WO2019120298A1 - N-(2-cyclohexylethyl)formamide derivative, preparation method therefor, and pharmaceutical use thereof - Google Patents

Abstract

The present application relates to an N-(2-cyclohexylethyl)formamide derivative, a preparation method therefor, and pharmaceutical use thereof. Specifically, a compound of formula (I) or a pharmaceutically acceptable salt, a stereoisomer or a solvate compound thereof, a preparation method therefor and use thereof are disclosed.

Description

N-(2-cyclohexylethyl)carboxamide derivative, its preparation method and medical use Technical field

The invention relates to the technical field of medicine, in particular to an N-(2-cyclohexylethyl)formamide derivative, a preparation method thereof and application as an IDO inhibitor, and pharmaceutical compositions and pharmaceutical compositions prepared therefrom .

Background technique

Indoleamine 2,3-dioxygenase (IDO) is a protease involved in the metabolism of tryptophan. Tryptophan is one of the eight essential amino acids. In vivo, tryptophan can be used to synthesize proteins. Tryptophan can also be used as a precursor substrate to synthesize serotonin and melatonin through the methoxypurine metabolic pathway (N- Acetyl-5-methoxytryptamine). Serotonin and melatonin are neurotransmitters and neuroendocrine hormones that are involved in the regulation of various neurological and physiological processes in the body. In addition, tryptophan can also produce metabolites such as kynurenine through the kynurenine metabolic pathway. The first step in the kynurenine metabolic pathway is the tryptophan L-color ammonia catalyzed by indoleamine 2,3-dioxygenase or tryptophan 2,3-dioxygenase (TDO). Acid degradation to N-formyl-kynurenine, N-formyl-kynurenine forms kynurenine under the catalysis of kynurenine carboxamide, and kynurenine can be further metabolized. 3-hydroxyanthranilic acid, quinolinic acid, picolinic acid. Quinolinic acid is neurotoxic, while picolinic acid has neuroprotective effects. Canine uridine and 3-hydroxyanthranilic acid are involved in the regulation of lymphocyte activity leading to inhibition of the immune system.

In addition to placental tissue, indoleamine 2,3-dioxygenase is not expressed in most tissue cells under normal health conditions. In the region of inflammation, inflammatory cytokines such as interferon gamma can induce an increase in the expression of indoleamine 2,3-dioxygenase. The results of various experiments prove that the high expression of indoleamine 2,3-dioxygenase in tissue cells can lead to inhibition of the immune system of the tissue microenvironment, or immune suppression or immune checkpoint. . High expression of placental tissue indoleamine 2,3-dioxygenase prevents immune rejection of the fetus. The high expression of indoleamine 2,3-dioxygenase in the inflammatory region prevents excessive immune responses and prevents excessive damage to cellular tissues. One of the mechanisms leading to inhibition of immunity is that high expression of indoleamine 2,3-dioxygenase causes local L-tryptophan depletion, which is sensed by surrounding lymphocytes through mechanisms such as GCN2, causing CD8+ cytotoxic T cells. Cell cycle arrest or apoptosis occurs. Another mechanism that leads to inhibition of immunity is the high expression of indoleamine 2,3-dioxygenase, which causes an increase in kynurenine. After kynurenine formation, it leaves the cell and enters the extracellular matrix, and then enters the nearby lymph. The cells regulate CD8+ T cells and regulatory Treg cells by binding to AHR transcription factors, and the activity of CD8+ cytotoxic T cells is inhibited, while the number of regulatory Treg cells is increased and activated, resulting in inhibition of immunity.

In many different types of tumors, indoleamine 2,3-dioxygenase is abnormally highly expressed, including hematological tumors and solid tumors such as colorectal cancer, liver cancer, lung cancer, pancreatic cancer, and throat cancer. The abnormally high expression of indoleamine 2,3-dioxygenase was positively correlated with poor tumor prognosis. Tumor cell escape immune monitoring is a key step in the further development of cancer and cancer. The abnormally high expression of indoleamine 2,3-dioxygenase in tumors may be the escape of tumor cells.

A major mechanism of immunological monitoring, inhibition of the activity of indoleamine 2,3-dioxygenase may activate the suppressed immune system to inhibit tumor growth, so indoleamine 2,3-dioxygenase Inhibitors, as an immune checkpoint inhibitor, have attracted a lot of interest in the medical community. There are two kinds of indoleamine 2,3-dioxygenase (IDO), IDO-1 and IDO-2. The main inhibitory of the above immunity is IDO-1. The role of IDO-2 in immune suppression is not yet very clear. Tryptophan 2,3-dioxygenase (TDO) is also abnormally highly expressed in many types of tumors, and some tumors also exhibit double positive IDO and TDO, so some people think that it can also suppress TDO immune checkpoints. The purpose of cancer treatment. Because normal liver cells express TDO, it is unclear whether TDO inhibitors affect liver function and normal tryptophan metabolism, but there is no abnormality in the mouse model of TDO knockout, indicating that TDO inhibitors may not affect liver function and normality. The metabolism of tryptophan. The mechanism by which IDO and TDO cause immune suppression is basically the same, so IDO/TDO bispecific inhibitors have also attracted interest in the pharmaceutical industry. IDO/TDO bispecific inhibitors will be suitable for IDO positive, TDO positive, IDO/TDO double positive. Patient.

Many metabolites of the kynurenine metabolic pathway of tryptophan are associated with schizophrenia, depression, and neuronal degeneration, and indoleamine 2,3-dioxygenase inhibitors may also be useful in the treatment of these diseases. Canine uridine can be converted to canine urinary quinolinic acid under the catalysis of kynurenine aminotransferase. Canine urinary quinolinic acid is an NMDA antagonist, which is common in the central nervous system of patients with schizophrenia. Canine urinary quinolinic acid levels. Quinolinic acid is neurotoxic and can cause neuronal apoptosis and neurodegeneration. Indoleamine 2,3-dioxygenase is not only involved in the metabolism of tryptophan, but also involved in the metabolism of tryptamine. The serotonin can be converted to 5-- under the catalysis of indoleamine 2,3-dioxygenase. Hydroxamic acid, a decrease in serotonin may be one of the factors leading to depression.

At present, indoleamine 2,3-dioxygenase inhibitors are still in the early stage of development, and it is of great clinical significance to develop IDO inhibitors with better activity and lower toxicity on the existing basis.

Summary of the invention

It is an object of the present invention to provide a novel structure of IDO inhibitors, methods for their preparation and uses.

The first aspect of the invention provides a compound of the formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

In the formula,

A is a 4 to 6 membered saturated monoheterocyclic ring or a 5 to 6 membered monocyclic heteroaryl ring;

n is 1, 2 or 3;

Z ₁ is N or CR ₅ ; Z ₂ is N or CR ₆ ; Z ₃ is N or CR ₇ ; Z ₁ , Z ₂ and Z _{3 are} not N at the same time;

R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ are each independently hydrogen, halogen, C _1-10 alkyl, C _1-10 alkoxy, halo C _1-10 alkane a C _3-10 cycloalkyl group, a halogenated C _1-10 alkoxy group, NR _a0 R _b0 or a -C(O)C _1-10 alkyl group;

The 4- to 6-membered saturated monoheterocyclic ring or 5- to 6-membered monocyclic heteroaryl ring is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of NR _a0 R _b0 , halogen, Cyano, acetyl, hydroxy, hydroxymethyl, hydroxyethyl, carboxy, C _1-8 alkyl, C _1-8 alkoxy, halo C _1-8 alkyl, C _3-8 cycloalkyl, C _3-8 cycloalkoxy, halo C _1-8 alkoxy, -C(O)C _1-10 alkyl, -C(O)OC _1-10 alkyl, -OC(O)C _{1 -10} alkyl, -CONR _a0 R _b0 ; R _a0 , R _b0 are each independently hydrogen or C _1-8 alkyl.

In another preferred embodiment, the 4- to 6-membered saturated monoheterocyclic ring is selected from the group consisting of azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, piperazine, morpholine. , thiomorpholine, thiomorpholine-1,1-dioxide or tetrahydropyran.

In another preferred embodiment, the 4- to 6-membered saturated monoheterocyclic ring is selected from the following structures:

The above 4- to 6-membered saturated monoheterocyclic ring is optionally substituted with 1, 2 or 3 substituents selected from the group A1.

In another preferred embodiment, the 5- to 6-membered monocyclic heteroaryl ring is selected from the group consisting of a thiophene ring, an N-alkylcyclopyrrole ring, a furan ring, a thiazole ring, an imidazole ring, an oxazole ring, a pyrrole ring, and a pyrazole. Ring, triazole ring, 1,2,3-triazole ring, 1,2,4-triazole ring, 1,2,5-triazole ring, 1,3,4-triazole ring, tetrazole ring, Isoxazole ring, oxadiazole ring, 1,2,3-oxadiazole ring, 1,2,4-oxadiazole ring, 1,2,5-oxadiazole ring, 1,3,4-oxa An oxadiazole ring, a thiadiazole ring, a pyridine ring, a pyridazine ring, a pyrimidine ring or a pyrazine ring.

In another preferred embodiment, the 5- to 6-membered monocyclic heteroaryl ring is selected from the following structures:

The above 5- to 6-membered monocyclic heteroaryl ring is optionally substituted with 1, 2 or 3 substituents selected from the group A1.

In another preferred embodiment, the 5- to 6-membered monocyclic heteroaryl ring is selected from the following structures

In another preferred embodiment, the substituent of the group A1 is: halogen, C _1-8 alkyl (preferably C _1-6 alkyl, more preferably C _1-3 alkyl), C _3-8 cycloalkyl (preferably C _3-6 cycloalkyl), halogenated C _1-8 alkyl (preferably halogenated C _1-6 alkyl, more preferably halogenated C _1-3 alkyl), -C(O) OC _1-6 alkyl, acetyl, C _1-8 alkoxy (preferably C _1-6 alkoxy, more preferably C _1-3 alkoxy), halogenated C _1-8 alkoxy ( It is preferably a halogenated C _1-6 alkoxy group, more preferably a halogenated C _1-3 alkoxy group).

In another preferred embodiment, n is 1 or 2.

In another preferred embodiment, two of Z ₁ , Z ₂ and Z ₃ are not N.

In another preferred embodiment, Z ₁ is N; Z ₂ is CR ₆ ; Z ₃ is CR ₇ ; and R ₆ and R _{7 are} as defined in claim 1.

In another preferred embodiment, Z ₁ is CR ₅ ; Z ₂ is N; Z ₃ is CR ₇ ; and R ₅ and R _{7 are} as defined in claim 1.

In another preferred embodiment, Z ₁ is CR ₅ ; Z ₂ is CR ₆ ; Z ₃ is N; and R ₅ and R _{6 are} as defined in claim 1.

In another preferred embodiment, A is a thiophene ring; the thiophene ring is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of NR _a0 R _b0 , halogen, cyano, acetyl , hydroxy, hydroxymethyl, hydroxyethyl, carboxyl, C _1-3 alkyl, C _1-3 alkoxy, halo C _1-3 alkyl, C _3-6 cycloalkyl, C _3-6 ring Alkoxy, halo C _1-3 alkoxy, -C(O)C _1-3 alkyl, -C(O)OC _1-3 alkyl, -OC(O)C _1-3 alkyl, -CONR _a0 R _b0 ; R _a0 , R _b0 are each independently hydrogen or C _1-3 alkyl.

In another preferred embodiment, A is a thiophene ring; the thiophene ring is unsubstituted or substituted by 1, 2 or 3 (preferably 1) substituents selected from the group consisting of halogen (preferably Cl), cyano group.

In another preferred embodiment, R ₁ is hydrogen or halogen; and R ₂ , R ₃ , and R ₄ are hydrogen.

In another preferred embodiment, R ₅ , R ₆ , and R ₇ are each independently hydrogen, halogen, C _1-3 alkyl, C _1-3 alkoxy, halo C _1-3 alkyl, C _{3 - 6} cycloalkyl, halo C _1-3 alkoxy, NR _a0 R _b0 or -C(O)C _1-3 alkyl.

In another preferred example,

For the next group structure:

In another preferred example,

For the next group structure:

In another preferred embodiment, the compound of formula (I) is of the formula (II) or formula (III):

Wherein m is 0 or 1; A, Z ₁ , Z ₂ , Z ₃ , R ₁ , R ₂ , R ₃ and R _{4 are} as defined in claim 1.

In another preferred embodiment, A is a thiophene ring; the thiophene ring is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of NR _a0 R _b0 , halogen, cyano, acetyl , hydroxy, hydroxymethyl, hydroxyethyl, carboxyl, C _1-3 alkyl, C _1-3 alkoxy, halo C _1-3 alkyl, C _3-6 cycloalkyl, C _3-6 ring Alkoxy, halo C _1-3 alkoxy, -C(O)C _1-3 alkyl, -C(O)OC _1-3 alkyl, -OC(O)C _1-3 alkyl, -CONR _a0 R _b0 ; R _a0 , R _b0 are each independently hydrogen or C _1-3 alkyl.

In another preferred embodiment, R ₁ is hydrogen or halogen; and R ₂ , R ₃ , and R ₄ are hydrogen.

In another preferred embodiment, Z ₁ is N; Z ₂ is CR ₆ ; Z ₃ is CR ₇ ; R ₆ and R ₇ are each independently hydrogen, halogen, C _1-3 alkyl, C _1-3 alkoxy A halogenated C _1-3 alkyl group, a C _3-6 cycloalkyl group, a halogenated C _1-3 alkoxy group, NR _a0 R _b0 or a —C(O)C _1-3 alkyl group.

In another preferred example,

For the next group structure:

And A is a thiophene ring.

In another preferred embodiment, the compound of formula (I) is selected from the group consisting of structures.

In another preferred embodiment, the Group A structure is selected from the group consisting of:

A second aspect of the present invention provides a pharmaceutical composition comprising the compound of the first aspect of the present invention or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier .

A third aspect of the invention provides a compound according to the first aspect of the invention, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to the second aspect of the invention, in the preparation of a medicament In use, the medicament is for inhibiting the activity of indoleamine 2,3-dioxygenase or for inhibiting immunosuppression in a patient.

In another preferred embodiment, the medicament is for treating or preventing cancer or tumor, viral infection, depression, neurodegenerative disorder, trauma, age-related cataract, organ transplant rejection or autoimmune disease in a patient; preferably, wherein The cancer or tumor is selected from the group consisting of lung cancer, bone cancer, stomach cancer, pancreatic cancer, skin cancer, head and neck cancer, uterine cancer, ovarian cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, Vulvar cancer, rectal cancer, colon cancer, anal cancer, breast cancer, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, urinary tract cancer, penile cancer, prostate cancer, pancreatic cancer, brain Cancer, testicular cancer, lymphoma, transitional cell carcinoma, bladder cancer, renal or ureteral cancer, renal cell carcinoma, renal pelvic cancer, Hodgkin's disease, non-Hodgkin's lymphoma, soft tissue sarcoma, solid tumor of children, lymphocytic Lymphoma, central nervous system (CNS) tumor, primary central nervous system lymphoma, tumor angiogenesis, spinal tumor, brain stem glioma, pituitary adenoma, black Tumor, Kaposi's sarcoma, epidermoid cancer, squamous cell cancer, T-cell lymphoma, chronic leukemia and acute, or a combination of the cancer.

In another preferred embodiment, said use refers to a therapeutically effective amount of a compound of the above formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above, and an anti-CTLA-4 antibody , anti-PD-1 antibody, anti-PD-L1 antibody, antiviral agent, chemotherapeutic agent, immunosuppressant, radiation, anti-tumor vaccine, anti-viral vaccine, cytokine therapy or tyrosine kinase inhibitor for combination; preferred The cytokine is preferably IL-2, IL-3, IL-4 or IL-5, the chemotherapeutic agent is preferably a cytotoxic agent, and the anti-PD-1 antibody is preferably a Keytruda antibody.

A fourth aspect of the invention provides a method of modulating the activity of guanamine 2,3-dioxygenase comprising administering a therapeutically effective amount of a compound of the above formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof Or the aforementioned pharmaceutical composition is contacted with indoleamine 2,3-dioxygenase. Preferably, the adjustment is preferably an inhibitory effect.

A fifth aspect of the invention provides a method of inhibiting immunosuppression in a patient, comprising administering a therapeutically effective amount of a compound of the above formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a combination thereof Give the patient a dose.

A sixth aspect of the invention provides a method of treating cancer, which comprises administering to a patient a therapeutically effective amount of a compound of the formula (I) of the present invention or a tautomer, a mesogen thereof, a foreign body A form of a rot, an enantiomer, a diastereomer or a mixture thereof, or a pharmaceutically acceptable salt thereof.

In another preferred embodiment, the cancer or tumor is selected from the group consisting of lung cancer, bone cancer, gastric cancer, pancreatic cancer, skin cancer, head and neck cancer, uterine cancer, ovarian cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, Cervical cancer, vaginal cancer, vulvar cancer, rectal cancer, colon cancer, anal cancer, breast cancer, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, urinary tract cancer, penile cancer, Prostate cancer, pancreatic cancer, brain cancer, testicular cancer, lymphoma, transitional cell carcinoma, bladder cancer, kidney cancer or ureteral cancer, renal cell carcinoma, renal pelvic cancer, Hodgkin's disease, non-Hodgkin's lymphoma, soft tissue sarcoma, Child solid tumor, lymphocytic lymphoma, central nervous system (CNS) tumor, primary central nervous system lymphoma, tumor angiogenesis, spinal tumor, brain stem glioma, pituitary adenoma, melanoma, card Combination of Posey sarcoma, epidermoid carcinoma, squamous cell carcinoma, T-cell lymphoma, chronic or acute leukemia, and said cancer.

It is to be understood that within the scope of the present invention, the various technical features of the present invention and the various technical features specifically described hereinafter (as in the embodiments) may be combined with each other to constitute a new or preferred technical solution.

Detailed ways

The inventors have unexpectedly discovered a class of N-(2-cyclohexylethyl)carboxamide derivatives which have better inhibitory activity and lower toxicity after long-term and intensive research.

Definition of Terms

As used herein, "alkyl" refers to both straight-chain and branched saturated aliphatic hydrocarbon groups, and _C1-10- alkyl is alkyl having from 1 to 10 carbon atoms, preferably _C1-8 alkyl, more preferably Preferred are C _1-6 alkyl groups, most preferably C _1-3 alkyl groups, the definitions are similar; non-limiting examples of alkyl groups include: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl , tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl Base, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3 - dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl , n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethyl Hexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-decyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n- Base, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched isomers thereof.

As used herein, "cycloalkyl" refers to a saturated or partially unsaturated monocyclic cyclic hydrocarbon group, and "C _3-10 cycloalkyl" refers to a cyclic hydrocarbon group containing from 3 to 10 carbon atoms, preferably C _3-8 A cycloalkyl group, more preferably a C _3-6 cycloalkyl group, is similarly defined; non-limiting examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexene A group, a cyclohexadienyl group, a cycloheptyl group, a cycloheptatrienyl group, a cyclooctyl group or the like is preferably a cyclopropyl group, a cyclopentyl group or a cyclohexenyl group.

As used herein, "C _1-10 alkoxy" refers to -O-(C _1-10 alkyl), wherein alkyl is as defined above. A C _1-8 alkoxy group is preferred, a C _1-6 alkoxy group is more preferred, and a C _1-3 alkoxy group is most preferred. Non-limiting examples include methoxy, ethoxy, propoxy, isopropoxy, butoxy, tert-butoxy, isobutoxy, pentyloxy and the like.

As used herein, "C _3-8 cycloalkoxy" refers to -O-(C _3-8 cycloalkyl), wherein cycloalkyl is as defined above. Preference is given to C _3-6 cycloalkoxy. Non-limiting examples include cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy, and the like.

As used herein, "C _6-10 aryl" refers to an all-carbon monocyclic or fused polycyclic ring (ie, a ring that shares a pair of adjacent carbon atoms) having a conjugated π-electron system, meaning 6 to 10 An aryl group of a carbon atom; preferably a phenyl group and a naphthyl group, most preferably a phenyl group.

As used herein, "a bond" refers to the attachment of two groups attached thereto through a covalent bond.

As used herein, "halogen" refers to fluoro, chloro, bromo or iodo.

As used herein, "halo" means that one or more (eg 1, 2, 3, 4 or 5) hydrogens in the group are replaced by a halogen.

For example, "halo C _1-8 alkyl" refers to an alkyl group substituted with one or more (eg 1, 2, 3, 4 or 5) halo, wherein alkyl is as defined above. It is selected as a halogenated C _1-6 alkyl group, more preferably a halogenated C _1-3 alkyl group. Examples of halogenated C _1-8 alkyl groups include, but are not limited to, monochloromethyl, dichloromethyl, trichloromethyl, monochloroethyl, 1,2-dichloroethyl, trichloroethyl, Monobromoethyl, monofluoromethyl, difluoromethyl, trifluoromethyl, monofluoroethyl, difluoroethyl, trifluoroethyl, and the like.

As another example, "halo C _1-8 alkoxy" means that the alkoxy group is substituted by one or more (eg 1, 2, 3, 4 or 5) halogens, wherein the alkoxy group is as defined above. It is preferably a halogenated C _1-6 alkoxy group, more preferably a halogenated C _1-3 alkoxy group. Examples of halo C _1-8 alkoxy include, but are not limited to, trifluoromethoxy, trifluoroethoxy, monofluoromethoxy, monofluoroethoxy, difluoromethoxy, difluoroethane. Oxyl and the like.

As another example, "halo C _3-8 cycloalkyl" refers to a cycloalkyl group substituted with one or more (eg, 1, 2, 3, 4, or 5) halo, wherein cycloalkyl is as defined above. Preferred is a halogenated C _3-6 cycloalkyl group. Examples of halogenated C _3-8 cycloalkyl groups include, but are not limited to, trifluorocyclopropyl, monofluorocyclopropyl, monofluorocyclohexyl, difluorocyclopropyl, difluorocyclohexyl, and the like.

As used herein, "deuterated C _1-8 alkyl" refers to an alkyl group substituted with one or more (eg 1, 2, 3, 4 or 5) deuterium atoms, wherein alkyl is as defined above. It is preferably a deuterated C _1-6 alkyl group, more preferably a deuterated C _1-3 alkyl group. Examples of deuterated C _1-8 alkyl groups include, but are not limited to, monodeuterated methyl, monodeuterated ethyl, dideuterated methyl, diterpene ethyl, triterpene methyl, triterpenoid B Base.

As used herein, "amino" refers to NH _2, "cyano" refers to the CN, "Nitro" refers to NO _2, "benzyl" refers to -CH ₂ - phenyl, "oxo" refers to = O, "carboxy" refers to -C (O) OH, "acetyl" means a -C (O) CH _3, "hydroxymethyl group" refers to -CH ₂ OH, "hydroxyethyl" refers to -CH ₂ CH ₂ OH, "hydroxy" means - OH, "thiol" refers to SH, "cyclopropylene" structure is:

As used herein, "heteroaryl ring" and "heteroaryl" are used interchangeably and mean having 5 to 10 ring atoms, preferably 5 or 6 membered monocyclic heteroaryl or 8 to 10 membered bicyclic heteroaryl. The ring array shares 6, 10 or 14 π electrons; and has a group of 1 to 5 hetero atoms in addition to carbon atoms. "Hetero atom" means nitrogen, oxygen or sulfur.

As used herein, "4- to 6-membered saturated monocyclic" refers to a saturated or partially unsaturated, all-carbon monocyclic ring containing from 4 to 6 ring atoms. Examples of 3- to 6-membered saturated or partially unsaturated monocyclic rings include, but are not limited to, cyclopropyl ring, cyclobutyl ring, cyclopentyl ring, cyclopentenyl ring, cyclohexyl ring, cyclohexenyl ring, ring. A hexadienyl ring, a cycloheptyl ring, a cycloheptatrienyl ring, a cyclooctyl ring or the like.

As used herein, "4 to 6 membered saturated monoheterocycle" means that 1, 2 or 3 carbon atoms in a 4 to 6 membered monocyclic ring are selected from nitrogen, oxygen or S(O) _t (where t is an integer 0) The heteroatoms to 2) are substituted, but do not include the ring moiety of -OO-, -OS- or -SS-, and the remaining ring atoms are carbon; preferably 4 to 6 members, more preferably 5 to 6 members. Examples of 4- to 6-membered saturated monoheterocycles include, but are not limited to, azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, pyrroline, oxazolidine, piperazine , dioxolane, dioxane, morpholine, thiomorpholine, thiomorpholine-1,1-dioxide, tetrahydropyran, and the like.

As used herein, a "5- to 6-membered monocyclic heteroaryl ring" refers to a monoheteroaryl ring containing from 5 to 6 ring atoms, including, for example, but not limited to, a thiophene ring, an N-alkylcyclopyrrole ring, Furan ring, thiazole ring, imidazole ring, oxazole ring, pyrrole ring, pyrazole ring, triazole ring, 1,2,3-triazole ring, 1,2,4-triazole ring, 1,2,5- a triazole ring, a 1,3,4-triazole ring, a tetrazole ring, an isoxazole ring, an oxadiazole ring, a 1,2,3-oxadiazole ring, a 1,2,4-oxadiazole ring, 1,2,5-oxadiazole ring, 1,3,4-oxadiazole ring, thiadiazole ring, pyridine ring, pyridazine ring, pyrimidine ring or pyrazine ring, and the like.

As used herein, "8- to 10-membered bicyclic heteroaryl ring" refers to a bi-heteroaryl ring containing from 8 to 10 ring atoms, including, for example, but not limited to, benzofuran, benzothiophene, anthracene, Isoindole, quinoline, isoquinoline, carbazole, benzothiazole, benzimidazole, quinazoline, quinoxaline, porphyrin, pyridazine.

As used herein, "substituted" refers to one or more hydrogen atoms in the group, preferably 1 to 5 hydrogen atoms are independently substituted with each other by a corresponding number of substituents, more preferably 1 to 3 hydrogen atoms are independent of each other. The ground is replaced by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art will be able to determine (by experiment or theory) substitutions that may or may not be possible without undue effort. For example, an amino group or a hydroxyl group having a free hydrogen may be unstable when combined with a carbon atom having an unsaturated (e.g., olefinic) bond.

As used herein, any of the above groups may be substituted or unsubstituted. When the above group is substituted, the substituent is preferably a group of 1 to 5 or less, independently selected from NR _a0 R _b0 , halogen, cyano, acetyl, hydroxy, hydroxymethyl, hydroxyethyl, carboxyl, C _{1 -8} alkyl, C _1-8 alkoxy, halo C _1-8 alkyl, C _3-8 cycloalkyl, C _3-8 cycloalkoxy, halogenated C _1-8 alkoxy, - C(O)C _1-10 alkyl, -C(O)OC _1-10 alkyl, -OC(O)C _1-10 alkyl, -CONR _a0 R _b0 ; R _a0 , R _b0 are each independently Hydrogen or C _1-8 alkyl.

The various substituent groups described herein above are themselves themselves replaceable by the groups described herein.

When the 4 to 6 membered saturated monoheterocycles described herein are substituted, the positions of the substituents may be at their possible chemical positions, and representative substitutions for the exemplary monoheterocycles are as follows:

Wherein "Sub" each independently represents a substituent of the type described herein;

Represents a connection to another atom.

Where the cycloalkyl groups described herein are substituted, the positions of the substituents may be at their possible chemical positions, and representative substitutions for the exemplary monoheterocycles are as follows:

Wherein "Sub" each independently represents a substituent of the type described herein;

Represents a connection to another atom.

As used herein, a compound of formula (I) may be present in one or more crystalline forms, and the active compounds of the invention include various crystalline forms and mixtures thereof.

The "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

"Pharmaceutically acceptable acid addition salt" means a salt formed with an inorganic or organic acid which retains the bioavailability of the free base without any other side effects.

"Pharmaceutically acceptable base addition salts", including but not limited to salts of inorganic bases such as sodium, potassium, calcium and magnesium salts, and the like. These include, but are not limited to, salts of organic bases such as ammonium salts, triethylamine salts, lysine salts, arginine salts and the like.

As used herein, "solvate" refers to a complex of a compound of the invention with a solvent. They either react in a solvent or precipitate out of the solvent or crystallize out. For example, a complex formed with water is referred to as a "hydrate." Solvates of the compounds of formula (I) are within the scope of the invention.

The compounds of formula (I), formula (II) or (III) of the present invention may contain one or more chiral centers and exist in different optically active forms. When the compound contains a chiral center, the compound contains the enantiomer. The invention includes mixtures of the two isomers and isomers, such as racemic mixtures. Enantiomers can be resolved by methods known in the art, such as crystallization and chiral chromatography. When a compound of formula (I), formula (II) or (III) contains more than one chiral center, diastereomers may be present. The present invention includes resolved optically pure specific isomers as well as mixtures of diastereomers. Diastereomers can be resolved by methods known in the art, such as crystallization and preparative chromatography.

The invention includes prodrugs of the above compounds. Prodrugs include known amino protecting groups and carboxy protecting groups which are hydrolyzed under physiological conditions or released via an enzymatic reaction to give the parent compound. Specific prodrug preparation methods can be referred to (Saulnier, MG; Frennesson, DB; Deshpande, MS; Hansel, SB and Vysa, DM Bioorg. Med. Chem Lett. 1994, 4, 1985-1990; and Greenwald, RB; Choe, YH; Conover, CD; Shum, K.; Wu, D.; Royzen, MJ Med. Chem. 2000, 43, 475.).

In general, a compound of the invention, or a pharmaceutically acceptable salt thereof, or a solvate thereof, or a stereoisomer, or prodrug thereof, can be administered in a suitable dosage form with one or more pharmaceutically acceptable carriers. These dosage forms are suitable for oral, rectal, topical, intraoral, and other parenteral administration (eg, subcutaneous, intramuscular, intravenous, etc.). For example, dosage forms suitable for oral administration include capsules, tablets, granules, and syrups and the like. The compound of the present invention contained in these preparations may be a solid powder or granule; a solution or suspension in an aqueous or non-aqueous liquid; a water-in-oil or oil-in-water emulsion or the like. The above dosage forms can be prepared from the active compound with one or more carriers or excipients via conventional pharmaceutical methods. The above carriers need to be compatible with the active compound or other excipients. For solid formulations, commonly used non-toxic carriers include, but are not limited to, mannitol, lactose, starch, magnesium stearate, cellulose, glucose, sucrose, and the like. Carriers for liquid preparations include water, physiological saline, aqueous dextrose, ethylene glycol, polyethylene glycol, and the like. The active compound can form a solution or suspension with the above carriers.

The compositions of the present invention are formulated, quantified, and administered in a manner consistent with medical practice. The "therapeutically effective amount" of a given compound will be determined by the particular condition being treated, the individual being treated, the cause of the condition, the target of the drug, and the mode of administration.

As used herein, "therapeutically effective amount" refers to a compound of the invention that will elicit a biological or medical response to an individual, such as reducing or inhibiting the activity of an enzyme or protein or ameliorating a condition, alleviating a condition, slowing or delaying the progression of a disease, or preventing a disease, and the like. the amount.

The therapeutically effective amount of the compound of the present invention or a pharmaceutically acceptable salt thereof, or a solvate thereof, or a stereoisomer thereof, contained in the pharmaceutical composition of the present invention is preferably 0.1 mg to 5 g/kg (body weight).

As used herein, "pharmaceutically acceptable carrier" means a non-toxic, inert, solid, semi-solid substance or liquid filler, diluent, encapsulating or auxiliary formulation or any type of excipient that is compatible with the patient, most Preferably, it is a mammal, more preferably a human, which is suitable for delivering the active agent to a target of interest without terminating the activity of the agent.

As used herein, "patient" refers to an animal, preferably a mammal, and more preferably a human. The term "mammal" refers to warm-blooded vertebrate mammals including, for example, cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, rats, pigs, and humans.

As used herein, "treating" refers to alleviating, delaying progression, attenuating, preventing, or maintaining an existing disease or condition (eg, cancer). Treatment also includes curing, preventing, or alleviating one or more symptoms of the disease or condition to some extent.

Preparation

The present invention provides a process for the preparation of a compound of formula (I). The compounds of the present invention can be prepared by a variety of synthetic procedures, and exemplary methods of preparation of such compounds can include, but are not limited to, the schemes described below.

Preferably, the compounds of formula (I) of the present invention can be prepared by the following schemes and exemplary methods described in the Examples and related publications used by those skilled in the art.

The steps in the method can be extended or merged as needed during the specific operation.

Route 1

Step: The compound of the formula (I-1) and the compound of the formula (I-2) is subjected to amidation to form a compound of the formula (I). The synthesis of the amide can be carried out by using an active ester method, a carbodiimide condensing agent method, or a sulfonium salt condensing agent. A method such as a method and an organophosphorus condensing agent, and the condensing agent used includes, but not limited to, a carbodiimide type condensing agent (e.g., N, N'-dicyclohexylcarbodiimide DCC, diisopropylcarbodimide) Amine DIC, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide EDCI, etc.) in combination with activators HOSu, HOBt, HOAt, HOOBt, etc., carbonium salt condensing agents (eg TBTU) , HCTU, HBTU, TNTU, HATU, HAPyU, HBPyU, TSTU, etc.), phosphonium salt condensing agents (such as BOP, PyBOP, PyAOP, etc.), organophosphorus condensing agents (such as DPP-Cl, DECP, DPPA, MPTA, BOP-Cl, etc.), triphenylphosphine-polyhalomethane, triphenylphosphine-hexachloroacetone, triphenylphosphine-NBS, and the like.

Route 2

Step: Conversion of the compound of the formula (I-3-1) to the corresponding cyclohexyl compound (I-3-2) can be carried out under acidic conditions using a metal (may be but not limited to iron powder, zinc powder) or stannous chloride Reduction is carried out; or hydrogenation reduction under palladium carbon catalysis. The compound of formula (I-3-2) can be converted into the corresponding primary amine compound (I-3) by using AlCl ₃ /LiAlH 4 , NaBH ₄ /CoCl ₂ .6H ₂ O, Raney Ni/NH ₂ NH ₂ .HCOOH, I ₂ The reaction is carried out with a reducing agent such as /NaBH ₄ .

Route 3

Step: The compound of the formula (I-1-1) is converted into a compound of the formula (I-1-2) by a Tosmic reaction in the presence of p-toluenesulfonylmethyl isonitrile, and the compound of the formula (I-1-2) is Reduction of the compound of the formula (I-1-3) in the presence of a reducing agent of NiCl and NaBH ₄ , and finally removal of Boc in the presence of an acid gives the compound of the formula (I-1).

The starting materials of the starting materials (I-1-1) and (I-3-1) in the route may be commercially available depending on the specific structure, or may be prepared by a method known to those skilled in the art.

The reactions in the above various steps are conventional reactions known to those skilled in the art. Unless otherwise stated, the reagents and starting materials used in the synthetic route are either commercially available or can be prepared by those skilled in the art according to known methods of designing different compound structures.

The main advantage of the present invention over the prior art is that it has better IDO inhibitory activity and lower toxicity.

The invention is further illustrated below in conjunction with specific embodiments. It is to be understood that the examples are not intended to limit the scope of the invention. The experimental methods in the following examples which do not specify the specific conditions are usually carried out according to conventional conditions or according to the conditions recommended by the manufacturer. Percentages and parts are by weight unless otherwise stated. Unless otherwise defined, the terms used herein have the same meaning as commonly understood by those skilled in the art. In addition, any methods and materials similar or equivalent to those described can be applied to the present invention.

As used herein, room temperature means about 20-25 °C.

As used herein, DMB is 2,4-dimethoxybenzyl, THF is tetrahydrofuran, EA is ethyl acetate, PE is petroleum ether, Ac ₂ O is acetic anhydride, and NBS is N-bromosuccinimide. DCM is dichloromethane, AIBN is azobisisobutyronitrile, Pd(dppf)Cl ₂ is 1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, TFA is trifluoroacetic acid, TBSCl Is tert-butyldimethylchlorosilane, NCS is N-chlorosuccinimide, DHP is dihydrotetrahydropyran, LiAlH ₄ is lithium aluminum hydride, PMB is p-methoxybenzyl, LiHMDS is two Lithium (trimethylsilyl)amide, Pd ₂ (dba) ₃ is tris(dibenzylideneacetone)dipalladium, RuPhos is 2-dicyclohexylphosphorin-2',6'-diisopropoxy-1 , 1 '-biphenyl, DMAP is 4-dimethylaminopyridine, THP is tetrahydrotetrahydropyran, n-BuLi is n-butyl lithium, TMsOTf is trimethylsilyl trifluoromethanesulfonate, TEBAC is three Ethylbenzylammonium chloride, HATU is 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, DMF is dimethylformamide DMSO is dimethyl sulfoxide, DIEA is N,N-diisopropylethylamine, and BINAP is (2R,3S)-2,2'-bisdiphenylphosphino-1,1'-binaphthyl. TosMic is p-toluenesulfonyl isocyanide.

Example 1

Step 1: a solution of compound 1.1 (6.74 g, 38.04 mmol), THF (60.00 mL) and DMPU (13.28 g, 103.74 mmol), NaH (1.45 g, 36.31 mmol, 60% purity) was added portionwise at 0 °C. Compound 1-1 (5.40 g, 34.58 mmol). The mixture was stirred at room temperature for 15 h under a nitrogen atmosphere and was then observed by LC-MS. Water was added to the system, the mixture was extracted with EA, the organic layer was _washed with saturated NaHC03 solution, dried over anhydrous Na ₂ SO4 and concentrated to give compound 1-2 (6.10g, 34.04mmol).

Step 2: Compound 1-2 (1.40g, 7.72mmol) and pyridine H p-toluenesulfonate (3.88g, 15.44mmol) in ₂ O (15.00mL), acetone (15.00 mL) added and the mixture stirred at room temperature 15h. Reaction was followed by TLC to completion .EA water added to the system, the water layer was extracted with EA, the combined organic layers were washed with saturated aqueous sodium chloride solution, dried over anhydrous Na ₂ SO4, and concentrated to give compound 1-3 (950.00mg, 6.93mmol).

Step 3: Compound 1-3 (950.00 mg, 6.93 mmol) in DCM (1 <RTI ID=0.0></RTI></RTI><RTIgt; After 20 minutes, compound 1.2 (3.91 g, 13.86 mmol) was added to the mixture. The mixture was stirred at room temperature under nitrogen for 1 h and then was taken to EtOAc. Joining DCM, washed with water and the mixture are saturated saline, dried over anhydrous Na ₂ SO4, and concentrated to give compound 1-4 (1.80g, 6.69mmol).

Step 4: A solution of compound 1-4 (1.80 g, 6.69 mmol), 1,4-dioxane (15.00 mL) and potassium acetate (1.89 g, 20.07 mmol), then Pd(dppf)Cl ₂ (489.04) The mixture was stirred at 0&lt;0&gt;C for 15 h under N.sub.2. The mixture was concentrated and purified by silica gel column chromatography (EtOAc: EtOAc: EtOAc)

Step 5: In a solution of compound 1-5 (500.00 mg, 2.02 mmol), 1,4-dioxane (9.98 mL) and Pd(dppf)Cl ₂ (147.66 mg, 202.00 umol), Na ₂ CO was added successively. ₃ (642.36 mg, 6.06 mmol), compound 1.4 (630.39 mg, 3.03 mmol) and H ₂ O (525.26 uL). The mixture was stirred at 80 ° C for 15 h under nitrogen atmosphere, and LC-MS was followed until the reaction was completed. The mixture was concentrated, EtOAc (EtOAc m.

Step 6: Compound 1-6 (450.00 mg, 1.81 mmol), EtOAc (EtOAc m. The mixture was filtered, and the filtered cake was washed with EtOAc.

Step 7: Compound 1-7 (440.00 mg, 1.76 mmol), EtOAc (EtOAc (EtOAc) LC-MS was followed until the reaction was complete. The mixture was filtered, and the filtered cake was washed with EtOAc EtOAc EtOAc

Step 8: DIPEA (116.86 mg, 904.19 umol) was added to a solution of compound 1a (147.01 mg, 904.19 umol), DCM (10.00 mL), HATU (343.59 mg, 904.19 umol). After the mixture was stirred at room temperature for 30 min, compound 1-8 (230.00 mg, 904.19 umol) was added. The mixture was stirred at room temperature for 2 h and was taken up by LC-MS until complete. The reaction solution was concentrated and purified by column chromatography (PE: EA = 1:1) to obtain compound z-1, which was purified by prep-HPLC (H ₂ O/CH ₃ CN 30%-40%-20 min) to obtain Z-1 (27.2). Mg, 66.07 umol, 3.82% yield). MS (ESI) m/z: 399 (M+H) ⁺ . ¹ H NMR (400 MHz, DMSO) δ 9.12 (d, J = 5.2 Hz, 1H), 8.64 (dd, J = 12.4, 7.2 Hz, 1H) ), 8.49 (d, J = 8.4 Hz, 1H), 8.22 (d, J = 8.4 Hz, 1H), 8.02 (t, J = 7.6 Hz, 1H), 7.87 (dt, J = 17.4, 6.0 Hz, 2H) ), 7.66 (d, J = 4.0 Hz, 1H), 7.18 (d, J = 4.0 Hz, 1H), 3.58 (d, J = 28.8 Hz, 1H), 3.33 - 3.28 (m, 2H), 1.96 - 1.51 (m, 11H).

Preparation of Compound Z-2 of Example 2

The preparation method is the same as the compound Z-1 except that the compound 1.4 in the step 5 of the Z-1 process is replaced with the compound 3-bromoquinoline to obtain the compound Z-2. MS (ESI) m/z: 399 (M + +) ⁺ . ¹ H NMR (400 MHz, DMSO) δ 9.26-9.24 (m, 1H), 8.61 (s, 1H), 8.07 (d, J = 8.4 Hz , 1H), 7.91 (t, J = 8.0 Hz, 1H), 7.75-7.59 (m, 4H), 7.19 (d, J = 4.0 Hz, 1H), 3.32-3.27 (m, 2H), 2.89-2.68 ( m, 1H), 2.00-1.91 (m, 2H), 1.75-1.42 (m, 8H), 1.19-1.10 (m, 1H).

Preparation of Compound Z-3 of Example 3

The preparation method is the same as the compound Z-1, except that the compound 1.4 in the step 5 of the Z-1 process is replaced with the compound 4-bromoisoquinoline to obtain the compound Z-3. MS (ESI) m/z: 399 (M + +) ⁺ . ¹ H NMR (400 MHz, DMSO) δ 9.16 (s, 1H), 8.60 (s, 1H), 8.43 (d, 1H), 8.16 (dd) , 2H), 7.83-7.79 (m, 1H), 7.69-7.64 (m, 2H), 7.18 (d, 1H), 3.30 (s, 2H), 3.07-3.05 (m, 1H), 1.94 (d, 2H) ), 1.87-1.65 (m, 6H), 1.62 (d, 1H), 1.51 (d, 1H), 1.29 (s, 1H).

Preparation of Compound Z-4 of Example 4

The preparation method is the same as the compound Z-1 except that the compound 1.4 in the step 5 of the Z-1 process is replaced with the compound 1-bromoisoquinoline to obtain the compound Z-4. MS (ESI) m / z: 399 (M + 1) +. ^{1 H NMR (400MHz, DMSO)} δ = 8.60 (t, 1H), 8.42 (m, 1H), 8.31 (d, 1H), 7.94 (d, 1H), 7.74 (dd, 1H), 7.70-7.65 (m , 0.7H), 7.66 - 7.59 (m, 2H), 7.17 (d, 1H), 3.67 (m, 1H), 3.30 - 3.22 (m, 2H), 2.09 - 1.45 (m, 10H).

Preparation of Compound Z-5 of Example 5

A solution of compound Z-1 (350.00 mg, 877.30 umol), EtOAc (10.00 mL) LC-MS was followed until the reaction was complete. The mixture was filtered, and the filtered cake was washed with EtOAc EtOAc EtOAc EtOAc EtOAc MS (ESI) m / z 365 (M + H) +. 1 H NMR (400MHz, DMSO) δ9.17-9.16 (m, 1H), 8.57-8.51 (m, 2H), 8.26 (d, H), 8.07(t,1H),7.92-7.86(m,2H),7.78-7.72(m,2H),7.15-7.13(m,1H),3.63-3.60(m,1H),3.36-3.29(m,2H ), 1.92-1.61 (m, 10H), 1.55-1.48 (m, 1H).

Preparation of Compound 6-6 of Example 6

The preparation method is the same as the compound Z-5, except that the compound Z-1 in the Z-5 process is replaced with the compound Z-2 to obtain the compound Z-6. MS (ESI) m/z </RTI>^</RTI><RTIID=0.0></RTI></RTI><RTIgt;^</RTI><RTIgt;^</RTI><RTIgt;^</RTI><RTIgt;^</RTI></RTI></RTI></RTI></RTI></RTI></RTI><RTIgt; , 2H), 7.75 (ddt, 2H), 7.69 (ddt, 1H), 7.58 (ddd, 1H), 7.15 (ddd, 1H), 3.34-3.26 (m, 2H), 2.97-2.64 (m, 1H), 1.94 (d, 2H), 1.85-1.58 (m, 6H), 1.57-1.42 (m, 2H), 1.16 (dd, 1H).

Preparation of Compound 13 of Example 13

Step 1: The preparation method is the same as the compound 1-8 except that the compound 1-7 in the 1-8 process is replaced with the compound 13-1.

Step 2: The preparation method is the same as the compound Z-1 except that the compound 1-8 in the Z-1 process is replaced with the compound 13-2.

Step 3: A solution of compound 13-3 (372.91 mg, 1.00 mmol), EtOAc (EtOAc EtOAc) , 1.39 mmol) was used directly in the next reaction.

Step 4: toluene (10.00 mL) Compound 13-4 (365.54mg, 1.34mmol) was added at room temperature Pd _{2 (dba) 3 (61.32mg,} 67umol), BINAP (83.43mg, 134umol), Cs 2 CO ₃ (1.31 g, 4.02 mmol) and compound 13.1 (278.79 mg, 1.34 mmol). The mixture was stirred at 100 ° C for 15 h under nitrogen atmosphere. LC-MS was followed until the end of the reaction. The reaction mixture was concentrated and purified by prep-TLC (DCM:MEOH=15:1) and prep-HPLC (H ₂ O/CH ₃ CN 22%-32%-20 min) to obtain compound Z-13 (93.40 mg, 15.61% yield ). MS (ESI) m / z 400 (M + H) ⁺ . ^{1 H NMR (400MHz, DMSO)} δ8.65 (d, 1H), 8.60 (t, 1H), 8.16 (s, 1H), 7.98 (d, 1H), 7.93 (d, 1H), 7.77-7.59 (m , 2H), 7.58-7.47 (m, 1H), 7.18 (d, 1H), 6.95 (d, 1H), 3.53 (d, 2H), 3.33 (dd, 2H), 2.79 (t, 2H), 1.89 ( d, 2H), 1.55 (m, 5H).

Preparation of Compound Z-15 of Example 15

Step 1: The preparation method is the same as that of the compound 1-6, except that the compound 1-5 and the compound 1.4 in the 1-6 process are replaced with the compound 15.1 and the compound 15-1. Purification via combiflash to give oily compound 15-2 (3.6g, 92% yield), MS m / z (ESI ): 286 [M + H] +.

Step 2: The preparation method is the same as the compound 1-7, except that the compound 1-6 in the 1-7 process is replaced with the compound 15-2. MS m/z (ESI): 288[M+H] ⁺ .

Step 3: To a solution of EtOAc (EtOAc m. LCMS was followed until the end of the reaction. The reaction mixture was stirred with EtOAc EtOAc. MS m/z (ESI): 244 [M+H] ⁺ .

Step 4: A solution of compound 15.2 ( 1.36 g, 7. <RTI ID=0.0></RTI><RTIgt;</RTI><RTIgt;</RTI><RTIgt;</RTI><RTIgt; A solution of the compound 15-4 (1.7 g, 7.0 mmol) in THF (20 mL) was evaporated. LCMS was followed until the end of the reaction. Water was added to the system and extracted with EA. The organic layer was concentrated to give compound 15-5. MS m/z (ESI): 266 [M+H] ⁺ .

Step 5: The preparation method is the same as the compound 1-7 except that the compound 1-6 in the 1-7 process is replaced with the compound 15-5. MS m/z (ESI): 266 [M+H] ⁺ .

Step 6: compound of THF 15-6 (50mL) was added LiH ₄ Al (623mg, 16.42mmol) at -10 ℃. The mixture was stirred at -10 ° C for 1 h, then warmed to 0 ° C and stirred for 3 h. LCMS was followed until the end of the reaction. 10H ₂ O·Na ₂ SO ₄ was added to the system, and the mixture was stirred at room temperature for 1 hr, then filtered and concentrated to give compound 15-7. MS m/z (ESI): 266 [M+H] ⁺ .

Step 7: A solution of compound 15-7 (0.96 g, <RTI ID=0.0></RTI></RTI><RTIgt; The mixture was stirred at room temperature for 1 h. LCMS was followed until the end of the reaction. The mixture was concentrated and purified by prep-HPLC to afford compound Z-15 (38 </RTI><RTIgt; MS m/z (ESI): 417 [M+H] ⁺ .

Preparation of Compound 16 of Example 16

Step 1: To a solution of the compound TosMic (603 mg, 3.09 mmol) in EtOAc (EtOAc (EtOAc) ) was added to the reaction system and stirring was continued at room temperature overnight. LCMS was followed until the end of the reaction. The mixture was concentrated under reduced pressure and purified by combiflash to afford 200 mg of Compound 16-1. MS m/z (ESI): 255.2 [M+H] ⁺ .

Step 2: Compound 16-1 (200mg, 0.8mmol) in methanol (50mL) was added _{(Boc) 2 O (349.2mg,} 1.6mmol), NiCl (311mg, 2.4mmol) and NaBH ₄ (152mg, 4mmol). The mixture was stirred at room temperature for 2 h. LCMS was followed until the end of the reaction. The mixture was concentrated under reduced pressure and purified by combiflash to yield compound (di). MS m/z (ESI): 359.3 [M+H] ⁺ .

Step 3: The preparation method is the same as that of the compound 8-4, except that the compound 8-3 in the 8-4 process is replaced with the compound 16-2. MS m/z (ESI): 259.3 [M+H] ⁺ .

Step 4: The preparation method is the same as the compound Z-15 except that the compound 15-7 in the Z-15 process is replaced with the compound 16-3. Purification by prep-HPLC gave compound Z-16 (54 mg, 40.6% yield). MS m/z (ESI): 403.1 [M+H] ⁺ . The obtained compound Z-16 was separated by column AY-H (250*4.6 mm 5 um) (mobile phase: hexane (0.1% DEA): ethanol (0.1% DEA) = 80:20) to obtain 4.89 mg of compound Z-27, respectively. And 4.56 mg of compound Z-28.

Example 17 Preparation of Compound Z-17

Step 1: 4-Chloro-6-fluoroquinoline (2.50 g, 13.81 mmol), 1,4-dioxa-spiro[4,5]indole-7-ene-8-boronic acid pinacol ester (4.00 g, 15.19 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (1.0 g, 1.38 mmol) and sodium carbonate (4.20 g, 41.43 mmol) dissolved in dioxane In a mixed solution of a ring (60 mL) and water (15 mL), the reaction mixture was then heated to 100 ° C and stirred for 18 hours. When the reaction was completed, the reaction was cooled to room temperature, then filtered, evaporated, evaporated, evaporated. MS (ESI) 286.1 [M+H]+; ¹ H NMR (400 MHz, dmso) δ 8.78 (d, J = 4.4 Hz, 1H), 8.09 - 8.01 (m, 1H), 7.67 - 7.57 (m, 2H) ), 7.30 (d, J = 4.4 Hz, 1H), 5.69 - 5.62 (m, 1H), 3.93 (d, J = 3.3 Hz, 4H), 2.48 - 2.44 (m, 2H), 2.41 (d, J = 3.1Hz, 2H), 1.86 (t, J = 6.4Hz, 2H).

Step 2: 6-Fluoro-4-(1,4-dioxaspiro[4.5]indole-7-en-8-yl)quinoline (3.10 g, 10.88 mmol) was dissolved in THF (60 mL) Palladium on charcoal (10%, 50% w/w, 600 mg) was added, and the reaction mixture was stirred with EtOAc. MS (ESI) 288.1 [M+H]+.

Step 3: 6-Fluoro-4-(1,4-dioxaspiro[4.5]decane-8-yl)quinoline (3.10 g, 10.80 mmol) was dissolved in THF (10 mL). 2M, 20 mL, 40 mmol), and the mixture was stirred at room temperature for 1 hour, then the mixture was stirred with EtOAc EtOAc EtOAc The filtrate was evaporated to dryness to dryness crystall MS (ESI) 244.1 [M+H]+.

Step 4: 2-Ethoxyphosphorylacetonitrile (1.36 g, 7.70 mmol) was dissolved in THF (50 mL). N,N-dimethylpropenyl urea (2.70 g, 21.00 mmol) The liquid was cooled to 0 ° C, NaH (60%, 336 mg, 8.40 mmol) was added portionwise, stirred at 0 ° C for 30 min, and finally 4-(6-fluoro-4-quinolinyl)-cyclohexanone (1.70) A solution of g, 7.00 mmol) in THF (20 mL) was added dropwise to the reaction mixture at 0 ° C. After the dropwise addition was completed, the reaction mixture was stirred at 0 ° C for 1 hour, and then allowed to react to room temperature for 1 hour. When the reaction mixture was completed, the reaction mixture was poured into EtOAc EtOAc (EtOAc) 100%). MS (ESI) 267.0 [M+H]+.

Step 5: Add 2-(4-(6-fluoroquinolin-4-yl)cyclohexylene)acetonitrile (2.10 g, 7.89 mmol) and palladium on carbon (10%, 50% w/w, 500 mg) to methanol (20 mL) and a mixture of acetic acid (10 mL), and the mixture was stirred under H2HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH The organic phase was dried over anhydrous sodium sulfate (MgSO4).

MS (ESI) 269.1 [M+H]+.

Step 6: Dissolve 2-(4-(6-fluoroquinolin-4-yl)cyclohexyl)acetonitrile (1.10 g, 4.10 mmol) in THF (50 mL). Lithium aluminum hydride (623 mg, 16.42 mmol) was added, and the reaction solution was stirred at -10 ° C for 1 hour, and then further stirred at 0 ° C for 3 hours. When the starting material was completely reacted, sodium sulfate decahydrate was added portionwise to the reaction mixture. The mixture was stirred for 1 hour, and the insoluble material was filtered to remove the insoluble material. The filtrate was evaporated to dryness to give a crude oily product (1.07 g, 96%). MS (ESI) 273.3 [M+H]+.

Step 7: Dissolving 2-(4-(6-fluoroquinolin-4-yl)cyclohexyl)ethyl-1-amine (50 mg, 0.18 mmol) and diisopropylethylamine (71 mg, 0.55 mmol) In dichloromethane (10 mL), 3-chlorobenzoyl chloride (38 mg, 0.22 mmol) was added to the reaction mixture and stirred at room temperature for 1 hour. When the starting material was completely reacted, the reaction solution was sub-steamed and dried. Phase Preparation gave the white solid product Z-17 (6.2 mg, 8%). MS (ESI) 411.2 [M+H] +; ¹ H NMR (400 MHz, DMSO) δ 8.80 (d, J = 4.6 Hz, 1H), 8.60 (t, J = 5.4 Hz, 1H), 8.09 (dd, J = 9.2, 5.8 Hz, 1H), 7.98 (dd, J = 11.0, 2.7 Hz, 1H), 7.90 (t, J = 1.7 Hz, 1H), 7.83 (d, J = 7.8 Hz, 1H), 7.70 - 7.63 (m, 1H), 7.63 - 7.57 (m, 1H), 7.51 (t, J = 7.8 Hz, 1H), 7.44 (d, J = 4.6 Hz, 1H), 3.31 - 3.14 (m, 3H), 1.91 (s, 4H), 1.64–1.40 (m, 5H), 1.39–1.20 (m, 2H).

Preparation of Example 18 Compound Z-18

Step 1: Dissolve Boc-D-proline (95 mg, 0.44 mmol) and diisopropylethylamine (142 mg, 1.10 mmol) in dichloromethane (10 mL), then add 2-(7-azobenzene) And triazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (168 mg, 0.44 mmol) and stirred at room temperature for 1 hour, finally adding 2-(4-(6-fluoroquinoline) 4-yl)cyclohexyl)ethyl-1-amine (100 mg, 0.37 mmol) was stirred at room temperature for 2 hr., then, the reaction mixture was diluted with dichloromethane (30 mL) and then water (10 mL) The organic phase was washed with brine (10 mL). MS (ESI) 470.2 [M+H]+.

Step 2: The crude product (R)-2-((2-(4-(6-fluoroquinolin-4-yl)cyclohexyl)ethyl)carboxamide)pyrrolidine-1-carboxylic acid tert-butyl ester (312 mg, 0.67 mmol) was dissolved in dichloromethane (5 mL). Hydrochloric acid in dioxane (4M, 5 mL, 20 mm ol) was added and stirred for 1 hour at room temperature. Evaporation and purification by high-performance liquid to give white solid product HY 803183 (14 mg, 10%, two steps). MS (ESI) 370.2 [M+H] +; ¹ H NMR (400 MHz, DMSO) δ 9. s (s, 2H), 8.85 (s, 1H), 8.46 (t, J = 5.3 Hz, 1H), 8. 8.12 (m, 2H), 7.87 (s, 1H), 7.73 (d, J = 10.1 Hz, 1H), 4.15 - 4.05 (m, 1H), 3.51 (s, 1H), 3.38 - 3.06 (m, 4H) , 2.35–2.22 (m, 1H), 2.02–1.20 (m, 14H).

Preparation of Compound 20 of Example 20

1-Methylpyrazole-4-carboxylic acid (56 mg, 0.44 mmol) and diisopropylethylamine (142 mg, 1.10 mmol) were dissolved in dichloromethane (10 mL) and then 2-(7-azobenzene) And triazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (168 mg, 0.44 mmol) and stirred at room temperature for 1 hour, finally adding 2-(4-(6-fluoroquinoline) 4-yl)cyclohexyl)ethyl-1-amine (100 mg, 0.37 mmol) and stirred at room temperature for 2 hours. When the starting material was completely reacted, the reaction mixture was evaporated to dryness in vacuo. Product HY803185 (20 mg, 14%). MS (ESI) 381.3 [M+H] +; ¹ H NMR (400 MHz, DMSO) δ 8.84 (t, J=4.2 Hz, 1H), 8.12 (dd, J = 15.5, 8.1 Hz, 2H), 8.06– 7.96 (m, 2H), 7.84 (s, 1H), 7.70 (td, J = 8.8, 2.7 Hz, 1H), 7.50 (dd, J = 18.2, 4.6 Hz, 1H), 3.85 (d, J = 4.1 Hz) , 3H), 3.44 - 3.20 (m, 3H), 1.90 (d, J = 10.5 Hz, 3H), 1.70 (d, J = 7.0 Hz, 3H), 1.62 - 1.38 (m, 3H), 1.31 (dd, J=22.7, 10.6Hz, 2H).

The compounds Z-19, Z-21 to Z-25, Z-29 to Z-30 were prepared by the method of the compound Z-20. The compounds Z-26, Z-31, Z-32, Z-33 and Z-40 were prepared by the method of the compound Z-17. Z-34, Z-35, Z-41 and Z-42 were prepared by the method of the compound Z-17, and the obtained racemic product was prepared by high pressure liquid phase.

Preparation of Example 36-37 Compound Z-36/Z-37

2-(4-(6-Fluoroquinolin-4-yl)cyclohexyl)ethyl-1-amine (0.96 g, 3.53 mmol) and triethylamine ( 1.43 g, 14.12 mmol). In 35 mL), 5-chloro-2- acylchlorothiophene ( 0.70 g, 3.88 mmol) was added and stirred at room temperature for 1 hour. When the reaction was completed, the reaction mixture was evaporated to dryness and evaporated The solid product, the trans product was compound Z-36 (213 mg, 14%). MS (ESI) 417.1 [M+H] +; ¹ H NMR (400 MHz, dmso) δ 8.77 (d, J = 4.5 Hz, 1H), 8.52 (s, 1H), 8.05 (dd, J = 9.2, 5.8 Hz, 1H), 7.94 (dd, J = 11.0, 2.7 Hz, 1H), 7.67 - 7.58 (m, 2H), 7.41 (d, J = 4.5 Hz, 1H), 7.15 (d, J = 4.0 Hz, 1H) ), 3.25 (s, 2H), 1.87 (d, J = 11.3 Hz, 4H), 1.42 (ddd, J = 76.8, 20.2, 12.1 Hz, 8H). The cis product is compound Z-37 (168 mg, 11%) ). MS (ESI) 417.1 [M+H] +; ¹ H NMR (400 MHz, dmso) δ 8.81 (d, J = 4.5 Hz, 1H), 8.57 (t, J = 5.6 Hz, 1H), 8.08 (dd, J=9.2, 5.9 Hz, 1H), 7.96 (dd, J=11.0, 2.8 Hz, 1H), 7.69–7.61 (m, 2H), 7.48 (d, J=4.5 Hz, 1H), 7.18 (d, J) =4.0 Hz, 1H), 3.34 (d, J = 4.2 Hz, 1H), 3.30 - 3.25 (m, 2H), 1.91 - 1.64 (m, 11H).

Preparation of Example 38-39 Compound Z-38/Z-39

2-((1r,4r)-4-(quinolin-4-yl)cyclohexyl)ethyl-1-amine and 2-((1s,4s)-4-(quinolin-4-yl)cyclohexyl Preparation of ethyl-1-amine

2-(4-(Quinolin-4-yl)cyclohexyl)acetonitrile (1.60 g, 6.40 mmol) was dissolved in THF (70 mL), and the reaction mixture was reduced to -10 ° C, and lithium aluminum hydride ( 973 mg) was added portionwise. , 25.60 mmol), then the reaction solution was stirred at -10 ° C for 1 hour, and then increased to 0 ° C and stirred for 3 hours. When the starting material was completely reacted, sodium sulfate decahydrate was added to the reaction solution in portions until no bubbles were generated. The mixture was stirred for 1 hour, and the undissolved material was removed by filtration. The filtrate was evaporated to dryness to give a crude oily product, and then the crude product was purified by high pressure liquid to give two white solid products, the trans product 2-((1r, 4r)-4-(Quinolin-4-yl)cyclohexyl)ethyl-1-amine (270 mg, 11%). MS (ESI) 255.3 [M + H] +; 1 H NMR (400MHz, DMSO) δ8.84 (d, J = 4.5Hz, 1H), 8.44 (s, 2H), 8.22 (d, J = 8.3Hz, 1H), 8.03 (d, J = 8.4 Hz, 1H), 7.80 - 7.70 (m, 1H), 7.68 - 7.59 (m, 1H), 7.43 (d, J = 4.5 Hz, 1H), 3.43 (s, 1H) ), 2.88–2.75 (m, 2H), 1.94–1.54 (m, 11H). The cis product is 2-((1s,4s)-4-(quinolin-4-yl)cyclohexyl)ethyl-1 -Amine (140 mg, 6%). MS (ESI) 255.3 [M+H]+; ¹ H NMR (400 MHz, DMSO) δ 8.84 (d, J = 4.5 Hz, 1H), 8.38 (s, 2H), 8.22 (d, J = 8.4 Hz, 1H), 8.03 (d, J = 8.3 Hz, 1H), 7.79 - 7.71 (m, 1H), 7.67 - 7.58 (m, 1H), 7.42 (d, J = 4.5 Hz, 1H), 3.52 - 3.33 (m , 1H), 2.88–2.76 (m, 1H), 2.04–1.39 (m, 12H).

Preparation of Compound Z-38

Dissolving 2-((1r,4r)-4-(quinolin-4-yl)cyclohexyl)ethyl-1-amine (270 mg, 1.06 mmol) and triethylamine ( 429 mg, 4.25 mmol) in dichloro Add methane (15 mL), and add 5-chloro-2-acylchlorothiophene ( 230 mg, 1.28 mmol) and stir at room temperature for 1 hour. When the starting material is completely reacted, the reaction mixture is evaporated to dryness under reduced pressure and purified by high-purity liquid. Product Z-38 (117 mg, 28%). MS (ESI) 399.2 [M+H] +; ¹ H NMR (400 MHz, dmso) δ 8.78 (d, J = 4.5 Hz, 1H), 8.53 (t, J = 5.4 Hz, 1H), 8.18 (d, J=8.0 Hz, 1H), 7.98 (dd, J=8.4, 0.9 Hz, 1H), 7.71 (ddd, J=8.3, 6.9, 1.3 Hz, 1H), 7.63–7.56 (m, 2H), 7.36 (d) , J=4.6Hz, 1H), 7.15 (d, J=4.0Hz, 1H), 3.32 (dd, J=23.6, 9.2Hz, 3H), 1.89 (d, J=10.5Hz, 4H), 1.63–1.15 (m, 7H).

Preparation of Compound Z-39

Dissolving 2-((1s,4s)-4-(quinolin-4-yl)cyclohexyl)ethyl-1-amine (140 mg, 0.55 mmol) and triethylamine ( 223 mg, 2.20 mmol) in dichloro Add methane (12 mL), add 5-chloro-2- acylchlorothiophene ( 119 mg, 0.66 mmol) and stir at room temperature for 1 hour. When the starting material is completely reacted, the reaction mixture is evaporated to dryness under reduced pressure and purified by high-purity liquid. Product Z-39 (51 mg, 23%). MS (ESI) 399.1 [M+H] +; ¹ H NMR (400 MHz, dmso) δ 8.79 (d, J = 4.5 Hz, 1H), 8.54 (t, J = 5.5 Hz, 1H), 8.18 (d, J = 8.1 Hz, 1H), 7.98 (dd, J = 8.4, 0.9 Hz, 1H), 7.71 (ddd, J = 8.3, 6.9, 1.2 Hz, 1H), 7.65 - 7.54 (m, 2H), 7.40 (d) , J = 4.6 Hz, 1H), 7.14 (d, J = 4.0 Hz, 1H), 3.39 (s, 1H), 3.27 - 3.21 (m, 2H), 1.91 - 1.60 (m, 11H).

Test Example 1 Inhibitory activity test of Hela cells

Reagent

Hela cells were from ATCC; DMEM phenol-free erythrocyte medium was from Gibco, product number: 21063-029; INF-γ was from Life Technologies, product number: PHC4031 100ug; fetal bovine serum was from Gibco, product number: 10099-141; 0.25% pancreas Protease from GIBCO, product number: 25200-072; phosphate buffer from Hyclone, product number: SH30256.01B; 6.1N trichloroacetic acid from Sigma, product number: T0699; p-dimethylaminobenzaldehyde (pDMAB) from Sigma, product ID: 15647-7; L-tryptophan from Sigma, product number: T0254-25G; DMSO from Sigma, product number: D5879-1L; 96-well cell culture plate from BD Falcon, product number: 353072.

Reagent preparation

1, 1 gram of pDMAB is dissolved in 50 ml of acetic acid (protected from light, now available);

2. Configure L-tryptophan in PBS to a concentration of 200 ug/ml for use;

3. Configure INF-γ with phenol-free red medium to 250 ng/ml for use;

Experimental procedure

1. On the first day, Hela cells were seeded in a cell culture plate at 4E3 cells per well in a volume of 70 ul per well, and incubated in a cell culture incubator for 24 hours;

2, the next day, each well was added 10ul concentration of 10X test compound, 10ul L-tryptophan and 10ul INF-γ, DMSO reaction concentration of 0.5%, L-tryptophan reaction concentration of 20ug / ml, INF- The γ reaction concentration is 25 ng/ml;

3. The cell culture plate is cultured in a cell culture incubator for 48 hours;

4. On the fourth day, take 70 ul of cell culture supernatant and another reaction plate, add 5 ul of 6.1 N trichloroacetic acid to each well, and react at 50 ° C for 30 minutes;

5. Centrifuge the reaction plate at 2500 rpm for 10 minutes, take 50 ul of supernatant from each well to a new reaction plate; add 50 ul of pDMAB (2%) to shake the shaker gently; read the absorbance at 480 nm with a microplate reader, and calculate with XLfit software. Compound IC50 value. The test results are shown in Table 1.

Table 1 Inhibitory activity of exemplary compounds of the invention on Hela cells

Compound number	Hela/μM	Compound number	Hela/μM
Z-1	0.011	Z-5	0.024
Z-15	0.013	Z-16	0.025
Z-17	0.684	Z-18	11.035
Z-19	0.903	Z-20	0.508
Z-21	0.330	Z-22	0.394
Z-23	0.381	Z-24	0.523
Z-25	0.573	Z-27	0.449
Z-28	0.024	Z-29	0.441
Z-30	0.162	Z-31	0.433
Z-32	0.220	Z-36	0.198
Z-37	0.006	Z-38	0.393
Z-39	0.008	Z-40	0.162
Z-41	0.064	Z-42	0.504

*Compounds Z-27 and Z-28 are a pair of enantiomers isolated from compound z-16, and compounds Z-36 and Z-37 are a pair of enantiomers isolated from compound z-15. Compounds Z-38 and Z-39 are a pair of enantiomers isolated from compound z-1.

As can be seen from Table 1, the compound of the present invention has an excellent inhibitory activity against HeLa cells up to 50 nM or less. In particular, when Z ₁ is N, Z ₂ and Z ₃ are CH, and R ₁ is F, R ₂ , R ₃ and R ₄ are both hydrogen, and A is a thiophene ring, the isotopic configuration of the compound of the present invention is different. The activity of the construct can even reach below 10 nM.

Test Example 2 Inhibitory activity test of HEK293-hIDO1 cells

Reagent

HE293-hIDO1-7 stable cell line was from TGZ0172; DMEM phenol-free erythrocyte medium was from Gibco, product number: 21063-029; fetal bovine serum was from Gibco, product number: 10099-141; 0.25% trypsin from Gibco, product number :25200-072; phosphate buffer from Hyclone, product number: SH30256.01B; 6.1N trichloroacetic acid from Sigma, product number: T0699; p-dimethylaminobenzaldehyde (pDMAB) from Sigma, product number: 15647-7; L-tryptophan was from Sigma, product number: T0254-25G; DMSO was from Sigma, product number: D5879-1L; 96-well cell culture plate was from BD Falcon, product number: 353072.

Reagent preparation

1, 1 gram of pDMAB is dissolved in 50 ml of acetic acid (protected from light, now available);

2. Configure L-tryptophan in PBS to a concentration of 200 ug/ml for use;

Experimental procedure

1. On the first day, 293-hIDO1 cells were planted in 4E4 cells per well in a cell culture plate at a volume of 80 ul per well;

2, 10ul of 10X concentration of test compound and 10ul of L-tryptophan were added to each well, the final concentration of DMSO was 0.5%, and the final concentration of L-tryptophan was 20ug/ml;

3. The cell culture plate is cultured in a cell culture incubator for 48 hours;

4. On the third day, take 70 ul of cell culture supernatant and another reaction plate, add 5 ul of 6.1 N trichloroacetic acid per well, and let react at 50 ° C for 30 minutes;

5. Centrifuge the reaction plate at 2500 rpm for 10 minutes, take 50 ul of supernatant from each well to a new reaction plate; add 50 ul of pDMAB (2%) to shake the shaker gently; read the absorbance at 480 nm with a microplate reader, and calculate with XLfit software. Compound IC50 value. The test results are shown in Table 2.

Table 2 Inhibitory activity of exemplary compounds of the invention on HEK-293 cells

Compound number	HEK-293/μM	Compound number	HEK-293/μM
Z-28	0.021	Z-30	0.034
Z-32	0.265	Z-33	0.052
Z-34	0.062	Z-35	0.046
Z-36	0.114	Z-37	0.004
Z-38	0.300	Z-39	0.004

As can be seen from Tables 1 and 2, the exemplified compounds of the present invention have good inhibitory activities against Hela and HEK-293 cells.

All documents mentioned in the present application are hereby incorporated by reference in their entirety in their entireties in the the the the the the the the In addition, it should be understood that various modifications and changes may be made by those skilled in the art in the form of the appended claims.

Claims (18)

1. A compound of the formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

   

   In the formula,

   A is a 4 to 6 membered saturated monoheterocyclic ring or a 5 to 6 membered monocyclic heteroaryl ring;

   n is 1, 2 or 3;

   Z ₁ is N or CR ₅ ; Z ₂ is N or CR ₆ ; Z ₃ is N or CR ₇ ; Z ₁ , Z ₂ and Z _{3 are} not N at the same time;

   R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ are each independently hydrogen, halogen, C _1-10 alkyl, C _1-10 alkoxy, halo C _1-10 alkane a C _3-10 cycloalkyl group, a halogenated C _1-10 alkoxy group, NR _a0 R _b0 or a -C(O)C _1-10 alkyl group;

   The 4- to 6-membered saturated monoheterocyclic ring or 5- to 6-membered monocyclic heteroaryl ring is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of NR _a0 R _b0 , halogen, Cyano, acetyl, hydroxy, hydroxymethyl, hydroxyethyl, carboxy, C _1-8 alkyl, C _1-8 alkoxy, halo C _1-8 alkyl, C _3-8 cycloalkyl, C _3-8 cycloalkoxy, halo C _1-8 alkoxy, -C(O)C _1-10 alkyl, -C(O)OC _1-10 alkyl, -OC(O)C _{1 -10} alkyl, -CONR _a0 R _b0 ; R _a0 , R _b0 are each independently hydrogen or C _1-8 alkyl.
2. The compound according to claim 1 or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the 4- to 6-membered saturated monocyclic ring is selected from the group consisting of azetidine and oxetane , tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, piperazine, morpholine, thiomorpholine, thiomorpholine-1,1-dioxide or tetrahydropyran.
3. The compound according to claim 1 or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the 4- to 6-membered saturated monocyclic ring is selected from the following structures:

   

   

   The above 4- to 6-membered saturated monoheterocyclic ring is optionally substituted with 1, 2 or 3 substituents selected from the group A1.
4. The compound of claim 1 or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the 5- to 6-membered monocyclic heteroaryl ring is selected from the group consisting of: a thiophene ring, an N-alkylcyclopyrrole ring , furan ring, thiazole ring, imidazole ring, oxazole ring, pyrrole ring, pyrazole ring, triazole ring, 1,2,3-triazole ring, 1,2,4-triazole ring, 1,2,5 - triazole ring, 1,3,4-triazole ring, tetrazole ring, isoxazole ring, oxadiazole ring, 1,2,3-oxadiazole ring, 1,2,4-oxadiazole ring 1,2,5-oxadiazole ring, 1,3,4-oxadiazole ring, thiadiazole ring, pyridine ring, pyridazine ring, pyrimidine ring or pyrazine ring.
5. The compound according to claim 1 or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the 5- to 6-membered monocyclic heteroaryl ring is selected from the following structures:

   

   The above 5- to 6-membered monocyclic heteroaryl ring is optionally substituted with 1, 2 or 3 substituents selected from the group A1.
6. The compound according to claim 1 or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the 5- to 6-membered monocyclic heteroaryl ring is selected from the following structures:

   
7. The compound according to claim 1 or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein n is 1 or 2.
8. The compound according to claim 1 or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein two of Z ₁ , Z ₂ and Z ₃ are not N.
9. The compound according to claim 1 or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein Z ₁ is N; Z ₂ is CR ₆ ; Z ₃ is CR ₇ ; R ₆ , R _{7 are} as defined Requirement 1 is defined.
10. The compound according to claim 1 or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein Z ₁ is CR ₅ ; Z ₂ is N; Z ₃ is CR ₇ ; R ₅ , R _{7 are} as defined Requirement 1 is defined.
11. The compound according to claim 1 or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein Z ₁ is CR ₅ ; Z ₂ is CR ₆ ; Z ₃ is N; and R ₅ and R _{6 are} as defined. Requirement 1 is defined.
12. The compound of claim 1 or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein

    

    For the next group structure:

    
13. The compound according to claim 1, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the compound of the formula (I) is selected from the group A structure.
14. A pharmaceutical composition comprising the compound of any one of claims 1 to 13 or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.
15. The use of a compound according to any one of claims 1 to 13 or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 14, for the preparation of a medicament It is used to inhibit the activity of indoleamine 2,3-dioxygenase or to inhibit immunosuppression in patients.
16. A method of modulating guanamine 2,3-dioxygenase activity comprising administering a therapeutically effective amount of a compound of claim 1, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a method according to claim 14. The pharmaceutical composition is contacted with indoleamine 2,3-dioxygenase. Preferably, the adjustment is preferably an inhibitory effect.
17. A method of inhibiting immunosuppression in a patient, the method comprising administering to a patient a therapeutically effective amount of the compound of claim 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 14. .
18. A method of treating cancer, the method comprising administering to a patient a therapeutically effective amount of a compound of claim 1 or a tautomer, a mesophil, a racemate, an enantiomer, a diastereomer Isomer or a mixture thereof, or a pharmaceutically acceptable salt thereof.