AKR1C3 INHIBITOR AND MEDICAL USE

Abstract

Described are compounds that act as AKR1C3 inhibitors or pharmaceutically acceptable salts or solvates or isotopically substituted compounds thereof and methods thereof.

Claims

1. A compound of the following formula: ##STR00097## wherein R.sub.1 and R.sub.2 are each independently hydrogen, deuterium, aryl or Z-substituted aryl, heteroaryl or Z-substituted heteroaryl, C.sub.1-C.sub.6 alkyl or Z-substituted alkyl, C.sub.2-C.sub.6 alkenyl or Z-substituted alkenyl, C.sub.2-C.sub.6 alkynyl or Z-substituted alkynyl, or C.sub.3-C.sub.8 cycloalkyl or Z-substituted cycloalkyl; R.sub.3 is hydrogen, halogen, cyano or isocyanato, hydroxyl, mercapto, amino, oximido, hydrazono, OTs, OMs, C.sub.1-C.sub.6 alkyl or Z-substituted alkyl, C.sub.2-C.sub.6 alkenyl or Z-substituted alkenyl, C.sub.2-C.sub.6 alkynyl or Z-substituted alkynyl, C.sub.3-C.sub.8 cycloalkyl or Z-substituted cycloalkyl, C.sub.6-C.sub.10 aryl or Z-substituted aryl, 4-15-membered heterocyclic radical or Z-substituted heterocyclic radical, 5-15-membered heteroaryl or Z-substituted heteroaryl, or C.sub.1-C.sub.6 alkoxy or Z-substituted C.sub.1-C.sub.6 alkoxy; or R.sub.3 is —CONR.sup.6R.sup.7, —SO.sub.2NR.sup.6R.sup.7, —SO.sub.2R.sup.6, —OCO—R.sup.6, —OCOO—R.sup.6, —COOR.sup.6, —NR.sup.6COR.sup.7, —NR.sup.6SO.sub.2R.sup.7, or —NR.sup.6CONR.sup.6R.sup.7; and R.sup.6 and R.sup.7 together with N to form or not to form a 4-8-membered Z-substituted heterocyclic ring, or two R.sub.3 together with the atom on a benzene ring to which they are bonded to form a 7-15-membered fused ring or a Z-substituted fused ring; R.sup.6 and R.sup.7 are each independently hydrogen, cyano or isocyanato, C.sub.1-C.sub.6 alkyl or Z-substituted alkyl, C.sub.2-C.sub.6 alkenyl or Z-substituted alkenyl, C.sub.2-C.sub.6 alkynyl or Z-substituted alkynyl, C.sub.3-C.sub.8 cycloalkyl or Z-substituted cycloalkyl, C.sub.6-C.sub.10 aryl or Z-substituted aryl, 4-15-membered heterocyclic radical or Z-substituted heterocyclic radical, 5-15-membered heteroaryl or Z-substituted heteroaryl, or C.sub.1-C.sub.6 alkoxy or Z-substituted C.sub.1-C.sub.6 alkoxy, or R.sup.6 and R.sup.7 together with the atom to which they are bonded to form a 3-7-membered heterocyclic radical or Z-substituted 3-7-membered heterocyclic radical; a is 0, 1, 2 or 3; X is C or N; Y is O or S; Cx is selected from the group consisting of C.sub.6-C.sub.10 aryl or Z-substituted aryl, 4-15-membered heterocyclic radical or Z-substituted 4-15-membered heterocyclic radical, 5-15-membered heteroaryl or Z-substituted heteroaryl, 7-15-membered fused ring or Z-substituted fused ring, and —CONR.sup.6R.sup.7, —SO.sub.2NR.sup.6R.sup.7, —SO.sub.2R.sup.6, —OCOO—R.sup.6, —COOR.sup.6, —NR.sup.6COR.sup.7, —OCOR.sup.6, —NR.sup.6SO.sub.2R.sup.7, —NR.sup.6SO.sub.2NR.sup.6R.sup.7, —COR.sup.6, —NR.sup.6CONR.sup.6R.sup.7 substituted C.sub.6-C.sub.10 aryl, 4-15-membered heterocyclic radical, 5-15-membered heteroaryl, and 7-15-membered fused ring; and R.sup.6 and R.sup.7 together with N to form or not to form a 4-8-membered Z-substituted heterocycle; L is selected from the group consisting of —O—, —S—, —OCOO—, —NR.sup.6CO—, —OCO—, —NR.sup.6SO.sub.2—, —OCONR.sup.6—, quaternary ammonium, and sulfonate group —OSO.sub.2—; Cy is selected from the group consisting of hydrogen, deuterium, C.sub.6-C.sub.10 aryl or Z-substituted aryl, 4-15-membered heterocyclic or Z-substituted heterocyclic, 5-15-membered heteroaryl or Z-substituted heteroaryl, 7-15-membered fused ring or Z-substituted fused ring, C.sub.1-C.sub.6 alkyl or Z-substituted alkyl, C.sub.2-C.sub.6 alkenyl or Z-substituted alkenyl, C.sub.2-C.sub.6 alkynyl or Z-substituted alkynyl, and C.sub.3-C.sub.8 cycloalkyl or Z-substituted C.sub.3-C.sub.8 cycloalkyl; or Cy is selected from the group consisting of ##STR00098## or Cy is selected from the group consisting of a 5-10-membered ring group formed by the two OR.sup.6 and the P atom in ##STR00099## a 5-10-membered ring group formed by the OR.sup.6 and the NR.sup.6R.sup.7 together with the P atom in ##STR00100## and a 5-10-membered ring group formed by the two NR.sup.6R.sup.7 and the P atom in ##STR00101## and -L-Cy excludes the residues of these phosphoramidate alkylating agents after losing a H atom: —P(Z.sup.1)(NR.sup.9CH.sub.2CH.sub.2X.sup.1).sub.2, —P(Z.sup.1)(NR.sup.9.sub.2)(N(CH.sub.2CH.sub.2X.sup.1)2), —P(Z.sup.1)(N(CH.sub.2CH.sub.2X.sup.1)).sub.2 or —P(Z.sup.1)(N(CH.sub.2CH.sub.2X.sup.1).sub.2).sub.2, each R.sup.9 is independently hydrogen or C.sub.1-C.sub.6 alkyl, or two R.sup.9 together with the nitrogen atom to which they are bonded to form a 5-7-membered heterocyclyl, Z.sup.1 is O or S, X.sup.1 is Cl, Br or OMs, and -L-Cy excludes —OH or —SH; the substituent Z is a halogen atom, cyano or isocyanato, hydroxyl, mercapto, amino, oximido, hydrazono, OTs, OMs, C.sub.1-C.sub.3 alkyl or substituted alkyl, C.sub.1-C.sub.3 alkoxy or substituted alkoxy, C.sub.2-C.sub.3 alkenyl or substituted alkenyl, C.sub.2-C.sub.3 alkynyl or substituted alkynyl, C.sub.3-C.sub.8 cycloalkyl or substituted cycloalkyl, an aromatic ring, heterocyclic ring, a heteroaromatic ring and fused ring or a substituted aromatic ring, heterocyclic ring, heteroaromatic ring and fused ring, and the pattern of substitution being mono-substitution or geminal di-substitution; Cz group is a C-, P-, S-containing group which can be hydrolyzed by hydrolytic enzymes such that corresponding C—N, P—N or S—N bond is cleaved, or a pharmaceutically acceptable salt, or a solvate, or an isotopically substituted compound thereof.

2. The compound according to claim 1, wherein the compound is a compound of formula I-1 to I-5, ##STR00102## wherein, I-5 is a prodrug that can be converted in vivo to the above compound I-3, R.sub.1 and R.sub.2 are each independently hydrogen, deuterium, aryl or Z-substituted aryl, heteroaryl or Z-substituted heteroaryl, C.sub.1-C.sub.6 alkyl or Z-substituted alkyl, C.sub.2-C.sub.6 alkenyl or Z-substituted alkenyl, C.sub.2-C.sub.6 alkynyl or Z-substituted alkynyl, or C.sub.3-C.sub.8 cycloalkyl or Z-substituted cycloalkyl; R.sub.3 is hydrogen, halogen, cyano or isocyanato, hydroxyl, mercapto, amino, oximido, hydrazono, OTs, OMs, C.sub.1-C.sub.6 alkyl or Z-substituted alkyl, C.sub.2-C.sub.6 alkenyl or Z-substituted alkenyl, C.sub.2-C.sub.6 alkynyl or Z-substituted alkynyl, C.sub.3-C.sub.8 cycloalkyl or Z-substituted cycloalkyl, C.sub.6-C.sub.10 aryl or Z-substituted aryl, 4-15-membered heterocyclic radical or Z-substituted heterocyclic radical, 5-15-membered heteroaryl or Z-substituted heteroaryl, or C.sub.1-C.sub.6 alkoxy or Z-substituted C.sub.1-C.sub.6 alkoxy; or R.sub.3 is —CONR.sup.6R.sup.7, —SO.sub.2NR.sup.6R.sup.7, —SO.sub.2R.sup.6, —OCO—R.sup.6, —OCOO—R.sup.6, —COOR.sup.6, —NR.sup.6COR.sup.7, —NR.sup.6SO.sub.2R.sup.7, or —NR.sup.6CONR.sup.6R.sup.7, and R.sup.6 and R.sup.7 together with the N to form or not to form a 4-8-membered Z-substituted heterocyclic ring; R.sub.4 and R.sub.5 are each independently hydrogen, halogen, cyano or isocyano, hydroxy, mercapto, amino, oximido, hydrazono, OTs, OMs, C.sub.1-C.sub.6 alkyl or Z-substituted alkyl, C.sub.2-C.sub.6 alkenyl or Z-substituted alkenyl, C.sub.2-C.sub.6 alkynyl or Z-substituted alkynyl, C.sub.3-C.sub.8 cycloalkyl or Z-substituted cycloalkyl, C.sub.6-C.sub.10 aryl or Z-substituted aryl, 4-15-membered heterocyclic radical or Z-substituted heterocyclic radical, 5-15-membered heteroaryl or Z-substituted heteroaryl, or C.sub.1-C.sub.6 alkoxy or Z-substituted C.sub.1-C.sub.6 alkoxy; or R.sub.4 and R.sub.5 are each —CONR.sup.6R.sup.7, —SO.sub.2NR.sup.6R.sup.7, —SO.sub.2R.sup.6, —OCOO—R.sup.6, —COOR.sup.6, —NR.sup.6COR.sup.7, —OCOR.sup.6, —NR.sup.6SO.sub.2R.sup.7, or —NR.sup.6CONR.sup.6R.sup.7, or R.sub.4 and R.sub.5 together with the atom on a benzene ring to which they are bonded to form a 7-15-membered fused ring or a Z-substituted fused ring, and R.sub.6 and R.sub.7 together with N to form or not to form a 4-8-membered Z-substituted heterocyclic ring; R.sub.6 and R.sub.7 are each independently hydrogen, cyano or isocyanato, C.sub.1-C.sub.6 alkyl or Z-substituted alkyl, C.sub.2-C.sub.6 alkenyl or Z-substituted alkenyl, C.sub.2-C.sub.6 alkynyl or Z-substituted alkynyl, C.sub.3-C.sub.8 cycloalkyl or Z-substituted cycloalkyl, C.sub.6-C.sub.10 aryl or Z-substituted aryl, 4-15-membered heterocyclic radical or Z-substituted heterocyclic radical, 5-15-membered heteroaryl or Z-substituted heteroaryl, or C.sub.1-C.sub.6 alkoxy or Z-substituted C.sub.1-C.sub.6 alkoxy; or R.sup.6 and R.sup.7 groups together with the atom to which they are bonded to form a 3-7-membered heterocyclyl or a Z-substituted 3-7-membered heterocyclyl, and R.sup.6 and R.sup.7 together with N to form or not to form a 4-8-membered Z-substituted heterocyclic ring; Y is O or S; Cx is selected from the group consisting of C.sub.6-C.sub.10 aryl or Z-substituted aryl, 4-15-membered heterocyclic radical or Z-substituted 4-15-membered heterocyclic radical, 5-15-membered heteroaryl or Z-substituted heteroaryl, 7-15-membered fused ring or Z-substituted fused ring, and —CONR.sup.6R.sup.7, —SO.sub.2NR.sup.6R.sup.7, —SO.sub.2R.sup.6, —OCOO—R.sup.6, —COOR.sup.6, —NR.sup.6COR.sup.7, —OCOR.sup.6, —NR.sup.6SO.sub.2R.sup.7, —NR.sup.6SO.sub.2NR.sup.6R.sup.7, —COR.sup.6, —NR.sup.6CONR.sup.6R.sup.7 substituted C.sub.6-C.sub.10 aryl, 4-15-membered heterocyclic radical, 5-15-membered heteroaryl, 7-15-membered fused ring, and R.sup.6 and R.sup.7 together with N to form or not to form a 4-8-membered Z-substituted heterocycle; L is selected from the group consisting of —O—, —S—, —OCOO—, —NR.sup.6CO—, —OCO—, —NR.sup.6SO.sub.2—, —OCONR.sup.6—, quaternary ammonium, and sulfonate group —OSO.sub.2—; Cy is selected from the group consisting of hydrogen, deuterium, C.sub.6-C.sub.10 aryl or Z-substituted aryl, 4-15-membered heterocyclic radical or Z-substituted heterocyclic radical, 5-15-membered heteroaryl or Z-substituted heteroaryl, 7-15-membered fused ring or Z-substituted fused ring, C.sub.1-C.sub.6 alkyl or Z-substituted alkyl, C.sub.2-C.sub.6 alkenyl or Z-substituted alkenyl, C.sub.2-C.sub.6 alkynyl or Z-substituted alkynyl, or C.sub.3-C.sub.8 cycloalkyl or Z-substituted C.sub.3-C.sub.8 cycloalkyl; or Cy is selected from the group consisting of ##STR00103## or Cy is selected from the group consisting of a 5-10-membered ring group formed by the two OR.sup.6 and the P atom in ##STR00104## a 5-10-membered ring group formed by the OR.sup.6 and the NR.sup.6R.sup.7 together with the P atom in ##STR00105## and a 5-10-membered ring group formed by the two NR.sup.6R.sup.7 and the P atom in ##STR00106## the substituent Z is a halogen atom, cyano or isocyanato, hydroxyl, mercapto, amino, oximido, hydrazono, OTs, OMs, C.sub.1-C.sub.3 alkyl or substituted alkyl, C.sub.1-C.sub.3 alkoxy or substituted alkoxy, C.sub.2-C.sub.3 alkenyl or substituted alkenyl, C.sub.2-C.sub.3 alkynyl or substituted alkynyl, C.sub.3-C.sub.8 cycloalkyl or substituted cycloalkyl, an aromatic ring, heterocyclic ring, a heteroaromatic ring and fused ring or a substituted aromatic ring, heterocyclic ring, heteroaromatic ring and fused ring, and the pattern of substitution being mono-substitution or geminal di-substitution; Cz group is a C-, P-, S-containing group which can be hydrolyzed by hydrolytic enzymes such that corresponding C—N, P—N or S—N bond is cleaved.

3. The compound according to claim 1, wherein, R.sub.1 and R.sub.2 are each independently hydrogen, deuterium, C.sub.1-C.sub.6 alkyl or Z-substituted alkyl, C.sub.2-C.sub.6 alkenyl or Z-substituted alkenyl, C.sub.2-C.sub.6 alkynyl or Z-substituted alkynyl, or C.sub.3-C.sub.8 cycloalkyl or Z-substituted cycloalkyl.

4. The compound according to claim 3, wherein R.sub.1 and R.sub.2 are each independently hydrogen, deuterium, or methyl.

5. The compound according to claim 2, wherein: R.sub.3, R.sub.4 and R.sub.5 are each independently hydrogen, Cx is —CONR.sup.6R.sup.7 substituted phenyl, and R.sup.6 and R.sup.7 together with N to form or not to form a 4-8-membered Z-substituted heterocyclic ring, L is selected from —O— or —S, or Cy is selected from the group consisting of C.sub.6-C.sub.10 aryl or halo-substituted aryl, 4-15-membered heterocyclic or halo-substituted heterocyclic, 5-15-membered heteroaryl or halo-substituted heteroaryl, and 7-15-membered fused ring or halo-substituted fused ring.

6. (canceled)

7. (canceled)

8. (canceled)

9. The compound according to claim 5, wherein Cy is selected from the group consisting of fluorophenyl, difluorophenyl, and trifluorophenyl.

10. The compound according to claim 1, wherein Cy is selected from the group consisting of ##STR00107## or Z-substituted ##STR00108##

11. The compound according to claim 1, wherein Cz is selected from —COR.sup.6, or —COOR.sup.6.

12. The compound according to claim 1, wherein —NH-Cz is a phosphamido group.

13. The compound according to claim 1, wherein the compound is: ##STR00109## ##STR00110## ##STR00111## ##STR00112##

14. The compound according to claim 1, wherein the compound is the pharmaceutically acceptable salt thereof being a basic salt or an acid salt, or wherein the compound is the solvate thereof being a hydrate or an alcoholate.

15. A medicament comprising the compound according to claim 1, or the pharmaceutically acceptable salt, or the solvate, or the isotopically substituted compound thereof, and a pharmaceutically acceptable adjuvant or excipient.

16. The medicament according to claim 15, wherein the medicament comprises a therapeutically effective amount of the compound according to claim 1.

17. (canceled)

18. A method for the treatment or prevention of a disease/disorder in a subject, the method comprising the step of administering to the subject a therapeutically effective amount of at least one compound of claim 1, wherein the disease/disorder is: endometriosis, uterine leiomyoma, uterine hemorrhagic disorders, dysmenorrhea, prostatic hyperplasia, acne, seborrhea, alopecia, premature sexual maturation, polycystic ovary syndrome, chronic obstructive pulmonary disease COPD, obesity, inflammatory pain, cancer, inflammation, or cancer pain.

19. A method for enhancing sensitivity to radiation therapy for cancers or tumors in a subject, the method comprising the step of administering to the subject a therapeutically effective amount of at least one compound of claim 1, wherein the method increases the efficacy of radiation therapy when the subject has a cancer or a tumor that is resistant to radiation therapy.

20. A method for enhancing sensitivity to immunotherapy for cancers or tumors in a subject, the method comprising the step of administering to the subject a therapeutically effective amount of at least one compound of claim 1, wherein the method enhances the efficacy of radiation therapy when the subject has a cancer or a tumor that is resistant to immunotherapy.

21. A method for enhancing sensitivity to chemotherapy for cancers or tumors in a subject, the method comprising the step of administering to the subject a therapeutically effective amount of at least one compound of claim 1, wherein the method enhances the efficacy of chemotherapy when the subject has a cancer or a tumor that is resistant to chemotherapy.

22. A method for enhancing sensitivity to chemotherapy for cancers or tumors in a subject, the method comprising the step of administering to the subject a therapeutically effective amount of at least one compound of claim 1, wherein the method increases the efficacy of chemotherapy when the subject has a cancer or a tumor that is resistant to chemotherapy, and wherein the chemotherapy contains daunorubicin or cytarabine.

23. A method for enhancing the sensitivity to radiation therapy for a cancer or a tumor in a subject, the method comprising the step of administering to the subject a therapeutically effective amount of at least one compound of claim 1, wherein the method enhances the sensitivity to chemotherapy for the cancer or the tumor for a treatment using a medicament containing daunorubicin or cytarabine.

24. A method for the treatment or prevention of a disease/disorder in a subject, the method comprising the step of administering to the subject a therapeutically effective amount of at least one compound of claim 1, wherein the disease/disorder is treated by the compound acting as an AKR1C3 enzyme inhibitor.

25.-29. (canceled)

Description

DESCRIPTION OF EMBODIMENTS

[0128] The present application will be described below with reference to specific examples. It will be appreciated by those skilled in the art that these examples are intended to describe the present application only and are not intended to limit the scope of the present application in any way.

[0129] The experimental methods in the following examples, unless otherwise specified, are all conventional methods. The raw materials of the medicaments, the reagent materials, and the like used in the following examples are all commercially available products unless otherwise specified. “Patient” and “subject” are used interchangeably to refer to a mammal in need of treatment for cancer.

[0130] Typically, the patient is a human. Typically, the patient is a human diagnosed with cancer. In certain embodiments, a “patient” or “subject” may refer to a non-human mammal, such as non-human primate, dog, cat, rabbit, pig, mouse or rat, used in screening, characterizing, and evaluating drugs and therapies. “Prodrug” refers to a compound that, upon feeding or administration, is metabolized or otherwise converted to a biologically active or more active compound (or drug) with respect to at least one property. A prodrug is chemically modified relative to the drug in a manner that renders it less active or inactive relative to the drug, but the chemical modification is such that the corresponding drug is generated by metabolic or other biological processes following administration of the prodrug. A prodrug may have altered metabolic stability or transport characteristics, fewer side effects or lower toxicity, or improved flavor relative to the active drug (see, for example, reference Nogrady, 1985, Medicinal Chemistry A Biochemical Approach, Oxford University Press, New York, pages 388-392, which is incorporated herein by reference). A prodrug may be synthesized using reactants other than the corresponding drug.

[0131] “Solid tumor” refers to solid tumors including, but not limited to, metastatic tumors in bone, brain, liver, lung, lymph nodes, pancreas, prostate, skin, and soft tissue (sarcoma).

[0132] “Therapeutically effective amount” of a drug refers to an amount of a drug that, when fed or administered to a patient with cancer, will have the intended therapeutic effect, e.g., alleviation, amelioration, palliation or elimination of one or more clinical manifestations of the cancer in the patient. A therapeutic effect does not necessarily occur by feeding or administration of one dose, and may occur only after feeding or administration of a series of doses. Thus, a therapeutically effective amount may be fed or administered in one or more administrations.

[0133] “Treatment” of a condition or patient refers to taking steps to obtain beneficial or desired results including clinical results. For purposes of this application, beneficial or desired clinical results include, but are not limited to, alleviation or amelioration of one or more symptoms of a cancer; diminishment of extent of disease; delay or slowing of disease progression; amelioration, palliation, or stabilization of the disease state; or other beneficial results. In some cases, treatment of a cancer may result in partial response or stable disease.

[0134] “Tumor cell” refers to a tumor cell of any appropriate species (e.g., mammalian, such as murine, canine, feline, equine, or human).

[0135] Radiation therapy, i.e., radiation therapy of a cancer, has a variety of embodiments, and the most commonly used being:

[0136] Photon (γ-ray) beam emission, in which a number of photon (γ-ray) beams are emitted by a linear accelerator;

[0137] Neutron beam radiation, which can be used to treat certain carcinomas with narrow tissue margins;

[0138] Electron beam radiation, which is useful in the treatment of skin or superficial carcinomas due to its very shallow depth of penetrating tissues;

[0139] Proton radiation, which, although has limitation in use, provides a sharp ray perimeter for a very narrow field of illumination requiring a certain depth;

[0140] Brachytherapy, in which a powerful source of radiation is implanted through a needle into the tumor tissue itself (e.g., a prostate or a lung) to achieve an effect with a small range and a high dose;

[0141] Systemic radionuclide therapy, which can be used for organ receptors with nuclide uptake (such as thyroid cancer) or receptors that inhibit systemic skeletal sites (such as radioactive strontium to treat metastatic prostate cancer);

[0142] Curative radiation therapy, which generally entails inclusion of the tumor locally or its local area within the field of irradiation;

[0143] The damage of radioactive irradiation to cells is non-selective and non-specific and produces complex effects on DNA, with the superior efficacy depending on how much the damage to cells exceeds the ability thereof to repair. In general, a normal tissue is repaired more efficiently than cancer cells, so that radiation therapy can benefit the patients.

[0144] Chemotherapy, which is a short term for chemodrug therapy, is used to kill cancer cells using chemotherapeutic drugs for therapeutic purposes. Chemotherapy is currently one of the most effective means to treat cancers. Chemotherapy, surgery and radiotherapy together are referred to as the three major treatments for cancers. Surgery and radiotherapy are local treatments that are only effective against tumors at the site of treatment, and effective treatments are difficult to exert for potential metastatic lesions (cancer cells actually have metastasized, but not yet clinically discovered and detectable because of limitations of current technical means) and cancers that have already developed clinical metastases. Whereas chemotherapy is a means of systemic treatment, chemotherapy drugs circulate throughout the vast majority of organs and tissues throughout the body with the blood, whatever route of administration is used (oral, intravenous and body cavity administration, etc.). Therefore, chemotherapy is the primary treatment for some tumors that have a tendency to spread systemically as well as mid-late tumors that have metastasized. Chemotherapeutic drugs in general are cytotoxic drugs that kill cells directly, but due to certain different characteristics between cancer cells and normal cells, chemotherapeutic drugs often have a greater killing effect for cancer cells, which can benefit the patients. However, chemotherapy still suffers from serious toxic side effects, including digestive system reactions, bone marrow suppression, hair loss, and some of these toxic side effects may not continue to receive chemotherapy or some chemotherapeutic agent.

[0145] Regarding immunotherapy, normally, the body's immune system can recognize and clear tumor cells from the tumor microenvironment, but tumor cells, in order to survive and grow, can adopt different strategies such that the body's immune system is suppressed and unable to kill tumor cells properly, thereby surviving various stages of an anti-tumor immune response. The process by which tumor cells are recognized and cleared by the human immune system (tumor-immune cycle) has multiple stages, and any of the stages is inhibited, it will lead to the result that tumor cells cannot be cleared normally. Different tumors can inhibit the effective recognition and killing of tumor cells by the immune system through abnormalities in different stages and thus produce immune tolerance, and even the development of tumors can be promoted.

[0146] Tumor immunotherapy is a treatment that restores the body's normal anti-tumor immune response to control and clear tumors by reinitiating and maintaining the tumor-immune cycle. Tumor immunotherapy includes, among others, monoclonal antibody-based immune checkpoint inhibitors, therapeutic antibodies, cancer vaccines, cell therapies, and small molecule inhibitors. In recent years, there has been ongoing good news for tumor immunotherapy, and at present, it has already shown strong anti-tumor activity in the treatment of a variety of tumors such as melanoma, non-small cell lung cancer, renal cancer and prostate cancer, and a number of tumor immunotherapy drugs have been approved for clinical use.

[0147] The foregoing description of specific embodiments of the application does not limit the application, and those skilled in the art can make various changes and modifications in light of the application without departing from the spirit of the application, which shall fall within the scope of the appended claims.

[0148] The following is specific tests and examples of the present application.

[0149] The following tests will reveal the in vitro inhibitory activity of AKR1C3 inhibitors developed by the applicant, who hereby state that the right to the following experimental data belongs to the applicant.

[0150] The applicant states that some of the specific compounds disclosed in the tests below can be synthesized based on specific synthetic methods and synthetic routes of the compounds disclosed herein, with reference to patent publications (such as Application No. PCT/US2016/021581, Publication No. WO2016/145092; Application No. PCT/US2016/062114, Publication No. WO 2017/087428) or other publications (although the substrates are different and the yields are either high or low, but the technicians who have mastered the skills of organic synthesis in this field, such as pharmaceutical chemistry and organic chemistry, can still synthesize them), and the applicant has confirmed the structure by NMR and MS. This application provides synthesis methods, NMR data and in vitro inhibitory activities against AKR1C3 enzyme of representative compounds.

[0151] Compound Synthesis Experiments

[0152] Overview of Synthetic Methods

##STR00025## ##STR00026##

Description of English Abbreviations

[0153] MTBE, methyl tert-butyl ether; DMAP, 4-dimethylaminopyridine; T.sub.3P, propylphosphonic anhydride; THF, tetrahydrofuran; DCM, dichloromethane; EA or EtOAC, ethyl acetate; TEA, triethylamine; HPLC, high performance liquid chromatography; DBAD, di-tert-butyl azodicarboxylate; TFA, trifluoroacetic acid; LCMS, liquid chromatography-mass spectrometer; EtOH, ethanol; T-BuOH, tert-butanol; DMF, dimethylformamide; PE, petroleum ether; eq, equivalent i.e. molar ratio; TBAF, tetrabutylammonium fluoride; DIPEA, N, N-diisopropylethylamine; reflux, backflow; rt, room temperature.

[0154] The chemical reagents and drugs whose sources are not indicated in the synthesis process are analytically pure or chemically pure, and they are all purchased from commercial reagent companies.

[0155] Reference shall be made to the interpretations in the field of organic chemistry for other English abbreviations.

[0156] Synthesis of Compound #2

##STR00027##

[0157] Under the protection of nitrogen, 2-A1 (500 mg, 1.51 mmol, synthesized with reference to the synthesis method of Compound #29) and 2-A2 (336 mg, 2.27 mmol, commercially available) were dissolved in anhydrous THF (10 mL). The reactants were cooled to 0° C., triphenylphosphonium (991 mg, 3.78 mmol) was added, then a solution of di-tert-butyl azodicarboxylate (870 mg, 3.78 mmol) in THF (5 mL) was slowly added dropwise, with the temperature being kept at 0° C. for half an hour. Then the reactants were stirred at room temperature for 2.5 hours. After the reaction was completed, water (5 mL) was added dropwise at 0° C. The reactants were extracted with DCM (10 mL×3), washed with water (3 mL×2), dried and concentrated, followed by high performance liquid chromatography to obtain pure Compound #2 (340 mg, 48.9%) as a pale yellow oil. .sup.1H-NMR (400M, CD.sub.3OD): 58.01 (d, J=8.4 Hz, 1H), 7.52-7.47 (m, 2H), 7.45 (s, 2H), 7.30 (d, J=1.2 Hz, 1H), 7.25-7.23 (m, 1H), 7.10-7.07 (m, 2H), 5.54-5.52 (m, 1H), 3.08 (s, 3H), 2.98 (s, 3H), 1.61 (d, J=6.4 Hz, 3H).

[0158] Synthesis of Compound #3

##STR00028##

[0159] Under the protection of nitrogen, 3-A1 (200 mg, 0.632 mmol, synthesized with reference to the synthesis method of Compound #29) and 3-A2 (140 mg, 0.948 mmol, commercially available) were dissolved in anhydrous THF (5 mL). The reactants were cooled to 0° C., triphenylphosphonium (414 mg, 1.58 mmol) was added, then a solution of di-tert-butyl azodicarboxylate (363 mg, 1.58 mmol) in THF (2 mL) was slowly added dropwise, with the temperature being kept at 0° C. for half an hour. Then the reactants were stirred at room temperature for 2.5 hours. After the reaction was completed, water (5 mL) was added dropwise at 0° C. The reactants were extracted with DCM (5 mL×3), washed with water (2 mL×2), dried and concentrated, followed by high performance liquid chromatography to obtain pure Compound #3 (160 mg, with a yield of 56.7%) as a pale yellow oil. .sup.1H-NMR (400M, CDCl.sub.3): S7.95 (d, J=8.4 Hz, 1H), 7.38-7.32 (m, 2H), 7.25 (s, 1H), 7.20-7.16 (m, 3H), 7.05-7.02 (m, 2H), 4.94 (s, 2H), 3.02 (s, 3H), 2.90 (s, 3H).

[0160] Synthesis of Compound #4

##STR00029##

[0161] Under the protection of nitrogen, phosphorus oxychloride (580 mg, 3.78 mmol) was added dropwise to anhydrous DCM (5 mL). The reactants were cooled to −40° C., a solution of 4-A1 (500 mg, 1.51 mmol, synthesized with reference to the synthesis method of Compound #29) in DCM (3 mL) was added dropwise, then a solution of TEA (397 mg, 3.9 mmol) in DCM (2 mL) was added dropwise, with the temperature being kept at −40° C. for 6 h until the raw material disappeared. Methylamine (1.6 g, 12.8 mmol, 25% in THF) was added dropwise, then a solution of TEA (1.29 g, 12.85 mmol) in DCM (5 mL) was added dropwise. The temperature was kept at −40° C. for half an hour and then is naturally increased to an ambient temperature, and the reactants were stirred overnight. After the reaction was completed, the reactants were cooled to 0° C., a potassium carbonate solution (1 g, 10 mL) was added dropwise. The reactants were extracted with DCM (10 mL×3), washed with water (5 mL×2), dried and concentrated, followed by high performance liquid chromatography to obtain the pure product (380 mg, with a yield of 57.7%) as a pale yellow viscous oil. .sup.1H-NMR (400 MHz, CDCl.sub.3): δ7.95 (d, J=8.4 Hz, 1H), 7.42 (t, J=7.9 Hz, 1H), 7.21-7.18 (m, 2H), 7.12-7.07 (m, 2H), 7.04 (s, 1H), 5.47-5.44 (m, 1H), 3.08 (s, 3H), 2.97 (s, 3H), 2.59 (d, J=12.2 Hz, 3H), 2.51 (m, 2H), 2.38 (d, J=12.2 Hz, 3H), 1.53 (d, J=6.5 Hz, 3H). MS: Calculated 436.2, found 437.1 ([M+H].sup.+).

[0162] Synthesis of Compound #5

##STR00030##

[0163] Under the protection of nitrogen, phosphorus oxychloride (242 mg, 1.58 mmol) was added dropwise to anhydrous DCM (5 mL). The reactants were cooled to −40° C., a solution of 5-A1 (200 mg, 0.63 mmol, synthesized with reference to the synthesis method of Compound #29) in DCM (3 mL) was added dropwise, then a solution of TEA (166 mg, 1.64 mmol) in DCM (2 mL) was added dropwise, with the temperature being kept at −40° C. for 6 h until the raw material disappeared. Methylamine (670 g, 5.40 mmol, 25% in THF) was added dropwise, then a solution of TEA (546 g, 5.4 mmol) in DCM (5 mL) was added dropwise. The temperature was kept at −40° C. for half an hour and then is naturally increased to an ambient temperature, and the reactants were stirred overnight. After the reaction was completed, the reactants were cooled to 0° C., a potassium carbonate solution (1 g, 10 mL) was added dropwise. The reactants were extracted with DCM (10 mL×3), washed with water (5 mL×2), dried and concentrated, followed by high performance liquid chromatography to obtain pure Compound #5 (130 mg, with a yield of 48.9%) as a pale yellow oil. .sup.1H-NMR (400M, CDCl.sub.3): δ7.99 (d, J=8.4 Hz, 1H), 7.47-7.44 (m, 2H), 7.24 (d, J=1.6 Hz, 1H), 7.07-7.04 (m, 3H), 4.94 (d, J=7.9 Hz, 2H), 3.11-3.02 (m, 6H), 2.62 (s, 3H), 2.59 (s, 3H). MS: Calculated 422.1, found 423.2 ([M+H].sup.+).

[0164] Synthesis of Compound #6

##STR00031##

[0165] Under the protection of nitrogen, tris(dimethylamino)phosphine (179 mg, 1.1 mmol, commercially available) and tetrazole (0.64 mg, 0.009 mmol) were added to acetonitrile (3 mL), then a solution of 6-A1 (300 mg, 0.914 mmol, synthesized with reference to the synthesis method of Compound #29) in acetonitrile (2 mL) was added dropwise. The reactants were stirred at room temperature for two hours, and then TEA (277 mg, 2.74 mmol) and tert-butanol peroxide (329 mg, 3.66 mmol) were added dropwise. The reactants were stirred at room temperature for three hours, until the reaction was completed. An aqueous sodium thiosulfate solution (4 mL) was added dropwise, and the reactants were extracted with DCM (5 mL×3), dried and concentrated, followed by high performance liquid chromatography to separate and obtain Compound #6 (120 mg, with a yield of 28.3%) as a pale yellow oil. .sup.1H-NMR (400M, CDCl.sub.3):57.97 (d, J=8.4 Hz, 1H), 7.42-7.38 (m, 1H), 7.23-7.20 (m, 2H), 7.10-7.05 (m, 3H), 5.41-5.38 (m, 1H), 3.08 (s, 3H), 2.98 (s, 3H), 2.64 (d, J=10.0 Hz, 6H), 2.43 (d, J=10.0 Hz, 6H), 1.52 (d, J=6.5 Hz, 3H). MS: Calculated 464.2, found 465.2 ([M+H].sup.+).

[0166] Synthesis of Compound #7

##STR00032##

[0167] Under the protection of nitrogen, tris(dimethylamino)phosphine (124 mg, 0.76 mmol, commercially available) and tetrazole (0.4 mg, 0.0057 mmol) were added to acetonitrile (3 mL), then a solution of 7-A1 (200 mg, 0.63 mmol, synthesized with reference to the synthesis method of Compound #29) in acetonitrile (2 mL) was added dropwise. The reactants were stirred at room temperature for two hours. TEA (192 mg, 1.90 mmol) and tert-butanol peroxide (228 mg, 2.53 mmol) were added dropwise. The reactants were stirred at room temperature for three hours, until the reaction was completed. An aqueous sodium thiosulfate solution (4 mL) was added dropwise, and the reactants were extracted with DCM (5 mL×3), dried and concentrated, followed by high performance liquid chromatography to separate and obtain Compound #7 (90 mg, with a yield of 31.6%) as a pale yellow oil. .sup.1H-NMR (400M, CDCl.sub.3): 58.00 (d, J=8.4 Hz, 1H), 7.46-7.42 (m, 1H), 7.28-7.23 (m, 2H), 7.12-7.08 (m, 3H), 4.97-4.95 (m, 2H), 3.11 (s, 3H), 3.00 (s, 3H), 2.60 (d, J=10.0 Hz, 12H). MS: Calculated 450.2, found 451.2 ([M+H]).

[0168] Synthesis of Compound #8

##STR00033##

[0169] Under the protection of nitrogen, 8-A1 (300 mg, 0.91 mmol, synthesized with reference to the synthesis method of Compound #29) and 8-A2 (202 mg, 1.36 mmol) were dissolved in anhydrous THF (5 mL). The reactants were cooled to 0° C. Triphenylphosphonium (600 mg, 2.30 mmol) was added, then a solution of di-tert-butyl azodicarboxylate (530 mg, 2.30 mmol) in THF (3 mL) was slowly added dropwise. The reactants were kept at 0° C. for half an hour and then the reactants were stirred at room temperature for 2.5 hours. After the reaction was completed, water (5 mL) was added dropwise at 0° C., and the reactants were extracted with DCM (5 mL×3), washed with water (2 mL×), dried and concentrated, followed by high performance liquid chromatography to obtain pure Compound #8 (230 mg, with a yield of 54.9%) as a pale yellow solid. .sup.1H-NMR (400M, DMSO-d6): δ8.13 (d, J=8.4 Hz, 1H), 7.68 (s, 2H), 7.56 (dd, J=8.4, 2.0 Hz, 1H), 7.47 (dd, J=6.8, 2.0 Hz, 2H), 7.33 (d, J=1.6 Hz, 1H), 7.05-7.03 (m, 2H), 5.52-5.50 (m, 1H), 2.95 (s, 6H), 1.57 (d, J=6.5 Hz, 3H).

[0170] Synthesis of Compound #9

##STR00034##

[0171] Under the protection of nitrogen, 9-A1 (300 mg, 0.95 mmol, synthesized with reference to the synthesis method of Compound #29) and 2,4,6-trifluorophenol (210 mg, 1.40 mmol) were dissolved in dry THF (5 mL). Triphenylphosphine (622 mg, 2.40 mmol) was added, and the temperature was reduced to 0° C. A solution of di-tert-butyl azodicarboxylate (550 mg, 2.40 mmol) in THF (4 mL) was added dropwise to react for 2.5 h, and after the reaction was completed, water (5 mL) was added. The reactants were extracted with DCM (3×15 mL), washed with saline water and dried to remove the solvent, followed by high performance liquid chromatography to obtain Compound #9 (210 mg, with a yield of 49.5%) as an off-white solid. .sup.1H-NMR (400M, CDCl.sub.3): δ8.02 (d, J=8.4 Hz, 1H), 7.46 (d, J=8.8 Hz, 2H), 7.40 (d, J=8.4 Hz, 1H), 7.32 (s, 2H), 7.24 (s, 1H), 7.07 (d, J=8.8 Hz, 2H), 5.01 (s, 2H), 3.11 (s, 3H), 3.02 (s, 3H).

[0172] Synthesis of Compound #10

##STR00035##

[0173] Under the protection of nitrogen, phosphorus oxychloride (348 mg, 2.30 mmol) was added dropwise to anhydrous DCM (5 mL). The reactants were cooled to −40° C., a solution of 10-A1 (300 mg, 0.91 mmol, synthesized with reference to the synthesis method of Compound #29) in DCM (3 mL) was added dropwise, then a solution of TEA (240 mg, 2.40 mmol) in DCM (2 mL) was added dropwise, with the temperature being kept at −40° C. for 6 h until the raw material disappeared. Methylamine (960 mg, 7.70 mmol, 25% in THF) was added dropwise, then a solution of TEA (782 mg, 7.70 mmol) in DCM (5 mL) was added dropwise. The temperature was kept at −40° C. for half an hour and then is naturally increased to an ambient temperature, and the reactants were stirred overnight. The reactants were cooled to 0° C., and a potassium carbonate solution (1 g, 10 mL) was added dropwise. The reactants were extracted with DCM (10 mL×3), washed with water (5 mL×2), dried and concentrated, followed by high performance liquid chromatography to obtain pure Compound #10 (60 mg, with a yield of 15.2%) as a pale yellow viscous oil. .sup.1H-NMR (400 MHz, CDCl.sub.3): δ7.97 (d, J=8.4 Hz, 1H), 7.46 (d, J=8.5 Hz, 2H), 7.24-7.22 (m, 1H), 7.09 (s, 1H), 7.05 (d, J=8.5 Hz, 2H), 5.48-5.44 (m, 1H), 3.07 (s, 6H), 2.61 (d, J=12.1 Hz, 3H), 2.42 (d, J=12.1 Hz, 3H), 1.54 (d, J=6.4 Hz, 3H). MS: Calculated 436.2, found 437.2 ([M+H].sup.+).

[0174] Synthesis of Compound #11 Under the protection of nitrogen, phosphorus oxychloride (242 mg, 1.58 mmol) was added dropwise to anhydrous DCM (5 mL). The reactants were cooled to −40° C., a solution of 11-A1 (200 mg, 0.63 mmol, synthesized with reference to the synthesis method of Compound #29) in DCM (3 mL) was added dropwise, then a solution of TEA (166 mg, 1.64 mmol) in DCM (2 mL) was added dropwise, with the temperature being kept at −40° C. for 6 h until the raw material disappeared. Methylamine (670 mg, 5.40 mmol, 25% in THF) was added dropwise, then a solution of TEA (546 mg, 5.4 mmol) in DCM (5 mL) was added dropwise. The temperature was kept at −40° C. for half an hour and then is naturally increased to an ambient temperature, and the reactants were stirred overnight. The reactants were cooled to 0° C., and a potassium carbonate solution (1 g, 10 mL) was added dropwise. The reactants were extracted with DCM (10 mL×3), washed with water (5 mL×2), dried and concentrated, followed by high performance liquid chromatography to obtain pure Compound #11 (95 mg, with a yield of 35.7%) as a pale yellow viscous oil. .sup.1H-NMR (400, MeOD): δ8.04 (d, J=8.4 Hz, 1H), 7.49 (d, J=8.7 Hz, 2H), 7.40 (d, J=8.5 Hz, 1H), 7.27 (s, 1H), 7.09 (d, J=8.6 Hz, 2H), 4.99 (d, J=7.9 Hz, 2H), 3.12 (s, 3H), 3.07 (s, 3H), 2.50 (d, J=12.4 Hz, 6H). MS: Calculated 422.1, found 423.2 ([M+H].sup.+).

[0175] Synthesis of Compound #12

##STR00036##

[0176] Under the protection of nitrogen, tris(dimethylamino)phosphine (179 mg, 1.1 mmol, commercially available) and tetrazole (0.64 mg, 0.009 mmol) were added to acetonitrile (3 mL), then a solution of 12-A1 (300 mg, 0.91 mmol, synthesized with reference to the synthesis method of Compound #29) in acetonitrile (2 mL) was added dropwise. The reactants were stirred at room temperature for two hours, and then TEA (277 mg, 2.74 mmol) and tert-butanol peroxide (329 mg, 3.66 mmol) were added dropwise. The reactants were stirred at room temperature for three hours, until the reaction was completed. An aqueous sodium thiosulfate solution (4 mL) was added dropwise, and the reactants were extracted with DCM (5 mL×3), dried and concentrated, followed by high performance liquid chromatography to separate and obtain Compound #12 (76.5 mg, with a yield of 18.2%) as a pale yellow oil. .sup.1H-NMR (400M, CDCl.sub.3): δ7.98 (d, J=8.4 Hz, 1H), 7.45 (d, J=8.6 Hz, 2H), 7.24 (d, J=1.5 Hz, 1H), 7.09 (d, J=1.4 Hz, 1H), 7.04 (d, J=8.6 Hz, 2H), 5.43-5.39 (m, 1H), 3.07 (s, 6H), 2.65 (d, J=10.0 Hz, 6H), 2.44 (d, J=10.0 Hz, 6H), 1.52 (d, J=6.5 Hz, 3H). MS: Calculated 464.2, found 465.2 ([M+H].sup.+).

[0177] Synthesis of Compound #13

##STR00037##

[0178] Under the protection of nitrogen, tris(dimethylamino)phosphine (124 mg, 0.76 mmol, commercially available) and tetrazole (0.4 mg, 0.0057 mmol) were added to acetonitrile (3 mL), then a solution of 13-A1 (200 mg, 0.63 mmol, synthesized with reference to the synthesis method of Compound #29) in acetonitrile (2 mL) was added dropwise. The reactants were stirred at room temperature for two hours, and then TEA (192 mg, 1.90 mmol) and tert-butanol peroxide (228 mg, 2.53 mmol) were added dropwise. The reactants were stirred at room temperature for two hours, until the reaction was completed. An aqueous sodium thiosulfate solution (4 mL) was added dropwise, and the reactants were extracted with DCM (5 mL×3), dried and concentrated, followed by high performance liquid chromatography to separate and obtain Compound #13 (31 mg, with a yield of 9.0%) as a pale yellow oil. .sup.1H-NMR (400M, CDCl.sub.3): δ7.99 (d, J=8.4 Hz, 1H), 7.45 (d, J=8.8 Hz, 2H), 7.24 (m, 1H), 7.06-7.04 (m, 3H), 4.93 (d, J=7.6 Hz, 2H), 3.11 (s, 3H), 3.02 (s, 3H), 2.62 (s, 6H), 2.59 (s, 6H). MS: Calculated 450.2, found 451.1 ([M+H].sup.+).

[0179] Synthesis of Compound #14

##STR00038##

[0180] Under the protection of nitrogen, phosphorus oxychloride (246 mg, 0.949 mmol) was added dropwise to anhydrous DCM (10 mL). The reactants were cooled to −40° C., a solution of 14-A1 (200 mg, 0.633 mmol, synthesized with reference to the synthesis method of Compound #29) in DCM (4 mL) was added dropwise, then TEA (96 mg, 0.949 mmol) was added dropwise, and the temperature was kept at −40° C. to −35° C. for 2 h. As shown by the follow-up detection of HPLC, 14-A1 was exhausted and converted into an intermediate. At −40° C., a solution of N,N′-dimethyl-1,3-propanediamine (130 mg, 1.27 mmol, commercially available) in DCM (2 mL) was added dropwise, and then a solution of TEA (130 mg, 1.266 mmol) in DCM (2 ml) was added dropwise, with the temperature being kept at −40° C. for one hour until the conversion of the intermediate was completed. The temperature was naturally raised to 0° C., and a saturated aqueous solution of ammonium chloride (5 mL) was added dropwise. The reactants were extracted with DCM (10 mL×3), washed with purified water (3 ml×3), dried and concentrated, followed by HPLC to obtain Compound #14 (8.5 mg, with a yield of 2.9%) as a pale yellow oily liquid. .sup.1H-NMR (400 MHz, CDCl.sub.3): δ7.98 (d, J=8.4 Hz, 1H), 7.45 (d, J=8.4 Hz, 2H), 7.23 (d, J=8.5 Hz, 1H), 7.11-7.02 (m, 3H), 4.92 (d, J=7.5 Hz, 2H), 3.15-2.86 (m, 10H), 2.65 (s, 3H), 2.63 (s, 3H), 2.03-1.97 (m, 1H), 1.52-1.50 (m, 1H). MS: Calculated 462.2, found 463.1 ([M+H].sup.+).

[0181] Synthesis of Compound #15

##STR00039##

[0182] Under the protection of nitrogen, phosphorus oxychloride (53 mg, 0.348 mmol) was added dropwise to anhydrous DCM (10 mL). The reactants were cooled to −40° C., a solution of 15-A1 (100 mg, 0.316 mmol, synthesized with reference to the synthesis method of Compound #29) in DCM (2 mL) was added dropwise, then TEA (35 mg, 0.348 mmol) was added dropwise, and the temperature was kept at −40° C. to −35° C. for 2 h.

[0183] As monitored by HPLC and LC-MS, 15-A1 was exhausted and converted into an intermediate. At −40° C., a solution of 3-amino-1-propanol (26 mg, 0.348 mmol) in DCM (2 mL) was added dropwise, and then a solution of TEA (96 mg, 0.948 mmol) in DCM (2 ml) was added dropwise, with the temperature being kept at −40° C. for one hour until the conversion of the intermediate was completed. The temperature was naturally raised to 0° C., and a saturated aqueous solution of ammonium chloride (5 mL) was added dropwise. The reactants were extracted with DCM (10 mL×3), washed with purified water (3 ml×3), dried and concentrated, followed by HPLC to obtain Compound #15 (44.5 mg, with a yield of 32.3%) as a white solid. .sup.1H-NMR (400M, CDCl.sub.3): 57.99 (d, J=8.4 Hz, 1H), 7.46 (dd, J=9.0, 2.2 Hz, 2H), 7.27-7.25 (m, 1H), 7.08 (s, 2H), 7.06 (d, J=2.5 Hz, 1H), 5.03-5.00 (m, 2H), 4.45-4.30 (m, 1H), 4.23-4.16 (m, 1H), 3.30-3.24 (m, 1H), 3.12-3.07 (m, 7H), 2.07-1.98 (m, 2H), 1.67-1.62 (m, 1H). MS: Calculated 435.1, found 436.1 ([M+H].sup.+).

[0184] Synthesis of Compound #16

##STR00040##

[0185] Under the protection of nitrogen, phosphorus oxychloride (54 mg, 0.35 mmol) was added dropwise to anhydrous DCM (10 mL). The reactants were cooled to −40° C., a solution of 16-A1 (100 mg, 0.32 mmol, synthesized with reference to the synthesis method of Compound #29) in DCM (2 mL) was added dropwise, then TEA (36 mg, 0.35 mmol) was added dropwise, and the temperature was kept at −40° C. for 3 h. At −40° C., a solution of 3-(methylamino)-1-propanol (31 mg, 0.35 mmol, commercially available) in DCM (2 mL) was added dropwise, and then a solution of TEA (107 mg, 1.05 mmol) in DCM (2 ml) was added dropwise, with the temperature being kept at −40° C. for 3 h. The temperature was naturally raised to room temperature, and the reactants were allowed to react overnight. At 0° C., a saturated aqueous solution of ammonium chloride (5 mL) was added dropwise. The reactants were extracted with DCM (8 mL×3), washed with purified water (3 ml×3), dried and concentrated, followed by HPLC to obtain Compound #16 (33 mg, with a yield of 22.9%) as a semisolid. .sup.1H-NMR (400M, CDCl.sub.3): δ7.99 (d, J=8.4 Hz, 1H), 7.46 (d, J=8.4 Hz, 2H), 7.23-7.25 (m, 1H), 7.06-7.08 (m, 3H), 4.94-5.12 (m, 2H), 4.22-4.31 (m, 1H), 4.05-4.12 (m, 1H), 2.97-3.12 (m, 8H), 2.65 (d, J=10.8 Hz, 3H), 2.10-2.19 (m, 1H), 1.71-1.76 (m, 1H). MS: Calculated 449.1, found, 450.1 ([M+H].sup.+).

[0186] Synthesis of Compound #17

##STR00041##

[0187] Under the protection of nitrogen, phosphorus oxychloride (53.4 mg, 0.348 mmol) was added dropwise to anhydrous DCM (5 mL). The reactants were cooled to −40° C., a solution of 17-A1 (100 mg, 0.316 mmol, synthesized with reference to the synthesis method of Compound #29) in DCM (2 mL) was added dropwise, then TEA (35.2 mg, 0.348 mmol) was added dropwise, and the temperature was kept at −40° C. for 1.5 h until the raw material is completely converted into an intermediate. At −40° C., a solution of 1,3-propanediol (26.5 mg, 0.348 mmol) in DCM (2 mL) was added dropwise, and then a solution of TEA (106 mg, 1.043 mmol) in DCM (2 ml) was added dropwise, with the temperature being kept at −40° C. until the reaction was completed. At 0° C., a saturated aqueous solution of ammonium chloride (3 mL) was added dropwise. The reactants were extracted with DCM (5 mL×3), washed with purified water (3 ml×3), dried and concentrated, followed by HPLC to obtain Compound #17 (33 mg, with a yield of 22.9%) as a pale yellow oily liquid. .sup.1H-NMR (400M, CDCl.sub.3): δ8.00 (d, J=8.4 Hz, 1H), 7.47 (d, J=8.5 Hz, 2H), 7.27-7.25 (m, 1H), 7.11-7.04 (m, 3H), 5.10 (d, J=8.5 Hz, 2H), 4.47-4.35 (m, 2H), 4.27 (m, 2H), 3.07 (s, 6H), 2.27 (dd, J=10.9, 4.4 Hz, 1H), 1.80 (m, 1H), 1.34-1.18 (m, 2H). MS: Calculated 436.1, found 437.1 ([M+H].sup.+).

[0188] Synthesis of Compound #19

##STR00042##

[0189] Under the protection of nitrogen, 19-A1 (2.0 g, 11.69 mmol) and 19-A2 (6.4 g, 46.76 mmol, commercially available) were dissolved in DMF (10 mL), then potassium carbonate (6.5 g, 46.76 mmol) was added, and the reactants was stirred overnight at 40° C. After the reaction was complete, the temperature was reduced to room temperature. Water (20 ml) was added dropwise, followed by extraction with EA (20 mL×3). The substance obtained was washed with water (8 ml×5) and saline water (8 mL×3), dried and concentrated, followed by column separation (300-400 mesh silica gel, n-heptane: EA, 25%-35% EA) to obtain 19-A3 (1.05 g, 31.3%) as a pale yellow solid. .sup.1H-NMR (400 MHz, CDCl.sub.3): δ8.02 (d, J=8.4 Hz, 1H), 7.99-7.95 (m, 2H), 7.32-7.28 (m, 1H), 7.18 (d, J=0.6 Hz, 1H), 7.06-7.00 (m, 2H), 4.77 (s, 2H), 2.58 (s, 3H). MS: Calculated 287.1, found 288.0 ([M+H].sup.+).

[0190] Under the protection of nitrogen, phosphorus oxychloride (59 mg, 0.382 mmol) was added dropwise to anhydrous DCM (10 mL). The reactants were cooled to −40° C., a solution of 19-A3 (100 mg, 0.347 mmol) in DCM (2 ml) was added dropwise, then TEA (39 mg, 0.382 mmol) was added dropwise, and the temperature was kept at −40° C. to −35° C. for 2 h. As monitored by HPLC and LC-MS, 19-A3 was exhausted and converted into an intermediate. At −40° C., a solution of N,N′-dimethyl-1,3-propanediamine (39 mg, 0.382 mmol, commercially available) in DCM (2 mL) was added dropwise, and then a solution of TEA (105 mg, 1.041 mmol) in DCM (2 ml) was added dropwise, with the temperature being kept at −40° C. for one hour until the conversion of the intermediate was completed. The temperature was naturally raised to 0° C., and a saturated aqueous solution of ammonium chloride (5 mL) was added dropwise. The reactants were extracted with DCM (10 mL×3), washed with purified water (3 ml×3), dried and concentrated, followed by HPLC to obtain Compound #19 (34.5 mg, with a yield of 20.8%) as a pale yellow oily liquid. .sup.1H-NMR (400 MHz, CDCl.sub.3): δ8.02 (d, J=8.4 Hz, 1H), 7.97 (dd, J=11.2, 2.4 Hz, 2H), 7.30 (dd, J=8.4, 1.0 Hz, 1H), 7.15 (s, 1H), 7.04 (dd, J=11.2, 2.4 Hz, 2H), 4.95 (d, J=7.6 Hz, 2H), 3.10-3.04 (m, 2H), 3.01-2.88 (m, 2H), 2.65 (s, 3H), 2.63 (s, 3H), 2.58 (s, 3H), 2.04-2.01 (m, 1H), 1.51-1.47 (m, 1H). MS: Calculated 433.1, found 434.1 ([M+H].sup.+).

[0191] Synthesis of Compound #20

##STR00043##

[0192] Under the protection of nitrogen, phosphorus oxychloride (59 mg, 0.382 mmol) was added dropwise to anhydrous DCM (10 mL). The reactants were cooled to −40° C., a solution of 20-A1 (100 mg, 0.347 mmol, i.e., 19-A3) in DCM (2 ml) was added dropwise, then TEA (39 mg, 0.382 mmol) was added dropwise, and the temperature was kept at −40° C. to −35° C. for 2 h. As monitored by HPLC and LC-MS, 20-A1 was exhausted and converted into an intermediate. At −40° C., a solution of 3-amino-1-propanol (29 mg, 0.382 mmol, commercially available) in DCM (2 mL) was added dropwise, and then a solution of TEA (105 mg, 1.041 mmol) in DCM (2 ml) was added dropwise, with the temperature being kept at −40° C. for one hour until the conversion of the intermediate was completed. The temperature was naturally raised to 0° C., and a saturated aqueous solution of ammonium chloride (5 mL) was added dropwise. The reactants were extracted with DCM (10 mL×3), washed with purified water (3 ml×3), dried and concentrated, followed by HPLC to obtain the product (13.3 mg, with a yield of 9.4%) as a pale yellow oily liquid. .sup.1H-NMR (400M, CDCl.sub.3): δ8.03 (d, J=8.4 Hz, 1H), 7.98 (d, J=8.8 Hz, 2H), 7.35 (dd, J=8.4, 1.1 Hz, 1H), 7.18 (s, 1H), 7.04 (d, J=8.8 Hz, 2H), 5.05 (d, J=7.6 Hz, 2H), 4.33-4.41 (m, 1H), 4.19-4.26 (m, 1H), 3.14-3.34 (m, 2H), 2.58 (s, 3H), 1.97-2.09 (m, 1H), 1.61-1.65 (m, 1H). MS: Calculated 406.1, found 407.1 ([M+H].sup.+).

[0193] Synthesis of Compound #21

##STR00044##

[0194] Under the protection of nitrogen, phosphorus oxychloride (59 mg, 0.382 mmol) was added dropwise to anhydrous DCM (10 mL). The reactants were cooled to −40° C., a solution of 21-A1 (100 mg, 0.347 mmol, i.e., 19-A3) in DCM (2 ml) was added dropwise, then TEA (39 mg, 0.382 mmol) was added dropwise, and the temperature was kept at −40° C. to −35° C. for 2 h. At −40° C., a solution of 3-(methylamino)-1-propanol (34 mg, 0.382 mmol) in DCM (2 mL) was added dropwise, and then a solution of TEA (105 mg, 1.041 mmol) in DCM (2 ml) was added dropwise, with the temperature being kept at −40° C. for 0.5 h. The temperature was naturally raised to room temperature, and the reactants were allowed to react overnight. At 0° C., a saturated aqueous solution of ammonium chloride (5 mL) was added dropwise. The reactants were extracted with DCM (8 mL×3), washed with purified water (3 ml×3), dried and concentrated, followed by HPLC to obtain Compound #21 (41 mg, with a yield of 27.2%) as a semisolid. .sup.1H-NMR (400M, CDCl.sub.3): δ8.03 (d, J=8.4 Hz, 1H), 7.98 (d, J=8.8 Hz, 2H), 7.30 (dd, J=8.4, 1.6 Hz, 1H), 7.16 (d, J=1.6 Hz, 1H), 7.05 (d, J=8.8 Hz, 2H), 4.97-5.12 (m, 2H), 4.23-4.33 (m, 1H), 4.09-4.16 (m, 1H), 2.97-3.16 (m, 2H), 2.65 (d, J=10.8 Hz, 3H), 2.59 (s, 3H), 2.08-2.24 (m, 1H), 1.71-1.77 (m, 1H). MS: Calculated 420.1, found 421.1 ([M+H].sup.+).

[0195] Synthesis of Compound #23

##STR00045##

[0196] Under the protection of nitrogen, phosphorus oxychloride (59 mg, 0.382 mmol) was added dropwise to anhydrous DCM (10 mL). The reactants were cooled to −40° C., a solution of 23-A1 (100 mg, 0.347 mmol, i.e., 19-A3) in DCM (2 ml) was added dropwise, and then TEA (39.0 mg, 0.382 mmol) was added dropwise. The temperature was kept at −40° C. for 1.5 h, and the raw materials were completely converted into an intermediate. At −40° C., a solution of 4-amino-1-butanol (34.0 mg, 0.382 mmol, commercially available) in DCM (2 mL) was added dropwise, and then a solution of TEA (116 mg, 1.145 mmol) in DCM (2 ml) was added dropwise, with the temperature being kept at −40° C. The reaction was completely after 5 h. At 0° C., a saturated aqueous solution of ammonium chloride (3 mL) was added dropwise. The reactants were extracted with DCM (5 mL×3), washed with purified water (3 ml×3), dried and concentrated to obtain Compound #23 (15.7 mg, 10.7%) as a pale yellow oily liquid. .sup.1H-NMR (400 MHz, CDCl.sub.3): δ8.03 (d, J=8.4 Hz, 1H), 7.99 (t, J=5.7 Hz, 2H), 7.33 (d, J=8.5 Hz, 1H), 7.18 (s, 1H), 7.04 (d, J=8.7 Hz, 2H), 5.07 (m, 2H), 4.33-4.23 (m, 1H), 4.13 (m, 1H), 3.10-2.99 (m, 1H), 2.90-2.83 (m, 1H), 2.59 (s, 3H), 1.91-1.75 (m, 4H). MS: Calculated 420.1, found 421.1 ([M+H]+).

[0197] Synthesis of Compound #24

##STR00046##

[0198] 24-A1 (100 mg, 0.32 mmol) was dissolved in DCM (3 ml). The temperature was reduced to 0° C., thionyl chloride (57.12 mg, 4.8 mmol, 1.5 eq) was added dropwise with stirring at room temperature. The reaction was completed after 1.5 h. The temperature was reduced to 0-5° C., a saturated solution of sodium bicarbonate was added dropwise, and the pH was adjusted to be weakly alkaline (pH=7-8). The reactants were extracted with DCM (10 mL×2), dried and concentrated to obtain 24-A2 (130 mg) as a yellow oil, which was used directly into the next step. 24-A2 (130 mg, 0.388 mmol) was dissolved in DMF (3 ml). 3-hydroxypyridazine (74.57 mg, 0.776 mmol, 2 eq) was added, and cesium carbonate (316 mg, 0.96 mmol, 2.5 eq) was added with stirring at room temperature. The reaction was completed after 40 min. EA (a volume of 20 times) was added. The reactants were washed with saturated sodium carbonate (5 ml×3), then with saline water, and dried and concentrated to obtain the product (26 mg, 17.0%) as a yellow oil. .sup.1H-NMR (400 MHz, CDCl.sub.3): δ8.05 (s, 1H), 8.00 (d, J=8.2 Hz, 1H), 7.94 (s, 1H), 7.49 (d, J=7.8 Hz, 2H), 7.13 (d, J=8.2 Hz, 1H), 7.09 (d, J=7.8 Hz, 2H), 6.94 (s, 1H), 6.72 (s, 1H), 5.18 (s, 2H), 3.15 (s, 3H), 3.05 (s, 3H). MS: Calculated 394.1, found 394.9 ([M+H].sup.+).

[0199] Synthesis of Compound #25

##STR00047##

[0200] 24-A1 (50 mg, 0.16 mmol) was dissolved in DCM (3 ml). 29-B1 (141.8 mg, 0.96 mmol, 6 eq) was added, and the temperature was reduced to 0° C. TEA (95.9 mmol, 0.96 mmol, 6 eq) was added dropwise, DMAP (4.85 mg, 0.04 mmol, 0.25 eq) was added, and the temperature was raised to 40° C. with stirring overnight. After the reaction was completed, the solvent was removed, and HPLC was performed to obtain a product of 14.1 mg as a white solid, with a yield of 20.5%. .sup.1H-NMR (400M, CDCl.sub.3): δ7.99 (d, J=8.4 Hz, 1H), 7.46 (d, J=8.4 Hz, 2H), 7.21-7.23 (m, 1H), 7.06 (d, J=8.4 Hz, 2H), 7.01 (m, 1H), 5.11 (s, 2H), 3.46-3.65 (m, 8H), 3.11 (s, 3H), 3.02 (s, 3H). MS: Calculated 429.2, found 429.9 ([M+H].sup.+).

[0201] Synthesis of Compound #26

##STR00048##

[0202] Triphosgene (3 g, 10.1 mmol) was dissolved in DCM (300 ml) at 0° C., and then 26-A1 (1.52 g, 20.2 mmol, 2 eq) was dissolved in DCM (60 ml) followed by dropwise addition into the system over 20 mm. After 4 h of reaction, the system was spin-dried and slurried with MTBE (20 ml). After suction filtration, the mother liquid was spin-dried to obtain 1.2 g of 26-A2 crude product as a light brown liquid, which was directly used in the next step reaction.

[0203] 26-A2 (300 mg, 0.949 mmol) and 26-A3 (652.5 mg, 4.744 mmol, 5 eq,

##STR00049##

synthesized with reference to the synthesis method of Compound #29) were dissolved in DCM (12 ml). After the temperature was reduced to 0° C., TEA (480 mg, 4.744 mmol, 5 eq) and DMAP (29.1 mg, 0.237 mmol, 0.25 eq) were added to the system. The system was warmed to 35° C. and the reaction was completed after approximately 12 h. As monitored by HPLC and LCMS, after the reaction was completed, the temperature was reduced to room temperature. Saturated NaHCO.sub.3 (15 ml×2) was added for washing. The organic phase was washed with water (10 ml×2), and the aqueous phase was extracted with DCM (20 ml×1). The organic phase was spin-dried, followed by HPLC to obtain Compound #26 (150 mg, with a yield of 37.8%) as a yellow oily liquid. .sup.1H-NMR (400M, CDCl.sub.3): δ7.94 (d, J=8.4 Hz, 1H), 7.43 (t, J=8.4 Hz, 2H), 7.19 (d, J=8.4 Hz, 1H), 7.02 (t, J=2.4 Hz, 3H), 5.24 (s, 1H), 5.06 (s, 2H), 3.42 (t, J=4.8 Hz, 2H), 3.34 (d, J=2.8 Hz, 2H), 3.32 (d, J=5.6 Hz, 3H), 3.08 (s, 3H), 3.00 (s, 3H). MS: Calculated 417.2, found 418.0 ([M+1].sup.+).

[0204] Synthesis of Compound #27

##STR00050##

[0205] Under the protection of nitrogen, 27-A1 (120 mg, 0.358 mmol, synthesized with reference to the synthesis method of Compound #29) was dissolved in DMF (5 mL). 2,4 difluorothiophenol (104 mg, 0.716 mmol, 2 eq) was added, and Cs.sub.2CO.sub.3 (291.6 mg, 0.895 mmol, 2.5 eq) was added. The reactants were stirred at ambient temperature, and the reaction was completed after 1 h. EA (50 mL) was added. The reactants were washed with aqueous saturated sodium carbonate (20 ml×3) and saline water (10 ml×2), and dried and concentrated to obtain the product (41 mg, with a yield of 25.7%) as a yellow oily liquid. .sup.1H-NMR (400M, CDCl.sub.3): δ7.89 (d, J=8.4 Hz, 1H), 7.44 (d, J=8.4 Hz, 2H), 7.20-7.24 (m, 1H), 7.06-7.08 (m, 1H), 6.92 (d, J=8.4 Hz, 2H), 6.79-6.82 (m, 3H), 3.93 (s, 2H), 3.12 (s, 3H), 3.03 (s, 3H). MS: Calculated 444.1, found 444.9 ([M+1].sup.+).

[0206] Synthesis of Compound #28

##STR00051##

[0207] 24-A1 (100 mg, 0.32 mmol, synthesized with reference to the synthesis method of Compound #29) and pyridine (55.7 mg, 0.70 mmol, 2.2 eq) were dissolved in DCM (3 ml), then cooled to 0° C. Isopropyl chloroformate (129.6 mg, 1.2 mmol, 3.6 eq, commercially available) was added to the system. The reactants were reacted at 20° C. for 18 h, and cooled to 0° C.-5° C. 1N of hydrochloric acid (3 ml) was added dropwise, followed by extraction with DCM (10 mL×2). The organic phase was washed with hydrochloric acid (1N, 10 mL×5), water (5 mL×3) and saline water (5 mL×2), and dried over sodium sulfate, concentrated and neutralized to obtain a product (27 mg, 21.2%) as an off-white solid. .sup.1H-NMR (400M, CDCl.sub.3): δ7.98 (d, J=8.4 Hz, 1H), 7.46 (d, J=8.4 Hz, 2H), 7.26 (s, 1H), 7.04-7.07 (m, 3H), 5.11 (s, 2H), 4.87 (q, J=6.4 Hz, 1H), 3.11 (s, 3H), 3.03 (s, 3H), 1.29 (d, J=6.4 Hz, 6H). MS: Calculated 402.1, found 403.0 ([M+1].sup.+).

[0208] Synthesis of Compound #29

##STR00052##

[0209] 29-A1 (1 g, 5.84 mmol) and 3-hydroxy-N, N-dimethylbenzamide (2.8 g, 17.53 mmol, 3 eq) were dissolved in MeCN (30 ml), then K.sub.2CO.sub.3 (2.8 g, 20.45 mmol, 3.5 eq) was added to the system, and the system was warmed to 65° C. The reaction was completed after 3 h, and then the system was cooled to 20° C. H.sub.2O (15 ml) was added, followed by extraction with EA (30 mL×3). The organic phase was washed with 1N of NaOH solution (50 ml×3), spin-dried, and the sample was mixed, followed by 200-300 silica gel (EA: PE, 1: 3) to obtain 29-A2 (m=950 mg, with a yield of 51.4%). .sup.1H-NMR (400M, CDCl.sub.3): δ7.96 (d, J=8.4 Hz, 1H), 7.39-7.43 (m, 1H), 7.19-7.22 (m, 2H), 7.09-7.11 (m, 1H), 7.04-7.07 (m, 2H), 4.69 (s, 2H), 3.00 (brs, 6H), 1.97 (s, 1H). MS: Calculated 316.1, found 317.1 [(M+1).sup.+].

[0210] 29-A2 (50 mg, 0.158 mmol) and piperidine-1-formyl chloride (141.9 mg, 0.948 mmol, 6 eq) were dissolved in DCM (2 ml), then TEA (32.0 mg, 0.316 mmol, 2 eq) and DMAP (4.9 mg, 0.04 mmol, 0.25 eq) were added to the system, at which moment the system is pale yellow. The system was warmed to 40° C. and the reaction was completed after 12 h. The solvent was removed, and HPLC was carried out to obtain 16.5 mg of the pure product, with a yield of 24.3%. .sup.1H-NMR (400M, CDCl.sub.3): δ7.97 (d, J=8.4 Hz, 1H), 7.40-7.44 (m, 1H), 7.18-7.23 (m, 2H), 7.08-7.10 (m, 1H), 7.01-7.06 (m, 2H), 5.11 (s, 2H), 3.64 (s, 4H), 3.46 (s, 4H), 3.11 (s, 3H), 2.98 (s, 3H). MS: Calculated 429.2, found 330.0 ([M+H].sup.+).

[0211] Synthesis of Compound #30

##STR00053##

[0212] Under the protection of nitrogen, 30-A1 (1.76 g, 7.72 mmol) and 3,3-difluoroazetidine hydrochloride (1.0 g, 7.72 mmol, 1 eq) were dissolved in DCM (20 ml). A solution of T.sub.3P in EA (10 g, 15.4 mmol, 2 eq, 50%) was added and the system was cooled to 5° C. DIEA was added dropwise, followed by stirring at room temperature. As monitored by HPLC, after the reaction has completed for 2 h, water (5 ml) was added to the system, followed by extraction with DCM (15 ml×3). The organic phase was washed with water (10 ml×2), spin-dried to obtain a crude product (m=2.5 g, with a yield of 100%) as a pale yellow solid. .sup.1H-NMR (400M, DMSO-d6): δ7.45-7.47 (m, 2H), 7.38-7.42 (m, 3H), 7.31-7.35 (m, 1H), 7.24-7.26 (m, 2H), 7.18-7.21 (m, 1H), 5.16 (s, 2H), 4.47-4.70 (m, 4H). Calculated 303.1, found 304.1 ([M+H].sup.+).

[0213] Under the protection of nitrogen, 30-A2 (2.5 g, 8.242 mmol) was dissolved in EtOH (37 ml), and then Pd/C (630 mg, 25% A2) was added to the system. The system was flushed with N.sub.2 for three times to fill the system with N.sub.2, then flushed with H.sub.2 for three times to fill the system with H.sub.2, followed by stirring at ambient temperature. As monitored by HPLC, the reaction was completed after 8 h. Under the protection of nitrogen, H.sub.2 was drawn off, then Pd/C was filtered off with suction. The system was washed with DCM (30 ml×3) and the mother liquid was spin-dried to obtain 30-A3 crude product (1.5 g, with a yield of 85.4%) as an off-white solid. .sup.1H-NMR (400M, DMSO-d6): δ7.24-7.28 (m, 1H), 7.05-7.09 (m, 2H), 6.92-6.94 (m, 1H), 4.46-4.69 (m, 4H). Calculated 213.1, found 214.1 ([M+H].sup.+).

[0214] 30-A3 (1.5 g, 7.04 mmol) and methyl 3-fluoro-4-nitrobenzoate (2.8 g, 14.06 mmol, 2 eq) were dissolved in ACN (15 ml), and then K.sub.2CO.sub.3 (2.9 g, 21.01 mmol, 3 eq) was added to the system. The system was fluxed at 60° C. overnight. The reaction was completed the next day as monitored by HPLC, and the temperature was reduced to room temperature. Water (10 ml) was added, and the system was extracted with DCM (20 mL×4).

[0215] The organic phase was washed with water (10 ml×2) and saline water, then spin-dried to obtain 30-A4 crude product (m=1.8 g, with a yield of 65.2%) as yellow solid. .sup.1H-NMR (400M, CDCl.sub.3): δ7.99 (d, J=8.4 Hz, 1H), 7.92 (dd, J.sub.1=8.4 Hz, J.sub.2=1.6 Hz, 1H), 7.70 (d, J=1.6 Hz, 1H), 7.43-7.51 (m, 2H), 7.29-7.30 (m, 1H), 7.20-7.23 (m, 1H), 4.50-4.56 (m, 4H), 3.92 (s, 3H). Calculated MS: 392.1, found 393.0 ([M+H]).

[0216] 30-A4 (500 mg, 1.27 mmol) was dissolved in THF (2 ml) and then the mixture was cooled to 0° C. NaBH.sub.4 (193 mg, 5.10 mmol, 4 eq) was added in batches to the system and the system was warmed to 60° C. to reflux.

[0217] The reaction was completed after 15 h as monitored by HPLC. The temperature was reduced to 0° C. and the reaction was quenched with water (20 ml). The system was extracted with DCM (20 ml×3), spin-dried and concentrated, and the sample was mixed, followed by 300-400 silica gel passed through Flash column chromatography to obtain the pure product 30-A5 (140 mg, with a yield of 30.2%) as an off-white solid. .sup.1H-NMR (400M, CDCl.sub.3): δ7.96 (d, J=8.4 Hz, 1H), 7.34-7.38 (m, 1H), 7.15-7.18 (m, 2H), 7.04-7.06 (m, 2H), 6.97 (dd, J.sub.1=8.4 Hz, J.sub.2=2.0 Hz, 1H), 4.63-4.68 (m, 4H), 3.67-3.76 (m, 2H). Calculated 364.1, found 365.1 ([M+H].sup.+).

[0218] 30-A5 (140 mg, 0.384 mmol) and piperidine-1-carbonyl chloride (287.4 mg, 1.92 mmol, 5 eq) were dissolved in DCM (6 ml). After cooling to 0° C., TEA (194.3 mg, 1.92 mmol, 5 eq) and DMAP (11.8 mg, 0.096 mmol, 0.25 eq) were added to the system and then the system was warmed to 35° C. The reaction was completed after about 12 h. After the reaction was completed as monitored by HPLC and LCMS, the temperature was reduced to room temperature. Saturated NaHCO.sub.3 (10 ml×2) was added for washing, the organic phase was washed with water (10 ml×2) and the aqueous phase was extracted with DCM (10 ml). The organic phase was spin-dried and then separated by HPLC under neutral conditions to obtain the pure product (22.5 mg, with a yield of 12.3%) as a white solid. .sup.1H-NMR (400M, CDCl.sub.3): δ8.00 (d, J=8.8 Hz, 1H), 7.40-7.48 (m, 2H), 7.19-7.26 (m, 3H), 7.02 (s, 1H), 5.13 (s, 2H), 4.49-4.55 (m, 4H), 3.64 (brs, 4H), 3.46 (s, 4H). Calculated 477.1, found 478.0 ([M+H].sup.+).

[0219] Synthesis of Compound #31

##STR00054##

[0220] PSCl.sub.3 (4.5 g, 26.6 mmol) was dissolved in chloroform (50 ml), then 31-A1 (2 g, 26.6 mmol) was dissolved in chloroform (40 ml) and added dropwise to the system over 1 h. After stirring for 1 h, DIEA (3.44 g, 26.6 mmol) was dissolved in chloroform (10 ml) and added dropwise to the system, and the system was warmed to 20° C. overnight. Water (20 ml×4) was added for extraction and the organic phase was spin-dried to obtain 31-A2 crude product (2.4 g) as an off-white solid.

[0221] 31-A2 (1.8 g, 10.5 mmol) was added to H.sub.2O (18 ml) to produce a white suspension, and then NaOH (923 mg, 23.1 mmol, 2.2 eq) was slowly added in batches to the system, after which the system gradually became clear. There was a desired product as monitored by LCMS. After reacting overnight, MeCN (20 ml) was added and mixed, and then moisture was partially removed by spin-drying. The remaining mixture was adjusted to acidic with 12 N of HCl, and a large amount of solids were precipitated. After suction filtration, the resultant was slurried with MeCN (10 ml) and the moisture was removed. After suction filtration, MeCN (10 ml) was added again and spin-dried to obtain 800 mg of 31-A3 in total as a white solid, which was directly used in the next reaction.

[0222] Under the protection of nitrogen, 31-A4 (100 mg, 0.299 mmol,

##STR00055##

synthesized with reference to the synthesis method of Compound #29) was dissolved in DMF (2 ml), then 31-A3 (91.5 mg, 0.597 mmol, 2 eq) and TEA (90.7 mg, 0.896 mmol, 3 eq) were added to the system, after which the system was pale orange yellow. The reaction was performed overnight at 20° C. until the reaction was completed. A saturated aqueous solution of NaHCO.sub.3 (5 ml) was added, followed by extraction with EA (5 ml×3). The organic phase was washed with water (10 ml×5) and the aqueous phase was washed with EA (10 ml×2). The organic phase was spin-dried, and subjected to HPLC in neutral conditions. After separation, the solution was lyophilized to obtain the pure product (67.4 mg, with a yield of 50.0%) as an off-white solid. .sup.1H-NMR (400M, CD.sub.3OD): δ7.99 (d, J=8.4 Hz, 1H), 7.48-7.50 (m, 2H), 7.40 (dd, J.sub.1=8.4 Hz, J.sub.2=1.6 Hz, 1H), 7.27 (d, J=1.6 Hz, 1H), 7.08-7.10 (m, 2H), 4.21-4.26 (m, 2H), 4.03-4.07 (m, 2H), 3.16-3.25 (m, 2H), 3.10 (s, 3H), 3.05 (s, 3H) 1.94-2.04 (m, 1H), 1.67-1.71 (m, 1H). MS: Calculated 451.1, found 452.1 ([M+H].sup.+).

[0223] Synthesis of Compound #32

##STR00056##

[0224] 31 (20 mg, 0.044 mmol) was dissolved in DMF (1 ml), and immediately K.sub.2CO.sub.3 (12.2 mg, 0.088 mmol, 2 eq) was added to the reaction system. The system was stirred at room temperature for 1 h. Then, Mel (22 mg, 0.132 mmol, 3 eq) was added dropwise to the system, after which there was no change after 45% conversion rate was monitored. Additional K.sub.2CO.sub.3 (18.3 mg, 0.132 mmol, 2 eq) and Mel (22 mg, 0.132 mmol, 3 eq) was added and the system was warmed to 35° C. The conversion rate reached 75% after overnight. After cooling to 10° C., a saturated aqueous NaHCO.sub.3 solution (2 ml) was added to the system. Extraction was carried out using EA (3 ml×3), and the organic phase was washed with saline water (2 ml×3), spin-dried and then separated by HPLC under neutral conditions to obtain 6.1 mg of 32 in total as a pale yellow solid. .sup.1H-NMR (400M, CD.sub.3OD): δ7.99 (d, J=8.4 Hz, 1H), 7.48-7.50 (m, 2H), 7.39-7.41 (m, 1H), 7.27 (d, J=1.6 Hz, 1H), 7.09-7.11 (m, 2H), 4.06-4.10 (m, 2H), 4.03-4.10 (m, 2H), 3.05-3.10 (m, 8H), 2.60 (d, J=7.2 Hz, 3H), 2.07-2.17 (m, 1H), 1.68-1.80 (m, 1H). MS: Calculated 465.1, found 466.1 ([M+1].sup.+).

[0225] Synthesis of Compound #33

##STR00057##

[0226] PSCl.sub.3 (4.5 g, 26.3 mmol) was dissolved in chloroform (50 ml), then 33-A1 (2 g, 26.3 mmol) was dissolved in chloroform (40 ml) and added dropwise to the system over 1 h. After stirring for 1 h, DIEA (3.4 g, 26.3 mmol) was dissolved in chloroform (10 ml) and added dropwise to the system. The system was warmed to 20° C. and reacted overnight. Water was added for extraction (20 ml×4) and the organic phase was spin-dried to obtain 33-A2 crude product (3.5 g) as an off-white solid, which was directly used in the next step reaction. 33-A2 (1 g, 5.86 mmol) was added to H.sub.2O (10 ml), then NaOH (281 mg, 7.03 mmol, 1.2 eq) was slowly added in batches to the system and reacted overnight. The desired product was monitored by LCMS. The system was adjusted to pH=3-4 with 12 N of HCl, no solid was precipitated. The moisture was removed to obtain 500 mg of 33-A3 crude product as a pale yellow liquid.

[0227] Under the protection of nitrogen, 33-A4 (100 mg, 0.299 mmol,

##STR00058##

synthesized by reference to the synthesis method of Compound #29) was dissolved in DMF (2 ml), and then 33-A3 (184.3 mg, 1.196 mmol, 4 eq) and TEA (121.0 mg, 1.196 mmol, 4 eq) were added to the system. After reacting overnight at room temperature, the reaction was complete as monitored by HPLC. Saturated NaHCO.sub.3 (10 mL) was added, and extraction was carried out with EA (5 ml×3). The organic phase was washed with water (5 ml×2) and the aqueous phase was extracted with EA (5 ml×3). The organic phase was spin-dried and subjected to HPLC under neutral conditions to obtain 29.7 mg of Compound #33 in total as a yellow solid, with a yield of 22.0%. .sup.1H-NMR (400M, CD.sub.3OD): δ8.00 (d, J=8.4 Hz, 1H), 7.48-7.51 (m, 2H), 7.42 (dd, J=8.4 Hz, 2.0 Hz, 1H), 7.28 (d, J=1.6 Hz, 1H), 7.10-7.11 (m, 2H), 4.34-4.41 (m, 4H), 4.15 (d, J=16.8 Hz, 2H), 3.10 (s, 3H), 3.05 (s, 3H), 2.22-2.27 (m, 1H), 1.79-1.85 (m, 1H). MS: Calculated 452.1, found 453.0 ([M+1].sup.+).

[0228] Synthesis of Compound #34

##STR00059##

[0229] PSCl.sub.3 (2 g, 11.8 mmol) was dissolved in chloroform (25 ml) at 0° C., then 34-A1 (11.8 ml, 23.6 mmol, 2 eq, 2 MinTHF) was added dropwise to the system and after which the system was stirred for 1 h. Then, DIEA (3.44 g, 26.6 mmol) was dissolved in chloroform (10 ml) and then added dropwise to the system. The system was warmed to 20° C. to react overnight. The solvent was removed to obtain 34-A2 crude product. Water (20 ml) was added and stirred for 30 min, then the spin drying was carried out to obtain 5 g of 34-A3 crude product as a colorless oily liquid.

[0230] Under the protection of nitrogen, 34-A4 (50 mg, 0.149 mmol,

##STR00060##

synthesized with reference to the synthesis method of Compound #29) was dissolved in DMF (1 mL), then 34-A3 (418.6 mg, 1.194 mmol, 8 eq) and TEA (120.8 mg, 1.194 mmol, 8 eq) were added to the system. The system was warmed to 30° C. and reacted overnight. After the reaction was completed as monitored by HPLC and LCMS, the temperature was reduced to room temperature. A saturated aqueous solution of NaHCO.sub.3 (5 mL) was added, followed by extraction with EA (5 mL×3). The organic phase was washed with water (5 mL×3) and saline water, dried over sodium sulfate, spin-dried, and subjected to HPLC under neutral conditions to obtain the pure product (20.0 mg, with a yield of 30.6%) as a light yellow solid. .sup.1H-NMR (400M, CD.sub.3OD): δ7.99 (d, J=8.4 Hz, 1H), 7.48-7.50 (m, 2H), 7.40 (dd, J=8.4 Hz, 2.0 Hz, 1H), 7.26 (d, J=1.6 Hz, 1H), 7.07-7.10 (m, 2H), 3.96 (d, J=12.8 Hz, 2H), 3.10 (s, 3H), 3.05 (s, 3H), 2.51 (s, 3H), 2.48 (s, 3H). MS: Calculated 438.1, found 439.1 ([M+1].sup.+).

[0231] Synthesis of Compound #35

##STR00061##

[0232] PSCl.sub.3 (4.56 g, 26.9 mmol) was dissolved in chloroform (50 ml), then a solution of 35-A1 (2 g, 26.9 mmol) in chloroform (40 ml) was added dropwise to the system over 30 mm. After stirring for 1 h, a solution of DIEA (3.48 g, 26.9 mmol) in chloroform (10 ml) was added dropwise and the system was warmed to 20° C. overnight. After spinning dry, 35-A2 crude product was obtained. Water (20 ml) was added and the system was stirred for 30 min. Water was removed to obtain a white solid. MeCN (3 ml) was used for slurry and then filtration was carried out with suction to obtain 1.2 g of dry 35-A3 as a white solid, which was directly used in the next step reaction.

[0233] Under the protection of nitrogen, 35-A4 (100 mg, 0.299 mmol,

##STR00062##

synthesized with reference to the synthesis method of Compound #29) was dissolved in DMF (2 ml), then 35-A3 (182 mg, 1.196 mmol, 4 eq) and TEA (121 mg, 1.196 mmol, 4 eq) were added to the system. The system was warmed to 25° C. and the reaction was complete after reacting overnight as monitored by HPLC. A saturated aqueous solution of NaHCO.sub.3 (5 ml) was added and extraction was carried out with EA (5 mL×3). The organic phase was washed with water (10 ml×2) and the aqueous phase was extracted with EA (5 ml×2). The organic phase was spin-dried and then separated by HPLC under neutral conditions to obtain 23.1 mg of Compound #35 in total as a pale yellow solid, with a yield of 17.2%. .sup.1H-NMR (400M, CD.sub.3OD): δ7.98 (d, J=8.4 Hz, 1H), 7.47-7.50 (m, 2H), 7.40 (dd, J.sub.1=8.4 Hz, J.sub.2=1.6 Hz, 1H), 7.28 (d, J=1.6 Hz, 1H), 7.07-7.10 (m, 2H), 3.99 (d, J=12.4 Hz, 2H), 3.05-3.24 (m, 10H), 1.69-1.82 (m, 1H), 1.60-1.65 (m, 1H). MS: Calculated 450.1, found 451.1 ([M+1].sup.+).

[0234] Compound #1

[0235] Compound #1 was synthesized in a similar manner to Compound #2/3 above.

[0236] .sup.1H-NMR (400 MHz, CDCl3): δ 7.91 (d, J=8.4 Hz, 1H), 7.41 (t, J=7.9 Hz, 1H), 7.33-7.27 (m, 1H), 7.21 (d, J=7.6 Hz, 1H), 7.15-6.98 (m, 3H), 4.55 (s, 2H), 3.54 (s, 3H), 3.36 (s, 3H), 3.09 (s, 3H), 2.97 (s, 3H).

[0237] In Vitro AKR1C3 Enzyme Activity Inhibition Experiments

[0238] Experimental Apparatus:

[0239] Waters Acquity I Class UPLC Ultra Performance Liquid Chromatograph equipped with a Xevo G2-XSQ Tof HIRMS Quadrupole Time-of-flight High Resolution Mass Spectrometer.

[0240] Buffers and Materials:

[0241] 1. PBS phosphate buffered saline solution

[0242] 2. 20 mM NADPH in PBS phosphate buffered saline solution

[0243] 3. 250 μg/mL AKR1C3 in PBS phosphate buffered saline solution

[0244] 4.250 μM test compound in 50% MeOH/H.sub.2O

[0245] 5.250 μM progesterone in 50% MeOH/H.sub.2O

[0246] 6.1 μg/mL propranolol in 100% acetonitrile

[0247] Experimental Procedures

[0248] In step 1, the reaction mixtures were prepared into Eppendorf tubes in duplicate (n=2) according to the table below and mixed gently.

TABLE-US-00001 Negative controls Materials (μL) Samples (μL) PBS 68 66 NADPH (20 mM) 10 10 AKR1C3 (250 μg/mL) 10 10 Test compound (250 μM)  0  2

[0249] In step 2, the above mixtures in duplicate were pre-incubated at 37° C. for 30 minutes.

[0250] In step 3, an additional 10 μL of 20 mM NADPH in PBS phosphate buffered salt solution and 2 μL of 250 μM progesterone in 50% MeOH/H.sub.2O were added to each Eppendorf tube and gently mixed.

[0251] In step 4, 50 μL of the mixture from the above step was immediately transferred to 100 μL of 1 μg/mL propranolol (internal standard, IS) in 100% acetonitrile.

[0252] In step 5, the remaining sample was incubated at 37° C. for 30 minutes and 100 μL of 1 μg/mL propranolol (internal standard, IS) in 100% acetonitrile was added.

[0253] In step 6, for all the samples, 100 μL of reagent water was added, vortex-mixed at 1100 rpm for 5 minutes, and centrifuged at 15,000 rpm for 10 minutes at room temperature.

[0254] In step 7, all the samples were loaded on LC/MS to determine the content of reduced progesterone, i.e. 20α-dihydroprogesterone.

[0255] The test conditions for the LC-MS apparatus were as follows:

TABLE-US-00002 Items Conditions Apparatus: Waters Acquity I Class Liquid Chromatograph Chromatographic Acquity UPLC BEH C18 Chromatographic column: Column (50*2. 1 mm, 1.7 μm) Flow rate: 0.4 mL/min Injection volume: 3 μL The composition A: 0.1% (V/V) aqueous formic acid solution of the mobile phase: B: 0.1% (V/V) formic acid in acetonitrile The temperature 40° C. of the column oven: Detector: Quadrupole Time-of-flight Mass Spectrometer Q-TOFMS

[0256] The elution gradient of the liquid phase

TABLE-US-00003 Time (min) A (%) B (%) 0.00 90.0 10.0 1.5 5.0 95.0 2.00 5.0 95.0 2.30 90.0 10.0 3.00 90.0 10.0

[0257] Parameters of the Quadrupole Time-of-flight Mass Spectrometer

TABLE-US-00004 Items Parameters Capillary Voltage (Capilary kV) 2.5 Sampling Cone Voltage (Sampling Cone V) 40 Source Temperature (° C.) 100 Flow Rate of Sampling Cone Gas (L/h) 50 Flow Rate of Desolvation Gas (L/h) 600 InterfaceType ES, Positive Analyser Mode Sensitivity Scan Range 50-800 m/z

[0258] In step 9, calculation of reduced progesterone (20α-dihydroprogesterone): the peak areas of the reduced progesterone i.e., 20α-dihydroprogesterone and propranolol, in each sample were determined by LC/MVS. The ratio of the peak area of reduced progesterone to that of propranolol was calculated and the ratio was set to 0% when the time is 0.

[0259] AKR1C3 activity (%)=[(the amount of reduced progesterone after normalization of the sample).sub.30 min−(the amount of reduced progesterone after normalization of the sample).sub.0 min]/[(the amount of reduced progesterone after normalization of the negative control group).sub.30 min−(the amount of reduced progesterone after normalization of the negative control group).sub.0 min ]*100.

[0260] The AKR1C3 relative activity % results for the compounds in the table below at a concentration of 5 μM/L were calculated according to the above formula.

TABLE-US-00005 Relative Serial Activity No. The structures of the compound %  1 [00063]   7%  2 [00064]   78%  3 [00065]   34%  4 [00066]   58%  5 [00067]   9%  6 [00068]   58%  7 [00069]   5%  8 [00070]   69%  9 [00071]   60% 10 [00072]   96% 11 [00073]   35% 12 [00074]   15% 13 [00075] 68.3% 14 [00076] ND 15 [00077] 57.3% 16 [00078] 43.4% 17 [00079] 41.4% 19 [00080] 89.1% 20 [00081] 51.1% 21 [00082] 61.4% 23 [00083] 41.4% 24 [00084] 53.2% 25 [00085] 28.0% 26 [00086] 43.8% 27 [00087] 28.4% 28 [00088] 38.6% 29 [00089] 15.6% 30 [00090] 44.4% 31 [00091] 58.6% 32 [00092] 43.7% 33 [00093] 41% 34 [00094] 71.1% 35 [00095] 80.1% 36 [00096] 54.2% 37 Indomethacin 92.4% Remarks: ND, not detected.

[0261] The activity data of the above compounds were obtained from four tests: 2019/5/29, 2019/5/31, 2019/6/13, 2019/6/14, for which the corresponding control compound AST-3424 and indomethacin also have four values, and the arithmetic mean of these four values is used in the above table. The results of the tests and the time for test are specified as follows:

TABLE-US-00006 Compounds and Test Concentrations 2019 May 29 2019 May 31 2019 Jun. 13 2019 Jun. 14 Average AST-3424, 5 68.1 73.8 39.9 35 54.2 μM/L Indomethacin, 86.9 95 96.7 91 92.4 5 μM/L

[0262] Indomethacin is a typical AKR1C3 inhibitor and is used clinically for the relief and treatment of pain including cancer pain. The compound disclosed in the present application has a higher ability to inhibit AKR1C3 enzyme at a concentration of 5 μM/L than indomethacin, showing that the compound disclosed in the present application is a more potent inhibitor of AKR1C3.