0 mg, 81%).

[0338] LC-MS (ESI): m / z = 577.2 [M+H] + .

[0339] Step 6: Dissolve compound 15G (210.0 mg, 0.36 mmol) in methanol / tetrahydrofuran (v / v, 10 mL), add lithium hydroxide aqueous solution (34.0 mg, 1.4 mmol), react at room temperature for 14 h, concentrate, add dilute hydrochloric acid to acidify, filter and dry to obtain compound 15 (70.0 mg, 35%).

[0340] LC-MS (ESI): m / z = 549.2 [M+H] + .

[0341] 1 H NMR(400MHz,DMSO-d6)δ8.67(s,1H),7.63–7.59(m,1H),7.35–7.23(m,4H),7.04(d,2H),6.20(s,1H),4.35(s,2H),3.97(s ,2H),3.59–3.52(m,4H),2.84(t,2H),2.54(s,2H),2.30(m,2H),2.00–1.80(m,2H),1.00–0.84(m,2H),0.72–0.59(m,2H).

[0342] 19 F NMR(377MHz,DMSO-d6)δ-113.84(s).

[0343] Example 16

[0344]

[0345] Step 1: Dissolve compound 15F (185.0 mg, 0.45 mmol) in 1,4-dioxane (10 mL), add 11C (110.0 mg, 0.54 mmol) and DIPEA (230.0 mg, 1.8 mmol), heat to 90 °C for 14 h, concentrate, and perform column chromatography (dichloromethane / methanol = 10 / 1) to obtain compound 16A (250.0 mg, 96%).

[0346] LC-MS (ESI): m / z = 579.2 [M+H] + .

[0347] Step 2: Dissolve compound 16A (210.0 mg, 0.36 mmol) in methanol / tetrahydrofuran (v / v, 10 mL), add aqueous lithium hydroxide solution (34.0 mg, 1.4 mmol), react at room temperature for 14 h, concentrate, add dilute hydrochloric acid to acidify, filter and drain to obtain compound 16 (60.0 mg, 30%).

[0348] LC-MS (ESI): m / z = 551.2 [M+H] + .

[0349] 1 H NMR(400MHz,DMSO-d6)δ8.69(s,1H),8.54(s,2H),7.62(d,1H),7.42–7.22(m,2H),7.18(s,1H),4.44(s,2H),3.96(s,2H), 3.57(d,5H),2.86–2.83(m,2H),2.69(s,2H),2.37–2.24(m,2H),2.01–1.88(m,1H),1.09–0.92(m,2H),0.92–0.74(m,2H).

[0350] 19 F NMR(377MHz,DMSO-d6)δ-113.88(s).

[0351] Embodiment 17:

[0352]

[0353] Step 1: Compound 17A (1.0 g, 3.8 mmol), cyclopropylboronic acid (0.5 g, 5.7 mmol), palladium acetate (86 mg, 0.4 mmol), tricyclohexylphosphine (110 mg, 0.4 mmol), potassium phosphate (2.4 g, 11.4 mmol), and finally toluene (20 mL) and water (1 mL) were added, and nitrogen was replaced three times. After heating to 100 ° C for 3 h, the reaction was filtered, washed with ethyl acetate, concentrated, and column chromatography (petroleum ether / ethyl acetate = 10 / 1) was performed to obtain compound 17B (0.5 g, 58.7%).

[0354] 1 H NMR (400MHz, CDCl3) δ5.36(s,1H),3.84(d,2H),3.46(t,2H),1.94(d,2H),1.46(s,9H),1.40–1.29(m,1H),0.62–0.52(m,2H),0.51–0.37(m,2H).

[0355] Step 2: Compound 17B (500 mg, 2.2 mmol) was dissolved in dichloromethane (10 mL), and trifluoroacetic acid (2.0 mL) was added. After reacting at room temperature for half an hour, the mixture was concentrated to obtain compound 17C (0.5 g, crude product), which was used directly in the next step.

[0356] LC-MS (ESI): m / z = 124.1 [M+H] + .

[0357] Step 3: Compound 15F (0.2 g, 0.5 mmol) was dissolved in 1,4-dioxane (20 mL), and 17C (0.2 g, crude product) and DIPEA (0.25 g, 4.0 mmol) were added. The mixture was heated to 100 °C for 4 h, concentrated, and subjected to column chromatography (dichloromethane / methanol = 10 / 1) to give compound 17D (0.19 g, 79%).

[0358] LC-MS (ESI): m / z = 501.6 [M+H] + .

[0359] Step 4: Dissolve compound 17D (190 mg, 0.38 mmol) in methanol / tetrahydrofuran / water (v / v / v=1 / 1 / 1, 9 mL), add lithium hydroxide aqueous solution (46.0 mg, 1.9 mmol), react at room temperature for 14 h, concentrate, add dilute hydrochloric acid to acidify, filter and dry to obtain compound 17 (120 mg, 67%).

[0360] LC-MS (ESI): m / z = 473.2 [M+H] + .

[0361] 1 H NMR(400MHz, CDCl3)δ8.65(s,1H),7.70–7.59(m,1H),7.20(t,1H),7.15–7.11(m,1H),5.46(s,1H),4.19(s,2H),3.89(s,2H),3.68(s, 2H),3.40–3.33(m,2H),2.84(t,2H),2.47–2.40(m,2H),2.04(s,2H),1.42–1.34(m,1H),1.26(s,1H),0.65–0.55(m,2H),0.46(q,2H).

[0362] 19 F NMR(376MHz, CDCl3)δ-113.63(s).

[0363] Embodiment 18:

[0364]

[0365] Step 1: Dissolve compound 18A (5.0 g, 32 mmol) in dichloromethane (50 mL), and then add triethylamine (4.9 g, 48 mmol), di-tert-butyl dicarbonate (8.4 g, 38 mmol), and DMAP (0.4 g, 3.2 mmol) in sequence. After reacting at room temperature overnight, concentrate and purify on a silica gel column (PE / EA=3 / 1) to obtain compound 18B (7.0 g, 85%).

[0366] LC-MS (ESI): m / z = 257.2 [M+H] + .

[0367] Step 2: Dissolve compound 18B (2.0 g, 7.8 mmol) in DMF (20 mL), add sodium hydrogen sulfide (0.2 g, 9.3 mmol), stir at room temperature for 30 min, add iodomethane (1.4 g, 9.3 mmol), react at room temperature overnight, concentrate, and purify on a silica gel column (PE / EA=2 / 1) to obtain compound 18C (1.9 g, 90%).

[0368] LC-MS (ESI): m / z = 271.3 [M+H] + .

[0369] Step 3: Compound 18C (1.9 g, 7.0 mmol) was dissolved in methanol (20 mL), palladium carbon (0.1 g) was added, and the mixture was replaced with a hydrogen balloon for 3 times. After reacting at room temperature overnight, the mixture was filtered and concentrated to obtain compound 18D (1.7 g, 90%).

[0370] LC-MS (ESI): m / z = 241.3 [M+H] + .

[0371] Step 4: Compound 15D (0.5 g, 2.3 mmol) was dissolved in 1,4-dioxane (20 mL), and Pd2dba3 (95 mg, 0.1 mmol), Xantphos (120 mg, 0.2 mmol), 18D (0.5 g, 2.0 mmol) and sodium carbonate (550 mg, 5.2 mmol) were added, and nitrogen was replaced three times. The mixture was heated to 90 ° C. and reacted for 14 h, then passed through a short silica gel column, washed with ethyl acetate, concentrated, and column chromatography (dichloromethane / methanol = 15 / 1) was performed to obtain compound 18E (700 mg, 79%).

[0372] LC-MS (ESI): m / z = 425.9 [M + H] + .

[0373] Step 5: Compound 18E (0.7 g, 1.6 mmol) was dissolved in dichloromethane (10 mL), and m-CPBA (710 mg, 4.1 mmol) was added. After reacting at room temperature for 14 h, the mixture was concentrated and subjected to column chromatography (dichloromethane / methanol = 10 / 1) to obtain compound 18F (0.6 g, 80%).

[0374] LC-MS (ESI): m / z = 457.9 [M+H] + .

[0375] Step 6: Compound 18F (0.5 g, 1.1 mmol) was dissolved in 1,4-dioxane (20 mL), and 15C (260 mg, 1.3 mmol) and DIPEA (560 mg, 4.4 mmol) were added. The mixture was heated to 100 °C for 14 h, concentrated, and subjected to column chromatography (di-PE / EA=1 / 1) to give compound 18G (300 mg, 68%).

[0376] LC-MS (ESI): m / z = 620.7 [M+H] + .

[0377] Step 7: Compound 18G (260 mg, 0.36 mmol) was dissolved in DCM (3 mL), and trifluoroacetic acid (0.5 mL) was added. After reacting at room temperature for 20 min, the mixture was concentrated to obtain compound 18H (300 mg, crude product).

[0378] LC-MS (ESI): m / z = 420.6 [M + H] + .

[0379] Step 8: Compound 18H (300 mg, crude product) was dissolved in DCM (10 mL), and trichloroacetyl isocyanate (80 mg, 0.4 mmol) was added. After reacting at room temperature for 1 h, the mixture was concentrated to obtain compound 18I (380 mg, crude product).

[0380] LC-MS (ESI): m / z = 709.1 [M+H] + .

[0381] Step 9: Compound 18I (380 mg, crude product) was dissolved in ammonia methanol solution (7M, 10 mL), reacted at 70° C. for 12 h, concentrated, and purified by silica gel column (methanol / dichloromethane=10 / 1) to obtain compound 18 (120 mg).

[0382] LC-MS (ESI): m / z = 563.7 [M+H] + .

[0383] 1H NMR(400MHz, CDCl3)δ8.80(s,1H),7.85–7.80(m,1H),7.30(s,1H),7.28(s,2H),7.22(m,1H),7.24–7.20(m,2H),6.10(s,1H),4.40(s,4H),4.07 (s,2H),3.46–3.30(m,2H),3.24(s,3H),2.92(t,2H),2.62(s,2H),2.56 –2.41(m,2H),1.93–1.85(m,1H),1.01–0.92(m,2H),0.74–0.63(m,2H).

[0384] 19 F NMR(376MHz, CDCl3)δ-116.17(s).

[0385] Embodiment 19:

[0386]

[0387] Step 1: Compound 15G (200 mg, crude product) was dissolved in ammonia methanol solution (7M, 10 mL), reacted at 70° C. for 12 h, concentrated, and purified by silica gel column (methanol / dichloromethane=10 / 1) to obtain compound 19 (120 mg, 63.2%).

[0388] LC-MS (ESI): m / z = 548.2 [M+H] + .

[0389] 1 H NMR(400MHz, CDCl3)δ8.69(s,1H),7.75–7.57(m,1H),7.40–7.15(m,4H),7 .04(d,2H),6.08(s,1H),5.50(s,2H),4.50–4.25(m,2H),4.15–4.00(m,2H ),3.58(s,2H),3.45–3.25(m,2H),2.95–2.80(m,2H),2.68–2.55(m,2H),2 .54–2.40(m,2H),1.96–1.82(m,1H),1.02–0.92(m,2H),0.78–0.62(m,2H).

[0390] 19 F NMR(376MHz, CDCl3)δ-113.54(s).

[0391] Biological Testing:

[0392] 1. Effects of compounds on PDE4B2 activity

[0393] The effect of the compound on PDE4B2 activity was detected using a fluorescence polarization kit (BPS Bioscience, Catalog #60343). According to the kit instructions, a final concentration of 0.1 μM FAM-Cyclic-3', 5'-AMP, 1 ng / well PDE4B2 (negative control added PDE buffer), gradient diluted compounds (positive control wells added PDE buffer containing 10% DMSO) were fully mixed and reacted at room temperature for 1 hour. Use Binding Agent Diluent (cAMP) to dilute the Binding Agent at 1:100 for later use, take 50 μl / well of the Binding Agent dilution and add it to the test plate, incubate at room temperature with slow shaking for 20 minutes. After the incubation, Envision was used for FP detection, Excitation 480nm, Emission 535nm. FP is usually expressed by mP value.

[0394]

[0395] I II (S535): Fluorescence intensity in the parallel direction

[0396] I ┴ (P535): Fluorescence intensity in the vertical direction

[0397] G: G factor = 1

[0398] Calculation of inhibition rate (%Inhibition):

[0399] %Inhibition=[1-(mP (样品) -mP (阴性对照) ) / (mP (阳性对照) -mP (阴性对照) )]x100%

[0400] mxD (样品) : mP of the test compound reaction well

[0401] mxD (阴性对照) : Negative control well mP

[0402] mxD (阳性对照) :Positive control well mP

[0403] According to the calculated inhibition rate at each concentration, the IC of each compound was calculated using GraphPad Prism 8 software. 50 value.

[0404] The IC values ​​of the compounds of the present invention, such as the compounds in the examples, on PDE4B2 are 50 The value is <300 nM, preferably some compounds are <100 nM, more preferably some compounds are <50 nM, and further preferably some compounds are <10 nM.

[0405] IC of some specific compounds 50 The values ​​are as shown in Table 1, where A<10nM, 10nM≤B<50nM, 50nM≤C<100nM.

[0406] Table 1 PDE4B2 activity

[0407]

[0408]

[0409] As can be seen from the above, the compounds of the present invention, such as the examples, have good inhibitory activity. For example, the IC50 values ​​of compound 1-1 and compound 16 are 3.71 nM and 4.20 nM, respectively.

[0410] 2. Effects of compounds on PDE4D2 activity

[0411] The effect of the compound on PDE4D2 activity was detected using a fluorescence polarization kit (BPS Bioscience, Catalog #60345). During the reaction, 12.5 μL of 0.272 ng / well enzyme (final concentration 0.068 ng / well) was pre-incubated with 12.5 μL of graded diluted compound (DMSO concentration 4%) at room temperature for 15 minutes. The same volume and concentration of enzyme and 12.5 μL of PDE buffer containing 4% DMSO were added to the positive control wells, and 25 μL of PDE buffer containing 2% DMSO was added to the negative control wells. After completion, 25 μL of 0.2 μM FAM-Cyclic-3',5'-AMP (final concentration 0.1 μM) was added to each well, mixed thoroughly, and incubated at room temperature with slow shaking for 30 minutes. Dilute the Binding Agent with Binding Agent Diluent (cAMP) at 1:100 for later use. Take 100 μl of the Binding Agent diluent per well and add it to all wells of the test plate. Incubate at room temperature with slow shaking for 1 hour. After the incubation, use a BMG LRBTECH microplate reader to detect FP, with Excitation 485nm and Emission 520nm. FP is usually expressed by mP value.

[0412]

[0413] I II(S520): Fluorescence intensity in parallel direction

[0414] I ┴ (P520): Fluorescence intensity in the vertical direction

[0415] G: G factor = 1

[0416] Calculation of inhibition rate (%Inhibition):

[0417] %Inhibition=[1-(mP (样品) -mP (阴性对照) ) / (mP (阳性对照) -mP (阴性对照) )]x100%

[0418] mxD (样品) : mP of the test compound reaction well

[0419] mxD (阴性对照) : Negative control well mP

[0420] mxD (阳性对照) :Positive control well mP

[0421] According to the calculated inhibition rate at each concentration, the IC50 value of each compound was calculated using GraphPad Prism 8 software.

[0422] IC of the compounds of the present invention, such as the compounds in the examples, against PDE4D2 50 The value is less than 300 nM, and the preferred compound IC 50 The IC value of some specific compounds of the present invention is less than 100 nM, more preferably less than 50 nM, and further preferably less than 10 nM. 50 The values ​​are as shown in Table 2, where A<10nM, 10nM≤B<50nM, 50nM≤C<100nM.

[0423] Table 2 PDE4D2 activity

[0424] Compound <![CDATA[PDE4D2 IC 50 (nM)]]> Compound <![CDATA[PDE4D2 IC 50 (nM)]]> Compound 1-1 A Compound 1-2 A Compound 2-1 A Compound 2-2 A Compound 3-1 B Compound 3-2 B Compound 4-1 A Compound 4-2 B Compound 5-1 A Compound 5-2 A Compound 6-1 B Compound 6-2 A Compound 7-1 C Compound 7-2 B Compound 9-1 C Compound 9-2 C Compound 10-1 C Compound 10-2 C Compound 11-1 B Compound 12-1 B Compound 13-1 B Compound 13-3 A Compound 14-1 B Compound 15 B Compound 16 B Compound 17 B Compound 18 C Compound 19 C

[0425] The compounds of the present invention, such as the examples, have good selective inhibitory activity against PDE4B. For example, the inhibitory activity of compounds 16, 18 and 19 against PDE4B is 6-9 times that against PDE4D.

[0426] 3. Detection of the inhibitory activity of compounds on the release of tumor necrosis factor-α (TNF-α) from human peripheral blood mononuclear cells induced by lipopolysaccharide (LPS) in vitro

[0427] Normal human peripheral anticoagulated blood (citrate anticoagulation) was collected and hPBMCs were prepared using Ficoll-Paque PLUS (Cytiva, Cat#17144002, density 1.077 g / mL). The hPBMC cell concentration was adjusted to 0.25 x 10 6 / mL, and inoculated in a 96-well plate with 50,000 cells per well. Subsequently, different concentrations of drugs were added for pre-incubation for 1 hour (the final concentration of DMSO was 0.1%, and the positive and negative control wells were equal volumes of RPMI1640 containing 0.1% DMSO). After the pre-incubation was completed, 100 ng / mL LPS (SIGMA, L2630) was added to the compound and positive control wells, and the negative control wells were equal volumes of RPMI1640 containing 0.1% DMSO. They were cultured in an incubator at 37°C and 5% CO2 for 4 hours. The cell supernatant was collected, and the human TNF-α Elisa quantitative detection kit (Sino Biological, Cat#KIT10602) was used to detect the TNF-α content in the supernatant sample. GraphPad Prism software was used to calculate the IC 50 value.

[0428] IC values ​​of the inhibitory activity of the compounds of the present invention, such as the compounds in the examples, on LPS-induced TNF-α in vitro 50 The value is less than 300 nM, and the preferred compound IC 50 The value is less than 100 nM, more preferably less than 50 nM, and further preferred compounds are less than 10 nM.

[0429] 4. Pharmacokinetics test in mice

[0430] 4.1 Experimental animals: Male ICR mice, 20-25 g, 6 mice / compound, purchased from Chengdu Dashuo Experimental Animal Co., Ltd.

[0431] 4.2 Experimental design: On the day of the experiment, ICR mice were randomly divided into groups according to body weight. They were fasted but not watered for 12-14 hours one day before administration and fed 4 hours after administration.

[0432] Before and after drug administration, 0.06 mL of blood was collected from the eye socket under isoflurane anesthesia, placed in an EDTAK2 centrifuge tube, and centrifuged at 5000 rpm at 4°C for 10 min to collect plasma. The blood collection time points for the intravenous group and the gavage group were: 0, 5, 15, 30 min, 1, 2, 4, 6, 8, 24 h. Before analysis and testing, all samples were stored at -80°C and quantitatively analyzed by LC-MS / MS.

[0433] Conclusion: The compounds of the present invention, such as the examples, showed excellent pharmacokinetic properties in the mouse PK test.

[0434] 5. Pharmacokinetic test in rats

[0435] 5.1 Experimental animals: Male SD rats, about 220 g, 6 to 8 weeks old, 6 rats per compound, purchased from Chengdu Dashuo Experimental Animal Co., Ltd.

[0436] 5.2 Experimental design: On the day of the experiment, SD rats were randomly divided into groups according to body weight. The rats were fasted but not watered for 12-14 hours one day before administration and were fed 4 hours after administration.

[0437] Before and after drug administration, 0.15 ml of blood was collected from the eye socket under isoflurane anesthesia, placed in an EDTAK2 centrifuge tube, and centrifuged at 5000 rpm and 4°C for 10 min to collect plasma. The blood collection time points for the intravenous group and the gavage group were: 0, 5, 15, 30 min, 1, 2, 4, 6, 8, 24 h. Before analysis and testing, all samples were stored at -80°C and quantitatively analyzed by LC-MS / MS.

[0438] Conclusion: The compounds of the present invention, such as the examples, showed excellent pharmacokinetic properties in rat PK tests.

[0439] 6. Beagle dog pharmacokinetic test

[0440] 6.1 Experimental animals: Male beagle dogs, about 8-11 kg, 6 per compound, purchased from Beijing Mas Biotechnology Co., Ltd.

[0441] 6.2 Test method: On the day of the test, beagle dogs were randomly divided into groups according to body weight. They were fasted but not watered for 12-14 hours one day before administration and were fed 4 hours after administration.

[0442] Before and after administration, 1 ml of blood was collected from the jugular vein or limb vein and placed in an EDTAK2 centrifuge tube. The blood was centrifuged at 5000 rpm and 4°C for 10 min to collect plasma. The blood collection time points for the intravenous group and the gavage group were: 0, 5, 15, 30 min, 1, 2, 4, 6, 8, 10, 12, 24, 48, 72 h. Before analysis and testing, all samples were stored at -80°C and quantitatively analyzed by LC-MS / MS.

[0443] Conclusion: The compounds of the present invention, such as the examples, showed excellent pharmacokinetic properties in the beagle dog PK test.

[0444] 7. Detection of anti-TNF-α secretion activity in LPS-induced mouse lung inflammation model

[0445] In mice, intratracheal administration of lipopolysaccharides (LPS, Sigma, L2880) can induce an increase in the concentration of TNF-α in lung tissue. In this experiment, a certain number of mice (BABL / c, male, weight 18-22g) were randomly divided into groups, 8 mice in each group, and a certain dose of the test compound was orally administered (the test compound was suspended with 0.5% MC and prepared into a certain concentration, and administered orally in a volume of 10ml / kg body weight). After 0.5h, a 0.8mg / mL dose of LPS was administered intratracheally using a lung quantitative atomization needle according to the animal's body weight of 2.5uL / g. After 24h, the mice were anesthetized intraperitoneally with 20% urethane (10mL / kg), and then killed by cervical dislocation. The lung tissue was then taken, and the lower half of the left lobe was fully homogenized. The TNF-α content in the supernatant of the homogenate was detected using the Mouse TNFα ELISA Kit (RD, SMTA00B).

[0446] The inhibition rate of the test compound on TNF-α level (% Inh) = (model group - test group) / model group * 100%.

Claims

1. A compound represented by formula (I) or (II), a stereoisomer or a pharmaceutically acceptable salt thereof, in, Cy2 is selected from 3-6 membered cycloalkyl; Cy1 is selected from or Indicates the site where Cy1 and Cy2 are linked; Or Cy1-Cy2 together form R2 is selected from C 2-6 Alkynyl, 3-5 membered cycloalkyl or C 1-4 Alkoxy; q is selected from 1 or 2; R 12 is selected from 4-10 membered heterocycloalkylene, 3-10 membered cycloalkylene, 6-10 membered arylene, 5-10 membered heteroarylene, hydroxy-C 1-6 Alkyl, C 1-4 Alkyl -CN, -CRaRb=NOC 1-4 Alkyl, -C(NRaRb)=N-CN, -C(C 1-4 alkyl)=N-CN、-C(NRaRb)=CRa-NO 2、 -C(CH3)2-CH2-OC(O)NH2, -C(CH3)2-CH2-NH-C(O)O(C 1-4 Alkyl), -C(CH3)2-CH2-OC(O)NHCH3, -CH(CH3)-CH2-OC(O)NH2, -C(CH3)2-CH(CH3)-OC(O)NH2, -CH(CH3)-CH2-NH-C(O)O(C 1-4 alkyl), -CH(CH3)-CH2-OC(O)NHCH3, -C(O)-Ra or -C(O)-(4-6 membered heterocycloalkyl), wherein the heterocycloalkylene, cycloalkylene, arylene, heteroarylene may be further substituted with 1-3 halogens, CN, =O, OH, -SF5, C 1-4 Alkyl, C 1-4 Alkyl-CN, hydroxyl C 1-4 Alkyl, C 1-4 Haloalkyl, -(CH2) t -OC(O)NH2, -CH(CH3)-OC(O)NH2, -(CH2) t -NRa-C(O)NH2, -(CH2) t -NRa-C(=NH)NH2, -(CH2) t -NRa-C(O)-O-(C 1-4 Alkyl), -(CH2) t -NRa-C(C 1-4 alkyl)=N-CN, -(CH2) t -NRa-C(NRaRb)=N-CN、-(CH2) t -NRa-C(NRaRb)=CRa-NO2, -(CH2) t -C(NRaRb)=N-CN、-(CH2) t -C(C 1-4 alkyl)=N-CN, -(CH2) t -C(C 1-4 alkyl)=NO-(C 1-4 alkyl), =NOC 1-4 Alkyl, 5-6 membered heteroaryl, -(CH2) t -OC 1-4 Alkyl, -(CH2) t -OC(O)NHCH3, -(CH2) t -OC 1-2 Haloalkyl, -(CH2) t -OC 3-6 Cycloalkyl, -(CH2) t -OC 1-4 Alkyl-OC 3-6 Cycloalkyl, -(CH2) t -COOH, -(CH2) t -NRaRb, -(CH2) t -C(O)NRaRb, -(CH2) t -NRa-C(O)CH3 group substitution; Ra and Rb are each independently selected from H, halogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy or 4-6 membered heterocycloalkyl containing 1-2 heteroatoms selected from N and O; Alternatively, Ra, Rb and the atoms to which they are attached together form C 3-6 Cycloalkyl; t is selected from 0, 1, 2 or 3; The condition is that the compound of formula (I) or (II) is not the following structure:

2. The compound according to claim 1, its stereoisomer or pharmaceutically acceptable salt, wherein R 12 Selected from 3-10 membered cycloalkylene, 6-10 membered arylene, hydroxy-C 1-6 Alkyl, C 1-4 Alkyl-CN, -C(CH3)2-CH2-OC(O)NH2, -C(CH3)2-CH2-NH-C(O)O(C 1-4 alkyl), -C(CH3)2-CH(CH3)-OC(O)NH2, wherein the cycloalkylene and arylene groups are optionally further substituted with 1-3 halogens, C 1-4 Alkyl-CN, hydroxyl C 1-4 Alkyl, -(CH2) t -OC(O)NH2, -CH(CH3)-OC(O)NH2, -(CH2) t -NRa-C(O)NH2, -(CH2) t -COOH, -(CH2) t -C(O)NRaRb group substitution.

3. The compound according to claim 1, its stereoisomer or pharmaceutically acceptable salt, wherein Selected from: R2 is selected from cyclopropyl, ethynyl, propynyl or ethoxy.

4. The compound according to claim 1, its stereoisomer or pharmaceutically acceptable salt, wherein: Cy1 is selected from Indicates the site where Cy1 and Cy2 are linked; Cy2 is selected from R2 is selected from cyclopropyl; Selected from: The condition is that the compound of formula (I) or (II) is not 5. The compound according to claim 1, its stereoisomer or pharmaceutically acceptable salt, wherein the compound is selected from one of the structures in Table A.

6. A pharmaceutical composition or pharmaceutical preparation comprising the compound according to claims 1 to 5, its stereoisomers or pharmaceutically acceptable salts, and a pharmaceutically acceptable carrier and / or excipient.

7. The pharmaceutical composition or pharmaceutical preparation according to claim 6, comprising 1-1500 mg of the compound according to claims 1-5, its stereoisomer or pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier and / or excipient.

8. Use of the compound according to claims 1-5, its stereoisomer or pharmaceutically acceptable salt, or the composition according to claims 6-7 in the preparation of a medicament for treating / preventing a PDE4B-mediated disease.

9. The use according to claim 8, wherein the PDE4B-mediated disease is selected from cancer, COPD, idiopathic pulmonary fibrosis, and interstitial lung disease.

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