Quinolone analogs and their uses

Abstract

The present application provides a quinolone analogue represented by formula (I), or a pharmaceutically acceptable salt, stereoisomer, racemate, deuterated compound, or solvent compound thereof, and a pharmaceutical composition thereof, as well as their use in preparing a drug for treating tumors, autoimmune diseases, or diseases caused by bacteria, fungi, or viruses. #imgabs0#

Description

[0001] Citation of Related Applications

[0002] This application claims priority and benefits of Chinese Patent Application No. 202310438329.5 filed with the State Intellectual Property Office of China on April 23, 2023, and the contents disclosed in the application are incorporated herein by reference in their entirety. Technical Field

[0003] The present application relates to a class of quinolone analogs and preparation methods and uses thereof, and belongs to the field of medicinal chemistry. Background Art

[0004] Tetracyclic quinolone compounds interact with the quadruplex-forming regions of nucleic acids and regulate ribosomal RNA transcription. Specifically, tetracyclic quinolone compounds can stabilize DNA G-quadruplexes (G4s) in cancer cells, thereby inducing synthetic lethality in cancer cells. In addition, tetracyclic quinolone compounds selectively inhibit rRNA synthesis by Pol I in the nucleolus.

[0005] DNA usually exists in double-stranded form, but in some regions where guanine is repeated in tandem, guanine is connected by Hoogsteen hydrogen bonds, 4 Gs form a ring-shaped plane, and two or more layers of tetrads form a special higher-order structure - a four-stranded structure through π-π stacking. Quadruplex DNA is enriched at the ends of chromosomes and the promoter regions of genes. Quadruplex DNA at the ends of chromosomes has the effect of inhibiting telomerase, so stabilizing the quadruplex DNA at the ends of chromosomes can inhibit cancer growth. The promoter regions of many oncogenes, such as c-Myc, KRAS, and c-Kit, are rich in quadruplex DNA structures. There is evidence that stabilizing the quadruplex DNA in these regions can reduce oncogene expression, thereby inhibiting cancer growth.

[0006] Quadruplex DNA may usually be a temporary structure, which often occurs when double-stranded DNA is unwound during DNA replication and translation. There are also many unwinding enzymes in the cell that can unwind the higher-order structure of quadruplex DNA. However, when quadruplex DNA is combined with stabilizer drugs, making it difficult for unwinding enzymes to unwind the quadruplex structure, this stable structure will inevitably hinder DNA replication and translation, and even cause DNA breakage. The homologous recombination DNA repair pathway (HR) mediated by BRCA1 and BRCA2, and the non-homologous end joining repair pathway (NHEJ) are important pathways for repairing broken DNA, and may also be responsible for bypassing the quadruplex structure during DNA replication and translation to ensure the smooth progress of DNA replication and translation. Many literatures have reported that BRCA1 / 2-deficient cancers are highly sensitive to quadruplex DNA stabilizers. In addition, compared with normal cells, cancer cells have more quadruplex DNA structures. Quadruplex DNA stabilizers are expected to become clinical drugs for the treatment of BRCA1 / 2-deficient, HR-deficient and other DNA damage repair defective tumors.

[0007] CX-5461 was once considered an RNA polymerase I inhibitor that inhibits rDNA translation. There are also literatures showing that CX-5461 has the effect of inhibiting topoisomerase and stabilizing quadruplex DNA.

[0008] Although CX-5461 has shown preliminary efficacy in HR-deficient solid tumors, the dosage of CX-5461 in clinical trials is large. In the NCT02719977 clinical trial for the treatment of solid tumors in humans, the recommended dosage is 475 mg / m 2 , and observed side effects such as phototoxicity, neutropenia, extremity redness and swelling, and nausea. In addition, the administration method of CX-5461 in existing clinical trials is multiple intravenous drips, which need to be performed in hospitals, occupying a large amount of medical resources. It is not as convenient and economical as oral administration, and it also introduces the risks of infection, phlebitis, thrombosis, etc. related to intravenous drips. Considering the basic health status of cancer patients receiving CX-5461 treatment, it is very necessary to develop related oral drugs that do not occupy precious medical resources.

[0009] Currently, there is an urgent clinical need to enhance the tumor-killing activity of compounds and improve their metabolic stability, reduce their toxic side effects, and increase their bioavailability, so as to transform such compounds into oral preparations that are more convenient for patients to use. Summary of the invention

[0010] In one aspect, the present application provides a quinolone analogue as shown in the general formula (I) or a pharmaceutically acceptable salt, stereoisomer, racemate, deuterated compound, or solvent compound thereof:

[0011]

[0012] in:

[0013] R1, R2, R3, R4, R5, R6 are selected from H, deuterium, halogen, C1-C6 alkyl or one or more deuterated C1-C4 alkyl;

[0014] R7, R8, R9 are selected from H, deuterium, or one or more deuterated C1-C4 alkyl groups; they may be D, halogen, azido, -CN, -CF3, -CONR 10 R 11 、-NR 10 R 11 、-SR 11 、-OR 11 , -R 12 , -W, -MW, -V or -LN(R 13 )-V;

[0015] L is -(CH2) n -, n = 0-6;

[0016] R 10 Selected from H or C1-C6 alkyl, optionally substituted by one or more halogen or =O;

[0017] R 11 is selected from H, C1-C10 alkyl, C1-C10 heteroalkyl, C2-C10 alkenyl or C2-C10 heteroalkenyl, each of which may be optionally substituted with one or more halogen, =0 or an optionally substituted 3-7 membered carbocyclic or heterocyclic ring;

[0018] R 12 is selected from optionally substituted C1-C10 alkyl, C2-C10 alkenyl, C5-C10 aryl or C6-C12 arylalkyl or heterocycle, each of which may be optionally substituted with one or more halogen, =0 or an optionally substituted 3-6 membered carbocyclic or heterocyclic ring;

[0019] R 13 is selected from H, C1-C10 alkyl, C1-C10 heteroalkyl, C2-C10 alkenyl or C2-C10 heteroalkenyl, each of which may be optionally substituted with one or more halogen, =0 or an optionally substituted 3-7 membered carbocyclic or heterocyclic ring;

[0020] M is a linker selected from C1-C10 alkylene, C1-C10 heteroalkylene, C2-C10 alkenylene or C2-C10 heteroalkenylene, each of which may be optionally substituted with one or more selected from halogen, =0 or C1-C6 alkyl;

[0021] W is selected from an optionally substituted 4-7 membered nitrogen-containing heterocyclic ring, optionally containing a heteroatom of N, O or S as a ring member;

[0022] V is selected from an optionally substituted 3-4 membered carbon ring, or a C1-C6 alkyl group substituted by 1-4 fluorine atoms.

[0023] In some embodiments, in Formula (I):

[0024] R1, R2, R3, R4, R5, R6 are selected from H, deuterium, halogen, C1-C6 alkyl or one or more deuterated C1-C4 alkyl;

[0025] R7, R8, R9 are selected from H, deuterium, or one or more deuterated C1-C4 alkyl groups; they may be D, halogen, azido, -CN, -CF3, -CONR 10 R 11 、-NR 10 R 11 、-SR 11 、-OR 11 , -R 12 , -W, -MW or -V;

[0026] L is -(CH2) n -, n = 0-6;

[0027] R 10 Selected from H or C1-C6 alkyl, optionally substituted by one or more halogen or =O;

[0028] R 11 is selected from H, C1-C10 alkyl, C1-C10 heteroalkyl, C2-C10 alkenyl or C2-C10 heteroalkenyl, each of which may be optionally substituted with one or more halogen, =0 or an optionally substituted 3-7 membered carbocyclic or heterocyclic ring;

[0029] R 12 is selected from optionally substituted C1-C10 alkyl, C2-C10 alkenyl, C5-C10 aryl or C6-C12 arylalkyl or heterocycle, each of which may be optionally substituted with one or more halogen, =0 or an optionally substituted 3-6 membered carbocyclic or heterocyclic ring;

[0030] M is a linker selected from C1-C10 alkylene, C1-C10 heteroalkylene, C2-C10 alkenylene or C2-C10 heteroalkenylene, each of which may be optionally substituted with one or more selected from halogen, =0 or C1-C6 alkyl;

[0031] W is selected from an optionally substituted 4-7 membered nitrogen-containing heterocyclic ring, optionally containing a heteroatom of N, O or S as a ring member;

[0032] V is selected from an optionally substituted 3-4 membered carbon ring, or a C1-C6 alkyl group substituted by 1-4 fluorine atoms.

[0033] In some embodiments, in Formula (I):

[0034] R1, R2, R3, R4, R5, R6 are each independently selected from H, deuterium, halogen, C1-C6 alkyl or one or more deuterated C1-C4 alkyl;

[0035] R7, R8, and R9 are each independently selected from H, deuterium, halogen, azido, -CN, C1-C6 alkyl, C1-C6 haloalkyl, -O(C1-C6 alkyl), -S(C1-C6 alkyl), -CONR 10 R 11 、-NR 10 R 11 , 3-4 membered cycloalkyl, 4-7 membered nitrogen-containing heterocyclic ring or -(CH2) m -4-7 membered nitrogen-containing heterocyclic ring, wherein R 10 and R 11 Each is independently selected from H or C1-6 alkyl, the 4-7 membered nitrogen-containing heterocyclic ring contains 1, 2, 3 or 4 nitrogen atoms, and m=1-3;

[0036] L is -(CH2) n -, n = 1-6;

[0037] R 10 Selected from H or C1-C6 alkyl, wherein the C1-C6 alkyl is optionally substituted by one or more halogens;

[0038] R 11 is selected from H or C1-C6 alkyl, the C1-C6 alkane being optionally substituted by one or more halogens.

[0039] In some embodiments, R1, R2, R3, R4, R5, and R6 are each independently selected from H, deuterium, C1-C6 alkyl, or one or more deuterated C1-C4 alkyl.

[0040] In some embodiments, R1 is C1-C6 alkyl or one or more deuterated C1-C4 alkyl, and R2 to R6 are each independently H or deuterium.

[0041] In some embodiments, R1 is -CH3 or -CD3, and R2 to R6 are all H or all deuterium.

[0042] In some embodiments, R1 is -CH3 or -CD3, R2 to R6 are all H, or R4 is H and R2, R3, R5 and R6 are all deuterium.

[0043] In some embodiments, R1 is -CH3, and R2 to R6 are all H. In some embodiments, R1 is -CD3, and R2 to R6 are all H.

[0044] In some embodiments, R7, R8, and R9 are each independently selected from H, deuterium, halogen, azido, -CN, C1-C6 alkyl, C1-C6 haloalkyl, -O(C1-C6 alkyl), -S(C1-C6 alkyl), -CONR 10 R 11 、-NR 10 R 11 , 3-4 membered cycloalkyl, 5-6 membered nitrogen-containing heterocyclic ring or -CH2-5-6 membered nitrogen-containing heterocyclic ring, wherein R 10 and R 11 Each is independently selected from H or C1-6 alkyl, and the 5-6 membered nitrogen-containing heterocyclic ring contains 1, 2, 3 or 4 nitrogen atoms.

[0045] In some embodiments, one of R7, R8, and R9 is selected from H, halogen, azido, -CN, C1-C6 alkyl, C1-C6 haloalkyl, -O(C1-C6 alkyl), -S(C1-C6 alkyl), -CONR 10 R 11 、-NR 10 R 11 , 3-4 membered cycloalkyl, 5-6 membered nitrogen-containing heterocyclic ring or -CH2-5-6 membered nitrogen-containing heterocyclic ring, wherein R 10 and R 11 Each is independently selected from H or C1-6 alkyl, the 5-6 membered nitrogen-containing heterocyclic ring contains 1, 2, 3 or 4 nitrogen atoms; the remaining two are H. In some embodiments, R7, R8, and R9 are all deuterium.

[0046] In some embodiments, R8 is selected from H, halogen, azido, -CN, C1-C6 alkyl, C1-C6 haloalkyl, -O(C1-C6 alkyl), -S(C1-C6 alkyl), -CONR 10 R 11 、-NR 10 R 11 , 3-4 membered cycloalkyl, 5-6 membered nitrogen-containing heterocyclic ring or -CH2-5-6 membered nitrogen-containing heterocyclic ring, wherein R 10 and R 11 Each is independently selected from H or C1-6 alkyl, the 5-6 membered nitrogen-containing heterocyclic ring contains 1, 2, 3 or 4 nitrogen atoms; R7 and R9 are H.

[0047] In some embodiments, R7 is selected from H, halogen, azido, -CN, C1-C6 alkyl, C1-C6 haloalkyl, -O(C1-C6 alkyl), -S(C1-C6 alkyl), -CONR 10R 11 、-NR 10 R 11 , 3-4 membered cycloalkyl, 5-6 membered nitrogen-containing heterocyclic ring or -CH2-5-6 membered nitrogen-containing heterocyclic ring, wherein R 10 and R 11 Each is independently selected from H or C1-6 alkyl, the 5-6 membered nitrogen-containing heterocyclic ring contains 1, 2, 3 or 4 nitrogen atoms; R8 and R9 are H.

[0048] In some embodiments, R8 is H, F, Cl, -CF3, -CN, -N3, -CONHCH3, -CON(CH3)2, -OCH3, -SCH3, -NHCH3, -N(CH3)2, -C2H5, -CH2-1-pyrrolyl, -CH2-4-pyridinyl, 1-pyrrolyl, 2H-tetrazol-5-yl, cyclopropyl, -CH2CF3; R7 and R9 are H.

[0049] In some embodiments, R7 is H, F, Cl, -CF3, -CN, -N3, -CONHCH3, -CON(CH3)2, -OCH3, -SCH3, -NHCH3, -N(CH3)2, -C2H5, -CH2-1-pyrrolyl, -CH2-4-pyridyl, 1-pyrrolyl, 2H-tetrazol-5-yl, cyclopropyl, -CH2CF3; R8 and R9 are H. In some embodiments, R7 is H, F, Cl, -CF3, -CN, -N3, -CONHCH3, -CON(CH3)2, -OCH3, -SCH3, -NHCH3, -N(CH3)2, -C2H5, cyclopropyl, -CH2CF3; R8 and R9 are H. In some embodiments, R7 is H, F, -CN, -CONHCH3, -OCH3, -C2H5, cyclopropyl, -CH2CF3; R8 and R9 are H.

[0050] In some embodiments, L is -(CH2) n -, n = 1-3. In some embodiments, L is -CH2-.

[0051] In some embodiments, R 10 and R 11 Each is independently H or -CH3.

[0052] In some embodiments, the compound of formula (I) is selected from the following compounds:

[0053]

[0054]

[0055]

[0056] Furthermore, the compound also includes pharmaceutically acceptable salts, stereoisomers, racemates, deuterated compounds, or solvent compounds thereof.

[0057] The "pharmaceutically acceptable salt" of the compound shown in the general formula (I) of the present application is usually present in the form of a free acid. The acid addition salt of the free amino compound of the present application can be prepared by methods known in the art. Suitable acids include, but are not limited to, phosphoric acid, hydrochloric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, sulfuric acid, nitric acid, maleic acid, fumaric acid, benzoic acid, ascorbic acid, succinic acid, methanesulfonic acid, formic acid, acetic acid, trifluoroacetic acid, oxalic acid, propionic acid, tartaric acid, salicylic acid, citric acid, gluconic acid, lactic acid, mandelic acid, phenylacetic acid, aspartic acid, stearic acid, palmitic acid, glycolic acid, glutamic acid, p-toluenesulfonic acid and benzenesulfonic acid. In addition, the groups containing basic nitrogen atoms can be quaternized with the following reagents: lower alkyl halides, such as methyl, ethyl and butyl chloride, bromide and iodide; dialkyl sulfates, such as dimethyl, diethyl, dibutyl and diamyl sulfate; long chain halides, such as decyl, dodecyl, tetradecyl and octadecyl chloride, bromide and iodide; aralkyl halides, such as benzyl and phenethyl bromide and others. Water-soluble or oil-soluble or dispersible products are thus obtained. Therefore, "pharmaceutically acceptable salts" should include all acceptable salt forms.

[0058] The "stereoisomers" and "racemates" of the compounds represented by the general formula (I) in the present application generally refer to compounds of the formula (I) which may have chiral centers and may exist in the form of racemates, racemic mixtures, and individual enantiomers or diastereomers, and all isomeric forms are included in the present application, including mixtures thereof.

[0059] In addition, some of the compounds represented by the general formula (I) may also form solvates with water or other organic solvents, such as hydrates and tert-butyl alcohol solvates. Such solvates are also similarly included in the scope of the present application.

[0060] It will be understood by those skilled in the art that any compound may contain an atomic isotope of a non-natural ratio on one or more atoms constituting the compound. In the context of the present application, the deuterated compound refers to the deuterium atom contained in the relevant site of the compound in excess of the natural ratio (i.e., in excess of the natural abundance of deuterium). Therefore, any compound shown in the general formula (I) containing deuterium atoms in a ratio higher than the natural abundance of deuterium at the relevant site is within the scope of protection of the present application. For example, it will be appreciated that the compound shown in the corresponding general formula (I) obtained by introducing deuterium atoms using a commercially available deuterated reagent by the same or similar chemical synthesis means as shown in the embodiments of the present application with a corresponding deuterated rate or deuterium content is within the scope of protection of the present application. The chemical synthesis means and deuterated reagents herein are not limited to those exemplified in the embodiments, but should be understood as all synthetic methods or routes that can be used in the art to obtain the compounds of the present application, as well as all deuterated reagents that can be used to introduce deuterium atoms into the target molecule in conjunction with the aforementioned synthetic methods or routes.

[0061] On the other hand, the present application provides a pharmaceutical composition comprising the compound described herein or a pharmaceutically acceptable salt, stereoisomer, racemate, deuterated compound, or solvate thereof as an active ingredient.

[0062] In another aspect, the present application provides a pharmaceutical composition comprising a therapeutically effective amount of the compound described herein or a pharmaceutically acceptable salt, stereoisomer, racemate, deuterated compound, or solvent compound thereof, and one or more pharmaceutically acceptable carriers or excipients.

[0063] In yet another aspect, the present application provides use of the compound described herein or its pharmaceutically acceptable salt, stereoisomer, racemate, deuterated compound, or solvent compound in medicine.

[0064] On the other hand, the present application provides the use of the compound described in the present application or its pharmaceutically acceptable salt, stereoisomer, racemate, deuterated compound, or solvent compound or the pharmaceutical composition described in the present application in the preparation of a drug for treating tumors, autoimmune diseases, or diseases caused by bacteria, fungi or viruses.

[0065] On the other hand, the present application provides compounds described herein or pharmaceutically acceptable salts, stereoisomers, racemates, deuterated compounds, or solvent compounds thereof, or pharmaceutical compositions described herein for use in treating tumors, autoimmune diseases, or diseases caused by bacteria, fungi or viruses.

[0066] On the other hand, the present application provides a method for treating tumors, autoimmune diseases, or diseases caused by bacteria, fungi or viruses, which comprises administering to a patient a therapeutically effective amount of the compound described herein or its pharmaceutically acceptable salt, stereoisomer, racemate, deuterated compound, or solvent compound or the pharmaceutical composition described herein.

[0067] On the other hand, the present application provides the use of the compounds described herein or their pharmaceutically acceptable salts, stereoisomers, racemates, deuterated compounds, or solvent compounds, or the pharmaceutical compositions described herein for treating tumors, autoimmune diseases, or diseases caused by bacteria, fungi or viruses.

[0068] How to use

[0069] The present application provides the use of a quinolone analog of general formula (I) or a pharmaceutically acceptable salt, stereoisomer, racemate, deuterated compound, or solvent compound thereof in the preparation of a drug.

[0070] The present application provides the use of the above-mentioned compound or its pharmaceutically acceptable salt in the preparation of a treatment for cancers with BRCA-1 or BRCA-2 mutations (pure or heterozygous mutations), homologous recombination repair defective cancers, non-homologous end joining (NHEJ) repair defective cancers, or other DNA damage repair defective cancers, as well as the use of the above-mentioned compound or its pharmaceutically acceptable salt in the preparation of a treatment for cancers with overexpression of c-Myc, N-Myc or L-Myc. It includes single use and combined use. The combined use can be administered simultaneously or sequentially.

[0071] The present application provides the use of the above-mentioned compound or its pharmaceutically acceptable salt in the preparation of a treatment for a disease that is improved or resistant to PARP inhibitors, or the combination with a PARP inhibitor drug. The PARP inhibitor drug used in combination is selected from Olaparib, Niraparib, Rucaparib, Talazoparib, Fluzopari, Pamipari, or other PARP inhibitors.

[0072] The present application provides the combination of the above-mentioned compound or a pharmaceutically acceptable salt thereof with a topoisomerase I and topoisomerase II inhibitor, an ATM inhibitor, an ATR inhibitor, and a DNA-pK inhibitor.

[0073] Further, the cancer is breast cancer, ovarian cancer, endometrial cancer, cervical cancer, oral cancer, pancreatic cancer, prostate cancer, brain cancer, lung cancer, liver cancer (HCC), leukemia, lymphoma, myeloma, multiple myeloma, skin cancer, peritoneal cancer, colorectal cancer, glioblastoma, melanoma, osteosarcoma, cervical cancer, Ewing's sarcoma, lymph node cancer, gastrointestinal malignancies, head and neck cancer, kidney cancer, heart cancer, or other cancers.

[0074] The present application provides the use of the above-mentioned compound or its pharmaceutically acceptable salt in the preparation of a drug for treating autoimmune deficiency diseases such as multiple sclerosis, and a combined drug. The combined drug can be administered simultaneously or sequentially.

[0075] The present application provides the use of the above-mentioned compound or its pharmaceutically acceptable salt in the preparation of treatment or its pharmaceutically acceptable salt in the preparation of treatment for diseases caused by bacteria, fungi, and viruses. Viral infections include but are not limited to hepatitis B, hepatitis C virus, rhinovirus, herpes zoster virus, herpes simplex virus, cytomegalovirus, pox virus, encephalitis virus, Hantavirus, arbovirus, human papillomavirus (HPV), Sironi virus, HIV, influenza virus, Epstein-Barr virus, respiratory syncytial virus, coronavirus (SARS-CoV, SARS-CoV-2, MERS-CoV), etc. Including single use and combination therapy. The combination therapy can be administered simultaneously or sequentially.

[0076] preparation

[0077] The present application is a preparation prepared from a compound as described in the general formula (I) or a pharmaceutically acceptable salt, stereoisomer, racemate, deuterated compound, or solvent compound thereof as an active ingredient, and a pharmaceutically acceptable excipient or auxiliary ingredient. The pharmaceutically acceptable auxiliary ingredient has certain physiological activity, but the addition of the ingredient does not change the dominant position of the above-mentioned pharmaceutical composition in the disease treatment process, but only plays an auxiliary effect, which is only the utilization of the known activity of the ingredient, and is a conventional auxiliary treatment method in the medical field. If the above-mentioned auxiliary ingredients are used in conjunction with the pharmaceutical composition of the present application, it should still fall within the scope of protection of the present application.

[0078] "Pharmaceutically acceptable carriers" refer to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers, wetting agents, colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc., any type of non-toxic solid, semi-solid and liquid fillers, diluents, encapsulating materials and formulation adjuvants.

[0079] Furthermore, the present application provides a pharmaceutical composition, which contains a therapeutically effective amount of the quinolone compound or a pharmaceutically acceptable salt, stereoisomer, racemate, deuterated compound, or solvent compound thereof as an active ingredient and a pharmaceutically acceptable carrier.

[0080] Pharmaceutical composition

[0081] The pharmaceutical composition of the present application may be in solid or liquid form. On the one hand, the carrier is a microparticle, so that the composition is, for example, in the form of a tablet or a powder. The carrier may be a liquid, and the composition is, for example, an oral syrup, an injectable liquid, or an aerosol suitable for, for example, inhalation administration. When oral administration is desired, the pharmaceutical composition is preferably in solid or liquid form, wherein semi-solid, semi-liquid, suspension and gel forms are included in the form considered as solid or liquid herein. For oral solid compositions, the pharmaceutical composition can be formulated into forms such as powders, granules, compressed tablets, pills, capsules, chewable tablets, powder tablets, etc. Such solid compositions typically contain one or more inert diluents or edible carriers. In addition, one or more of the following substances may be present: binders, such as carboxymethyl cellulose, ethyl cellulose, microcrystalline cellulose, xanthan gum or gelatin; excipients, such as starch, lactose or dextrin; disintegrants, such as alginic acid, sodium alginate, corn starch, etc.; lubricants, such as magnesium stearate or hydrogenated vegetable oils; glidants, such as colloidal silicon dioxide; sweeteners, such as sucrose or saccharin; flavoring agents, such as mint, methyl salicylate or orange flavoring; and coloring agents.

[0082] The pharmaceutical composition can be administered parenterally, intracisternal, intravaginal, intraperitoneal, topically (e.g., powders, ointments, drops, and transdermal patches), rectally, or buccally. The term "parenteral" as used herein refers to a model of administration including intravenous, intramuscular, intraperitoneal, intrasternal, subcutaneous, and intraarticular injection and infusion. Pharmaceutical compositions for parenteral injection include pharmaceutically acceptable sterile water, or non-aqueous solutions, dispersions, suspensions, and emulsions, as well as sterile powders for reconstitution into sterile injections or dispersions immediately prior to use. Suitable aqueous and non-aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, etc.), carboxymethylcellulose, and suitable mixtures thereof, vegetable oils (e.g., olive oil), and injectable organic esters (e.g., ethyl oleate). In the case of dispersions, suitable fluidity can be maintained by using packaging materials such as lecithin to maintain the desired particle size, and by using surfactants.

[0083] When the pharmaceutical composition is in capsule form, such as a gelatin capsule, it may contain a liquid carrier, such as polyethylene glycol or oil, in addition to the above-mentioned types of substances. The pharmaceutical composition may be in liquid form, such as a tincture, syrup, solution, emulsion or suspension. When taken orally, the preferred composition contains one or more of a sweetener, a preservative, a dye / colorant and a flavor enhancer in addition to the compounds of the present application. In compositions intended for injection, one or more of a surfactant, a preservative, a wetting agent, a dispersant, a suspending agent, a buffer, a stabilizer and an isotonic agent may be included.

[0084] The pharmaceutical compositions of the present application may include a variety of materials that modify the physical form of the solid or liquid dosage unit.

[0085] Solid or liquid pharmaceutical compositions of the present application may include an agent that binds to the compounds of the present application and thereby facilitates the delivery of the compounds. Suitable agents with this capability include monoclonal or polyclonal antibodies, proteins or liposomes.

[0086] The pharmaceutical composition of the present application can be made using methods well known in the pharmaceutical field. For example, a pharmaceutical composition to be administered by injection can be prepared by combining the compound of the present application with sterile distilled water to form a solution. A surfactant can be added to form a uniform solution or suspension. A surfactant is a compound that non-covalently interacts with the compound of the present application, thereby promoting the dissolution or uniform suspension of the compound in an aqueous delivery system.

[0087] The compounds of the present invention as shown in general formula (I) or their pharmaceutically acceptable salts, stereoisomers, racemates, deuterated compounds, or solvates are administered in a therapeutically effective amount, which will vary depending on a variety of factors, including the activity of the specific compound used; the metabolic stability and duration of action of the compound; the patient's age, weight, general health status, sex and diet, mode and time of administration, excretion rate, drug combination, severity of a specific disease or condition; and the individual undergoing treatment.

[0088] The compound shown in the general formula (I) of the present application or its pharmaceutically acceptable salt, stereoisomer, racemate, deuterated compound, or solvent compound can also be administered at the same time, before or after administering one or more other therapeutic agents. This combination therapy includes administering a single pharmaceutical preparation containing the compound of the present application and one or more other active agents, and administering a compound shown in the general formula I of the present application and a separate pharmaceutical preparation of each active agent.

[0089] A medical composition, which is composed of a compound as shown in the general formula I of the present application or a pharmaceutically acceptable salt, stereoisomer, racemate, deuterated compound, or solvent compound thereof, and a combination of an immunomodulator, an antibiotic (such as penicillins, aminoglycosides, quinolones, macrolides, etc.), and an antiviral drug (such as a neuraminic acid inhibitor, a cap-dependent nuclease inhibitor, an RNA-dependent RNA polymerase inhibitor, an M2 protein inhibitor, etc.).

[0090] Definition and Description

[0091] The definitions of groups and terms recorded in this application, including their definitions as examples, exemplary definitions, preferred definitions, definitions of specific compounds in examples, etc., can be arbitrarily combined and coupled with each other.

[0092] A specific term should not be considered as uncertain or unclear without special definition, but should be understood according to the ordinary meaning in the art. When a trade name appears in this article, it is intended to refer to its corresponding product or its active ingredient.

[0093] Unless otherwise defined, the following terms used in the specification and claims have the following meanings.

[0094] "Stereoisomers" refer to isomers resulting from different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers and diastereomers.

[0095] "Pharmaceutical composition" refers to a mixture of one or more of the compounds of the present application with other chemical components, such as a pharmaceutically acceptable carrier. The purpose of a pharmaceutical composition is to facilitate the process of administration to an animal.

[0096] "Pharmaceutical carrier" refers to an inactive ingredient in a pharmaceutical composition that does not cause significant irritation to an organism and does not interfere with the biological activity and properties of the administered compound, such as but not limited to: calcium carbonate, various sugars (such as lactose, mannitol, etc.), starch, cyclodextrin, magnesium stearate, cellulose, gelatin, water, polyethylene glycol, propylene glycol, ethylene glycol, castor oil or hydrogenated castor oil or polyethoxylated hydrogenated castor oil, sesame oil, corn oil, peanut oil, etc.

[0097] The term "substituted" refers to any one or more hydrogen atoms on a particular atom being replaced by a substituent, as long as the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is an oxo (i.e., =O), it means that two hydrogen atoms are replaced, and the oxo does not occur on an aromatic group. Unless otherwise indicated, the term "substituted" refers to any level of substitution, such as mono-, di-, tri-, tetra- or penta-substitution, as long as such substitution is allowed. Substituents are independently selected, and substitutions can be located at any chemically accessible position. It should be understood that the substitution of a given atom is limited by valence. It should be understood that the substitution on a given atom produces a chemically stable molecule.

[0098] The term "optional" or "optionally" refers to that the event or situation described subsequently may or may not occur, and the description includes the occurrence of the event or situation and the non-occurrence of the event or situation. For example, ethyl is "optionally" substituted by halogen, meaning that ethyl can be unsubstituted (CH2CH3), monosubstituted (CH2CH2F, CH2CH2Cl etc.), polysubstituted (CHFCH2F, CH2CHF2, CHFCH2Cl, CH2CHCl2 etc.) or fully substituted (CF2CF3, CF2CCl3, CCl2CCl3 etc.). It will be appreciated by those skilled in the art that for any group comprising one or more substituents, any substitution or substitution pattern that may not exist and / or cannot be synthesized in space will not be introduced.

[0099] When any variable (e.g., B) occurs more than once in a compound's composition or structure, its definition is independent at each occurrence. b is replaced, then each R b All have independent options.

[0100] When a substituent's bond crosses two atoms in a ring, such substituent may be bonded to any atom in the ring.

[0101] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit-risk ratio.

[0102] The term "alkyl" used alone or in combination with other terms refers to a saturated hydrocarbon group that can be straight or branched. The term "Cn-Cm alkyl" refers to an alkyl group having n to m carbon atoms. In some embodiments, the alkyl group contains 1 to 10 carbon atoms, 1 to 6 carbon atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms. Examples of alkyl moieties include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, 2-methyl-1-butyl, n-pentyl, 3-pentyl, n-hexyl, 1,2,2-trimethylpropyl, etc.

[0103] The term "alkenyl" used alone or in combination with other terms refers to a straight or branched hydrocarbon group corresponding to an alkyl group having one or more carbon-carbon double bonds. The term "Cn-Cm alkenyl" refers to an alkenyl group having n to m carbons. In some embodiments, the alkenyl group contains 2 to 10, 2 to 6, 2 to 4, or 2 to 3 carbon atoms. Examples of alkenyl groups include, but are not limited to, vinyl, n-propenyl, isopropenyl, n-butenyl, sec-butenyl, etc.

[0104] The term "alkylene", used alone or in combination with other terms, refers to a divalent alkyl linking group. The term "Cn-Cm alkylene" refers to an alkylene group having n to m carbon atoms. In some embodiments, the alkylene group contains 1 to 10 carbon atoms, 1 to 6 carbon atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms. Examples of alkylene groups include, but are not limited to, methylene, ethylene, propylene, ethane-1,2-diyl, ethane-1,1-diyl, propane-1,3-diyl, propane-1,2-diyl, propane-1,1-diyl, butane-1,4-diyl, butane-1,3-diyl, butane-1,2-diyl, 2-methyl-propyl-1,3-diyl, etc.

[0105] The term "carbonyl," employed alone or in combination with other terms, refers to a -C(=O)- group, which may also be written C(O).

[0106] The term "azido" refers to -N3.

[0107] The term "cyano" or "nitrile" refers to a group of formula -C≡N, which may also be written as -CN.

[0108] The term "halo" or "halogen" used alone or in combination with other terms refers to fluorine, chlorine, bromine and iodine. In some embodiments, "halo" refers to a halogen atom selected from F or Cl. In some embodiments, the halo group is F.

[0109] As used herein, the term "haloalkyl" refers to an alkyl group in which one or more hydrogen atoms are replaced by a halogen atom. The term "Cn-Cm haloalkyl" refers to a Cn-Cm alkyl group having nm carbon atoms and at least one to a maximum of {2 (n to m) + 1} halogen atoms, the halogens being identical or different. In some embodiments, the halogen atom is a fluorine atom. In some embodiments, haloalkyl has 1 to 10, 1 to 6 or 1 to 4 carbon atoms. Examples of haloalkyl groups include CF3, C2F5, CH2CF3, CHF2, CH2F, CCl3, CHCl2, C2Cl5, etc. In some embodiments, haloalkyl is trifluoromethyl CF3.

[0110] The term "aryl" used alone or in combination with other terms refers to an aromatic hydrocarbon group, which can be monocyclic or polycyclic (e.g., having 2 fused rings). The term "Cn-Cm aryl" refers to an aryl group having n to m ring carbon atoms. Aryl includes, for example, phenyl, naphthyl, etc. In some embodiments, aryl has 5 to 10 carbon atoms. In some embodiments, aryl has 6 carbon atoms. In some embodiments, aryl has 10 carbon atoms. In some embodiments, aryl is phenyl.

[0111] The term "heterocycle" refers to a saturated, partially unsaturated, or fully unsaturated monocyclic or polycyclic heterocycle having at least one heteroatom ring member selected from sulfur, oxygen, and nitrogen. In some embodiments, the heterocycle has 1, 2, 3, or 4 heteroatom ring members independently selected from nitrogen, sulfur, and oxygen. In some embodiments, the heterocycle has 3-6, 3-7, 5-12, or 6-12 ring atoms (including carbon atoms) and 1, 2, 3, or 4 heteroatom ring members independently selected from nitrogen, sulfur, and oxygen. Heterocycles include heteroaromatic rings.

[0112] The term "heteroaryl" used alone or in combination with other terms refers to a monocyclic or polycyclic aromatic heterocycle having at least one heteroatom ring member selected from sulfur, oxygen and nitrogen. In some embodiments, the heteroaryl ring has 1, 2, 3 or 4 heteroatom ring members independently selected from nitrogen, sulfur and oxygen. In some embodiments, the heteroaryl has 5-14 or 5-12 ring atoms (including carbon atoms) and 1, 2, 3 or 4 heteroatom ring members independently selected from nitrogen, sulfur and oxygen. In some embodiments, the heteroaryl has 5-10 ring atoms (including carbon atoms) and 1, 2, 3 or 4 heteroatom ring members independently selected from nitrogen, sulfur and oxygen. In some embodiments, the heteroaryl has 5-6 ring atoms and 1 or 2 heteroatom ring members independently selected from nitrogen, sulfur and oxygen. In some embodiments, the heteroaryl is a five-membered or six-membered heteroaryl ring. In other embodiments, the heteroaryl is an eight-membered, nine-membered or ten-membered fused bicyclic heteroaryl ring. Examples of heteroaryl groups include, but are not limited to, thienyl, furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, tetrazolyl or thiadiazolyl, and the like, and benzo derivatives thereof, such as benzofuranyl, benzothienyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzotriazolyl, indazolyl, indolyl or isoindolyl, and the like; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl or triazinyl, and the like, and benzo derivatives thereof, such as quinolyl, quinazolinyl or isoquinolyl, and the like; or azinyl, indolizinyl, purinyl, and the like, and benzo derivatives thereof; or cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl or phenoxazinyl, and the like. The term "5-6 membered heteroaryl" refers to an aromatic ring system having 5 or 6 ring atoms and which contains 1-3, preferably 1-2 heteroatoms independently selected from N, O and S.

[0113] A five-membered heteroaryl ring is a heteroaryl group having five ring atoms, wherein one or more (eg, 1, 2, or 3) of the ring atoms are independently selected from N, O, and S.

[0114] A six-membered heteroaryl ring is a heteroaryl group having six ring atoms, wherein one or more (eg, 1, 2, or 3) of the ring atoms are independently selected from N, O, and S.

[0115] The term "carbocycle" used alone or in combination with other terms refers to a non-aromatic hydrocarbon ring system (monocyclic, bicyclic or polycyclic), including cyclized alkyl and alkenyl. The term "Cn-Cm carbocycle" refers to a cycloalkyl with n to m ring member carbon atoms. The carbocycle may have 3, 4, 5, 6 or 7 ring-forming carbons (C3-C7, C3-C6 or C3-C4). In some embodiments, the carbocycle has 3 to 7 ring members, 3 to 6 ring members or 3 to 4 ring members. In some embodiments, the carbocycle is monocyclic. In some embodiments, the carbocycle is monocyclic or bicyclic. Examples of carbocycles include but are not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl, etc. In some embodiments, the carbocycle is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. In some embodiments, the carbocycle is cyclopropyl.

[0116] The term "nitrogen-containing heterocycle" refers to an atom that contains at least one nitrogen heteroatom in addition to carbon atoms. In some embodiments, the nitrogen-containing heterocycle has 3 to 7 ring members, 3 to 6 ring members, or 3 to 4 ring members. Examples of nitrogen-containing heterocycles include, but are not limited to, pyrrole, imidazole, thiazole, pyridine, triazole, tetrazole, quinoline, and indole. In some embodiments, the 4-7-membered nitrogen-containing heterocycle is pyrrole, pyridine, or tetrazole.

[0117] The term "heteroalkyl" refers to an alkyl group in which one or more (preferably 1, 2 or 3) carbon atoms are replaced by oxygen, nitrogen, phosphorus, silicon or sulfur atoms (preferably oxygen, sulfur or nitrogen). For example, C1-C10 heteroalkyl, C1-C6 heteroalkyl, C1-C3 heteroalkyl. The term "heteroalkenyl" refers to an alkenyl group in which one or more (preferably 1, 2 or 3) carbon atoms are replaced by oxygen, nitrogen, phosphorus, silicon or sulfur atoms (preferably oxygen, sulfur or nitrogen). For example, C2-C10 heteroalkenyl, C2-C6 heteroalkenyl, C2-C3 heteroalkenyl.

[0118] As used herein, the expressions "ambient temperature" and "room temperature" are understood in the art and generally refer to a temperature, such as a reaction temperature, which is about the temperature of the room in which the reaction is carried out, for example, a temperature of about 20°C to about 30°C.

[0119] The present invention also includes pharmaceutically acceptable salts of the compounds described herein. The term "pharmaceutically acceptable salt" refers to derivatives of the disclosed compounds in which the parent compound is modified by converting an existing acid or base moiety into its salt form. Pharmaceutically acceptable salts of the present invention include non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. Pharmaceutically acceptable salts of the present invention can be synthesized from a parent compound containing a basic or acidic moiety by conventional chemical methods.

[0120] As used herein, the term "heteroatom" is understood to mean an atom other than carbon, such as O, N or S. BRIEF DESCRIPTION OF THE DRAWINGS

[0121] Figure 1 The curves show the effects of repeated administration of CX-5461, compound H-31 and solvent on the body weight of mice at a dose of 50 mg / kg.

[0122] Figure 2-1 and Figure 2-2 The curves show the effects of repeated administration of 55 mg / kg of CX-5461, H series compounds and solvents on the body weight of mice.

[0123] Figure 3 The results show the inhibitory effects of Olaparib, compound H-31 and solvent on tumors in the human triple-negative breast cancer MDA-MB-436 model. DETAILED DESCRIPTION

[0124] The following embodiments are used to further describe the present application, but these embodiments are not intended to limit the scope of the present application.

[0125] The experimental methods in the examples of this application without specifying specific conditions are usually based on conventional conditions, and the reagents without specifying specific sources are conventional reagents purchased from the market.

[0126] Unless otherwise defined, the following terms used in the specification and claims have the following meanings.

[0127] "Pharmaceutical composition" refers to a mixture of one or more of the compounds of the present application with other chemical components, such as a pharmaceutically acceptable carrier. The purpose of a pharmaceutical composition is to facilitate the process of administration to an animal.

[0128] "Pharmaceutical carrier" refers to an inactive ingredient in a pharmaceutical composition that does not cause significant irritation to an organism and does not interfere with the biological activity and properties of the administered compound, such as but not limited to: calcium carbonate, calcium phosphate, various sugars (such as lactose, mannitol, etc.), starch, cyclodextrin, magnesium stearate, cellulose, magnesium carbonate, acrylic acid polymers or methacrylic acid polymers, gelatin, water, polyethylene glycol, propylene glycol, ethylene glycol, castor oil or hydrogenated castor oil or polyethoxylated hydrogenated castor oil, sesame oil, corn oil, peanut oil, etc.

[0129] The present application is further described below in conjunction with embodiments:

[0130] Example 1 Preparation of Compound H-1

[0131] 1. Preparation of 2,5-dichloropyridine-3-carbonyl chloride

[0132]

[0133] Compound 2,5-dichloropyridine-3-carboxylic acid (5 g, 26.0 mmol, 1.0 eq) was added to thionyl chloride (40 mL) cooled in an ice bath, and then the system was heated to 80°C for overnight reaction (o / n, the same below). The thionyl chloride was removed in vacuo to obtain 2,5-dichloropyridine-3-carbonyl chloride (5.4 g, yellow oil, 98% yield). LC-MS (ESI): m / z 206.00 [M+1] + (Methanol quenches the acyl chloride, and MS shows the methyl ester of compound 1), 1 HNMR(400MHz, CDCl3)δ:8.40–8.30(m,1H),7.50–7.40(m,1H).

[0134] 2. Preparation of Compound 4

[0135]

[0136] Compound 3 (1.0 g, 4.5 mmol, 1.0 eq.) was dissolved in tetrahydrofuran (10 mL), and then anhydrous magnesium chloride (430 mg, 4.5 mmol, 1.0 eq.) was added to the system. Under nitrogen protection, the reaction solution was stirred at 0°C for half an hour. Then 2,5-dichloropyridine-3-carbonyl chloride (compound 2, 2.8 g, 13.5 mmol, 3.0 eq.) and triethylamine (2.3 g, 22.6 mmol, 5.0 eq.) were slowly added to the system. The mixture was reacted at room temperature overnight. The reaction solution was directly filtered, the filter cake was washed with water and methanol, and dried to obtain compound 4 (1.2 g, light yellow solid, 68% yield). LC-MS (ESI): m / z 359.2 [M+1] + , 1 HNMR(400MHz, CDCl3)δ:9.53(d,1H),8.84(d,1H),7.82–7.69(m,1H),7.64–7.52(m,2H),7.50–7.40(m,1H),4.58–4.42(m,2H),1.50–1.45(m,3H).

[0137] 3. Preparation of Compound 6

[0138]

[0139] Compound 4 (5 g, 13.9 mmol, 1.0 eq.) and potassium carbonate (9.6 g, 69.5 mmol, 5.0 eq.) were added to acetonitrile (80 mL). Then N-methyl homopiperazine (3.2 g, 27.9 mmol, 2.0 eq.) was added to the system. Under nitrogen protection, the reaction solution was reacted at 80 ° C overnight. After cooling to room temperature, the reaction solution was directly filtered, the filter cake was washed with water and methanol, and dried to obtain compound 6 (4.4 g, light yellow solid, 72% yield). LC-MS (ESI): m / z 437.2 [M+1] + , 1 HNMR(400MHz,DMSO-d6)δ:9.23(d,1H),8.15(d,1H),7.91–7.86(m,1H),7.46–7.32(m,2H),6.84(d,1H),4.28(q, 2H),3.90–3.50(m,4H),2.80–2.60(m,2H),2.50–2.40(m,2H),2.30–2.15(m,3H),2.00–1.85(m,2H),1.29(t,3H).

[0140] 4. Preparation of Compound 7

[0141]

[0142] Compound 6 (4.45 g, 10.2 mmol, 1.0 eq) was dissolved in tetrahydrofuran (30 mL) and water (30 mL), and lithium hydroxide monohydrate (1.23 g, 30.6 mmol, 3.0 eq) was added to the system. The reaction solution was reacted overnight at room temperature. Filtered directly. The filter cake was washed with ethyl acetate and vacuum dried to obtain compound 7 (4.15 g, light yellow solid, 98% yield). LC-MS (ESI): m / z 409.1 [M+1] + , 1 H NMR(400MHz,DMSO-d6)δ:9.40(d,1H),8.36(d,1H),7.87(d,1H),7.50–7.35(m,2H),7.10–6.90( m,1H),4.10–3.50(m,4H),2.75–2.65(m,2H),2.50–2.40(m,2H),2.24(s,3H),2.00–1.90(m,2H).

[0143] 5. Preparation of Compound H-1

[0144]

[0145] 2-(7-azabenzotriazole)-tetramethyluronium hexafluorophosphate (0.07 g, 0.184 mmol, 1.5 eq.) and N,N-diisopropylethylamine (0.065 g, 0.488 mmol, 4.0 eq.) were added to a solution of compound 7 (0.05 g, 0.122 mmol, 1.0 eq.) in N,N-dimethylformamide (3 mL). The mixture was stirred at 50 ° C for 0.5 hours under nitrogen protection. Then compound H1-1 (0.031 g, 0.184 mmol, 1.5 eq.) dissolved in N,N-dimethylformamide (1 mL) was added and the mixture was stirred at 50 ° C for 16 hours under nitrogen protection. The reaction solution was spin-dried to obtain a residue. The residue was slurried with methanol, filtered and the filter cake was washed, and dried to obtain compound H-1 (0.043 g, light yellow solid, yield 68%). LC-MS(ESI):m / z 518.4[M+1] + , 1 HNMR(400MHz,DMSO-d6)δ:11.11(d,1H),9.30–9.15(m,1H),8.95–8.85(m,2H),8.36(d,1H),7.95(d,1H),7.63–7.37(m,2H),7.15–6.95(m, 1H),4.85–4.70(m,2H),4.15–3.90(m,2H),3.90–3.60(m,2H),3.60–3 .50(m,2H),3.30–3.20(m,2H),2.80–2.60(m,3H),2.30–2.10(m,2H).

[0146] Example 2 Preparation of Compound H-2

[0147]

[0148] 2-(7-azabenzotriazole)-tetramethyluronium hexafluorophosphate (0.16 g, 0.42 mmol, 1.5 eq.) and N,N-diisopropylethylamine (0.15 mL, 0.84 mmol, 3.0 eq.) were added to a solution of compound 7 (0.114 g, 0.28 mmol, 1.0 eq.) in N,N-dimethylformamide (3 mL). The mixture was stirred at 50 ° C for 0.5 hours under nitrogen protection. Then compound H2-1 (0.06 g, 0.42 mmol, 1.5 eq.) dissolved in N,N-dimethylformamide (2 mL) was added and the mixture was stirred at 50 ° C for 16 hours under nitrogen protection. The reaction solution was spin-dried to obtain a residue. The residue was purified by preparative HPLC, and the preparative solution was lyophilized to obtain compound H-2 (0.0245 g, light yellow solid, yield 17%). LC-MS(ESI):m / z 534.2[M+1]+ , 1 H NMR(400MHz, DMSO-d6)δ:11.16(s,1H),9.50–9.35(m,1H),8.95–8.85(m,2H),8.45–8.30(m,1H),8.30–8.20(m,1H),8.05–7.90(m,1 H),7.60–7.40(m,2H),7.10–6.95(m,1H),4.85–4.70(m,2H),4.05–3.60(m,6H),2.85–2.65(m,2H),2.26(s,3H),2.05–1.90(m,2H).

[0149] Example 3 Preparation of Compound H-4

[0150]

[0151] Zinc cyanide (0.43 g, 3.62 mmol, 0.66 eq.), zinc (0.13 g, 1.97 mmol, 0.36 eq.), tris(dibenzylideneacetone)dipalladium (0.15 g, 0.16 mmol, 0.03 eq.) and 1,1'-bis(diphenylphosphino)ferrocene (0.19 g, 0.33 mmol, 0.06 eq.) were added to a solution of compound H4-1 (1.0 g, 5.48 mmol, 1.0 eq.) in DMA (40 mL), and the mixture was sealed at 110 ° C for 2 hours under nitrogen protection. The reaction solution was extracted with ethyl acetate and saturated brine, the organic phases were combined and dried, and the organic phases were spin-dried to obtain a residue. The residue was purified by silica gel column chromatography to obtain compound H4-2 (0.3 g, yellow oil, yield 30%). 1 H NMR(400MHz, CDCl3)δ:9.11(t,2H).

[0152]

[0153] To a solution of compound H4-2 (0.4 g, 2.31 mmol) in EtOH (20 mL) was added Raney-Ni (0.08 g) and NH3.H2O (0.2 mL). The mixture was stirred at room temperature for 2 hours under a hydrogen atmosphere (3.5 MPa). Filtered, the filtrate was spin-dried to obtain a residue. The residue was used directly in the next step without further purification. LC-MS (ESI): m / z 178.0 [M+1] +

[0154]

[0155] To a solution of crude compound H4-3 (0.22 g, 1.24 mmol, 1.0 eq.) in dichloromethane (6 mL) was added di-tert-butyl dicarbonate (0.33 g, 1.49 mmol, 1.2 eq.) and saturated sodium bicarbonate (4 mL) aqueous solution. The mixture was stirred at room temperature for 1 hour under nitrogen protection. The reaction solution was extracted with dichloromethane and water, the organic phases were combined and dried, and the organic phases were spin-dried to obtain a residue. The residue was purified by silica gel column chromatography to obtain compound H4-4 (0.07 g, yellow oil, yield 36%). LC-MS (ESI): m / z 279.2 [M+1] + , 1 H NMR (400MHz, CDCl3) δ: 8.93 (s, 2H), 5.57 (s, 1H), 4.66 (d, 2H), 1.46 (s, 9H).

[0156]

[0157] Add HCl-dioxane (4M, 3 mL) to compound H4-4 (0.07 g, 0.25 mmol). Stir the mixture at room temperature for 16 hours under nitrogen protection. The solvent was dried to obtain compound H4-5 (65 mg, gray solid, yield 100%). LC-MS (ESI): m / z 178.0 [M+1] + , 1 H NMR (400MHz, DMSO-d6)δ:9.36(s,2H),8.58(s,3H),4.41(s,2H).

[0158]

[0159] 2-(7-azabenzotriazole)-tetramethyluronium hexafluorophosphate (0.14 g, 0.37 mmol, 1.5 eq.) and N,N-diisopropylethylamine (0.13 g, 0.1 mmol, 4.0 eq.) were added to a solution of compound 7 (0.1 g, 0.25 mmol, 1.0 eq.) in N,N-dimethylformamide (4 mL). The mixture was stirred at 50 ° C for 0.5 hours under nitrogen protection. Then compound H4-5 (0.08 g, 0.37 mmol, 1.5 eq.) dissolved in N,N-dimethylformamide (2 mL) was added and the mixture was stirred at 50 ° C for 16 hours under nitrogen protection. The reaction solution was spin-dried to obtain a residue. The residue was separated and purified by preparative thin layer chromatography to obtain compound H-4 (0.06 g, light yellow solid, yield 44%). LC-MS (ESI): m / z 568.8 [M+1] + , 1H NMR(400MHz, DMSO-d6)δ:11.25–11.15(m,1H),9.45–9.30(m,1H),9.26(s,2H),8.32(d,1H),7.94(d,1H),7.55–7.45(m,1H),7.45-7. 35(m,1H),7.05–6.95(m,1H),4.90(d,2H),4.05–3.55(m,4H),2.80–2.65(m,2H),2.55–2.50(m,2H),2.26(s,3H),2.00–1.90(m,2H).

[0160] Example 4 Preparation of Compound H-5

[0161]

[0162] Potassium carbonate (616 mg, 4.459 mmol, 4.0 eq) and H5-1 (167 mg, 1.672 mmol, 1.5 eq) were added to a solution of compound 4 (400 mg, 1.114 mmol, 1.0 eq) in acetonitrile (8 mL). The reaction solution was stirred at 80 ° C for 16 hours under nitrogen protection. Directly spin-dried, water was added, and the mixture was washed with dichloromethane / methanol (10 / 1, v / v), and the organic phases were combined and spin-dried. The crude product was slurried with ethyl acetate overnight, and the filter residue was filtered to obtain the product H5-2 (320 mg, white solid, yield 68%). LC-MS (ESI): m / z 423.5 [M+1] +

[0163]

[0164] Potassium carbonate (265 mg, 1.917 mmol, 3.0 eq.) was added to a solution of compound H5-2 (270 mg, 0.639 mmol, 1.0 eq.) in N,N-dimethylformamide (7 mL), and iodomethane-d3 (70 mg, 0.479 mmol, 0.75 eq.) was added after stirring at 50 ° C for 10 minutes. The reaction solution was reacted at 50 ° C for 0.5 hours under a nitrogen atmosphere. Water was added to quench, extracted with ethyl acetate, the organic phases were combined, concentrated, and purified by column chromatography to finally obtain the product H5-3 (180 mg, white solid, yield 60%). LC-MS (ESI): m / z 440.6 [M+1] +

[0165]

[0166] LiOH.H2O (26 mg, 0.614 mmol, 3.0 eq.) was added to a solution of compound H5-3 (90 mg, 0.205 mmol, 1.0 eq) in tetrahydrofuran / methanol / water (3 mL / 1 mL / 3 mL). The mixture was stirred at room temperature overnight under nitrogen protection. The solvent was removed under reduced pressure and lyophilized to obtain compound H5-4 (91 mg, white solid, crude salt product). LC-MS (ESI): m / z 412.2 [M+1] + , 1 HNMR(400MHz,DMSO-d6)δ:9.50–9.35(m,1H),8.37(d,1H),7.95–7.85(m,1H),7.55–7.35(m,2 H),7.00(d,1H),4.10–3.60(m,4H),2.80–2.70(m,2H),2.55–2.50(m,2H),2.05–1.90(m,2H).

[0167]

[0168] 2-(7-azabenzotriazole)-tetramethyluronium hexafluorophosphate (125 mg, 0.328 mmol, 1.5 eq.) and N,N-diisopropylethylamine (142 mg, 1.094 mmol, 5.0 eq.) were added to a solution of compound H5-4 (91 g, 0.219 mmol, 1.0 eq.) in N,N-dimethylformamide (3 mL). The mixture was stirred at 50 ° C for 0.5 hours under nitrogen protection. Then compound H5-5 (72 mg, 0.437 mmol, 2.0 eq.) dissolved in N,N-dimethylformamide (1 mL) was added, and the mixture was stirred at 50 ° C for 16 hours under nitrogen protection. The reaction mixture was spin-dried to obtain a residue. The crude product was directly slurried with methanol (5 mL) for 30 minutes, filtered to obtain a filter residue, and separated and purified by preparative HPLC to finally obtain compound H-5 (12.8 mg, light yellow solid, two-step yield 11%). LC-MS(ESI):m / z 521.4[M+1] + , 1 H NMR(400MHz, DMSO-d6)δ:11.25–11.15(m,1H),9.50–9.40(m,1H),8.88(s,2H),8.45–8.30(m,1H),8.15–8.10(m,1H),7.65–7. 40(m,2H),7.10–6.95(m,1H),4.85–4.75(m,2H),4.05–3.60(m,4H),3.10–2.75(m,2H),2.70–2.55(m,2H),2.10–1.95(m,2H).

[0169] Example 5 Preparation of Compound H-6

[0170]

[0171] Potassium carbonate (4.3 g, 31.3 mmol, 5.0 eq.) was added to a solution of compound 4 (1.7 g, 6.27 mmol, 1.0 eq.) in acetonitrile (50 mL). Compound H6-1 (2.5 g, 6.89 mmol, 1.1 eq.) was then added, and the mixture was stirred at 80 ° C for 16 hours under nitrogen protection. The reaction mixture was then cooled to room temperature, washed with dichloromethane / methanol (10 / 1, v / v), dried and spin-dried to remove the solvent to obtain a residue. The residue was purified by silica gel column chromatography to obtain compound H6-2 (2.8 g, light yellow solid, yield 72%). LC-MS (ESI): m / z 521.2 [M+1] + , 1 H NMR(400MHz,DMSO-d6)δ:9.40–9.20(m,1H),8.23(d,1H),8.05–7.85(m,1H),7.55-7.35(m,2H ),7.30–7.10(m,5H),6.94(d,1H),4.29(q,2H),3.90–3.65(m,2H),3.59(s,2H),1.29(t,3H).

[0172]

[0173] N,N-diisopropylethylamine (1.1 g, 8.64 mmol, 2.5 eq.) was added to a solution of compound H6-2 (1.8 g, 3.46 mmol, 1.0 eq.) in 1,2-dichloroethane (30 mL). Compound H6-3 (0.99 g, 6.91 mmol, 2.0 eq.) was then added at 0 °C, and the mixture was stirred at room temperature for 1 hour under nitrogen protection. The reaction mixture was then dried and dissolved in methanol (30 mL). The mixture was stirred at 70 °C under nitrogen protection for 1 hour. The reaction solution was dried, dichloromethane / ethyl acetate (10 mL / 10 mL) was added, and the mixture was stirred at room temperature overnight. It was then filtered, the filter cake was washed with dichloromethane, and dried to obtain the crude product of compound H6-4 (1.64 g, yellow solid). LC-MS (ESI): m / z 431.0 [M+1] + , 1H NMR(400MHz,DMSO-d6)δ:9.30–9.15(m,2H),8.23(d,1H),8.00–7.90(m,1H),7.60–7.5 0(m,1H),7.50–7.40(m,1H),7.00(d,1H),4.29(q,2H),3.90–3.75(m,2H),1.30(t,3H).

[0174]

[0175] HCHO (12 mL) was added to a solution of HCOOH (12 mL) of compound H6-4 (1.13 g). The mixture was stirred at 50 ° C overnight under nitrogen protection. The pH of the solution was adjusted to a weak base with saturated sodium bicarbonate aqueous solution and the aqueous phase was extracted with dichloromethane / methanol (10: 1, v / v, 300 mL x 2). The combined organic layers were concentrated to obtain a residue. The residue was separated and purified by silica gel column chromatography to obtain compound H6-5 (0.85 g, light yellow solid, yield 72%). LC-MS (ESI): m / z 445.0 [M+1] + , 1 HNMR(400MHz,DMSO-d6)δ:9.40–9.20(m,1H),8.19(d,1H),8.00–7.90(m,1H),7.55–7.3 0(m,2H),7.00–6.80(m,1H),4.28(q,2H),3.80–3.50(m,2H),2.27(s,3H),1.30(t,3H).

[0176]

[0177] To a mixture of compound H6-5 (0.85 g, 1.91 mmol, 1.0 eq.) in tetrahydrofuran (10 mL), methanol (5 mL) and water (10 mL) was added LiOH·H2O (0.24 g, 5.74 mmol, 3.0 eq.). The reaction mixture was stirred at room temperature for 16 hours under nitrogen protection. The reaction mixture was concentrated to obtain a residue. The residue was washed with water and filtered. The filter cake was dried to obtain compound H6-6 (0.9 g, light yellow solid) for the next step reaction.

[0178] LC-MS(ESI):m / z 417.0[M+1] + , 1H NMR (400MHz, DMSO-d6)δ:9.50–9.35(m,1H),8.37(d,1H),7.88(d,1H),7.55–7.30(m,2H),6.99(d,1H),4.00–3.50(m,2H),2.25(s,3H).

[0179]

[0180] 2-(7-azabenzotriazole)-tetramethyluronium hexafluorophosphate (69 mg, 0.18 mmol, 1.5 eq.) and N,N-diisopropylethylamine (0.08 mL, 0.48 mmol, 4.0 eq.) were added to a solution of compound H6-6 (50 mg, 0.12 mmol, 1.0 eq.) in N,N-dimethylformamide (2 mL). The mixture was stirred at 50 ° C for 0.5 hours under nitrogen protection. Then, compound H6-7 (0.03 g, 0.18 mmol, 1.5 eq.) dissolved in N,N-dimethylformamide (1 mL) was added and the mixture was stirred at 50 ° C for 16 hours under nitrogen protection. The reaction solution was spin-dried to obtain a residue, and the residue was separated and purified by preparative HPLC, and the preparative solution was lyophilized to obtain compound H-6 (6.7 mg, light yellow solid, two-step yield 10%). LC-MS(ESI):m / z 526.2[M+1] + , 1 H NMR(400MHz,DMSO-d6)δ:11.25–11.15(m,1H),9.00–8.90(m,1H),8.95–8.85(m,2H),8.40–8.30(md,1H),8.25–8.15 (m,1H),8.05–7.95(m,1H),7.60–7.40(m,2H),7.10–6.95(m,1H),4.85–4.75(m,2H),3.85–3.65(m,2H),2.26(s,3H).

[0181] Example 6 Preparation of Compound H-7

[0182] 1. Preparation of Compound 9

[0183]

[0184] Ethyl 3-((tert-butyloxycarbonyl)(2-ethoxy-2-oxoethyl)amino)propanoate (1.0 g, 3.3 mmol, 1.0 eq.) was dissolved in deuterated methanol (15 mL), and 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) (8.3 g, 33.0 mmol, 10.0 eq.) was added to the solution. The mixture was stirred at 65°C for 16 hours. The reaction mixture was extracted with water and dichloromethane and concentrated to the next step. MS-ESI: m / z 286[M+1] +

[0185] 2. Preparation of Compound 10

[0186]

[0187] Compound 9 (500.0 mg, 1.8 mmol, 1.0 eq.) was dissolved in tetrahydrofuran (300 mL), and lithium aluminum deuteride (83 mg, 2.0 mmol, 1.1 eq.) was added to the solution. The mixture was stirred at 0°C for 2 hours. The reaction mixture was extracted with water and dichloromethane and concentrated to give compound 10 (400 mg, yellow oil, 97.5% yield). MS-ESI: m / z 228 [M+1] +

[0188] 3. Preparation of Compound 11

[0189]

[0190] Compound 10 (400.0 mg, 1.8 mmol, 1.0 eq.) was dissolved in dichloromethane (200 mL), and triethylamine (545.0 mg, 5.4 mmol, 3.0 eq.) and methanesulfonyl chloride (311.0 mg, 2.7 mmol, 1.5 eq.) were added to the solution. The mixture was stirred at room temperature for 3 hours. The reaction mixture was extracted with water and dichloromethane and concentrated to give compound 11 (400 mg, yellow oil, 58.1% yield). MS-ESI: m / z 384 [M+1] +

[0191] 4. Preparation of Compound 12

[0192]

[0193] Compound 11 (400.0 mg, 1.0 mmol, 1.0 eq.) was dissolved in benzylamine (200 mL). The mixture was stirred at room temperature for 16 hours. The reaction mixture was extracted with water and dichloromethane, concentrated, and purified by column chromatography to obtain compound 12 (200 mg, yellow solid, yield 64.7%). MS-ESI: m / z 299 [M+1]+

[0194] 5. Preparation of Compound 13

[0195]

[0196] Compound 12 (200 mg, 0.7 mmol, 1.0 eq.) was dissolved in a 1,4-dioxane (4 M) solution of hydrochloric acid. The mixture was stirred at room temperature for 18 hours. The reaction mixture was filtered and concentrated to give compound 13 (150 mg, yellow oil, 100% yield). MS-ESI: m / z 199 [M+1] +

[0197] 6. Preparation of Compound H7-1

[0198]

[0199] Compound 13 (150.0 mg, 0.6 mmol, 1.0 eq.) and compound 4 (215.0 mg, 0.6 mmol, 1.0 eq.) in the previous step were dissolved in acetonitrile (15 mL), and potassium carbonate (248.0 mg, 1.8 mmol, 3.0 eq.) was added to the solution. The mixture was stirred at 110° C. for 4 hours. The reaction mixture was extracted with water and dichloromethane and concentrated to give compound H7-1 (300 mg, yellow solid, 94.0% yield). MS-ESI: m / z 521 [M+1] + , 1 H NMR(400MHz,DMSO-d6)δ:9.40–9.20(m,1H),8.23(d,1H),8.00–7.90(m,1H),7.55–7.35(m,2H), 7.35–7.15(m,5H),7.00–6.85(m,1H),4.29(q,2H),3.85–3.60(m,2H),3.59(s,2H),1.30(t,3H).

[0200] 7. Preparation of Compound H7-3

[0201]

[0202] N,N-diisopropylethylamine (1.1 g, 8.64 mmol, 2.5 eq.) was added to a solution of compound H7-1 (1.8 g, 3.46 mmol, 1.0 eq.) in 1,2-dichloroethane (30 mL). Compound H7-2 (0.99 g, 6.91 mmol, 2.0 eq.) was then added at 0 °C, and the mixture was stirred at room temperature for 1 hour under nitrogen protection. The reaction mixture was then dried and dissolved in methanol (30 mL). The mixture was stirred at 70 °C under nitrogen protection for 1 hour. The reaction solution was dried, dichloromethane / ethyl acetate (10 mL / 10 mL) was added, and the mixture was stirred at room temperature overnight. It was then filtered, the filter cake was washed with dichloromethane, and dried to obtain the crude product of compound H7-3 (1.64 g, yellow solid). LC-MS (ESI): m / z 431.0 [M+1] + , 1 H NMR(400MHz,DMSO-d6)δ:9.35–9.15(m,2H),8.23(d,1H),8.00–7.90(m,1H),7.60–7 .50(m,1H),7.50–7.40(m,1H),7.00(d,1H),4.29(q,2H),3.83(s,2H),1.30(t,3H).

[0203] 8. Preparation of Compound H7-4

[0204]

[0205] Compound H7-3 (200.0 mg, 0.5 mmol, 1.0 eq.) was dissolved in acetonitrile (10 mL), potassium carbonate (352.0 mg, 2.6 mmol, 5.0 eq.) and iodomethane-d3 (81.0 mg, 0.6 mmol, 1.1 eq.) were added to the solution, and the mixture was stirred at 50°C for 2 hours. The reaction mixture was cooled to room temperature, the aqueous layer was extracted with dichloromethane, and concentrated to give compound H7-4 (60 mg, yellow solid, 23.0% yield). MS-ESI: m / z 448 [M+1] + , 1 H NMR(400MHz,DMSO-d6)δ:9.40–9.30(m,1H),8.37(d,1H),7.90–7.80(m,1H), 7.60–7.40(m,2H),6.94(d,1H),4.28(q,2H),3.85–3.55(m,2H),1.30(t,3H).

[0206] 9. Preparation of Compound H7-5

[0207]

[0208] Compound H7-4 (60.0 mg, 0.1 mmol, 1.0 eq.) was dissolved in tetrahydrofuran / water (1:1, v / v, 4 mL) and lithium hydroxide (10.0 mg, 0.4 mmol, 3.0 eq.) was added. The mixture was stirred at room temperature for 18 hours. Concentration gave compound H7-5, which was used directly in the next reaction (70 mg, yellow solid, 100.0% yield). MS-ESI: m / z 420 [M+1] + , 1 H NMR(400MHz, DMSO-d6)δ:9.40–9.30(m,1H),8.23(d,1H),8.00–7.90(m,1H),7.55–7.45(m,1H),7.45–7.35(m,1H),6.99(d,1H),3.90–3.60(m,2H).

[0209] 10. Preparation of H-7

[0210]

[0211] 2-(7-azabenzotriazole)-tetramethyluronium hexafluorophosphate (41 mg, 0.107 mmol, 1.5 eq) and N,N-diisopropylethylamine (28 mg, 0.215 mmol, 3.0 eq) were added to a solution of compound H7-5 (30 mg, 0.072 mmol, 1.0 eq.) in N,N-dimethylformamide (2 mL). The mixture was stirred at 50 ° C for 0.5 hours under nitrogen protection. Then 2-methylaminopyrimidine compound H7-6 (12 mg, 0.107 mmol, 1.5 eq.) dissolved in N,N-dimethylformamide (1 mL) was added, and the mixture was stirred at 50 ° C for 16 hours under nitrogen protection. The reaction mixture was spin-dried to obtain a residue, which was separated and purified by preparative HP LC, and the preparative solution was lyophilized to obtain compound H-7 (11 mg, yellow solid, yield 30%). LC-MS(ESI):m / z 511.2[M+1] + , 1 HNMR(400MHz,DMSO-d6)δ:11.25–11.10(m,1H),9.50–9.35(m,1H),8.88(s,2H),8.40–8.30(m,1H) ),8.05–7.90(m,1H),7.60–7.35(m,2H),7.10–6.95(m,1H),4.85–4.70(m,2H),3.95–3.70(m,2H).

[0212] Example 7 Preparation of Compound H-8

[0213]

[0214] Potassium carbonate (4.3 g, 31.3 mmol, 5.0 eq.) was added to a solution of compound 4 (1.7 g, 6.27 mmol, 1.0 eq.) in acetonitrile (50 mL). Compound 13 (2.5 g, 6.89 mmol, 1.1 eq.) was added, and the mixture was stirred at 80 ° C for 16 hours under nitrogen protection. The reaction mixture was then cooled to room temperature, washed with dichloromethane / methanol (10 / 1, v / v), dried and spin-dried to obtain a residue. The residue was separated and purified by silica gel column chromatography to obtain compound H8-1 (2.8 g, light yellow solid, yield 72%). LC-MS (ESI): m / z 521.2 [M+1] + , 1 H NMR(400MHz,DMSO-d6)δ:9.50 -9.25(m,1H),8.23(d,1H),8.00–7.90(m,1H),7.60–7.35(m,2H),7.30–7.10(m,5H),7.00–6.90(m,1H),4.29(q,2H),4.00 -3.50(m,4H),1.29(t,3H).

[0215]

[0216] N,N-diisopropylethylamine (1.1 g, 8.64 mmol, 2.5 eq.) was added to a solution of compound H8-1 (1.8 g, 3.46 mmol, 1.0 eq.) in 1,2-dichloroethane (30 mL). Compound H8-2 (0.99 g, 6.91 mmol, 2.0 eq.) was then added at 0 °C and the mixture was stirred at room temperature for 1 hour under nitrogen protection. The reaction mixture was then dried and dissolved in methanol (30 mL). The mixture was stirred at 70 °C under nitrogen protection for 1 hour. The reaction solution was dried, dichloromethane / ethyl acetate (10 mL / 10 mL) was added, and the mixture was stirred at room temperature overnight. It was then filtered, the filter cake was washed with dichloromethane, and dried to obtain a crude product of compound H8-3 (1.64 g, yellow solid). LC-MS (ESI): m / z 431.0 [M+1] +

[0217]

[0218] Potassium carbonate (0.58 g, 4.17 mmol, 3.0 eq.) was added to a solution of compound H8-3 (0.60 g, 1.39 mmol, 1.0 eq.) in N,N-dimethylformamide (10 mL). The mixture was stirred at 45 ° C for 0.5 hours, and then iodomethane-d3 (0.2 g, 1.39 mmol, 1.0 eq.) was added, and the mixture was stirred at 45 ° C for 0.5 hours under nitrogen protection. Then water was added to quench and the aqueous phase was extracted with dichloromethane / methanol (5 / 1, v / v, 150 mLx5). The combined organic layer was concentrated to obtain a residue. The residue was purified by silica gel column chromatography) to obtain compound H8-4 (recovered raw materials and re-reacted, a total of two reactions) (0.30 g, light yellow solid, two-step yield 40%). LC-MS (ESI): m / z 448.2 [M+1] + , 1 H NMR(400MHz,DMSO-d6)δ:9.50–9.35(m,1H),8.23(d,1H),7.95–7.85(m,1H),7.6 0–7.35(m,2H),7.00–6.90(m,1H),4.28(q,2H),4.00–3.60(m,2H),1.29(t,3H).

[0219]

[0220] LiOH·H2O (0.17 g, 4.02 mmol, 2.5 eq.) was added to a mixture of compound H8-4 (0.72 g, 1.61 mmol, 1.0 eq.) in tetrahydrofuran (8 mL), methanol (4 mL) and water (8 mL). The reaction mixture was stirred at room temperature for 16 hours under nitrogen protection. The reaction mixture was concentrated to obtain a residue, which was washed with water and filtered. The filter cake was dried to obtain compound H8-5 (0.78 g, light yellow solid), which was used directly in the next step. LC-MS (ESI): m / z 419.8 [M+1] + , 1 HNMR(400MHz, DMSO-d6)δ:9.50–9.35(m,1H),8.38(d,1H),7.95–7.85(m,1H),7.60–7.35(m,2H),7.10–6.95(m,1H),4.00–3.60(m,2H).

[0221]

[0222] 2-(7-azabenzotriazole)-tetramethyluronium hexafluorophosphate (0.13 g, 0.34 mmol, 1.5 eq.) and N,N-diisopropylethylamine (0.12 mL, 0.68 mmol, 3.0 eq.) were added to a solution of compound H8-6 (0.095 g, 0.27 mmol, 1.0 eq.) in N,N-dimethylformamide (2 mL). The mixture was stirred at 50 ° C for 0.5 hours under nitrogen protection. Then compound H8-7 (0.056 g, 0.34 mmol, 1.5 eq.) dissolved in N,N-dimethylformamide (1 mL) was added and the mixture was stirred at 50 ° C for 16 hours under nitrogen protection. The reaction solution was spin-dried to obtain a residue, and the residue was separated and purified by preparative HPLC, and the preparative solution was lyophilized to obtain compound H-8 (0.012 g, light yellow solid, 10% yield). LC-MS(ESI):m / z 529.2[M+1] + , 1 HNMR(400MHz,DMSO-d6)δ:11.25–11.10(m,1H),9.50–9.35(m,1H),8.88(s,2H),8.40–8.30(m,1H) ),8.05–7.90(m,1H),7.60–7.35(m,2H),7.10–6.95(m,1H),4.85–4.70(m,2H),3.95–3.70(m,2H).

[0223] Example 8 Preparation of Compound H-11

[0224]

[0225] Raney-Ni (0.08 g) and NH3.H2O (0.2 mL) were added to a solution of compound H11-1 (0.5 g, 2.72 mmol) in EtOH (20 mL). The mixture was stirred at room temperature for 2 hours under a hydrogen atmosphere (3.5 MPa). The filtrate was filtered and dried to obtain a residue. The residue was used directly in the next step without further purification. LC-MS (ESI): m / z 188.0 [M+1] +

[0226]

[0227] 2-(7-azabenzotriazole)-tetramethyluronium hexafluorophosphate (0.23 g, 0.6 mmol, 1.5 eq.) and N,N-diisopropylethylamine (0.16 g, 1.2 mmol, 3.0 eq.) were added to a solution of compound 7 (0.16 g, 0.4 mmol, 1.0 eq.) in N,N-dimethylformamide (5 mL). The mixture was stirred at 50 ° C for 0.5 hours under nitrogen protection. Then, compound H11-2 (0.3 g, 1.6 mmol, 4.0 eq.) dissolved in N,N-dimethylformamide (2 mL) was added and the mixture was stirred at 50 ° C for 16 hours under nitrogen protection. The reaction solution was spin-dried to obtain a residue, the residue was slurried with methanol, filtered and the filter residue was washed, and dried to obtain compound H11-3 (0.2 g, white solid, 56% yield). LC-MS: m / z 578.00,580.00[M+1] + , 1 H NMR(400MHz, DMSO-d6)δ:11.15–11.00(m,1H),9.35–9.15(m,1H),8.98(s,2H),8.45–8.30(m,1H),8.05–7.90 (m,1H),7.65–7.30(m,2H),7.10–6.95(m,1H),4.85–4.65(m,2H),4.20–3.70(m,4H),3.60–3.40(m,2H),3.10 -2.60(m,5H),2.30–2.15(m,2H).

[0228]

[0229] Zinc cyanide (0.037 g, 0.31 mmol, 1.5 eq.), zinc (0.007 g, 0.104 mmol, 0.5 eq.), tris(dibenzylideneacetone)dipalladium (0.02 g, 0.021 mmol, 0.1 eq.) and 1,1'-bis(diphenylphosphino)ferrocene (0.023 g, 0.04 mmol, 0.2 eq.) were added to a DMA (6 mL) solution of compound H11-3 (0.12 g, 0.21 mmol, 1.0 eq.). The mixture was sealed at 110 ° C for 2 hours under nitrogen protection. The reaction solution was concentrated, the residue was slurried with methanol, the filter residue was separated and purified by preparative HPLC, and the preparative solution was lyophilized to obtain compound H-11 (0.0199 g, light yellow solid, yield 19%). LC-MS (ESI): m / z 525.2 [M+1] + , 1H NMR(400MHz, DMSO-d6)δ:11.15–11.05(m,1H),9.29(s,2H),9.25–9.15(m,1H),8.27(d,1H),7.95–7.85(m,1H),7.55–7.30(m, 2H),7.05–6.85(m,1H),4.87(d,2H),4.05–3.85(m,2H),3.85–3.60(m,2H),3.15–2.85(m,4H),2.37(s,3H),2.15–2.05(m,2H).

[0230] Example 9 Preparation of Compound H-12

[0231]

[0232] Compound H12-1 (2 g, 15.557 mmol, 1.0 eq.) was dissolved in carbon tetrachloride (16 mL), and NBS (3.1 g, 17.113 mmol, 1.1 eq.) and AIBN (0.51 g, 3.111 mmol, 0.2 eq.) were added, and stirred at 80°C overnight. Ethyl acetate and water were added for washing, and the organic phase was dried and concentrated, and then separated and purified by column chromatography to finally obtain compound H12-2 (500 mg, yellow solid, yield 15%). 1 H NMR (400MHz, DMSO-d6)δ:8.79(d,1H),7.64(d,1H),4.64(s,2H).

[0233]

[0234] NH3.H2O (14 mL) was added to a solution of compound H12-2 (500 mg, 2.410 mmol, 1.0 eq.) in methanol (7 mL), and the mixture was stirred at room temperature for 1 hour. Dichloromethane / methanol (10 / 1, v / v) was added for washing, and the organic phase was dried and concentrated, and then purified by column chromatography to finally obtain compound H12-3 (450 mg, yellow solid, yield 90%). 1 H NMR (400MHz, DMSO-d6)δ:8.85(d,1H),8.42(s,2H),7.75(d,1H),4.30(s,2H).

[0235]

[0236] Saturated sodium bicarbonate aqueous solution (3.2 mL) and di-tert-butyl dicarbonate (396 mg, 1.811 mmol, 1.3 eq.) were added to a solution of compound H12-3 (200 mg, 1.393 mmol, 1.0 eq.) in dichloromethane (4 mL). The reaction solution was stirred at room temperature for 1 hour under a nitrogen atmosphere. Ethyl acetate and water were added for extraction, and the organic phase was dried and concentrated, and then purified by column chromatography to finally obtain compound H12-4 (150 mg, white solid, yield 44%).

[0237] LC-MS(ESI):m / z 244.7[M+1] +

[0238]

[0239] Zinc cyanide (51 mg, 0.431 mmol, 0.7 eq.) and zinc (16 mg, 246 mmol, 0.4 eq.) were added to a solution of compound H12-4 (150 mg, 0.615 mmol, 1.0 eq.) in N,N-dimethylformamide (5 mL), and then 1,1'-bis(diphenylphosphino)ferrocene (21 mg, 0.036 mmol, 0.06 eq.) was added, and finally tri(dibenzylideneacetone)dipalladium (17 mg, 0.018 mmol, 0.03 eq.) was added, and the mixture was stirred at 110°C overnight under nitrogen protection. Ethyl acetate and water were added for washing, and the organic phase was dried and concentrated, and then separated and purified by column chromatography to finally obtain compound H12-5 (70 mg, light yellow solid, yield 49%). LC-MS (ESI): 135.1 [M+1] + (without Boc group), 1 H NMR (400MHz, DMSO-d6)δ:9.07(d1H),7.98(d,1H),7.38(s,1H),4.34(d,2H),1.36(s,9H).

[0240]

[0241] Compound H12-5 (70 mg, 0.299 mmol, 1.0 eq) was dissolved in HCl / dioxane (3 mL) and stirred at room temperature overnight. The solvent was removed by concentration to obtain a crude product of compound H12-6 (50 mg, yellow solid).

[0242] LC-MS(ESI):m / z 135.1[M+1] + , 1H NMR (400MHz, DMSO-d6)δ:9.07(d,1H),8.62(s,2H),7.93(d,1H),4.37–4.34(m,2H).

[0243]

[0244] 2-(7-azabenzotriazole)-tetramethyluronium hexafluorophosphate (155 mg, 0.410 mmol, 1.5 eq) and N,N-diisopropylethylamine (105 mg, 0.808 mmol, 3.0 eq) were added to a solution of compound H12-6 (110 mg, 0.273 mmol, 1.0 eq.) in N,N-dimethylformamide (3 mL). The mixture was stirred at 50 ° C for 0.5 hours under nitrogen protection. Then compound 7 (55 mg, 0.410 mmol, 1.5 eq.) dissolved in N,N-dimethylformamide (1 mL) was added, and the mixture was stirred at 50 ° C for 16 hours under nitrogen protection. The reaction mixture was spin-dried to obtain a residue. Preparation HPLC separation and purification finally gave compound H-12 (12.9 mg, light yellow solid, two-step yield 10%). LC-MS (ESI): m / z 525.2 [M+1] + , 1 H NMR(400MHz, DMSO-d6)δ:11.20–11.10(m,1H),9.45–9.35(m,1H),9.13(d,1H),8.45–8.35(m,1H),8.30–8.20(m,1H),8.10–7.95(m,2H),7.60 –7.40(m,2H),7.10–7.00(m,1H),4.90–4.80(m,2H),3.80–3.40(m,4H) ,2.80–2.65(m,1H),2.60–2.50(m,2H),2.25(s,3H),2.05–1.90(m,2H).

[0245] Example 10 Preparation of Compound H-13

[0246]

[0247] Compound H13-1 (4.67 g, 33.81 mmol, 1.0 eq.) was dissolved in a mixed solution of tetrahydrofuran (70 mL) and dichloromethane (70 mL), EDCI (9.72 g, 50.713 mmol, 1.5 eq.), HOBT (6.85 g, 50.713 mmol, 1.5 eq.), N,N-diisopropylethylamine (13.10 g, 101.426 mmol, 3.0 eq.) was added, and finally methylamine hydrochloride (2.1 g, 67.617 mmol, 2.0 eq.) was added, and the mixture was stirred at room temperature overnight. The reaction solution was concentrated, separated and purified by column chromatography to obtain compound H13-2 (1.31 g, light yellow solid, yield 25%).

[0248] LC-MS(ESI):m / z 152.2[M+1] + , 1 H NMR (400MHz, DMSO-d6)δ:9.00(s,2H),8.70(d,1H),2.77(d,3H),2.63(s,3H).

[0249]

[0250] 4-Dimethylaminopyridine (0.529 g, 4.333 mmol, 0.5 eq.), di-tert-butyl dicarbonate (3.782 g, 17.331 mmol, 2.0 eq.) were added to a solution of compound H13-2 (1.31 g, 8.666 mmol, 1.0 eq.) in dichloromethane (35 mL), and triethylamine (4.384 g, 43.329 mmol, 5.0 eq.) was added, and the mixture was stirred at room temperature overnight. Dichloromethane and water were added for washing, and the organic phase was dried and concentrated, and then separated and purified by column chromatography to obtain compound H13-3 (1.77 g, light yellow solid, yield 81%).

[0251] LC-MS(ESI):m / z 252.3[M+1] + , 1 H NMR(400MHz, DMSO-d6)δ:8.76(s,2H),3.17(s,3H),2.64(s,3H),1.17(s,9H).

[0252]

[0253] N-bromosuccinimide (1.38 g, 7.748 mmol, 1.1 eq.), azobisisobutyronitrile (0.116 g, 0.7043 mmol, 0.1 eq.) and benzoyl peroxide (0.171 g, 0.7043 mmol, 0.1 eq.) were added to a solution of compound H13-3 (1.77 g, 7.043 mmol, 1.0 eq.) in carbon tetrachloride (50 mL). The reaction solution was stirred at 80 ° C overnight under a nitrogen atmosphere. Dichloromethane / methanol (10 / 1, v / v) and water were added for washing. After the organic phase was dried and concentrated, it was separated and purified by column chromatography to obtain compound H13-4 (690 mg, white oil, yield 30%). LC-MS (ESI): m / z 331.2 [M+1] + , 1 H NMR(400MHz, DMSO-d6)δ:8.91(s,1H),4.71(s,2H),3.18(s,3H),1.16(s,9H).

[0254]

[0255] H13-4 (690 mg, 2.09 mmol, 1.0 eq.) was dissolved in dichloromethane (5 ml), trifluoroacetic acid (2.38 g, 20.89 mmol, 10 eq.) was added, and stirred at room temperature overnight. The solvent was removed to obtain compound H13-5 (740 mg, dark yellow solid), which was used directly in the next step. LC-MS (ESI): m / z 252.3 [M+1] + , 1 H NMR (400MHz, DMSO-d6)δ:9.11(s,2H),4.70(s,2H),2.78(d,3H).

[0256]

[0257] NH3.H2O (15 mL) was added to a solution of compound H13-5 (740 mg, 3.216 mmol, 1.0 eq.) in methanol (10 mL). The mixture was stirred at room temperature overnight. The solvent was removed and lyophilized to obtain compound H13-6 (754 mg), which was used directly in the next step. LC-MS (ESI): m / z 167.2 [M+1] + , 1 H NMR (400MHz, DMSO-d6)δ:9.18(s,2H),8.87(d,1H),4.34(s,2H),2.80(d,3H).

[0258]

[0259] 2-(7-azabenzotriazole)-tetramethyluronium hexafluorophosphate (1.5 eq.) and N,N-diisopropylethylamine (4.0 eq.) were added to a solution of compound 7 (754 mg, 1.0 eq.) in N,N-dimethylformamide (15 mL). The mixture was stirred at 50 ° C for 0.5 hours under nitrogen protection. Then compound H13-6 (2.5 eq.) dissolved in N,N-dimethylformamide (5 mL) was added. The mixture was stirred at 50 ° C overnight under nitrogen protection. The reaction mixture was spin-dried to obtain a residue. Methanol (8 mL) was added and slurried overnight, filtered, the filter residue was collected, and dried to obtain compound H-13 (212.3 mg, light yellow solid, three-step yield 21%). LC-MS (ESI): m / z 557.2 [M+1] + , 1 H NMR(400MHz, DMSO-d6)δ:11.25–11.15(m,1H),9.45–9.35(m,1H),9.11(s,2H),8.80–8.75(m,1H),8.33(d,1H),8.00–7.90(m,1H),7.60–7.35(m ,2H),7.44(d,J=7.6Hz,1H),7.05–6.95(m,1H),4.84(d,2H),4.05–3.60 (m,4H),2.80(d,3H),2.75–2.65(m,2H),2.25(s,3H),2.05–1.90(m,2H).

[0260] Example 11 Preparation of Compound H-15

[0261]

[0262] Compound H15-1 (2 g, 14.48 mmol, 1.0 eq.) was dissolved in a mixed solution of N,N-dimethylformamide (0.01 mL) and dichloromethane (80 mL), and oxalyl chloride (2.76 g, 21.72 mmol, 1.5 eq.) was added at 0°C, and the mixture was returned to room temperature and stirred overnight. The reaction solution was concentrated to obtain a crude product of compound H15-2 (1.5 g, light yellow solid, yield 60%), which was used directly in the next step.

[0263] LC-MS(ESI):m / z 166.2[M+1] +

[0264]

[0265] N-bromosuccinimide (1.75 g, 9.85 mmol, 1.1 eq.) and azobisisobutyronitrile (441 mg, 2.69 mmol, 0.3 eq.) were added to a solution of compound H15-2 (1.48 g, 8.96 mmol, 1.0 eq.) in carbon tetrachloride (10 mL). The mixture was stirred at 80°C overnight. Dichloromethane and water were added for washing, and the organic phase was dried and concentrated, and then separated and purified by column chromatography to obtain compound H15-3 (240 mg, light yellow oil, yield 10%).

[0266] LC-MS(ESI):m / z 245.1[M+1] + , 1 H NMR(400MHz, DMSO-d6)δ:8.88(s,2H),4.69(s,2H),2.98(s,3H),2.93(s,3H).

[0267]

[0268] NH3.H2O (6 ml) was added to a methanol (3 mL) solution of compound H15-3 (200 mg, 0.82 mmol, 1.0 eq). The mixture was stirred at room temperature for 2 hours. The reaction solvent was removed and lyophilized to obtain compound H15-4 (180 mg, yellow solid), which was used directly in the next step. LC-MS (ESI): m / z 181.2 [M+1] + , 1 H NMR(400MHz, DMSO-d6)δ:8.96(s,2H),4.34(s,2H),2.99(s,3H),2.93(s,3H).

[0269]

[0270] 2-(7-azabenzotriazole)-tetramethyluronium hexafluorophosphate (148 mg, 0.39 mmol, 1.5 eq.) and N,N-diisopropylethylamine (101 mg, 0.78 mmol, 3.0 eq.) were added to a solution of compound 7 (106 mg, 0.26 mmol, 1.0 eq.) in N,N-dimethylformamide (2 mL). The mixture was stirred at 50 ° C for 0.5 hours under nitrogen protection. Then compound H15-4 (70 mg, 0.39 mmol, 1.5 eq.) dissolved in N,N-dimethylformamide (1 mL) was added, and the mixture was stirred at 50 ° C overnight under nitrogen protection. The reaction solvent was concentrated to obtain a residue. Methanol (3 ml) was added and slurried overnight, filtered, and the filter residue was collected and dried to obtain compound H-15 (19.9 mg, white solid, two-step yield 19%). LC-MS(ESI):m / z 571.4[M+1] + , 1 H NMR(400MHz, DMSO-d6)δ:11.25–11.10(m,1H),9.40–9.25(m,1H),8.88(s,2H),8.45–8.35(m,1H),8.05–7.95(m,1H),7.65–7.35(m,2H) ,7.10–7.00(m,1H),4.86(d,2H),4.05–3.60(m,4H),3.50–3.40(m,4H),2.99(s,3H),2.96(s,3H),2.65–2.50(m,3H),2.20–2.05(m,2H).

[0271] Example 12 Preparation of Compound H-16

[0272]

[0273] Raney-Ni (0.02 g) and NH3.H2O (0.1 mL) were added to a solution of compound H16-1 (0.15 g, 1.11 mmol) in ethanol (6 mL). The mixture was stirred at room temperature for 2 hours under a hydrogen atmosphere (3.5 MPa). The filtrate was filtered and dried to obtain a residue. The residue was used directly in the next step without further purification. LC-MS (ESI): m / z 140.0 [M+1] +

[0274]

[0275] 2-(7-azabenzotriazole)-tetramethyluronium hexafluorophosphate (103 mg, 0.27 mmol, 1.5 eq.) and N,N-diisopropylethylamine (0.1 mL, 0.54 mmol, 3.0 eq.) were added to a solution of compound 7 (74 mg, 0.18 mmol, 1.0 eq.) in N,N-dimethylformamide (3 mL). The mixture was stirred at 50 ° C for 0.5 hours under nitrogen protection. Then, crude compound H16-2 (100 mg, 0.72 mmol, 4.0 eq.) dissolved in N,N-dimethylformamide (1 mL) was added, and the mixture was stirred at 50 ° C for 16 hours under nitrogen protection. The reaction solution was spin-dried to obtain a residue, and the residue was separated and purified by preparative HPLC to obtain compound H-16 (125 mg, light yellow solid, two-step yield 20%). LC-MS (ESI): m / z 530.2 [M+1] + , 1 H NMR(400MHz, DMSO-d6)δ:11.15–11.05(m,1H),9.45–9.30(m,1H),8.54(s,2H),8.35–8.20(m,1H),8.00–7.85(m,1H),7.60–7.30(m, 2H),7.05–6.85(m,1H),4.80–4.60(m,2H),4.00–3.55(m,7H),2.80–2.60(m,2H),2.60–2.50(m,2H),2.24(s,3H),2.05–1.90(m,2H).

[0276] Example 13 Preparation of Compound H-17

[0277]

[0278] Compound H17-1 (100 mg, 0.74 mmol, 1.0 eq.) was dissolved in ethanol (5 mL), Raney-Ni (40 mg,) and aqueous ammonia (47%, 0.1 mL) were added, and stirred at room temperature for 2 hours under a hydrogen atmosphere (3 MPa). Filter and freeze-dry to obtain compound H17-2 (40 mg, red oil, yield 38%). LC-MS (ESI): m / z 140.2 [M+1] + , 1 H NMR(400MHz, DMSO-d6)δ:8.42(s,1H),6.74(s,1H),3.87(s,3H),3.72(s,2H).

[0279]

[0280] Compound 7 (59 mg, 0.14 mmol, 1.0 eq.) was dissolved in N, N-dimethylformamide (2 mL), and 2-(7-azabenzotriazole)-tetramethyluronium hexafluorophosphate (84 mg, 0.22 mmol, 1.5 eq.) and N, N-diisopropylethylamine (57 mg, 0.44 mmol, 3.0 eq.) were added thereto. The reaction solution was reacted at 50 ° C under nitrogen protection for 0.5 hours, and then compound H17-2 (40 mg, 0.28 mmol, 2.0 eq.) was added, and the reaction solution continued to react at 50 ° C for 16 hours. The reaction solution was concentrated, and after removing N, N-dimethylformamide, methanol (2 mL) was added. The reaction solution was stirred at room temperature overnight. Filtered, and then the filter cake was washed with methanol, and prepared by HPLC separation and purification to obtain compound H-17 (3.1 mg, light yellow solid, 3.9% yield). LC-MS(ESI):m / z 530.2[M+1] + , 1 H NMR(400MHz, DMSO-d6)δ:11.25–11.15(m,1H),9.50–9.40(m,1H),8.49(d,1H),8.40–8.30(m,2H),7.98(d,1H),7.60–7.40(m,2H),7. 10–6.96(m,1H),6.83(d,1H),4.67(d,2H),4.05–3.60(m,7H),2.80–2.65(m,2H),2.55–2.50(m,2H),2.26(s,3H),2.05–1.90(m,2H).

[0281] Example 14 Preparation of Compound H-18

[0282]

[0283] Compound H18-1 (100 mg, 0.544 mmol, 1.0 eq.) was dissolved in N,N-dimethylformamide (2 mL), CH3SNa (229 mg, 0.652 mmol, 1.2 eq.) was added, and the mixture was stirred at 0°C for 1 hour. Ethyl acetate and water were added for extraction, and the organic phase was dried and concentrated, and then purified by column chromatography to obtain compound H18-2 (77 mg, light yellow solid, yield 93%). 1 H NMR(400MHz,DMSO-d6)δ:8.86(s,2H),2.63(s,3H).

[0284]

[0285] To a solution of compound H18-2 (77 mg, 0.509 mmol, 1.0 eq.) in ethanol (3 mL) was added NH3.H2O (194 mg, 5.534 mmol, 11 eq.) and Raney-Ni (18 mg, 0.307 mmol, 0.6 eq.). The mixture was stirred at room temperature overnight under a hydrogen atmosphere. Filtered with diatomaceous earth, the filtrate was concentrated, and then lyophilized with water to obtain compound H18-3 (61 mg, light yellow solid, yield 34%). LC-MS (ESI): m / z 156.0 [M+1] +

[0286]

[0287] 2-(7-azabenzotriazole)-tetramethyluronium hexafluorophosphate (185 mg, 0.485 mmol, 1.5 eq.) and N,N-diisopropylethylamine (126 mg, 0.966 mmol, 3.0 eq.) were added to a solution of compound 7 (132 mg, 0.322 mmol, 1.0 eq.) in N,N-dimethylformamide (4 mL). The mixture was stirred at 50 ° C for 0.5 hours under nitrogen protection. Then compound H18-3 (150 mg, 0.966 mmol, 3.0 eq.) dissolved in N,N-dimethylformamide (1 mL) was added, and the mixture was stirred at 50 ° C for 16 hours under nitrogen protection. The reaction mixture was dried to obtain a residue. It was extracted with ethyl acetate and water, and the organic phase was dried and concentrated and separated and purified by preparative HPLC to finally obtain compound H-18 (21 mg, light yellow solid, yield 12%). LC-MS(ESI):m / z 546.1[M+1] + , 1 H NMR(400MHz, DMSO-d6)δ:11.15–11.05(m,1H),9.40–9.30(m,1H),8.75–8.65(m,2H),8.45–8.35(m,1H),8.05–7.90(m,1H),7.65–7.40(m,2H) ,7.10–7.00(m,1H),4.80–4.70(m,2H),4.25–3.95(m,2H),3.90–3.70( m,2H),3.60–3.45(m,4H),2.79(s,3H),2.58(s,3H),2.40–2.25(m,2H).

[0288] Example 15 Preparation of Compound H-19

[0289]

[0290] Compound H19-1 (50 mg, 0.209 mmol, 1.0 eq.) was dissolved in HCl / dioxane (2 mL) and stirred at room temperature overnight. The solvent was removed by concentration to obtain a crude product of compound H19-2 (35 mg, light yellow solid). LC-MS (ESI): m / z 139.2 [M+1] + , 1 H NMR (400MHz, DMSO-d6)δ:8.37(d,3H),8.11(s,2H),4.03(d,2H),2.70(s,3H).

[0291]

[0292] 2-(7-azabenzotriazole)-tetramethyluronium hexafluorophosphate (98 mg, 0.253 mmol, 1.5 eq) and N,N-diisopropylethylamine (67 mg, 0.514 mmol, 3.0 eq) were added to a solution of compound 7 (70 mg, 0.169 mmol, 1.0 eq.) in N,N-dimethylformamide (2 mL). The mixture was stirred at 50 ° C for 0.5 hours under nitrogen protection. Then compound H19-2 (35 mg, 0.253 mmol, 1.5 eq.) dissolved in N,N-dimethylformamide (1 mL) was added, and the mixture was stirred at 50 ° C for 16 hours under nitrogen protection. The solvent was concentrated to remove the solvent, and the mixture was separated and purified by preparative HPLC to finally obtain compound H-19 (6.1 mg, light yellow solid, two-step yield 7%). LC-MS (ESI): m / z 529.2 [M+1] + , 1 H NMR(400MHz,DMSO-d6)δ:11.00–10.90(m,1H),9.25–9.10(m,1H),8.30 -8.20(m,1H),8.11(s,2H),7.95–7.85(m,1H),7.55–7.35(m,2H),7.00–6.90(m,1H),6.10–6.00(m,1H),4.65 –4.55(m,2H),4.10–3.95(m,2H),3.80–3.40(m,4H),3.20–3.05(m,2H),2.75–2.60(m,6H),2.30–2.20(m,2H).

[0293] Example 16 Preparation of Compound H-20

[0294]

[0295] HCl-dioxane (2 mL) was added to compound H20-1 (0.03 g, 0.12 mmol). The mixture was stirred at room temperature for 2 hours under nitrogen protection. The solvent was removed by concentration to obtain compound H20-2 (25 mg, yellow solid, yield 97%). LC-MS (ESI): m / z 153.2 [M+1] +

[0296]

[0297] 2-(7-azabenzotriazole)-tetramethyluronium hexafluorophosphate (0.06 g, 0.16 mmol, 1.5 eq.) and N,N-diisopropylethylamine (0.075 mL, 0.42 mmol, 4.0 eq.) were added to a solution of compound 7 (0.044 g, 0.11 mmol, 1.0 eq.) in N,N-dimethylformamide (2 mL). The mixture was stirred at 50 ° C for 0.5 hours under nitrogen protection, and compound H20-2 (0.03 g, 0.16 mmol, 1.5 eq.) dissolved in N,N-dimethylformamide (1 mL) was added, and the mixture was stirred at 50 ° C for 16 hours under nitrogen protection. The reaction solution was concentrated to obtain a residue. The residue was separated and purified by preparative HPLC, and the preparative solution was lyophilized to obtain compound H-20 (0.029 g, light yellow solid, yield 50%). LC-MS(ESI):m / z 543.2[M+1] + , 1 H NMR(400MHz, DMSO-d6)δ:11.10–11.00(m,1H),9.45–9.35(m,1H),8.35–8.30(m,1H),8.28(s,2H),8.00–7.90(m,1H),7.60–7.35(m,2H) ,7.05–6.95(m,1H),4.63(d,2H),4.05–3.60(m,4H),2.92(s,6H),2.80–2.70(m,2H),2.55–2.45(m,2H),2.25(s,3H),2.05–1.90(m,2H).

[0298] Example 17 Preparation of Compound H-21

[0299]

[0300] HCl-dioxane (4M, 3 mL) was added to compound H21-1 (0.05 g, 0.25 mmol). The mixture was stirred at room temperature for 2 hours under nitrogen protection. The solvent was removed by concentration to obtain compound H21-2 (30 mg, gray solid, yield 97%). LC-MS (ESI): m / z 138.4 [M+1] + , 1 H NMR (400MHz, DMSO-d6)δ:8.73(s,2H),8.49(s,3H),4.21(q,2H),2.63(q,2H),1.18(t,3H).

[0301]

[0302] 2-(7-azabenzotriazole)-tetramethyluronium hexafluorophosphate (0.11 g, 0.29 mmol, 1.5 eq.) and N,N-diisopropylethylamine (0.14 mL, 0.77 mmol, 4.0 eq.) were added to a solution of compound 7 (0.079 g, 0.19 mmol, 1.0 eq.) in N,N-dimethylformamide (3 mL). The mixture was stirred at 50° C. for 0.5 hour under nitrogen protection. Then, compound H21-2 (0.05 g, 0.29 mmol, 1.5 eq.) dissolved in N,N-dimethylformamide (1 mL) was added, and the mixture was stirred at 50° C. for 16 hours under nitrogen protection. The reaction solution was concentrated to obtain a residue. The residue was extracted with dichloromethane / methanol (10:1, v / v) and water, the organic phases were combined and dried, concentrated, and then slurried with methanol, filtered and the residue was dried to obtain compound H-21 (0.048 g, light yellow solid, yield 47%). LC-MS (ESI): m / z 528.2 [M+1] + , 1 H NMR(400MHz, DMSO-d6)δ:11.20–11.10(m,1H),9.40–9.25(m,1H),8.68(s,2H),8.50–8.40(m,1H),8.05–7.95(m,1H),7.65–7.40(m,2 H),7.15–7.05(m,1H),4.75(d,2H),4.10–3.40(m,6H),2.81(s,3H),2.61(q,2H),2.50–2.45(m,2H),2.30–2.15(m,2H),1.19(t,3H).

[0303] Example 18 Preparation of Compound H-22

[0304]

[0305] To a solution of compound H22-1 (0.56 g, 4.21 mmol) in ethanol (25 mL) was added Raney-Ni (0.08 g) and NH3.H2O (0.2 mL). The mixture was stirred at room temperature for 2 hours under a hydrogen atmosphere (3.5 MPa). The filtrate was filtered and concentrated to obtain a residue. The residue was used directly in the next step without further purification. LC-MS (ESI): m / z 138.1 [M+1] +

[0306]

[0307] Compound H22-3 (2.1 g, 16.2 mmol, 1.2 eq.) was added dropwise to a mixture of compound H22-2 (2.0 g, 13.4 mmol, 1.0 eq.) and iron triacetylacetonate (0.95 g, 2.7 mmol, 0.2 eq.) maintained at -78°C. After the reaction solution was stirred at -78°C for 0.5 hours, it was gradually warmed to room temperature and reacted at room temperature for 1 hour. The reaction solution was quenched with saturated ammonium chloride solution, then extracted with ethyl acetate and water, the organic phases were combined and dried, and the organic phases were concentrated to obtain a residue. The residue was separated and purified by column chromatography to obtain compound H22-4 (0.7 g, light yellow oil, yield 40%). LC-MS (ESI): m / z 143.1 [M+1] + , 1 H NMR(400MHz, CDCl3)δ:8.47(d,1H),7.11(d,1H),2.78(q,2H),1.29(t,3H).

[0308]

[0309] Potassium cyanide (0.73 g, 11.22 mmol, 2.0 eq.) and 1,4-diazabicyclo[2.2.2]octane (1.26 g, 11.22 mmol, 2.0 eq.) were added to a solution of compound H22-4 (0.8 g, 5.61 mmol, 1.0 eq.) in dimethyl sulfoxide (48 mL) and water (10 mL). The mixture was reacted at 80 ° C for 2 hours under nitrogen protection. The mixture was extracted with ethyl acetate and water, the organic phases were combined and dried, and the organic phases were concentrated to obtain a residue. The residue was separated and purified by column chromatography to obtain compound H22-5 (0.56 g, light yellow oil, yield 75%). LC-MS (ESI): m / z 134.1 [M+1] +

[0310]

[0311] To a solution of compound H22-5 (0.15 g, 0.36 mmol, 1.0 eq.) in dichloromethane (2 mL) was added di-tert-butyl dicarbonate (0.1 g, 0.44 mmol, 1.2 eq.) and a saturated aqueous sodium bicarbonate solution (1 mL). The mixture was stirred at room temperature for 1 hour under nitrogen protection. The reaction solution was extracted with dichloromethane and water, the organic phases were combined and dried, and the organic phases were concentrated to obtain a residue. The residue was separated and purified by column chromatography to obtain compound H22-6 (0.03 g, white solid, yield 30%). LC-MS (ESI): m / z 238.2 [M+1] + , 1 H NMR (400MHz, CDCl3) δ8.51(d,1H),7.01(d,1H),5.80–5.65(m,1H),4.51(d,2H),2.74(q,2H),1.43(s,9H),1.26(t,3H).

[0312]

[0313] HCl-dioxane (4M, 2 mL) was added to compound H22-6 (0.03 g, 0.13 mmol). The mixture was stirred at room temperature for 2 hours under nitrogen protection. The reaction solvent was removed by concentration to obtain compound H22-7 (25 mg, white solid, yield 98%). LC-MS (ESI): m / z 138.1 [M+1] +

[0314]

[0315] 2-(7-azabenzotriazole)-tetramethyluronium hexafluorophosphate (0.055 g, 0.14 mmol, 1.5 eq.) and N,N-diisopropylethylamine (0.07 mL, 0.38 mmol, 4.0 eq.) were added to a solution of compound 7 (0.04 g, 0.1 mmol, 1.0 eq.) in N,N-dimethylformamide (1 mL). The mixture was stirred at 50 ° C for 0.5 hours under nitrogen protection, and compound H22-7 (0.025 g, 0.14 mmol, 1.5 eq.) dissolved in N,N-dimethylformamide (2 mL) was added, and the mixture was stirred at 50 ° C for 16 hours under nitrogen protection. The reaction solution was concentrated to obtain a residue. The residue was separated and purified by preparative HPLC, and the preparative solution was lyophilized to obtain compound H-22 (0.03 g, light yellow solid, yield 59%). LC-MS(ESI):m / z 528.2[M+1] + , 1H NMR(400MHz, DMSO-d6)δ:11.20–11.10(m,1H),9.45–9.30(m,1H),8.70–8.60(m,1H),8.35–8.25(m,1H),8.00–7.90(m,1H),7.55–7.35(m,2H),7. 29(d,1H),7.05–6.90(m,1H),4.73(d,2H),4.05–3.60(m,6H),2.80–2.70 (m,2H),2.55–2.45(m,2H),2.24(s,3H),2.55–1.90(m,2H),1.24(t,3H).

[0316] Example 19 Preparation of Compound H-26

[0317]

[0318] Compound H26-1 (0.11 g, 0.21 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide (6 mL), sodium azide (0.082 g, 1.26 mmol, 6.0 eq.) and ammonium chloride (0.22 g, 4.2 mmol, 20.0 eq.) were added to the reaction solution, and the mixture was reacted at 100 ° C for 4 hours in a sealed tube. The compound H-26 (0.0076 g, white solid, yield 7%) was obtained by separation and purification by preparative HPLC and freeze-drying the preparative solution. LC-MS (ESI): m / z 568.2 [M+1] + , 1 H NMR(400MHz, DMSO-d6)δ:11.15–11.05(m,1H),9.40–9.30(m,1H),9.23(s,2H),8.45–8.35(m,1H),8.00–7.90(m,1H),7.60 –7.40(m,2H),7.10–7.00(m,1H),4.81(d,2H),4.05–3.60(m,4H),3.05–2.95(m,2H),2.65–2.45(m,5H),2.20–2.05(m,2H).

[0319] Example 20 Preparation of Compound H-27

[0320]

[0321] Compound H27-1 (600 mg, 3.3 mmol, 1.0 eq.) was dissolved in toluene (12 mL), tris(dibenzylideneacetone)dipalladium (589 mg, 0.65 mmol, 0.2 eq.), XPhos (620 mg, 1.3 mmol, 0.4 eq.), K3PO4.3H2O (2.6 g, 9.8 mmol, 3.0 eq.) and H27-2 (560 mg, 6.5 mmol, 2.0 eq.) were added, and the mixture was sealed and reacted overnight at 60°C. The mixture was filtered, the filtrate was concentrated, and the mixture was separated and purified by column chromatography to obtain compound H27-3 (500 mg, light yellow oil, purity ~80%), which was used directly in the next step. LC-MS (ESI): m / z 146.2 [M+1] + , 1 H NMR (400MHz, DMSO-d6) δ8.75(s,2H),2.03(s,1H),1.15–1.11(m,2H),0.97(m,2H).

[0322]

[0323] Compound H27-3 (200 mg, 1.4 mmol, 1.0 eq.) was dissolved in methanol (3 mL), Raney-Ni (60 mg,) and concentrated hydrochloric acid (0.2 mL) were added, and stirred at room temperature overnight under a hydrogen atmosphere. Filtered, separated and purified by thin-plate chromatography to obtain compound H27-4 (35 mg, light yellow oil, two-step yield 18.0%). LC-MS (ESI): m / z 150.3 [M+1] + , 1 H NMR (400MHz, DMSO-d6) δ8.50(s,2H),3.86(s,2H),1.90(m,1H),0.99(m,2H),0.79–0.76(m,2H).

[0324]

[0325] Compound 7 (48 mg, 0.12 mmol, 1.0 eq.) was dissolved in N, N-dimethylformamide (2 mL), and 2-(7-azabenzotriazole)-tetramethyluronium hexafluorophosphate (88 mg, 0.23 mmol, 2.0 eq.) and N, N-diisopropylethylamine (47 mg, 0.36 mmol, 3.0 eq.) were added thereto. The reaction solution was reacted at 50 ° C for 0.5 hours under nitrogen protection, and compound H27-4 (35 mg, 0.23 mmol, 2.0 eq.) was added, and the reaction mixture was continued to react at 50 ° C for 16 hours. The reaction solution was concentrated to remove N, N-dimethylformamide, and methanol (2 mL) was added. The mixture was stirred at room temperature overnight. Filter, then wash the filter cake with methanol, and dry to obtain compound H-27 (5.0 mg, light yellow solid, 7.9% yield). LC-MS (ESI): m / z 540.1 [M+1] + , 1 H NMR(400MHz,DMSO-d6)δ:11.20–11.10(m,1H),9.45–9.35(m,1H),8.38(s, 2H),8.35–8.25(m,1H),8.00–7.90(m,1H),7.60–7.35(m,2H),7.05–6.90( m,1H),4.74(d,2H),4.00–3.60(m,4H),2.80–2.70(m,2H),2.60–2.50(m,2 H),2.27(s,3H),2.05–1.90(m,2H),1.10–0.95(m,2H),0.85–0.75(m,2H).

[0326] Example 21 Preparation of Compound H-28

[0327]

[0328] Compound H28-1 (500 mg, 3.4 mmol, 1.0 eq.) was dissolved in tetrahydrofuran (10 mL), and Pd(dppf)Cl2 (246 mg, 0.34 mmol, 0.1 eq.), K3PO4.3H2O (2.2 g, 8.4 mmol, 2.5 eq.) and H28-2 (289 mg, 3.4 mmol, 1.0 eq.) were added, and microwave reaction was performed at 90°C for 1 hour. The mixture was filtered, the filtrate was concentrated, and the mixture was separated and purified by column chromatography to obtain compound H28-3 (130 mg, light yellow oil, yield 25%). LC-MS (ESI): m / z 155.2 [M+1] + , 1H NMR(400MHz, CDCl3)δ:8.33(d,1H),7.06(d,1H),1.96(m,1H),1.22–1.11(m,4H).

[0329]

[0330] Compound H28-3 (130 mg, 0.84 mmol, 1.0 eq.) was dissolved in dimethyl sulfoxide (7 mL) and water (1 mL), potassium cyanide (109 mg, 1.68 mmol, 2.0 eq.) and 1,4-diazabicyclo[2.2.2]octane (188 mg, 1.68 mmol, 2.0 eq.) were added, and the reaction mixture was reacted at 80°C overnight. Water and ethyl acetate were added for extraction, the organic phases were combined, dried and concentrated, and then purified by column chromatography to obtain compound H28-4 (80 mg, light yellow oil, yield 65.6%). LC-MS (ESI): m / z 146.2 [M+1] + , 1 H NMR (400MHz, DMSO-d6)δ:8.71(d,1H),7.74(d,1H),2.21(s,1H),1.20–1.15(m,2H),1.06(m,2H).

[0331]

[0332] Compound H28-4 (80 mg, 0.55 mmol, 1.0 eq.) was dissolved in methanol (3 mL), Raney-Ni (20 mg) and aqueous ammonia (47%) (0.1 mL) were added, and stirred at room temperature for 2 hours under a hydrogen (3 MPa) atmosphere. Filter and freeze-dry to obtain compound H28-5 (43 mg, light yellow oil, yield 52.4%). LC-MS (ESI): m / z 150.3 [M+1] +

[0333]

[0334] Compound 7 (36 mg, 0.09 mmol, 1.0 eq.) was dissolved in N, N-dimethylformamide (2 mL), and 2-(7-azabenzotriazole)-tetramethyluronium hexafluorophosphate (52 mg, 0.14 mmol, 1.5 eq.) and N, N-diisopropylethylamine (35 mg, 0.27 mmol, 3.0 eq.) were added thereto. The reaction solution was reacted at 50 ° C for 0.5 hours under nitrogen protection, and compound H28-5 (40 mg, 0.27 mmol, 3.0 eq.) was added, and the reaction solution was continued to react at 50 ° C for 16 hours. The reaction solution was concentrated to remove N, N-dimethylformamide. Methanol (2 mL) was added to the residue, and the mixture was stirred at room temperature overnight. Filter, then wash the filter cake with methanol, and separate and purify the filter cake by preparative HPLC to obtain compound H-28 (19.6 mg, light yellow solid, 40.8% yield). LC-MS(ESI):m / z 540.2[M+1] + , 1 H NMR(400MHz, DMSO-d6)δ:11.15–11.05(m,1H),9.30–9.20(m,1H),8.55–8.50(m,1H),8.35–8.25(m,1H),7.95–7.85(m,1H),7.55–7.35(m,2H) ,7.35–7.25(m,1H),7.0–6.90(m,1H),4.66(d,2H),4.00–3.60(m,4H), 3.15–2.80(m,4H),2.55(s,3H),2.20–2.00(m,3H),1.20–1.00(m,4H).

[0335] Example 22 Preparation of Compound H-31

[0336] 1. Preparation of compound H31-2

[0337]

[0338] Compound H31-1 (1 g, 9.98 mmol, 1.0 eq.) was dissolved in dichloromethane (20 mL), and N,N-diisopropylethylamine (1.29 g, 9.98 mmol, 1.0 eq.) and 4-dimethylaminopyridine (1.22 g, 9.98 mmol, 1.0 eq.) were added, and then di-tert-butyl dicarbonate (2.39 g, 10.98 mmol, 1.1 eq.) was added, and stirred at room temperature for 1 hour. The reaction mixture was washed with dichloromethane / methanol (10 / 1, v / v), and the organic phases were combined, dried, concentrated, and purified by column chromatography to obtain compound H31-2 (600 mg, light yellow solid, yield 30%). LC-MS (ESI): m / z 201.3 [M+1]+ , 1 H NMR (400MHz, DMSO-d6) δ3.33–3.28(m,4H),2.67(m,4H),1.66–1.54(m,2H),1.35(s,9H).

[0339] 2. Preparation of compound H31-3

[0340]

[0341] Potassium carbonate (6.2 g, 45.0 mmol, 1.0 eq.) was added to a solution of compound H31-2 (3 g, 15.0 mmol, 1.0 eq.) in N,N-dimethylformamide (45 mL). After stirring at 50°C for 10 minutes, iodomethane-d3 (1.74 g, 12.0 mmol, 0.8 eq.) was added. 0.2 eq of iodomethane-d3 was added every 10 minutes and the addition was completed within 30 minutes. The reaction mixture was reacted at 50°C for 0.5 hours under a nitrogen atmosphere. Water was added to quench the reaction, and the mixture was extracted with dichloromethane / methanol (8 / 1, v / v, 80 mL x 5). The organic phases were combined, dried, concentrated, and purified by column chromatography to obtain the product H31-3 (1.72 g, light yellow oil, yield 53%). LC-MS (ESI): m / z 218.0 [M+1] +

[0342] 3. Preparation of compound H31-4

[0343]

[0344] Compound H31-3 (1.95 g, 9.0 mmol, 1.0 eq.) was dissolved in HCl / dioxane (4 M, 20 mL) and reacted overnight at room temperature. The reaction solvent was removed, and dichloromethane was added to help remove the residual dioxane, and finally the product H31-4 (1.7 g, light yellow oil, yield 100%) was obtained. LC-MS (ESI): m / z 118.1 [M+1] + , 1 H NMR (400MHz, DMSO-d6) δ11.38(s,1H),9.70(s,1H),9.42(s,1H),3.68–3.57(m,2H),3.51–3.24(m,4H),3.25–2.10(m,2H),2.16–2.03(m,2H).

[0345] 4. Preparation of compound H31-5

[0346]

[0347] Potassium carbonate (2.47 g, 17.88 mmol, 3.0 eq.) and compound H31-4 (1.7 g, 8.94 mmol, 1.5 eq.) were added to a solution of compound 4 (2.14 g, 5.96 mmol, 1.0 eq.) in acetonitrile (20 mL). The reaction solution was stirred at 80 ° C for 16 hours under a nitrogen atmosphere. The reaction solvent was removed by concentration, water was added to the residue, and the mixture was washed with dichloromethane / methanol (10 / 1, v / v), and the organic phases were combined and concentrated. The crude product was slurried with ethyl acetate overnight, filtered, and the filter residue was dried to obtain the product H31-5 (1.6 g, light yellow solid, yield 61%). LC-MS (ESI): m / z 440.6 [M+1] + , 1 H NMR(400MHz,DMSO-d6)δ:9.34(d,1H),8.25–8.15(m,1H),8.25–7.90(m,1H),7.55–7.40(m,2H),6.93(d,1 H),4.28(q,2H),4.05–3.60(m,4H),2.90–2.65(m,2H),2.60–2.50(m,2H),2.02–1.85(m,2H),1.29(t,3H).

[0348] 5. Preparation of compound H31-6

[0349]

[0350] LiOH·H2O (133 mg, 3.18 mmol, 2.0 eq.) was added to a solution of compound H31-5 (700 mg, 1.59 mmol, 1.0 eq.) in tetrahydrofuran / methanol / water (6 mL / 2 mL / 6 mL). The mixture was stirred at room temperature overnight under nitrogen protection. The reaction solvent was removed by concentration and freeze-dried to obtain compound H31-6 (1.1 g, containing lithium hydroxide). The crude product was used directly in the next step. LC-MS (ESI): m / z 412.5 [M+1] + , 1 H NMR(400MHz,DMSO-d6)δ9.42(d,1H),8.37(d,1H),7.95–7.80(m,1H),7.60–7.30(m,2H), 7.00(d,1H),4.05–3.50(m,4H),2.90–2.65(m,2H),2.60–2.50(m,2H),2.02–1.85(m,2H).

[0351] 6. Preparation of Compound H-31

[0352]

[0353] To a solution of compound H31-6 (500 mg, 1.2 mmol, 1.0 eq.) in N,N-dimethylformamide (6 mL), 2-(7-azabenzotriazole)-tetramethyluronium hexafluorophosphate (690 mg, 1.8 mmol, 1.5 eq.) and N,N-diisopropylethylamine (470 mg, 3.6 mmol, 3.0 eq.) were added. The mixture was stirred at 50°C for 0.5 hours under nitrogen protection. Compound H31-7 (200 mg, 1.8 mmol, 1.5 eq.) dissolved in N,N-dimethylformamide (2 mL) was added, and the mixture was stirred at 50°C for 16 hours under nitrogen protection. The reaction mixture was concentrated to obtain a residue. The residue was dissolved in dichloromethane / methanol (10 / 1, v / v), washed with water, extracted, and the organic phases were combined, dried, and concentrated. The crude product was slurried with methanol (5 mL) overnight, filtered, and dried to obtain compound H-31 (269.7 mg, light yellow solid, yield 44%). LC-MS (ESI): m / z 503.2 [M+1] + , 1 H NMR(400MHz, DMSO-d6)δ:11.20–11.10(m,1H),9.31(s,1H),8.81(d,2H),8.29(d,1H),7.99–7.87(m,1H),7.51–7.36 (m,3H),7.05–6.90(m,1H),4.77(d,2H),4.05–3.50(m,4H),2.90–2.65(m,2H),2.60–2.50(m,2H),2.02–1.85(m,2H).

[0354] Other compounds from compound H-1 to compound H-31 in Table 1 were prepared using a method similar to the above examples.

[0355] Table 1: Compounds H-1 to H-31

[0356]

[0357]

[0358]

[0359]

[0360]

[0361] Example 23: Biological Evaluation

[0362] The present application is further described and explained below in conjunction with test examples, but these embodiments are not meant to limit the scope of the present application.

[0363] The experimental methods in the test examples of this application that do not specify specific conditions are usually carried out under conventional conditions or under conditions recommended by product manufacturers, and the reagents that do not specify specific sources are conventional reagents purchased on the market.

[0364] Test Example 1. Inhibition of cancer cells by compounds

[0365] Testing cell viability assays

[0366] (1) Compound treatment

[0367] a) Preparation of mother solutions of compounds

[0368] The test compound was dissolved in DMSO at a concentration of 10 mM or 5 mM, and Olaparib was dissolved in DMSO at 10 mM.

[0369] b) Compound Storage

[0370] All compounds dissolved in DMSO were stored in a drying oven at room temperature for a short term and moved into a -20°C refrigerator for long term storage.

[0371] c) Preparation of working concentration of compounds

[0372] The test compound was diluted 3-fold in DMSO to form 10 concentration gradients starting from 10 μM.

[0373] Olaparib was diluted 3-fold in DMSO to 10 concentrations starting from 10 μM.

[0374] The 384-well plate with added compounds was centrifuged at 1000 rpm for 1 minute.

[0375] (2) Activity detection

[0376] a) Add the cells to the culture medium and culture them in a cell culture incubator at 37°C and 5% CO2.

[0377] b) 40 μL of BRCA1 or BRCA2 deficient cells were added to a 384-well plate and cultured overnight at 37° C. and 5% CO 2 .

[0378] c) The drug to be tested was prepared into different concentration gradients on the master plate, 80 nL of the compound to be tested and Olaparib were taken from the master plate and added to the 384-well plate containing cells, and cultured at 37° C. and 5% CO 2 for 6 days.

[0379] d) CellTiterGlo (30 uL per well) was added to a 384-well plate, incubated at room temperature and protected from light for 30 minutes, and then measured using an Envisionplate reader.

[0380] (3) Data processing

[0381] a) Calculation of IC using GraphPad Prism software 50 , and draw the activity curve as the concentration changes.

[0382] b) The inhibition of the compound on cell activity was calculated by the following formula:

[0383]

[0384] Test results:

[0385] Table 2. Study on the inhibition of quinolone analogs on the activity of BRCA2 gene knockout DLD1 cancer cells

[0386]

[0387]

[0388]

[0389] The compounds shown in the table above have significantly higher inhibitory effects on the activity of BRCA2 gene knockout DLD1 cancer cells than the PARP inhibitors Olaparib and CX-5461.

[0390] Table 3. Study on the inhibition of quinolone analogs on the activity of BRCA1 gene-deficient ovarian cancer cell UWB1.289

[0391]

[0392] The study of the activity of BRCA1 gene-deficient ovarian cancer cells UWB1.289 found that compound H-31 has a stronger ability to kill BRCA1-deficient cancer cells than the control compounds Olaparib and CX-5461, and the IC 50 Significantly lower than Olaparib and CX5461.

[0393] Test Example 2 Pharmacokinetic study of compounds in mice

[0394] The test compound was administered to mice orally (PO) and intravenously (IV). The drug was dissolved at room temperature. The mice used in the study were Balb / c nude, all female, aged 6-8 weeks, weighing 20-30 grams. Before administration, the mice were allowed to eat and drink freely.

[0395] Blood samples (about 0.03 mL / sample) were collected from the vein at appropriate time points. The samples were placed in tubes containing anticoagulants and stored on ice until centrifugation. Blood samples were collected at 0.083, 0.25, 0.5, 1, 2, 4, 8, 24 hours after intravenous administration and 0.25, 0.5, 1, 2, 4, 8, 24 hours after oral administration. The blood samples were centrifuged at 2-8 ° C for 5 minutes at about 4000 rpm. Plasma samples were then collected and stored frozen at about -80 ° C until analysis. Bioanalytical work was performed using an LC-MS / MS system.

[0396] Table 4. Bioavailability assay

[0397] Bioavailability average value CX-5461 33.8％ Compound H-1 60.7％ Compound H-2 47.2％ Compound H-12 49.1％ Compound H-17 74.6％ Compound H-21 66.8％ Compound H-22 40.6％ Compound H-27 59.2％ Compound H-28 41.6％ Compound H-31 43.1％

[0398] Based on the in vivo pharmacokinetic study on mice, the compounds in Table 4 have significantly improved oral bioavailability relative to CX5461, which is beneficial to the development of oral drug formulations and provides patients with a more convenient administration method.

[0399] Test Example 3. Cardiotoxicity test of compound herg

[0400] Detection method:

[0401] 1) HEK 293 cell line (Cat#K1236) with stable expression of hERG was purchased from Invitrogen. The cells were cultured in 85% DMEM, 10% FBS, 0.1 mM NEAA, 25 mM HEPES, 100 U / mL penicillin-streptomycin, 5 μg / mL Blasticidin, and 400 μg / mL Geneticin. The cells were cultured using TrypLE TM Express were passaged approximately three times per week and maintained between 40% and 80% confluency. Cells were plated on coverslips at 5 × 105 cells / 6-cm dish and induced with 1 μg / ml doxycycline for 48 hours before assay.

[0402] 2) Remove the coverslip from the cell culture dish and place it on the microscope stage. Find a desired cell using the ×10 objective. Use the ×10 objective to find the tip of the electrode under the microscope, focusing just above the plane of the cell. Once the tip is in focus, use the coarse control of the manipulator to advance the electrode down toward the cell while moving the objective to keep the tip in focus. When directly over the cell, switch to the ×40 objective and use the fine control of the manipulator to approach the cell surface in small steps.

[0403] 3) A blank control is first performed to establish a baseline. Once the hERG current is found to be stable for 5 minutes, the working solution is applied. In the presence of the test compound, the hERG current is recorded for about 5 minutes to reach a steady state, and then 5 scans are captured. For dose-response testing, 5 doses of the test compound are cumulatively applied to the cells from low to high concentrations. The positive control, dofetilide at a concentration of 150nM, was applied to each cell, and the hERG current was measured at the highest concentration of the test compound as a negative control to normalize the inhibition percentage. In order to ensure the good performance of the cultured cells and operations, the same batch of cells was also tested with 5 doses of dofetilide.

[0404] Table 5. hERG toxicity test results

[0405] Test compound <![CDATA[hERG IC 50 [μM]]]> CX-5461 5.1 Compound H-13 >30.0 Compound H-15 >30.0 Compound H-18 10.7

[0406] Test results: hERG toxicity test IC of compounds listed in Table 5 50 It is greater than CX-5461, indicating that the cardiotoxicity is improved compared with CX-5461.

[0407] Test Example 4. Testing the toxicity of compounds using mouse models

[0408] a. Effects of repeated administration of 50 mg / kg CX-5461, compound H-31, and solvent on the body weight of mice

[0409] The test compound was administered orally (PO) to mice. The drug was dissolved at room temperature. The mice used in the study were all female and aged 6-8 weeks. Prior to administration, the mice were allowed free access to food and water.

[0410] Solvent control: 50mM NaH2PO4, pH4.0, administered once every 3 days. CX-5461, compound H-31 administered once every 3 days, dose: 50mg / kg. If the weight loss of mice exceeds 15%, stop the drug until the weight is restored and then administer again. The effect of the compound on the weight of mice is shown in Figure 1 .

[0411] The above experiment shows that after repeated administration for 21 days at a dose of 50 mg / kg, CX-5461 significantly reduced the weight of mice (P<0.01, t-test). In addition to the weight loss, it was also observed that the entire group of animals in the CX-5461 group were in poor condition, slow in movement, and their tails and toes were bleeding and white. Since the average weight loss was greater than 10%, the entire group of animals was given a drug withdrawal treatment. After the administration of compound H-31, the weight of mice changed in a similar trend to that of the control group, and no abnormal clinical symptoms were observed. This shows that the toxic side effects of compound H-31 are less than those of CX-5461.

[0412] b. Effects of repeated administration of 55 mg / kg, CX-5461, H series compounds and solvents on the body weight of mice

[0413] The test compound was administered orally (PO) to mice. The drug was dissolved at room temperature. The mice (Balb / c) used in the study were all female and aged 6-8 weeks. Before administration, the mice were allowed to eat and drink freely.

[0414] Solvent control: 50mM NaH2PO4, pH4.0. CX-5461 and H-series compounds were administered once every three days, at a dose of 55mg / kg, for 4 times. The effects of the compounds on the body weight of mice are shown in Figure 2-1 and Figure 2-2 .

[0415] The above experiment showed that after repeated administration of CX-5461 at a dose of 55 mg / kg for 4 times, the mice's body weight was significantly reduced, and all died on the 13th day; while the weight of compounds H-4 and H-17 recovered after a slight decrease, and no mice died; the weight of mice remained unchanged after administration of H-12, H-21, H-27, and H-28. The results show that the toxic and side effects of compounds H-4, H-12, H-17, H-21, H-27, and H-28 are greatly improved compared to CX-5461.

[0416] Test Example 5. Inhibition of human cancer by compounds in mouse models

[0417] The anti-tumor efficacy of the compounds of the present application was evaluated in NOD SCID mice. Female mice aged 6-8 weeks were subcutaneously inoculated with MDA-MB-436 cancer cells to form tumors. The tumors grew to 100-150 mm 3 The mice were randomly divided into groups, with 6 mice in each group. The administration method was oral administration. Olaparib was administered once a day at a dose of 100 mg / kg for a total of 28 days. The solvent control group and compound H-31 were administered once every three days at a dose of 25 mg / kg, once every three days.

[0418] like Figure 3 As shown, compared with the control group, both the Olaparib treatment group and the compound H-31 treatment group significantly inhibited tumor growth (p<0.001), and the cancer inhibition rate of the Olaparib treatment group was 63.4%. The cancer inhibition rate of the compound H-31 treatment group was 82.2%, which was significantly higher than that of the Olaparib treatment group (p<0.001), and the dosage of compound H-31 (25 mg / kg.Q3D) was much lower than that of Olaparib (100 mg / kg,Q1D).

[0419] Test Example 6 Inhibition of viral DNA replication by compounds

[0420] Take a section of viral DNA containing a quadruplex DNA sequence as a template, the primer is fluorescently labeled and annealed to the 3' end of the template DNA, and TagDNA polymerase is added to the system to synthesize a complementary chain with a fluorescently labeled primer. If the synthesis of DNA is not inhibited, a full-length copy of the template will be synthesized. If the added compound binds non-specifically to the double-stranded DNA, the full-length product will be reduced and short-chain PCR products of different lengths will be formed. If the compound specifically binds to the quadruplex DNA in the viral DNA sequence, DNA replication will only stop in the quadruplex DNA region. The PCR product is detected by DNA electrophoresis, and the compound is analyzed to see whether it inhibits the replication of viral DNA compared with the solvent control.

[0421] The experimental results show that the IC of the compound of formula (I) inhibits viral DNA replication 50 The concentration of the compound in formula (I) is 0.01-1 μM, so the compound in formula (I) has a strong inhibitory effect on the replication of viral DNA containing a quadruplex DNA structure.

[0422] Test Example 7 Determination of the Antiviral Cell Protection Activity of Compounds

[0423] MRC5 cells were seeded at a certain density in a microplate and cultured overnight in a 37°C, 5% CO2 incubator. Compounds diluted in multiple ratios and GFP-labeled human herpes virus HCMV-AD169 were added. Cell controls (cells, no compound treatment and virus infection) and virus controls (cells infected with virus, no compound treatment) were set up. The test compounds and control compounds were tested at 8 concentrations, 3-fold serial dilutions, and triplicate wells. The toxicity of the test compounds and control compounds to the cells was also determined. The cells were cultured in a 37°C, 5% CO2 incubator for 7 days. Acumen Cellista was used to detect the GFP fluorescence intensity in each well. Cell viability was detected using cell viability detection reagents CCK-8 or CellTiter Glo, and the raw data were used to calculate the cytotoxicity of the test compounds. GraphPad Prism software was used to analyze the dose-response curves of the test compounds and calculate the virus inhibition EC 50 and cytotoxic CC 50 Value, and calculate the therapeutic index SI, SI = CC 50 / EC 50 , used to evaluate the therapeutic potential of drugs. The higher the SI value, the stronger the antiviral efficacy of the drug.

[0424] Experiments show that compound IC in formula (I) 50When the concentration is about 0.01-1.0 μM, it has a protective effect on the decrease in cell activity caused by virus infection, among which the inhibitory effect of H-31 on HCMV exceeds that of Ganciclovir.

[0425] Table 6. H-31 effectively inhibits human herpes virus HCMV

[0426]

[0427] Test Example 8 Inhibition of Multiple Sclerosis by Compounds of Formula I

[0428] The autoimmune encephalomyelitis (EAE) model was established as follows: C57BL / 6 male mice, aged 6-8 weeks, 6 mice per group,

[0429] Group 1 = EAE + solvent;

[0430] Group 2 = EAE + PRN2246;

[0431] Group 3 = EAE + test drug.

[0432] Dosing frequency: once a day, dosing cycle: 21 days. Dilute MOG35-55 with PBS, then mix the diluent with complete Freund's adjuvant, pump it with a glass syringe to an oil-in-water state to prepare an antigen emulsion for inducing EAE. On the day of immunization and the second day, mice were given intraperitoneal injections of pertussis toxin to induce the establishment of a mouse autoimmune encephalomyelitis model.

[0433] Neurological function scoring criteria:

[0434] The scoring standard is a universal 0-5 scale:

[0435] Level 0, no disease;

[0436] Grade 1, decreased tone or paralysis of the mouse tail;

[0437] Grade 2, mouse tail paralysis and hind limb weakness;

[0438] Grade 3, paralysis of the mouse hind limbs;

[0439] Grade 4: the mouse has quadriplegia and cannot be reset by manual turning over;

[0440] Level 5: dying state or death after onset of illness.

[0441] Scoring frequency: starting from day 11, once a day for 10 days. Body weight measurement: twice a week.

[0442] Experiments have shown that the compound of formula (I) has a delayed effect on the development of multiple sclerosis when the concentration is 0.1-10 μM.

[0443] The above describes the specific embodiments of the present application, but it should be understood that the present application is not limited to the above-mentioned specific implementation methods. Those skilled in the art can make various deformations and modifications without departing from the concept of the present application, which all belong to the protection scope of the present application.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof: in: R1 is C1-C6 alkyl or one or more deuterated C1-C4 alkyl, and R2 to R6 are each independently H or deuterium; R7, R8, and R9 are each independently selected from H, deuterium, halogen, azido, -CN, C1-C6 alkyl, C1-C6 haloalkyl, -O(C1-C6 alkyl), -S(C1-C6 alkyl), -CONR 10 R 11 、-NR 10 R 11 , 3-4 membered cycloalkyl, 5-6 membered nitrogen-containing heterocyclic ring or -CH2-5-6 membered nitrogen-containing heterocyclic ring, wherein R 10 and R 11 Each is independently selected from H or C1-6 alkyl, and the 5-6 membered nitrogen-containing heterocyclic ring contains 1, 2, 3 or 4 nitrogen atoms; L is -(CH2) n -, n=1-3.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: R1 is -CH3 or -CD3, R2 to R6 are all H or all deuterium; Alternatively, R1 is -CH3 or -CD3, R4 is H and R2, R3, R5 and R6 are all deuterium.

3. The compound according to claim 2 or a pharmaceutically acceptable salt thereof, wherein: R1 is -CH3 or -CD3, and R2 to R6 are all H.

4. The compound according to claim 2 or a pharmaceutically acceptable salt thereof, wherein: R1 is -CH3, and R2 to R6 are all H.

5. The compound according to claim 2 or a pharmaceutically acceptable salt thereof, wherein: R1 is -CD3, and R2 to R6 are all H.

6. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, wherein: R8 is selected from H, halogen, azido, -CN, C1-C6 alkyl, C1-C6 haloalkyl, -O(C1-C6 alkyl), -S(C1-C6 alkyl), -CONR 10 R 11 、-NR 10 R 11 , 3-4 membered cycloalkyl, 5-6 membered nitrogen-containing heterocyclic ring or -CH2-5-6 membered nitrogen-containing heterocyclic ring, wherein R 10 and R 11 Each is independently selected from H or C1-6 alkyl, the 5-6 membered nitrogen-containing heterocyclic ring contains 1, 2, 3 or 4 nitrogen atoms; R7 and R9 are H.

7. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, wherein: R7 is selected from H, halogen, azido, -CN, C1-C6 alkyl, C1-C6 haloalkyl, -O(C1-C6 alkyl), -S(C1-C6 alkyl), -CONR 10 R 11 、-NR 10 R 11 , 3-4 membered cycloalkyl, 5-6 membered nitrogen-containing heterocyclic ring or -CH2-5-6 membered nitrogen-containing heterocyclic ring, wherein R 10 and R 11 Each is independently selected from H or C1-6 alkyl, the 5-6 membered nitrogen-containing heterocyclic ring contains 1, 2, 3 or 4 nitrogen atoms; R8 and R9 are H.

8. The compound according to claim 6 or a pharmaceutically acceptable salt thereof, wherein: R8 is H, F, Cl, -CF3, -CN, -N3, -CONHCH3, -CON(CH3)2, -OCH3, -SCH3, -NHCH3, -N(CH3)2, -C2H5, -CH2-1-pyrrolyl, -CH2-4-pyridyl, 1-pyrrolyl, 2H-tetrazol-5-yl, cyclopropyl or -CH2CF3; R7 and R9 are H.

9. The compound according to claim 7 or a pharmaceutically acceptable salt thereof, wherein: R7 is H, F, Cl, -CF3, -CN, -N3, -CONHCH3, -CON(CH3)2, -OCH3, -SCH3, -NHCH3, -N(CH3)2, -C2H5, -CH2-1-pyrrolyl, -CH2-4-pyridyl, 1-pyrrolyl, 2H-tetrazol-5-yl, cyclopropyl or -CH2CF3; R8 and R9 are H.

10. The compound according to claim 7 or a pharmaceutically acceptable salt thereof, wherein: R7 is H, F, Cl, -CF3, -CN, -N3, -CONHCH3, -CON(CH3)2, -OCH3, -SCH3, -NHCH3, -N(CH3)2, -C2H5, cyclopropyl or -CH2CF3; R8 and R9 are H.

11. The compound according to claim 7 or a pharmaceutically acceptable salt thereof, wherein: R7 is H, F, -CN, -CONHCH3, -OCH3, -C2H5, cyclopropyl or -CH2CF3; R8 and R9 are H.

12. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, wherein: R7, R8 and R9 are all deuterium.

13. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, wherein: L is -CH2-.

14. The compound according to claim 6 or a pharmaceutically acceptable salt thereof, wherein: L is -CH2-.

15. The compound according to claim 7 or a pharmaceutically acceptable salt thereof, wherein: L is -CH2-.

16. The compound according to claim 8 or a pharmaceutically acceptable salt thereof, wherein: L is -CH2-.

17. The compound according to claim 9 or a pharmaceutically acceptable salt thereof, wherein: L is -CH2-.

18. The compound according to claim 10 or a pharmaceutically acceptable salt thereof, wherein: L is -CH2-.

19. The compound according to claim 11 or a pharmaceutically acceptable salt thereof, wherein: L is -CH2-.

20. The compound of claim 12 or a pharmaceutically acceptable salt thereof, wherein: L is -CH2-.

21. The compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in claim 1, wherein the compound is selected from the following structural formula:

22. A pharmaceutical composition comprising an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 21 and a pharmaceutically acceptable excipient.

23. Use of the compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 21 or the pharmaceutical composition according to claim 22 in the preparation of a medicament for treating tumors, autoimmune diseases, or diseases caused by viruses, wherein The autoimmune disease is multiple sclerosis; wherein the tumor is cancer, and the cancer is cancer with BRCA-1 or BRCA-2 mutation; and wherein the virus is selected from herpes zoster virus or herpes simplex virus.

24. Use of the compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 21 or the pharmaceutical composition according to claim 22 in the preparation of a medicament for treating a tumor, wherein the tumor is cancer, and the cancer is breast cancer, ovarian cancer, pancreatic cancer, lung cancer or head and neck cancer.

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