Spiro compound and use thereof in the field of medicine

Abstract

The present application relates to a spiro compound and use thereof in the field of medicine. Specifically, disclosed in the present application are a compound represented by formula (I), an isotope-labeled compound thereof, an optical isomer thereof, a tautomer thereof, or a pharmaceutically acceptable salt thereof, or a crystal form or solvate form of the compound or pharmaceutically acceptable salt thereof.

Description

Spirocyclic compounds and their applications in medicine

[0001] This application claims priority to Chinese invention patent application No. 202310998036.2 filed on August 8, 2023, and the entire disclosure of that prior application is incorporated herein by reference. Technical Field

[0002] The present invention relates to a class of substituted 2'H, 3H-spiro[imidazo[1,2-a]pyridine-2,1'-naphthalene] compounds and their application in the field of medicine. Background Art

[0003] Under normal physiological conditions, EPO (erythropoietin) is produced by peritubular interstitial cells in response to local tissue hypoxia. However, in renal cell carcinoma, EPO is produced in tumor cells. Statistics show that two-thirds of renal cell carcinoma patients have elevated EPO levels compared to normal levels, and 8% of these patients experience polycythemia (Reviews in Urology. 2002, 4(4), 163–170). In these patients, the elevated serum red blood cell concentration is believed to be mediated by EPO. Polycythemia vera can easily lead to venous thrombosis, bleeding, and vascular and nervous system problems, leading to stroke and heart disease; if untreated, the patient is in danger of death. Current treatments include long-term phlebotomy, cytoreductive therapies such as hydroxyurea and interferon α, intravenous injection of radioactive phosphorus, persulfate, and JAK inhibitors such as ruxolitinib (Chin. J. Hematol. 2022, vol 43, 537-541).

[0004] In addition, EPO levels are generally elevated in renal cell carcinoma, making it a potential tumor marker. Renal cell carcinoma patients with elevated EPO levels may be more sensitive to their EPO levels, making lowering EPO levels a potential and effective treatment for polycythemia vera and renal cell carcinoma.

[0005] Therefore, it is desirable to develop an EPO inhibitor that can significantly reduce EPO levels, thereby realizing its application in the treatment of various diseases, such as polycythemia vera, renal cell carcinoma, and other cancers.

[0006] Summary of the Invention

[0007] The present invention provides a method for preparing a spirocyclic compound, and the compound has good chemical stability.

[0008] The inventors have successfully prepared a new class of spirocyclic compounds with excellent chemical stability. These compounds have been shown to significantly reduce EPO levels in a variety of biochemical, biological, and cellular tests, and therefore have promising applications in treating a variety of diseases, such as polycythemia vera, renal cell carcinoma, and other cancers.

[0009] In a first aspect, the present invention provides a compound represented by formula (I), an isotope-labeled compound thereof, an optical isomer thereof, a tautomer thereof, or a pharmaceutically acceptable salt thereof, or a crystalline form or solvate of the compound or its pharmaceutically acceptable salt. The compound represented by formula (I), an isotope-labeled compound thereof, an optical isomer thereof, a tautomer thereof, or a pharmaceutically acceptable salt thereof, or a crystalline form or solvate of the compound or its pharmaceutically acceptable salt can be used to treat various diseases associated with EPO, such as polycythemia vera, renal cell carcinoma, and other cancers.

[0010] In formula (I), R 1 Selected from H, D, F, Cl, Br, CF3, CN;

[0011] R 2 Selected from H, D, CF3, CH3, OCH3, OH;

[0012] R 3 Selected from H, D, F, Cl, Br, CF3, CN, OCH3, OCH2F, OCHF2, OCF3;

[0013] R 4 Selected from H, D, F, Cl, Br, CH3, CF3, CN;

[0014] R 5 Selected from H, D, F, Cl, Br, I, CH3, CF3, CN, OCH3;

[0015] R 6 Selected from H, D, F, Cl, Br, I, CH3, CF3, CN, OCH3;

[0016] X is selected from N,CR 7 ; Among them, R 7 Selected from H, D, F, Cl, Br, CF3, CN, CH3.

[0017] Preferably, the R 1 Selected from CF3, CN.

[0018] Preferably, R 2 Selected from OCH3, OH.

[0019] Preferably, R 3 Selected from OCH3, OCH2F, OCHF2, OCF3.

[0020] Preferably, R 4 Selected from H, D, F, Cl, CH3.

[0021] Preferably, R 5 Selected from H, D, F, Cl, CH3, CF3, OCH3.

[0022] Preferably, R 6 Selected from F, Cl, CH3.

[0023] Preferably, X is selected from N, CR 7 ; Among them, R 7 Selected from H, D, F, Br, CF3, CN, CH3.

[0024] In some most preferred embodiments, the compound of formula (I) of the present application is selected from the specific compounds shown in the examples of the present application.

[0025] Unless otherwise indicated, the "compound represented by formula (I)", "compound of formula (I)", "compound of the present application", "compound of the present invention" or similar terms described herein are also intended to cover its isotopically labeled compounds, its optical isomers, its tautomers, or its pharmaceutically acceptable salts, or the crystal form or solvate form of the compound or its pharmaceutically acceptable salt.

[0026] The term "optical isomer" means that when a compound has one or more chiral centers, each chiral center can exist in the R configuration or the S configuration, and the various isomers thus formed are optical isomers. Optical isomers include all diastereomers, enantiomers, meso-isomers, racemates, or mixtures thereof. For example, optical isomers can be separated by chiral chromatography or by chiral synthesis.

[0027] The term "tautomer" refers to isomers that result from the rapid shift of an atom between two positions in a molecule. Those skilled in the art will appreciate that tautomers can transform into each other and, under certain conditions, may reach an equilibrium state and coexist.

[0028] Unless otherwise indicated, the compound represented by formula (I) is intended to encompass isotopically labeled compounds in which one or more atoms are replaced by isotopes thereof.

[0029] Examples of suitable isotopes for inclusion in the compounds of the present application include isotopes of hydrogen such as 2 H(D) and 3 H(T); isotopes of carbon, such as11 C. 13 C and 14 C; isotopes of chlorine, such as 36 Cl; isotopes of fluorine, such as 18 F; isotopes of iodine, such as 123 I and 125 I; isotopes of nitrogen, such as 13 N and 15 N; isotopes of oxygen, such as 15 O. 17 O and 18 O; and isotopes of sulfur, such as 35 S.

[0030] Isotope-labeled compounds (e.g., those containing radioactive isotopes) are useful in drug and / or substrate tissue distribution studies. Radioactive isotopes such as deuterium (i.e., D) and carbon-14 (i.e., C-14) are used for the distribution of substances in the tissues of interest. 14 C) is particularly useful for this purpose.

[0031] Substitution with heavier isotopes such as deuterium (i.e., D) can provide certain therapeutic benefits, such as greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements), and therefore can be preferred in certain circumstances. Therefore, in some embodiments, the compounds of the present application are isotopically labeled compounds wherein H is optionally replaced by D at each occurrence.

[0032] Using positron-emitting isotopes such as 11 C. 18 F. 15 O and 13 N) substitution can be used in positron emission topography (PET) studies to detect substrate receptor occupancy status.

[0033] Isotopically labeled compounds can generally be prepared by conventional techniques known to those skilled in the art, or by using an appropriate isotopically labeled reagent in place of the non-labeled reagent previously used.

[0034] The compounds of the present application may exist in the form of pharmaceutically acceptable salts.

[0035] The term "pharmaceutically acceptable" refers to a compound, carrier or molecule that is suitable for administration to a human. Preferably, the term refers to a compound, carrier or molecule that is approved by any national regulatory agency such as CFDA (China), EMEA (Europe), FDA (USA) for use in mammals, preferably humans.

[0036] The pharmaceutically acceptable salts include acid addition salts and base addition salts thereof. Suitable acid addition salts are formed from acids which form non-toxic salts. Examples include, but are not limited to, acetate, adipate, aspartate, benzoate, benzenesulfonate, bicarbonate / carbonate, bisulfate / sulfate, borate, camphorsulfonate, citrate, cyclamates, edisylate, formate, fumarate, glucoheptonate, gluconate, glucuronate, hexafluorophosphate, 2-(4-hydroxybenzyl)benzoate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, 2-hydroxyethanesulfonate, lactate, malate, maleate, malonate, methanesulfonate, methylsulfate, naphthoate, 2-naphthalenesulfonate, nicotinate, nitrate, orotate, oxalate, hexadecanoate, phosphate / hydrogenphosphate / dihydrogenphosphate, pyroglutamate, glucarate, stearate, salicylate, tannate, tartrate, toluenesulfonate, and trifluoroacetate. Suitable base addition salts are formed with bases which form non-toxic salts. Examples include, but are not limited to, aluminum, arginine, calcium, choline, diethylamine, diethanolamine, glycine, lysine, magnesium, meglumine, ethanolamine, potassium, sodium, tromethamine, and zinc salts. Hemi-salts of acids and bases, such as hemisulfate and hemicalcium salts, can also be formed. For a review of suitable salts, see Handbook of Pharmaceutical Salts: Properties, Selection and Use by Stahl and Wermuth (Wiley-VCH, 2002). Methods for preparing pharmaceutically acceptable salts of the compounds described herein are known to those skilled in the art.

[0037] In addition, the compounds of the present invention may exist in unsolvated forms as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, etc. The compounds may also exist in one or more crystalline states, i.e., polymorphs, or they may exist as amorphous solids. All of these forms are included within the scope of the present application.

[0038] Preparation method of compound

[0039] The compound of (I) of the present invention can be synthesized by a variety of methods familiar to those skilled in the art. Some exemplary synthetic methods are provided below, and these methods are well known to those skilled in the art, and various modifications can be made to these methods as needed. If necessary, the starting materials, intermediates and final products in the separation and purification reactions can be obtained using conventional techniques (including but not limited to related technologies such as filtration, distillation, crystallization or column chromatography). In addition, each specific embodiment herein also illustrates the method for synthesizing the compounds of the present invention.

[0040] The synthesis methods of the compounds of the present invention are schematically illustrated below using Compound Ia and Compound Ib as examples.

[0041] Option 1:

[0042] Intermediate Ia-2 is obtained by heating Ia-1 (synthesized by known methods or commercially available) with DAST in an organic solvent. The preferred solvent for this step is dichloromethane or dichloroethane; the preferred reaction temperature is 70°C to 90°C. Intermediate Ia-2 is reacted with a base to obtain intermediate Ia-3. The preferred base for this step is sodium hydroxide, potassium hydroxide, or cesium carbonate; the preferred solvent for this step is methanol. Intermediate Ia-3 undergoes ring closure in the presence of sulfonyl chloride and a base to obtain compound Ia. The preferred base for this step is triethylamine or diisopropylethylamine; the preferred reaction temperature is 70°C to 90°C.

[0043] Option 2:

[0044] Intermediate Ib-2 is obtained by reacting Ib-1 (synthesized according to known methods or commercially available) with a reducing agent in an organic solvent. The preferred solvent for this step is methanol or ethanol; the preferred reaction temperature is 0°C to 30°C. Intermediate Ib-2 is reacted with a dehydrating agent to obtain compound Ib. The preferred dehydrating agent for this step is Burgess reagent [methyl N(triethylammoniumsulfonyl)carbamate, CAS: 29684-56-8] or p-toluenesulfonic acid; the preferred reaction temperature is 70°C to 90°C.

[0045] With reference to the schematic synthetic routes of compounds Ia and Ib above and the synthetic routes shown in the specific examples below, those skilled in the art can synthesize other desired compounds of formula (I) with similar structures by appropriate modification.

[0046] Application of compounds

[0047] Experiments have shown that the compounds of the present invention have excellent inhibitory activity against EPO. Therefore, the compounds of the present invention can be used as EPO inhibitors to treat EPO-related diseases, especially those related to excessive EPO levels.

[0048] In a second aspect, the present application provides a pharmaceutical composition containing the compound of the present application as described above (i.e., the compound of formula (I)) or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, adjuvants or excipients.

[0049] The pharmaceutical composition of the present application can be prepared in a manner well known in the pharmaceutical field and can be applied by a variety of routes, depending on whether local treatment or systemic treatment is desired and on the site to be treated. Administration can be local (including ophthalmic and to mucous membranes, including intranasal, vaginal and rectal delivery), pulmonary (e.g., by inhalation or insufflation of powder or aerosol, including by a nebulizer; intratracheal, intranasal, epidermal and transdermal), ocular, oral or parenteral. Methods for ocular delivery can include topical administration (eye drops), subconjunctival, periocular or intravitreal injection or introduction by a balloon catheter or ophthalmic insert placed in the conjunctival sac with a surgical method. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; or intracranial (e.g., intrathecal or intraventricular) administration. Parenteral administration can be in the form of a single bolus dose, or can be, for example, by a continuous infusion pump. Pharmaceutical compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders.

[0050] If a solid carrier is used, the preparation can be tableted, placed in a hard gel capsule in the form of a powder or granules, or in the form of a lozenge or pastille. Solid carriers may include conventional excipients, such as adhesives, fillers, tableting lubricants, disintegrants, wetting agents, etc. If necessary, the tablets may be film-coated by conventional techniques. If a liquid carrier is used, the preparation may be in the form of a syrup, emulsion, ointment, soft gel capsule, a sterile carrier for injection, an aqueous or non-aqueous liquid suspension, or may be a dry product reconstituted with water or other appropriate carriers before use. Liquid preparations may include conventional additives, such as suspending agents, emulsifiers, wetting agents, non-aqueous carriers (including edible oils), preservatives, and flavorings and / or coloring agents. For parenteral administration, typically the carrier at least comprises sterile water for the most part, but saline solutions, glucose solutions, etc. may also be used. Injectable suspensions may also be used, in which case conventional suspending agents may be used. Conventional preservatives, buffering agents, etc. may also be added to parenteral dosage forms. Pharmaceutical compositions are prepared by conventional techniques appropriate to the desired formulation containing appropriate amounts of the active ingredient (ie, the compound of the present application).

[0051] Compositions suitable for parenteral injection can include physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions and sterile powders for sterile injectable solutions or dispersions. Examples of suitable aqueous and non-aqueous carriers, diluents, solvents include water, ethanol, polyols (propylene glycol, polyethylene glycol, glycerol, etc.), suitable mixtures thereof, vegetable oils (e.g., olive oil) and injectable organic esters (e.g., ethyl oleate).

[0052] These compositions can also include various excipients, for example, preservatives, wetting agents, emulsifiers and dispersants. The inhibition of the effect of microorganisms can be ensured by various antibacterial and antifungal agents (for example, parabens, chlorobutanol, phenol, sorbic acid, etc.). Isotonic agents can also be included, for example, sugar, sodium chloride, etc. The absorption of injectable pharmaceutical dosage forms can be extended by using delayed absorption agents (for example, aluminum monostearate and gel).

[0053] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dosage forms, the active compound is mixed with at least one inert excipient (or carrier) (e.g., sodium citrate or dicalcium phosphate) and may also include: (a) fillers or admixtures (e.g., starch, lactose, sucrose, glucose, mannitol and silicic acid); (b) binders (e.g., carboxymethylcellulose, alginate, gelatin, polyvinyl pyrrolidone, sucrose and gum arabic); (c) humectants (e.g., glycerol); (d) disintegrants (e.g., agar); (e.g., fat-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain synthetic silicates, sodium carbonate); (e) solution retarding agents (e.g., paraffin); (f) absorption accelerators (e.g., quaternary ammonium compounds); (i) wetting agents (e.g., cetyl alcohol and glycerol monostearate); (h) adsorbents (e.g., kaolin and bentonite); and (i) lubricants (e.g., talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate), or mixtures thereof.

[0054] Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using, for example, lactose and high molecular weight polyethylene glycols as excipients.

[0055] Solid dosage forms (e.g., tablets, dragees, capsules, pills, and granules) can be prepared using coatings and shells (e.g., enteric coatings and others known in the art). They may contain opacifiers, and they may also be compositions that release the active compound or compounds in a delayed manner in a certain part of the intestine. Examples of useful embedding compositions are polymeric substances and waxes. The active ingredient may also be in microencapsulated form, if appropriate, with one or more of the above-mentioned excipients.

[0056] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, dispersions, syrups, and elixirs. In addition to the active compound, the liquid dosage form may contain inert diluents (e.g., water or other solvents), solubilizers and emulsifiers (e.g., ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide), oils (specifically, cottonseed oil, peanut oil, corn oil, olive oil, castor oil, sesame oil), glycerol, tetrahydrofuran alcohol, polyethylene glycol, and fatty acid esters of sorbitan, or mixtures thereof, etc., commonly used in the art.

[0057] Besides such inert diluents, the composition may also include, for example, wetting agents, emulsifying and suspending agents, perfuming, flavoring, and perfuming agents.

[0058] In addition to the active compounds, suspensions may contain suspending agents such as, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol, sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, or mixtures of these substances.

[0059] The topical dosage forms of the compound of the present application include ointments, powders, sprays and inhalants. The active ingredient is mixed with a physiologically acceptable carrier and any required preservative, buffer or propellant under aseptic conditions. Ophthalmic formulations, eye ointments, powders and solutions are also included in the scope of the present application.

[0060] The amount of the compound of the present application in the pharmaceutical composition and dosage form can be appropriately determined by those skilled in the art as needed. For example, the compound of the present application can be present in the pharmaceutical composition or dosage form in a therapeutically effective amount.

[0061] In a third aspect, the present application provides a compound of the present application (i.e., a compound of formula (I)) or a pharmaceutically acceptable salt thereof for use as an EPO inhibitor drug, or the use of a compound of the present application (i.e., a compound of formula (I)) or a pharmaceutically acceptable salt thereof in the manufacture of an EPO inhibitor drug.

[0062] In a fourth aspect, the present application also provides a method for treating diseases or conditions associated with EPO (especially associated with excessive EPO), which comprises administering to a patient in need thereof a therapeutically effective amount of a compound of the present application (i.e., a compound of formula (I)) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above.

[0063] In the fifth aspect, the present application also provides the use of the compound of the present application (i.e., the compound of formula (I)) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating patients suffering from diseases or conditions associated with EPO (especially associated with excessive EPO).

[0064] The patient is preferably a mammal, more preferably a human patient. During the treatment process, the route of administration can be oral, external (including but not limited to external application, spraying, etc.), parenteral (including subcutaneous, intramuscular, cortical and intravenous) administration, bronchial administration or nasal administration, etc.

[0065] In some embodiments, the EPO-related disease or condition is polycythemia vera, renal cell carcinoma, and other cancers.

[0066] Exemplary other cancers include bladder cancer, breast cancer, cervical cancer, colorectal cancer, small intestine cancer, colon cancer, rectal cancer, anal cancer, endometrial cancer, head and neck cancer (e.g., cancer of the larynx, hypopharynx, nasopharynx, oropharynx, lip and oral cavity), liver cancer (e.g., hepatocellular carcinoma, cholangiocellular carcinoma), lung cancer (e.g., adenocarcinoma, small cell lung cancer and non-small cell lung cancer, small cell carcinoma and non-small cell carcinoma, bronchogenic carcinoma, bronchial adenoma, pleuropulmonary blastoma), ovarian cancer, prostate cancer, testicular cancer, uterine cancer, esophageal cancer, gallbladder cancer, pancreatic cancer (e.g., exocrine pancreatic cancer), thyroid cancer, parathyroid cancer, skin cancer (e.g., squamous cell carcinoma, Kaposi's sarcoma, Merkel cell skin cancer), and brain cancer (e.g., astrocytoma, medulloblastoma, ependymoma, neuroectodermal tumor, pineal tumor).

[0067] In some preferred embodiments, the EPO-related disease or disorder is polycythemia.

[0068] In some preferred embodiments, the EPO-related disease or disorder is renal cell carcinoma.

[0069] The "therapeutically effective amount" of the compounds of the present application for treating the above-mentioned diseases can be reasonably determined by experienced physicians or researchers based on factors such as the patient's condition, physical condition, severity of the disease, and route of administration.

[0070] The present application is further illustrated and described below in conjunction with specific embodiments. Specific embodiments

[0071] The present invention will be further described below with reference to specific examples. It should be understood that these examples are only intended to illustrate the present invention and are not intended to limit the scope of the present invention.

[0072] In the following examples, if no specific conditions are specified, the experimental methods are generally carried out according to conventional conditions for such reactions or conditions recommended by the manufacturers.

[0073] Unless otherwise specified, the experimental materials and reagents used in the following examples can be obtained from commercial channels or prepared according to the cited literature; the instruments and equipment used in the synthesis experiments and product analysis and detection are conventional instruments and equipment commonly used in organic synthesis and analysis; the main reagents, instruments, etc. of the biological experiments are given in the experimental procedures.

[0074] The following abbreviations are used in this paper: BINAP: 1,1'-binaphthyl-2,2'-bis(diphenylphosphine); Cp*: pentamethylphosphine; DAST: diethylaminosulfur trifluoride; DCM: dichloromethane; DMSO: dimethyl sulfoxide; DMA: N,N-dimethylacetamide; DMF: N,N-dimethylformamide; DMI: 1,3-dimethyl-2-imidazolidinone; dppf: diphenylphosphinoferrocene; ESI-MS: electrospray ionization mass spectrometry; KHMDS: hexamethyl Potassium silylate; MsCl: methanesulfonyl chloride; NCS: N-chlorosuccinimide; NFSI: N-fluorobisbenzenesulfonamide; NMP: N-methylpyrrolidone; Pd2dba3: tris(dibenzylideneacetone)dipalladium; PPTS: pyridine p-toluenesulfonate; Selectfluor: 1-chloromethyl-4-fluoro-1,4-diazabicyclo[2,2,2]octane bis(tetrafluoroborate); SFC: supercritical fluid chromatography; TBSOTf: tert-butyldimethylsilyl trifluoromethanesulfonate; THF: tetrahydrofuran.

[0075] Example 1: Preparation of 1′-chloro-8-difluoromethoxy-8′-fluoro-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline

[0076] Step 1: Synthesis of 1-chloro-3′-(3-(difluoromethoxy)-5-(trifluoromethyl)pyridin-2-yl)-8-fluoro-6H-spiro[isoquinolin-5,4′-oxazoline]-2′-one

[0077] 1-Chloro-3′-(3-(difluoromethoxy)-5-(trifluoromethyl)pyridin-2-yl)-6,7-dihydro-8-H-spiro[isoquinoline-5,4′-oxazoline]-2′,8-dione (synthesized according to WO 2021 / 220170, 1.01 g) was dissolved in DCM (10 mL). DAST (14 mL) was added at room temperature and stirred at 80°C for 4 h. The reaction solution was added dropwise to a saturated sodium bicarbonate solution, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain 79.7 mg of the title compound. ESI-MS m / z: 507.28 [M+H+CH3CN] +

[0078] Step 2: Synthesis of (1-chloro-5-((3-(difluoromethoxy)-5-(trifluoromethyl)pyridin-2-yl)-amino)-8-fluoro-5,6-dihydroisoquinolin-5-yl)methanol

[0079] Dissolve 1-chloro-3′-(3-(difluoromethoxy)-5-(trifluoromethyl)pyridin-2-yl)-8-fluoro-6H-spiro[isoquinoline-5,4′-oxazoline]-2′-one (57 mg) in MeOH (10 mL). Add 4M NaOH (0.3 mL) dropwise at 0°C and stir at room temperature for 1 hour. Extract three times with ethyl acetate, dry the mixture, and separate on a preparative plate to obtain the title compound (12 mg). ESI-MS m / z: 438.29 [MH] -

[0080] Step 3: Synthesis of 1′-chloro-8-difluoromethoxy-8′-fluoro-6-trifluoromethyl-3H,6′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinoline

[0081] The compound (1-chloro-5-((3-(difluoromethoxy)-5-(trifluoromethyl)pyridin-2-yl)-amino)-8-fluoro-5,6-dihydroisoquinolin-5-yl)methanol (12 mg) was dissolved in toluene (1 mL), and MsCl (21 μL) and Et3N (76 μL) were added. The mixture was stirred at 80°C for 1 hour. Saturated sodium bicarbonate solution was added, and the mixture was extracted three times with ethyl acetate. The mixture was then spin-dried to dryness, and preparative liquid phase separation was performed to obtain the title compound, Example 1 (5.0 mg). ESI-MS m / z: 422.34 [M+H] + , 1 H NMR(500MHz,DMSO-d6)δ8.33(d,1H),7.85(t,1H),7.35(t,1H),7.27(dd,1H),7.0 3(d,1H),5.84(ddd,1H),4.37(d,1H),3.94(d,1H),2.60(ddd,1H),2.51(ddd,1H).

[0082] Example 1-A and Example 1-B: Preparation of (S)-1′-chloro-8-difluoromethoxy-8′-fluoro-6-trifluoromethyl-3H, 6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline and (R)-1′-chloro-8-difluoromethoxy-8′-fluoro-6-trifluoromethyl-3H, 6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline

[0083] Racemic 1′-chloro-8-difluoromethoxy-8′-fluoro-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline (i.e., the compound of Example 1) was subjected to preparative SFC to obtain a pair of enantiomers; the enantiomer eluted first from the chiral column below was Example 1-A; the enantiomer eluted later was Example 1-B (note that the absolute configurations of the two compounds of Examples 1-A and 1-B have not yet been determined. For the convenience of description only, the chiral center of Example 1-A is arbitrarily designated as S, while the chiral center of Example 1-B is designated as R).

[0084] Example 1-A: (S)-1′-chloro-8-difluoromethoxy-8′-fluoro-6-trifluoromethyl-3H,6′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinoline: SFC (RT = 4.37 min, ee = 99.7%). Analysis: A DAICEL CHIRALPAK AD-H column (0.46 cm × 25 cm) was used, with a mobile phase consisting of isopropanol and n-hexane (30%:70% by volume) at a flow rate of 1 mL / min. ESI-MS m / z: 422.34 [M+H] + ; 1 H NMR(500MHz,DMSO-d6)δ8.40(d,1H),7.94-7.91(m,1H),7.42(t,1H),7.34(dd,1H),7 .10(d,1H),5.91(ddd,1H),4.44(d,1H),4.00(d,1H),2.67(ddd,1H),2.57(ddd,1H).

[0085] Example 1-B: (R)-1′-chloro-8-difluoromethoxy-8′-fluoro-6-trifluoromethyl-3H,6′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinoline: SFC (RT=4.90 min, ee=99.3%). Analytical method: Same as above. ESI-MS m / z: 422.34 [M+H] + ; 1 H NMR(500MHz,DMSO-d6)δ8.40(d,1H),7.94-7.91(m,1H),7.42(t,1H),7.34(dd,1H),7 .10(d,1H),5.91(ddd,1H),4.44(d,1H),4.01(d,1H),2.67(ddd,1H),2.57(ddd,1H).

[0086] Example 2: Synthesis of 1′-chloro-8-difluoromethoxy-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline

[0087] Step 1: Synthesis of 1′-chloro-3-(3-(difluoromethoxy)-5-(trifluoromethyl)pyridin-2-yl)-6′,7′-dihydrodispiro[oxazoline-4,5′-isoquinolin-8′,2″-[1,3]dioxolane]-2-one

[0088] 1′-Chloro-6′,7′-dihydrodispiro[oxazoline-4,5′-isoquinoline-8′,2″-[1,3]dioxolane]-2-one (500 mg) was dissolved in DMF (10 mL), and CuBr (240 mg), Cs2CO3 (1.1 g), and 2-chloro-3-difluoromethoxy-5-trifluoromethylpyridine (840 mg) were added. The atmosphere was purged with nitrogen and the reaction was stirred at 130°C. After the reaction was complete, the mixture was filtered, extracted with ethyl acetate, dried over anhydrous sodium sulfate, spin-dried, and purified by column chromatography to obtain the title compound (536 mg). ESI-MS m / z: 508.23 [M+H] + .

[0089] Step 2: Synthesis of (1-chloro-5-((3-(difluoromethoxy)-5-(trifluoromethyl)pyridin-2-yl)amino)-6,7-dihydro-5H-spiro[isoquinolin-8-2′-[1,3]dioxolane]-5-yl)methanol

[0090] 1′-Chloro-3-(3-(difluoromethoxy)-5-(trifluoromethyl)pyridin-2-yl)-6′,7′-dihydrodispiro[oxazoline-4,5′-isoquinolin-8′,2″-[1,3]dioxolane]-2-one (150 mg) was dissolved in MeOH (3 mL). 4M NaOH (0.7 mL) was added dropwise at 0°C and stirred at room temperature for 2 h. The mixture was extracted with ethyl acetate and dried by spin drying. The title compound (108 mg) was obtained by preparative plate separation. ESI-MS m / z: 482.18 [M+H] +

[0091] Step 3: Synthesis of (1-chloro-5-((3-(difluoromethoxy)-5-(trifluoromethyl)pyridin-2-yl)amino)-5-(hydroxymethyl)-6,7-dihydroisoquinolin-8(5H)-one

[0092] Dissolve (1-chloro-5-((3-(difluoromethoxy)-5-(trifluoromethyl)pyridin-2-yl)amino)-6,7-dihydro-5H-spiro[isoquinolin-8-2′-[1,3]dioxolane]-5-yl)methanol (107 mg) in THF (1 mL), add HCl (0.4 mL), and stir at 50°C for 1 hour. Extract with ethyl acetate and dry to obtain the title compound (105 mg). ESI-MS m / z: 438.21 [M+H] + .

[0093] Step 4: Synthesis of 1′-chloro-8-(difluoromethoxy)-6-(trifluoromethyl)-6′,7′-dihydro-3H,8′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinolin]-8′-one

[0094] (1-Chloro-5-((3-(difluoromethoxy)-5-(trifluoromethyl)pyridin-2-yl)amino)-5-(hydroxymethyl)-6,7-dihydroisoquinolin-8(5H)-one (105 mg) was dissolved in toluene (5 mL), and MsCl (186 μL) and Et3N (675 μL) were added, and the mixture was stirred at 80°C. The mixture was extracted with ethyl acetate, dried by spin drying, and separated on a preparative plate to obtain the title compound (89 mg). ESI-MS m / z: 420.27 [M+H] + .

[0095] Step 5: Synthesis of 1′-chloro-8-(difluoromethoxy)-6-(trifluoromethyl)-7′,8′-dihydro-3H,6′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinolin]-8′-ol

[0096] 1′-Chloro-8-(difluoromethoxy)-6-(trifluoromethyl)-6′,7′-dihydro-3H,8′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinolin]-8′-one (89 mg) was dissolved in MeOH (20 mL). NaBH4 (8 mg, 1 eq.) was added at 0°C and stirred at 0°C for 30 min. The mixture was extracted with ethyl acetate, dried by spin drying, and separated on a preparative plate to obtain the title compound (78 mg). ESI-MS m / z: 422.32 [M+H] + .

[0097] Step 6: Synthesis of 1′-chloro-8-difluoromethoxy-6-trifluoromethyl-3H,6′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinoline

[0098] 1′-Chloro-8-(difluoromethoxy)-6-(trifluoromethyl)-7′,8′-dihydro-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinolin]-8′-ol (45 mg) was dissolved in toluene (3 mL), and Burgess reagent (80 mg) was added. The mixture was stirred at 90°C for 30 min. The mixture was extracted with ethyl acetate, dried by rotary evaporation, and the title compound (12 mg) was obtained by preparative plate separation. ESI-MS m / z: 448.32 [M+HCO2 - ] - . 1 H NMR(500MHz,DMSO-d6)δ8.28(d,1H),7.91(s,1H),7.43(t,1H),7.30(d,1H),7 .09(d,1H),6.81(d,1H),6.35(dt,1H),4.36(d,1H),3.93(d,1H),2.62(d,2H).

[0099] Example 2-A and Example 2-B: Preparation of (R)-1′-chloro-8-difluoromethoxy-6-trifluoromethyl-3H, 6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline and (R)-1′-chloro-8-difluoromethoxy-6-trifluoromethyl-3H, 6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline

[0100] Racemic 1′-chloro-8-difluoromethoxy-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline (i.e., the compound of Example 2) was subjected to preparative SFC to obtain a pair of enantiomers; the enantiomer eluted first from the chiral column below was Example 2-A; the enantiomer eluted later was Example 2-B (note that the absolute configurations of the two compounds of Examples 2-A and 2-B have not yet been determined. For the convenience of description only, the chiral center of Example 2-A is arbitrarily designated as R, and the chiral center of Example 2-B is designated as S).

[0101] Example 2-A: (R)-1′-chloro-8-difluoromethoxy-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline: SFC (RT = 5.10 min, ee = 99.6%). Analysis: A DAICEL CHIRALPAK IE-3 column (0.46 cm × 25 cm) was used, with a mobile phase consisting of isopropanol and n-hexane (30%:70% by volume) at a flow rate of 1 mL / min. ESI-MS m / z: 404.5 [M+H] + ; 1H NMR(500MHz,DMSO-d6)δ8.27(d,1H),7.90(s,1H),7.42(t,1H),7.29(dd,1H),7.08(s ,1H),6.82-6.78(dq,1H),6.34(dt,1H),4.35(d,1H),3.92(d,1H),2.63-2.60(m,2H).

[0102] Example 2-B: (S)-1′-chloro-8-difluoromethoxy-6-trifluoromethyl-3H,6′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinoline: SFC (RT = 5.73 min, ee = 98.2%). Analysis: A DAICEL CHIRALPAK IE-3 column (0.46 cm × 25 cm) was used, with a mobile phase consisting of isopropanol and n-hexane (30%:70% by volume) at a flow rate of 1 mL / min. ESI-MS m / z: 404.5 [M+H] + ; 1 H NMR(500MHz,DMSO-d6)δ8.27(d,1H),7.91(s,1H),7.42(t,1H),7.29(dd,1H),7.08(s ,1H),6.82-6.78(dq,1H),6.34(dt,1H),4.35(d,1H),3.92(d,1H),2.63-2.59(m,2H).

[0103] Example 3: Synthesis of 1′-chloro-8′-fluoro-7-methoxy-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline

[0104] Step 1: Synthesis of 1′-chloro-3-(4-methoxy)-5-(trifluoromethyl)pyridin-2-yl)-6′,7′-dihydrodispiro[oxazoline-4,5′-isoquinolin-8′,2″-[1,3]dioxolane]-2-one

[0105] 1′-Chloro-6′,7′-dihydrodispiro[oxazoline-4,5′-isoquinolin-8′,2″-[1,3]dioxolane]-2-one (1.2 g) was dissolved in toluene (15 mL), and 2-chloro-4-methoxy-5-trifluoromethylpyridine (1.0 g), Pd2(dba)3 (0.296 g), BINAP (0.403 g), and Cs2CO3 (1.97 g) were added. The atmosphere was purged with nitrogen and the reaction was carried out at 100°C. Water was added, the mixture was extracted with ethyl acetate, and the mixture was dried over anhydrous sodium sulfate. The title compound (1.14 g) was obtained by column chromatography. ESI-MS m / z: 472.32 [M+H] +

[0106] Step 2: Synthesis of 1-chloro-3′-(4-methoxy-5-(trifluoromethyl)pyridin-2-yl)-6,7-dihydro-8H-spiro[isoquinoline-5,4′-oxazoline]-2′,8-dione

[0107] Dissolve 1′-chloro-3-(4-methoxy)-5-(trifluoromethyl)pyridin-2-yl)-6′,7′-dihydrodispiro[oxazoline-4,5′-isoquinolin-8′,2″-[1,3]dioxolane]-2-one (1.14 g, 1.0 eq.) in THF (10 mL), add 4N HCl (4 mL), react at 50°C for 3 h, dilute with water, extract with ethyl acetate, dry over anhydrous sodium sulfate, and concentrate to obtain the title compound (1.14 g). ESI-MS m / z: 428.39 [M+H] + .

[0108] Step 3: Synthesis of 1-chloro-8-fluoro-3′-(4-methoxy-5-(trifluoromethyl)pyridin-2-yl)-6H-spiro[isoquinolin-5,4′-oxazoline]-2′-one

[0109] 1-Chloro-3′-(4-methoxy-5-(trifluoromethyl)pyridin-2-yl)-6,7-dihydro-8H-spiro[isoquinoline-5,4′-oxazoline]-2′,8-dione (1.1 g) was dissolved in DCM (10 mL), and DAST (4.1 g) was added. The mixture was reacted at 80°C. The reaction solution was added dropwise to saturated sodium bicarbonate, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain the title compound (100 mg). ESI-MS m / z: 428.50 [MH] -

[0110] Step 4: Synthesis of (1-chloro-8-fluoro-5-((4-methoxy-5-(trifluoromethyl)pyridin-2-yl)amino)-5,6-dihydroisoquinolin-5-yl)methanol

[0111] Dissolve 1-chloro-8-fluoro-3′-(4-methoxy-5-(trifluoromethyl)pyridin-2-yl)-6H-spiro[isoquinolin-5,4′-oxazoline]-2′-one (80 mg) in EtOH (5 mL), add 4N NaOH (90 μL), react at room temperature for 1 h, extract with ethyl acetate, dry over anhydrous sodium sulfate, and concentrate to obtain the title compound (90 mg). ESI-MS m / z: 403.32 [M+H] +

[0112] Step 5: Synthesis of 1′-chloro-8′-fluoro-7-methoxy-6-trifluoromethyl-3H,6′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinoline

[0113] (1-Chloro-8-fluoro-5-((4-methoxy-5-(trifluoromethyl)pyridin-2-yl)amino)-5,6-dihydroisoquinolin-5-yl)methanol (90 mg) was dissolved in toluene (3 mL), and MsCl (0.172 mL) and Et3N (0.62 mL) were added. The mixture was reacted at 80°C for 2 h, extracted with ethyl acetate, and dried over anhydrous sodium sulfate. The title compound (28 mg) was obtained by preparative plate separation. ESI-MS m / z: 386.31 [M+H] + , 1 H NMR(500MHz, Acetonitrile-d3)δ8.38(d,J=4.8Hz,1H),8.22(s,1H),7.74(s,1H),7.34(dd,1H),6 .28(s,1H),5.82(ddd,1H),4.43(d,1H),3.97(d,1H),3.93(s,3H),2.80(ddd,2H),2.60(ddd,1H).

[0114] Example 4: Synthesis of 1′-chloro-8′-fluoro-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinolinol]-7-ol

[0115] 1′-Chloro-8′-fluoro-7-methoxy-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline (i.e., the compound of Example 3) (13 mg) was dissolved in DMA (3 mL), and LiCl (21 mg) was added. The mixture was reacted at 85°C, and dilute hydrochloric acid was added. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and separated on a preparative plate to obtain the title compound (10 mg). ESI-MS m / z: 371.18 [MH] - , 1 H NMR(500MHz,DMSO-d6)δ8.40(d,1H),8.22(s,1H),7.85(s,1H),7.32(dd,1H ),5.89(ddd,1H),5.32(s,1H),4.36(d,1H),4.15(d,1H),2.71-2.66(m,2H).

[0116] Example 5: Synthesis of 1′-chloro-8-difluoromethoxy-8′-methoxy-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline]

[0117] Dissolve 1′-chloro-8-(difluoromethoxy)-6-(trifluoromethyl)-6′,7′-dihydro-3H,8′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinolin]-8′-one (50 mg) in methanol (1 mL), add PPTS (6 mg) and (CH3O)3CH (126 mg), and react at 120°C for 14 hours. Cool to room temperature, evaporate the solvent, and purify on a preparative plate to obtain the title compound (11.3 mg). ESI-MS m / z: 478.31 [M-H+CH3CN] - , 1 H NMR(500MHz,DMSO-d6)8.33(d,1H),7.90(s,1H),7.45(t,1H),7.28(d,1H),7.09(s,1H) ,5.31(dd,1H),4.46(d,1H),3.88(d,1H),3.70(s,3H),2.57(dd,1H),2.48-2.43(m,1H).

[0118] Example 6: Synthesis of 1′-chloro-8-difluoromethoxy-8′-methyl-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline]

[0119] Step 1: Synthesis of 1′-chloro-8-difluoromethoxy-6-trifluoromethyl-3H,6′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinolinyl]-8′-yl trifluoromethanesulfonate

[0120] Dissolve 1′-chloro-8-(difluoromethoxy)-6-(trifluoromethyl)-6′,7′-dihydro-3H,8′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinolin]-8′-one (300 mg) in THF (6 mL) and cool to -78°C. Add KHMDS (0.858 mL) dropwise. After 0.5 h, dissolve PhNTf2 (306 mg) in 1 mL of THF and slowly add dropwise to the reaction mixture. Let react for 1 h. Add 2 mL of saturated NH4Cl, extract, and separate by column chromatography to obtain the title compound (195 mg).

[0121] Step 2: Synthesis of 1′-chloro-8-difluoromethoxy-8′-methyl-6-trifluoromethyl-3H,6′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinoline]

[0122] 1′-Chloro-8-difluoromethoxy-6-trifluoromethyl-3H,6′H-spiro[imidazo[1,2-a]pyridin-2,5′-isoquinolin]-8′-yl trifluoromethanesulfonate (20 mg), MeB(OH)2 (6.5 mg), K3PO4 (23 mg), Pd(dppf)Cl2 (5 mg), 1,4-dioxane (1 mL), and H2O (1 mL) were added to a reaction tube, the atmosphere was purged with nitrogen, and the reaction was carried out at 70°C for 1 h. Extraction was performed, the solvent was evaporated, and the title compound (10 mg) was purified on a preparative plate. ESI-MS m / z [M+H] + 418.32, 1 H NMR(500MHz,DMSO-d6)8.29(d,1H),7.90(s,1H),7.44(t,1H),7.30(s,1H),7. 10(s,1H),6.15(td,1H),4.40(s,1H),3.85(s,1H),2.46(s,2H),2.31(dd,3H).

[0123] Example 7: Synthesis of 1′,8′-dichloro-8-difluoromethoxy-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline]

[0124] 1′-Chloro-8-difluoromethoxy-6-trifluoromethyl-3H,6′H-spiro[imidazo[1,2-a]pyridin-2,5′-isoquinolin]-8′-yl trifluoromethanesulfonate (50 mg), LiCl (1.2 mg), [Cp*Ru(MeCN)3OTf] (0.6 mg), and NMP (1 mL) were added to a reaction tube, replaced with nitrogen, and reacted at 100°C for 3 h. Extraction, spin-drying of the solvent, and purification on a preparative plate gave the title compound (5.2 mg). ESI-MS m / z [M+H] + 438.19, 1 H NMR (500MHz, DMSO-d6)8.40(d,1H),7.94(s,1H),7.42(t,1H),7.36(d,1H),7.12(s,1H),6.59(t,1H),4.45(d,1H),4.02(d,1H),2.62(d,2H).

[0125] Example 8: Synthesis of 1′-chloro-8-difluoromethoxy-6,8′-bis(trifluoromethyl)-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline]

[0126] Step 1: Synthesis of 1′-chloro-8-difluoromethoxy-8′-iodo-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline]

[0127] 1′-Chloro-8-difluoromethoxy-6-trifluoromethyl-3H,6′H-spiro[imidazo[1,2-a]pyridin-2,5′-isoquinolin]-8′-yl trifluoromethanesulfonate (75 mg), NaI (105 mg), [Cp*Ru(MeCN)3OTf] (14.2 mg), and DMI (5 mL) were added to a reaction tube. The atmosphere was purged with nitrogen and the reaction was carried out at 100°C for 12 h. Extraction and evaporation of the solvent were performed, and the title compound (52.6 mg) was obtained by purification on a preparative plate.

[0128] Step 2: Synthesis of 1′-chloro-8-difluoromethoxy-6,8′-bis(trifluoromethyl)-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline]

[0129] 1′-Chloro-8-difluoromethoxy-8′-iodo-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline] (30.0 mg, 1.0 eq.), copper powder (25.4 mg), and diphenyl(trifluoromethyl)sulfonium trifluoromethanesulfonate (57.8 mg) were dissolved in DMF (2.0 mL), replaced with nitrogen, and reacted at 100°C for 2 h. Extraction, spin drying, and purification on a preparative plate afforded the title compound (0.9 mg). ESI-MS: [M+H] + 472.30, 1 H NMR (500 MHz, CD3CN)

[0130] 1 H NMR(500MHz, Acetonitrile-d3)8.38(d,1H),7.55(t,1H),7.39(d,1H),7.24 -7.20(m,1H),7.20(t,1H),6.99(d,1H),4.34(d,1H),3.90(d,1H),2.78-2.71(m,1H),2.70-2.63(m,1H).

[0131] Example 9: Synthesis of 1′,7′-dichloro-8-difluoromethoxy-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline]

[0132] Step 1: Synthesis of 1′,7′-dichloro-8-difluoromethoxy-6-trifluoromethyl-6′,7′-dihydro-3H,8′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinolin]-8′-one

[0133] 1′-Chloro-8-(difluoromethoxy)-6-(trifluoromethyl)-6′,7′-dihydro-3H,8′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinolin]-8′-one (127 mg), TsOH (52 mg), NCS (61 mg), and CH3CN (4 mL) were added to a reaction flask and reacted at 50°C for 12 h. The reaction mixture was spin-dried and purified on a preparative plate to obtain the title compound (36 mg).

[0134] Step 2: Synthesis of 1′,7′-dichloro-8-difluoromethoxy-6-trifluoromethyl-7′,8′-dihydro-3H,6′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinolin]-8′-ol

[0135] Dissolve 1′,7′-dichloro-8-difluoromethoxy-6-trifluoromethyl-6′,7′-dihydro-3H,8′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinolin]-8′-one (36 mg) in MeOH (2 mL). Add NaBH4 (3 mg) under ice-cooling and allow to react for 0.5 h. Add water, extract, and purify on a preparative plate to obtain the title compound (20 mg).

[0136] Step 3: Synthesis of 1′,7′-dichloro-8-difluoromethoxy-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline]

[0137] Dissolve 1′,7′-dichloro-8-difluoromethoxy-6-trifluoromethyl-7′,8′-dihydro-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinolin]-8′-ol (20 mg) in 1,4-dioxane (3 mL), add Burgess reagent (31 mg), and react at 100°C for 1 hour. Add water, extract, and purify on a preparative plate to obtain the title compound (0.73 mg). ESI-MS m / z: [M+H] + 438.18, 1 H NMR(500MHz,CD3CN)8.16(d,1H),7.46(t,1H),7.18(d,1H),7.06(t,1H),6.9 3(d,1H),6.87(d,1H),4.25(d,1H),3.82(d,1H),2.98(dd,1H),2.69(d,1H).

[0138] Example 10: Synthesis of 1′-chloro-8-difluoromethoxy-7′-fluoro-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline]

[0139] Step 1: Synthesis of 8′-((tert-butyldimethylsilyl)oxy)-1′-chloro-8-difluoromethoxy-7′-fluoro-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline]

[0140] Dissolve 1′-chloro-8-(difluoromethoxy)-6-(trifluoromethyl)-6′,7′-dihydro-3H,8′H-spiro[imidazo[1,2-a]pyridin-2,5′-isoquinolin]-8′-one (200 mg) and 2,6-lutidine (257 mg) in DCM (10 mL). Replace the atmosphere with nitrogen and add TBSOTf (380 mg) under ice-cooling. React at room temperature for 10 minutes and then at 40°C for 2 hours. Add water, extract the mixture, spin dry, and separate by column chromatography to obtain the title compound (221.6 mg).

[0141] Step 2: Synthesis of 1′-chloro-8-difluoromethoxy-7′-fluoro-6-trifluoromethyl-6′,7′-dihydro-3H,8′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinolin]-8′-one

[0142] Dissolve 8′-((tert-butyldimethylsilyl)oxy)-1′-chloro-8-difluoromethoxy-7′-fluoro-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline] (221.6 mg) in MeCN (10 mL), cool to 0°C, add Selectfluor (159.4 mg), and react at room temperature for 3 h. Add water, extract, and dry by spin drying. Separate by column chromatography to obtain the title compound (75.6 mg).

[0143] Step 3: Synthesis of 1′-chloro-8-difluoromethoxy-7′-fluoro-6-trifluoromethyl-3H,6′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinolinyl]-8′-yl trifluoromethanesulfonate

[0144] 1′-Chloro-8-difluoromethoxy-7′-fluoro-6-trifluoromethyl-6′,7′-dihydro-3H,8′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinolin]-8′-one (40.0 mg) was dissolved in THF (8 mL), cooled to -78°C, and KHMDS (0.11 mL) was added dropwise. The mixture was stirred at -78°C for 0.5 h. PhNTf2 (48.6 mg) was added, and the mixture was stirred at -78°C for 2 h. The mixture was quenched, dried by spin drying, and purified on a preparative plate to obtain the title compound (15.4 mg).

[0145] Step 4: Synthesis of 1′-chloro-8-difluoromethoxy-7′-fluoro-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline]

[0146] 1′-Chloro-8-difluoromethoxy-7′-fluoro-6-trifluoromethyl-3H,6′H-spiro[imidazo[1,2-a]pyridin-2,5′-isoquinolin]-8′-yl trifluoromethanesulfonate (15.4 mg), Pd(PPh3)4 (3.1 mg), and LiCl (9.3 mg) were dissolved in THF (2 mL). The atmosphere was replaced with nitrogen. Bu3SnH (11.9 mg) was added and reacted at room temperature for 3 h. The mixture was extracted and dried by spin drying. The title compound (2.0 mg) was obtained by purification on a preparative plate. ESI-MS m / z [M+H] + 422.26, 1 H NMR(500MHz,CD3CN)8.23(d,1H),7.59(p,1H),7.27(dd,J=5.0,0.7Hz,1H),7.18(t,1H ),7.00(d,1H),6.52(ddd,1H),4.41(d,1H),3.96(d,1H),3.04(dt,1H),2.73(dd,1H).

[0147] Example 10-A and Example 10-B: Preparation of (R)-1′-chloro-8-difluoromethoxy-7′-fluoro-6-trifluoromethyl-3H, 6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline] and (S)-1′-chloro-8-difluoromethoxy-7′-fluoro-6-trifluoromethyl-3H, 6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline]

[0148] The racemic 1′-chloro-8-difluoromethoxy-7′-fluoro-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline] (i.e., the compound of Example 10) was subjected to preparative SFC to obtain a pair of enantiomers; the enantiomer eluted first from the chiral column below was Example 10-A; the enantiomer eluted later was Example 10-B (note that the absolute configurations of the two compounds of Examples 10-A and 10-B have not yet been determined. For the convenience of description only, the chiral center of Example 10-A is arbitrarily designated as R, and that of Example 10-B is designated as S).

[0149] Example 10-A: (R)-1′-chloro-8-difluoromethoxy-7′-fluoro-6-trifluoromethyl-3H,6′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinoline]: SFC (RT = 2.81 min, ee = 99.8%). Analysis: A DAICEL CHIRALPAK IE column (0.46 cm × 25 cm) was used, with a mobile phase consisting of isopropanol and n-hexane (20%:80% by volume) at a flow rate of 1 mL / min. ESI-MS m / z: 422.4 [M+H] + ; 1 H NMR(500MHz,DMSO-d6)δ8.26(d,1H),7.96(s,1H),7.39(t,1H),7.31(dd,1H),7. 11(s,1H),6.46(dd,1H),4.44(d,1H),4.12(d,1H),3.00(dd,1H),2.89(dd,1H).

[0150] Example 10-B: (S)-1′-chloro-8-difluoromethoxy-7′-fluoro-6-trifluoromethyl-3H,6′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinoline]: SFC (RT = 3.68 min, ee = 99.9%). Analysis: A DAICEL CHIRALPAK IE column (0.46 cm × 25 cm) was used, with a mobile phase consisting of isopropanol and n-hexane (20%:80% by volume) at a flow rate of 1 mL / min. ESI-MS m / z: 422.4 [M+H] + ; 1 H NMR(500MHz,DMSO-d6)δ8.26(d,1H),7.96(s,1H),7.39(t,1H),7.31(dd,1H),7. 11(s,1H),6.46(dd,1H),4.44(d,1H),4.11(d,1H),3.00(dd,1H),2.89(dd,1H).

[0151] Example 11: Synthesis of 6′-bromo-8-difluoromethoxy-4′,5′-difluoro-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]

[0152] Step 1: Synthesis of N′-(6′-bromo-8-difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′,3′-dihydro-3H,4′H-spiro[imidazo[1,2-a]pyridine-2,1′-naphthalene]-4′-ylidene)-4-toluenesulfonylhydrazone

[0153] 6′-Bromo-8-difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′,3′-dihydro-3′H,4′H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]-4′-one (synthesis procedure, see Example 2, Steps 1-4) (200 mg), TsNHNH2 (110 mg), and TsOH (10 mg) were added to MeOH (5 mL) and reacted at 80°C for 12 h. The mixture was extracted and concentrated. Purification on a preparative plate afforded the title compound (140 mg).

[0154] Step 2: Synthesis of 6′-bromo-8-difluoromethoxy-4′,5′-difluoro-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]

[0155] N′-(6′-bromo-8-difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′,3′-dihydro-3H,4′H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]-4′-ylidene)-4-toluenesulfonylhydrazone (45 mg), NFSI (32.7 mg), K2CO3 (11.6 mg), and KF (27.6 mg) were added to dioxane (2 mL) and reacted at 80°C for 12 h. Water was added, the mixture was extracted, and the mixture was spin-dried. The title compound (3.1 mg) was obtained by preparative plate purification. ESI-MS m / z [M+H] + 483.20, 485.20; 1 H NMR(500MHz,DMSO-d6)7.90(d,1H),7.72(dd,1H),7.42(t,1H),7.08(m,2H) ,5.79(ddd,1H),4.40(d,1H),3.93(d,1H),2.67(dd,1H),2.58-2.52(m,1H).

[0156] Example 12: Synthesis of 6′-bromo-8-difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]

[0157] Step 1: Synthesis of 6′-bromo-8-difluoromethoxy-5′-fluoro-6-trifluoromethyl-3′,4′-dihydro-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]-4′-ol

[0158] 6′-Bromo-8-difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′,3′-dihydro-3′H,4′H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]-4′-one (60 mg) was dissolved in MeOH (2 mL), and NaBH4 (2 mg) was added. The mixture was reacted at room temperature for 0.5 h, and water was added. The mixture was extracted and dried by spin drying. The mixture was purified on a preparative plate to obtain 31 mg of the target compound.

[0159] Step 2: Synthesis of 6′-bromo-8-difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]

[0160] Dissolve 6′-bromo-8-difluoromethoxy-5′-fluoro-6-trifluoromethyl-3′,4′-dihydro-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]-4′-ol (31 mg) and Burgess reagent (46 mg) in Toluene (2 mL) and react at 90°C for 5 h. Add water, extract, and dry the mixture. Separate the mixture with preparative HPLC to obtain 5.6 mg of the title compound. ESI-MS m / z [M+H] + 465.07, 467.14; 1 H NMR(500MHz,DMSO-d6)7.89(t,1H),7.56(dd,1H),7.43(t,1H),7.05(s,1H ),6.75(d,1H),6.24(dt,1H),4.31(d,1H),3.84(d,1H),2.59-2.55(m,2H).

[0161] Example 13: Synthesis of 6′-bromo-8-difluoromethoxy-5′-fluoro-4′-iodo-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]

[0162] Step 1: Synthesis of 6′-bromo-8-difluoromethoxy-5′-fluoro-4′-hydrazinylidene-6-trifluoromethyl-3′,4′-dihydro-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]

[0163] Add 6′-bromo-8-difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′,3′-dihydro-3′H,4′H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]-4′-one (40 mg), hydrazine hydrate (2 mL) and EtOH (2 mL) to a reaction tube, react at 80°C for 1 h, add water, extract and concentrate to obtain the title compound (40 mg).

[0164] Step 2: Synthesis of 6′-bromo-8-difluoromethoxy-5′-fluoro-4′-iodo-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]

[0165] 6′-Bromo-8-difluoromethoxy-5′-fluoro-4′-hydrazinylidene-6-trifluoromethyl-3′,4′-dihydro-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene] (45 mg), I2 (25 mg), and Et3N (25 mg) were added to THF (2 mL) and reacted at room temperature for 0.5 h. Water was added, the mixture was extracted, and the mixture was dried by spin drying. Purification on a preparative plate afforded 2.5 mg of the title compound. ESI-MS m / z [M+H] + 591.02, 592.84; 1 HNMR (500MHz, DMSO-d6)7.91(q,1H),7.69(dd,1H),7.42(t,1H),7.08-7.04(m,3H),4.41(d,1H),3.88(d,1H),2.57-2.52(m,1H),2.41(dd,1H).

[0166] Example 14: Synthesis of 6′-bromo-4′-chloro-8-difluoromethoxy-5′-fluoro-4′-iodo-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]

[0167] Step 1: Synthesis of 6′-bromo-8-difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′H,3H-spiro[imidazo[1,2-a]pyridine-2,1′-naphthalene]-4′-yl trifluoromethanesulfonate

[0168] Dissolve 6′-bromo-8-difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′,3′-dihydro-3′H,4′H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]-4′-one (200 mg) in THF (5 mL) and cool to -78°C. Add KHMDS (0.5 mL) dropwise. After 0.5 h, add PhNTf2 (178 mg) and react for 1 h. Quench with saturated NH4Cl solution, extract, spin dry, and separate on a silica gel column to obtain the desired product (145 mg).

[0169] Step 2: Synthesis of 6′-bromo-4′-chloro-8-difluoromethoxy-5′-fluoro-4′-iodo-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]

[0170] 6′-Bromo-8-difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′H,3H-spiro[imidazo[1,2-a]pyridine-2,1′-naphthyl]-4′-yl trifluoromethanesulfonate (30 mg), LiCl (3.1 mg), and [Cp*Ru(MeCN)3OTf] (1.2 mg) were added to NMP (2 mL), replaced with nitrogen, and reacted at 100°C for 3 h. Water was added, the mixture was extracted, and the mixture was spin-dried. Purification on a preparative plate afforded the title compound (14.5 mg). ESI-MS m / z [M+H] + 499.14, 500.69; 1 H NMR (500MHz, DMSO-d6)7.90(t,1H),7.73(dd,1H),7.42(t,1H),7.10(d,1H),7.07(d,1H),6.43(dd,1H),4.40(d,1H),3.92(d,1H),2.66-2.55(m,2H).

[0171] Example 15: Synthesis of 8-difluoromethoxy-4′,5′-fluoro-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]

[0172] Step 1: Synthesis of N′-(8-difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′,3′-dihydro-3H,4′H-spiro[imidazo[1,2-a]pyridine-2,1′-naphthalene]-4′-ylidene)-4-toluenesulfonylhydrazone

[0173] 8-Difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′,3′-dihydro-3′H,4′H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]-4′-one (150 mg), TsNHNH2 (104 mg), and TsOH (2 mg) were added to MeOH (5 mL) and reacted at 80°C for 12 h. The mixture was extracted and concentrated. Purification on a preparative plate afforded the title compound (55 mg).

[0174] Step 2: Synthesis of 8-difluoromethoxy-4′,5′-difluoro-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]

[0175] N′-(8-Difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′,3′-dihydro-3H,4′H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]-4′-ylidene)-4-toluenesulfonylhydrazone (45 mg), NFSI (37 mg), K2CO3 (32.6 mg), and KF (13.7 mg) were added to dioxane (2 mL) and reacted at 80°C for 12 h. Water was added, the mixture was extracted, and the mixture was dried by spin drying. The title compound (3.1 mg) was obtained by preparative plate purification. ESI-MS m / z [M+H] + 405.00; 1 H NMR(500MHz,DMSO-d6)7.90(d,1H),7.72(dd,1H),7.42(t,1H),7.08(m,2H) ,5.79(ddd,1H),4.40(d,1H),3.93(d,1H),2.67(dd,1H),2.58-2.52(m,1H).

[0176] Example 15-A and Example 15-B: Preparation of (S)-8-difluoromethoxy-4′,5′-difluoro-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene] and (R)-8-difluoromethoxy-4′,5′-difluoro-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]

[0177] The racemic 8-difluoromethoxy-4′,5′-difluoro-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene] (i.e., the compound of Example 15) was subjected to preparative SFC to obtain a pair of enantiomers; the enantiomer eluted first from the chiral column below was Example 15-A; the enantiomer eluted later was Example 15-B (note that the absolute configurations of the two compounds of Examples 15-A and 15-B have not yet been determined. For the convenience of description only, the chiral center of Example 15-A was arbitrarily designated as S, and that of Example 15-B was designated as R).

[0178] Example 15-A: (S)-8-Difluoromethoxy-4′,5′-difluoro-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]: SFC (RT = 8.49 min, ee = 99.9%). Analysis: A DAICEL CHIRALPAK IG column (0.46 cm × 25 cm) was used, with a mobile phase consisting of ethanol and n-hexane (9%:95% by volume) and 0.1% diethylamine at a flow rate of 1 mL / min. ESI-MS m / z: 405.5 [M+H] + ;1 H NMR(500MHz,DMSO-d6)δ7.90(s,1H),7.43(t,1H),7.41(dt,1H),7.23 -7.17(m,1H),7.13(dd,1H),7.06(s,1H),5.69(ddd,1H),4.41(d,1H),3.90(d,1H),2.65(ddd,1H),2.56-2.52(m,1H).

[0179] Example 15-B: (R)-8-Difluoromethoxy-4′,5′-difluoro-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]: SFC (RT = 9.88 min, ee = 99.3%). Analysis: A DAICEL CHIRALPAK IG column (0.46 cm × 25 cm) was used, with a mobile phase consisting of ethanol and n-hexane (5%:95% by volume) and 0.1% diethylamine at a flow rate of 1 mL / min. ESI-MS m / z: 405.5 [M+H] + ; 1 H NMR(500MHz,DMSO-d6)δ7.89(s,1H),7.43(t,1H),7.42(dt,1H),7.19(dd,1H),7.13(dd,1H ),7.05(s,1H),5.69(ddd,1H),4.41(d,1H),3.90(d,1H),2.65(ddd,1H),2.56-2.52(m,1H).

[0180] Example 16: Synthesis of 8-difluoromethoxy-4′,5′-fluoro-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]

[0181] Step 1: Synthesis of 8-difluoromethoxy-5′-fluoro-6-trifluoromethyl-3′,4′-dihydro-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]-4′-ol

[0182] 8-Difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′,3′-dihydro-3′H,4′H-spiro[imidazole[1, 2-a]pyridine-2,1′-naphthalene]-4′-one (60 mg) was dissolved in MeOH (2 mL), and NaBH4 (6 mg) was added. The mixture was reacted at 0°C for 0.5 h, and water was added. The mixture was extracted and dried by spin drying. The mixture was purified on a preparative plate to obtain 21 mg of the target compound.

[0183] Step 2: Synthesis of 8-difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]

[0184] Dissolve 8-difluoromethoxy-5′-fluoro-6-trifluoromethyl-3′,4′-dihydro-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]-4′-ol (21 mg) and burgess reagent (37 mg) in Toluene (2 mL) and react at 90°C for 0.5 h. Add water, extract, and spin dry. Separate with preparative HPLC to obtain 4 mg of the title compound. ESI-MS m / z [M+H] + 387.31; 1 H NMR(500MHz,DMSO-d6)7.53-7.51(m,1H),7.42(t,1H),7.29-7.25(m,1H),7.15-7.11(m,1H),7. 04(ddd,1H),6.92(d,1H),6.82(dd,1H),4.31(d,1H),3.79(d,1H),2.65(dt,1H),2.53(dd,1H).

[0185] Example 16-A and Example 16-B: Preparation of (S)-8-difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′H, 3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene] and (R)-8-difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′H, 3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]

[0186] The racemic 8-difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene] (i.e., the compound of Example 16) was subjected to preparative SFC to obtain a pair of enantiomers; the enantiomer eluted first from the chiral column below was Example 16-A; the enantiomer eluted later was Example 16-B (note that the absolute configurations of the two compounds of Examples 16-A and 16-B have not yet been determined. For the convenience of description only, the chiral center of Example 16-A was arbitrarily designated as S, and that of Example 16-B was designated as R).

[0187] Example 16-A: (S)-8-Difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]: SFC (RT = 7.26 min, ee = 99.1%). Analytical method: A DAICEL CHIRALPAK IG column (0.46 cm × 25 cm) was used, with a mobile phase consisting of ethanol and n-hexane (5%:95% by volume) and 0.1% diethylamine at a flow rate of 1 mL / min. ESI-MS m / z: 387.5 [M+H] + ; 1 H NMR(500MHz,DMSO-d6)δ7.88(s,1H),7.45(t,1H),7.29 -7.25(m,1H),7.14-7.09(m,1H),7.08(d,1H),7.04(s,1H),6.75(dd,1H),6.16(ddd,1H),4.32(d,1H),3.82(d,1H),2.58-2.53(m,2H).

[0188] Example 16-B: (R)-8-Difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]: SFC (RT = 7.82 min, ee = 96.1%). Analytical method: A DAICEL CHIRALPAK IG column (0.46 cm × 25 cm) was used, with a mobile phase consisting of ethanol and n-hexane (5%:95% by volume) and 0.1% diethylamine at a flow rate of 1 mL / min. ESI-MS m / z: 387.5 [M+H] + ; 1 H NMR(500MHz,DMSO-d6)δ7.88(s,1H),7.45(t,1H),7.29 -7.25(m,1H),7.14-7.09(m,1H),7.08(d,1H),7.04(s,1H),6.75(dd,1H),6.16(ddd,1H),4.32(d,1H),3.82(d,1H),2.58-2.53(m,2H).

[0189] Example 17: Synthesis of 8-difluoromethoxy-1′-methyl-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline]

[0190] Step 1: Synthesis of 8-difluoromethoxy-1′-methyl-6-trifluoromethyl-6′,7′-dihydro-3H,8′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinolin]-8′-one

[0191] Dissolve 1′-chloro-8-difluoromethoxy-6-trifluoromethyl-6′,7′-dihydro-3H,8′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinolin]-8′-one (100 mg) in dioxane (5 mL). Add H2O (1 mL), Pd(dppf)Cl2 (35.1 mg), and Cs2CO3 (156.4 mg). Replace the atmosphere with nitrogen. Add trimethylboroxine (0.7 mL) and react at 100°C for 13 h. Add water, extract, spin dry, and purify on a preparative plate to obtain 60 mg of the title compound.

[0192] Step 2: 8-Difluoromethoxy-1′-methyl-6-trifluoromethyl-7′,8′-dihydro-3H,6′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinolin]-8′-ol

[0193] Dissolve 8-difluoromethoxy-1′-methyl-6-trifluoromethyl-6′,7′-dihydro-3H,8′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinolin]-8′-one (38 mg) in MeOH (3 mL), cool to 0°C, slowly add NaBH4 (3.6 mg), and stir for 30 minutes. Add water, extract, spin dry, and purify on a preparative plate to obtain 12 mg of the title compound.

[0194] Step 3: Synthesis of 8-difluoromethoxy-1′-methyl-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline]

[0195] 8-Difluoromethoxy-1′-methyl-6-trifluoromethyl-7′,8′-dihydro-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinolin]-8′-ol (12 mg) was dissolved in PhMe (2 mL), and Burgess reagent (21.5 mg) was added. The atmosphere was purged with nitrogen and the reaction was carried out at 100°C for 1 h. Water was added, the mixture was extracted, and the mixture was dried by spin drying. The mixture was then purified on a preparative plate to obtain the target compound Example 17 (5 mg). ESI-MS m / z: 384.39 [M+H] + ; 1 H NMR(500MHz, Acetonitrile-d3)δ8.20(d,1H),7.41-7.39(m,1H),7.13(t,1H),6.98(s,1H),6.81(d,1H ),6.69(dd,1H),6.09-6.04(m,1H),4.20(d,1H),3.66(d,1H),2.52(dt,1H),2.44(s,3H),2.42(dd,1H).

[0196] Example 18: Synthesis of 8-difluoromethoxy-5′-fluoro-6′-methyl-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]

[0197] Step 1: Synthesis of 8-difluoromethoxy-5′-fluoro-6′-methyl-6-trifluoromethyl-2′,3′-dihydro-3H,4′H-spiro[imidazo[1,2-a]pyridine-2,1′-naphthalene]-4′-one

[0198] 6′-Bromo-8-difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′,3′-dihydro-3H,4′H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]-4′-one (80 mg), Pd(dppf)Cl2 (24.3 mg), and Cs2CO3 (108.2 mg) were dissolved in dioxane (3 mL) and H2O (0.6 mL). The atmosphere was replaced with nitrogen, and trimethylboroxine (0.47 mL) was added. The mixture was reacted at 100°C for 1 h. Water was added, the mixture was extracted, and the mixture was spin-dried to dryness. Purification on a preparative plate afforded 59 mg of the title compound.

[0199] Step 2: Synthesis of N′-(8-difluoromethoxy-5′-fluoro-6′-methyl-6-trifluoromethyl-2′,3′-dihydro-3H,4′H-spiro[imidazo[1,2-a]pyridine-2,1′-naphthalene]-4′-ylidene)-4-methylbenzenesulfonylhydrazone

[0200] 8-Difluoromethoxy-5′-fluoro-6′-methyl-6-trifluoromethyl-2′,3′-dihydro-3H,4′H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]-4′-one (59.4 mg), TsNHNH2 (40 mg), and TsOH (29.6 mg) were added to THF (5 mL), replaced with nitrogen, and reacted at 40°C for 2 h. Water was added, the mixture was extracted, and the mixture was spin-dried and purified on a preparative plate to obtain 97 mg of the title compound.

[0201] Step 3: Synthesis of 8-difluoromethoxy-5′-fluoro-6′-methyl-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]

[0202] N′-(8-difluoromethoxy-5′-fluoro-6′-methyl-6-trifluoromethyl-2′,3′-dihydro-3H,4′H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]-4′-ylidene)-4-methylbenzenesulfonylhydrazone (40 mg) and Cs2CO3 (66.8 mg) were added to dioxane (3 mL) and stirred at 90°C for 5 h. Water was added, the mixture was extracted, and the mixture was spin-dried and purified on a preparative plate to obtain the target compound Example 18 (13 mg). ESI-MS m / z: 401.13 [M+H] + ; 1 H NMR(500MHz,DMSO-d6)δ7.87(s,1H),7.44(t,1H),7.14(t,1H),7.02(s,1H),6.95(d,1H),6.74(d,1 H),6.16-6.10(m,1H),4.28(d,1H),3.79(d,1H),2.57-2.52(m,1H),2.49-2.46(m,1H),2.21(s,3H).

[0203] Example 19: Synthesis of 8-difluoromethoxy-5′-fluoro-4′-iodo-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]

[0204] Step 1: Synthesis of 8-difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′H,3H-spiro[imidazo[1,2-a]pyridine-2,1′-naphthalene]-4′-yl trifluoromethanesulfonate

[0205] 8-Difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′,3′-dihydro-3H,4′H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]-4′-one (500 mg) was dissolved in THF (4 mL), the atmosphere was replaced with nitrogen, and the temperature was cooled to -78°C. KHMDS (1.5 mL) was then added dropwise to the reaction solution and allowed to react for 1 hour. PhNTf2 was dissolved in 2 mL of THF and slowly added dropwise to the reaction solution for 30 minutes. The reaction was quenched by adding saturated NH4Cl solution, extracted, dried, and purified on a silica gel column to obtain 306 mg of the target compound.

[0206] Step 2: Synthesis of 8-difluoromethoxy-5′-fluoro-4′-iodo-6-trifluoromethyl-2′H,3H-spiro[imidazole[1,2-a]pyridine-2,1′-naphthalene]

[0207] 8-Difluoromethoxy-5′-fluoro-6-trifluoromethyl-2′H,3H-spiro[imidazo[1,2-a]pyridine-2,1′-naphthyl]-4′-yl trifluoromethanesulfonate (135 mg), NaI (120 mg), and [Cp*Ru(MeCN)3]OTf (15 mg) were dissolved in DMI (2 mL). After nitrogen replacement, the temperature was raised to 100°C and the reaction was allowed to proceed for 16 hours. Water was added, the mixture was extracted, and the mixture was dried by spin drying. Purification on a silica gel column afforded 117 mg of the title compound. ESI-MS m / z: 513.20 [M+H] + ; 1 H NMR(500MHz,DMSO-d6)δ7.90(s,1H),7.44(t,1H),7.39(td,1H),7.17(ddd,1H),7.10(dd ,1H),7.06(s,1H),6.99(dd,1H),4.42(d,1H),3.87(d,1H),2.54(dd,1H),2.40(dd,1H).

[0208] Example 20: Synthesis of 8-difluoromethoxy-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline]

[0209] Step 1: Synthesis of 8-difluoromethoxy-6-trifluoromethyl-6′,7′-dihydro-3H,8′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinolin]-8′-one

[0210] Dissolve 1′-chloro-8-difluoromethoxy-6-trifluoromethyl-6′,7′-dihydro-3H,8′H-spiro[imidazole[1,2-a]pyridin-2,5′-isoquinolin]-8′-one (100 mg) in N,N-dimethylacetamide (3 mL). Add Pd(PPh3)4 (27.7 mg), HCOOH (16.6 mg), and TEA (38.5 mg). After nitrogen substitution, heat to 100°C and react for 5 hours. Add water, extract, spin dry, and purify on a silica gel column to obtain 90 mg of the title compound.

[0211] Step 2: Synthesis of N′-(8-difluoromethoxy-6-trifluoromethyl-6′,7′-dihydro-3H,8′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinolinyl]-8′-ylidene)-4-methylbenzenesulfonylhydrazone

[0212] 8-Difluoromethoxy-6-trifluoromethyl-6′,7′-dihydro-3H,8′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinolin]-8′-one (40 mg), TsNHNH2 (29 mg), and TsOH (2 mg) were added to THF (2 mL), replaced with nitrogen, and reacted at 30°C for 4 h. Water was added, the mixture was extracted, and the mixture was spin-dried and purified on a preparative plate to obtain 48 mg of the title compound.

[0213] Step 3: Synthesis of 8-difluoromethoxy-6-trifluoromethyl-3H,6′H-spiro[imidazole[1,2-a]pyridine-2,5′-isoquinoline]

[0214] N′-(8-Difluoromethoxy-6-trifluoromethyl-6′,7′-dihydro-3H,8′H-spiro[imidazo[1,2-a]pyridine-2,5′-isoquinolinyl]-8′-ylidene)-4-methylbenzenesulfonylhydrazone (38 mg) was dissolved in dioxane (3 mL), and K2CO3 (32 mg) was added. The atmosphere was replaced with nitrogen and the reaction was carried out at 90°C for 16 h. Water was added, the mixture was extracted, and the mixture was dried by spin drying. The title compound, Example 20 (9.7 mg), was obtained. ESI-MS m / z: 369.92 [M+H] + ; 1 H NMR(500MHz, Acetonitrile-d3)δ8.44(d,J=5.0Hz,1H),8.37(s,1H),7.54-7.51(m,1H),7.24(t,1H),7.24 (d,1H),6.94(d,1H),6.66(dd,1H),6.17(ddd,1H),4.31(d,1H),3.83(d,1H),2.67(dt,1H),2.57(dd,1H).

[0215] Biological testing

[0216] Hep3B cells were used to detect the EPO inhibition rate of each compound

[0217] 5×10 4Hep3B cells (Cell Bank of the Chinese Academy of Sciences, TCHu106) were plated on a plate at 500 nm / mL and incubated overnight at 37°C in a 5% carbon dioxide incubator (Thermo Scientific BB150). The next day, the culture medium was replaced with a culture medium containing 0.5% FBS (VivaCell C04001-500) to starve the cells for 6 hours, and then a DMSO solution of (6'-hydroxy-3'-(4-methoxyphenyl)-8'-oxo-8'H-spiro[cyclopentane-1,5'-indolizine]-7'-carbonyl)glycine or its analogue was added to make the final concentration 30 μM. The plate was placed in a 37°C, 5% carbon dioxide incubator for 24 hours, and then a DMSO solution of the test compound was added to make the final concentration of the compound 30000nM, 7500nM, 1875nM, 469nM, 117nM, 29nM, 7.3nM, 1.8nM, 0.5nM, 0nM, a total of 10 concentrations, and the effect lasted for 48 hours. The cell supernatant was collected for EPO ELISA detection was performed according to the instructions of the ELISA kit (Shanghai Sangon Biotech Co., Ltd., D711311-0096), and the values ​​were read at a wavelength of 450 nm (microplate reader, Thermo Scientific (Multiskan Go), 511191200).

[0218] The formula for calculating compound inhibition rate is: compound inhibition rate = 1-100%*(EPO value of the dosing well - EPO value of the blank well) / (EPO value of the control well - EPO value of the blank well). The inhibition rate is linearly fitted to calculate IC 50 Control wells: wells containing cells treated with a DMSO solution containing 6'-hydroxy-3'-(4-methoxyphenyl)-8'-oxo-8'H-spiro[cyclopentane-1,5'-indolizine]-7'-carbonyl)glycine or its analogs at a final concentration of 30 μM at a test compound concentration of 0 nM for 72 hours. The EPO content in this well is defined as 100%. Blank wells: wells containing cells without any compound added.

[0219] IC values ​​of positive control compounds and certain example compounds 50 The data are provided in Table 1 (due to instrument errors and different operating habits of operators, the IC values ​​of compounds measured under different experimental conditions are different. 50 There may be some differences, so the values ​​listed in the table should be understood as expressing relative sizes within a certain error range).

[0220] As shown in Table 1, compared with the positive control compound, the tested compounds in each example showed an IC 50 Equivalent or lower IC 50; This indicates that the tested compounds in each example have excellent EPO-lowering activity and thus have the potential to be used in the treatment of EPO-related diseases.

[0221] Table 1

[0222] While specific embodiments of the present invention have been illustrated and described, it is not intended that these embodiments illustrate and describe all possible forms of the present invention. Rather, the words used in this specification are merely descriptive and are not restrictive. It will be apparent to those skilled in the art that various other changes and modifications may be made without departing from the general scope of this disclosure. Therefore, the appended claims are intended to include all such changes and modifications within the scope of the present invention.

Claims

1. A compound represented by formula (I), an isotope-labeled compound thereof, an optical isomer thereof, a tautomer thereof, or a pharmaceutically acceptable salt thereof, or a crystalline form or a solvate form of the compound or a pharmaceutically acceptable salt thereof, in, R 1 Selected from H, D, F, Cl, Br, CF3, CN; R 2 Selected from H, D, CF3, CH3, OCH3, OH; R 3 Selected from H, D, F, Cl, Br, CF3, CN, OCH3, OCH2F, OCHF2, OCF3; R 4 Selected from H, D, F, Cl, Br, CH3, CF3, CN; R 5 Selected from H, D, F, Cl, Br, I, CH3, CF3, CN, OCH3; R 6 Selected from H, D, F, Cl, Br, I, CH3, CF3, CN, OCH3; X is selected from N,CR 7 ; Among them, R 7 Selected from H, D, F, Cl, Br, CF3, CN, CH3.

2. The compound according to claim 1, its isotope-labeled compound, its optical isomer, its tautomer, or its pharmaceutically acceptable salt, or a crystalline form or solvate form of the compound or its pharmaceutically acceptable salt, wherein R 1 Selected from CF3, CN.

3. The compound according to claim 1 or 2, its isotope-labeled compound, its optical isomer, its tautomer, or its pharmaceutically acceptable salt, or a crystalline form or solvate form of the compound or its pharmaceutically acceptable salt, wherein R 2 Selected from OCH3, OH, H.

4. A compound according to any one of claims 1 to 3, an isotope-labeled compound thereof, an optical isomer thereof, a tautomer thereof, or a pharmaceutically acceptable salt thereof, or a crystalline form or a solvate form of the compound or a pharmaceutically acceptable salt thereof, wherein R 3 Selected from OCH3, OCH2F, OCHF2, OCF3, H.

5. A compound according to any one of claims 1 to 4, an isotope-labeled compound thereof, an optical isomer thereof, a tautomer thereof, or a pharmaceutically acceptable salt thereof, or a crystalline form or a solvate form of the compound or a pharmaceutically acceptable salt thereof, wherein R 4 Selected from H, F, Cl, CH3.

6. A compound according to any one of claims 1 to 5, an isotope-labeled compound thereof, an optical isomer thereof, a tautomer thereof, or a pharmaceutically acceptable salt thereof, or a crystalline form or a solvate form of the compound or a pharmaceutically acceptable salt thereof, wherein R 5 Selected from H, F, Cl, I, CH3, CF3, OCH3.

7. A compound according to any one of claims 1 to 6, an isotope-labeled compound thereof, an optical isomer thereof, a tautomer thereof, or a pharmaceutically acceptable salt thereof, or a crystalline form or a solvate form of the compound or a pharmaceutically acceptable salt thereof, wherein R 6 Selected from F, Cl, CH3, H.

8. The compound according to any one of claims 1 to 8, its isotope-labeled compound, its optical isomer, its tautomer, or its pharmaceutically acceptable salt, or a crystalline form or solvate form of the compound or its pharmaceutically acceptable salt, wherein X is selected from N, CR 7 ,in, R 7 Selected from H, F, Br, CF3, CN, CH3.

9. The compound according to any one of claims 1 to 8, its isotope-labeled compound, its optical isomer, its tautomer, or its pharmaceutically acceptable salt, or the crystalline form or solvate form of the compound or its pharmaceutically acceptable salt, wherein: The R 1 is CF3; said R 2 is selected from OCH3, OH, H; said R 3 is OCH2F; said R 4 is selected from H, F, Cl, CH3; said R 5 is selected from H, F, Cl, I, CH3, CF3, OCH3; said R 6 is selected from F, Cl, CH3; X is selected from N, CR 7 , where R 7 Selected from H, F, Br.

10. The compound according to any one of claims 1 to 8, its isotope-labeled compound, its optical isomer, its tautomer, or its pharmaceutically acceptable salt, or the crystalline form or solvate form of the compound or its pharmaceutically acceptable salt, wherein: The R 1 is CF3; said R 2 is selected from OCH3, OH, H; said R 3 is OCHF2 or H; said R 4 is selected from H, F, Cl, CH3; said R 5 is selected from H, F, Cl, Br, I, CH3, CF3, OCH3; said R 6 is selected from H, F, Cl, CH3; X is selected from N, CR 7 , where R 7 Selected from H, F, Br, CH3.

11. The compound according to claim 1, its isotope-labeled compound, its optical isomer, its tautomer, or its pharmaceutically acceptable salt, or a crystalline form or solvate form of the compound or its pharmaceutically acceptable salt, wherein: The compound is selected from:

12. The compound according to claim 1, its isotope-labeled compound, its optical isomer, its tautomer, or its pharmaceutically acceptable salt, or a crystalline form or solvate form of the compound or its pharmaceutically acceptable salt, wherein: The compound is selected from:

13. A pharmaceutical composition comprising a compound according to any one of claims 1 to 12, an isotope-labeled compound thereof, an optical isomer thereof, a tautomer thereof, or a pharmaceutically acceptable salt thereof, or a crystalline form or a solvate form of the compound or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, adjuvants or excipients.

14. Use of a compound according to any one of claims 1 to 12, an isotope-labeled compound thereof, an optical isomer thereof, a tautomer thereof, or a pharmaceutically acceptable salt thereof, or a crystalline form or a solvate form of the compound or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 13 in the preparation of a medicament for treating a disease or condition associated with EPO.

15. The use according to claim 14, wherein the diseases and disorders associated with EPO are selected from the group consisting of polycythemia vera, renal cell carcinoma and other cancers, including but not limited to bladder cancer, breast cancer, cervical cancer, colorectal cancer, small intestine cancer, colon cancer, rectal cancer, anal cancer, endometrial cancer, head and neck cancer, liver cancer, lung cancer, ovarian cancer, prostate cancer, testicular cancer, uterine cancer, esophageal cancer, gallbladder cancer, pancreatic cancer, thyroid cancer, parathyroid cancer, skin cancer and brain cancer.

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