RU2780118C2 - Iminourea derivatives - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to a compound represented by the formula I or its pharmaceutically acceptable salt, which can be used for the treatment of diseases characterized by a pathological path of tryptophan metabolism, mediated by indolamine-2,3-dioxygenase. In the formula I, R₁ is selected from a group consisting of H, C_1-6alkyl, and C_1-6alkyl-C(O)-; n is equal to an integer selected from 0-6. The invention also relates to specific compounds, methods for synthesis of compounds represented by formulas II and II', a pharmaceutical composition for inhibition of indolamine-2,3-dioxygenase, containing the proposed compounds, and a method for treatment of a disease characterized by a pathological path of tryptophan metabolism, mediated by indolamine-2,3-dioxygenase.

EFFECT: obtaining compounds for the treatment of diseases.

13 cl, 11 tbl, 58 ex

Description

The field of technology to which the invention belongs

The present invention relates to the field of medicines, in particular to an iminourea derivative and to a method for its preparation and use.

BACKGROUND OF THE INVENTION

Indolamine-2,3-dioxygenase, a monomeric heme-containing enzyme, is able to catalyze the oxidative opening of the L-tryptophan indole ring to form kynurenine. High expression of indolamine-2,3-dioxygenase leads to the depletion of the local store of tryptophan in cells, which induces arrest of the T-cell cycle in the G1 phase, thereby inhibiting T-cell proliferation. On the other hand, the breakdown of indolamine-2,3-dioxygenase-dependent tryptophan leads to an increase in the level of kynurenine, and also induces T-cell apoptosis mediated by oxygen free radicals. Thirdly, increased expression of indolamine-2,3-dioxygenase in dendritic cells enhances local immunosuppression mediated by regulatory T cells (Treg) through the breakdown of the local store of tryptophan, thereby enhancing the body's peripheral immune tolerance to tumor-specific antigen. Indolamine-2,3-dioxygenase has become the most important small molecular weight regulatory target for antitumor immunotherapy.

In studies, indolamine-2,3-dioxygenase has been found to be associated with many physiological processes in the human body. In 1998 Munn et al. found that the fetus is able to survive in the body of a mother with a different genotype during pregnancy without rejection due to the fact that placental plasmoditrophoblastic cells synthesize indolamine-2,3-dioxygenase, which inhibits the rejection reaction of maternal T-cells in relation to the fetus through the bloodstream. After additional subcutaneous implantation in pregnant mice of a sustained-release capsule containing the indolamine-2,3-dioxygenase inhibitor 1-methyltryptophan, they found that the embryo was rejected and aborted (Munn DH, Zhou M, Attwood JT, et al. Prevention of allogeneic fetal rejection by tryptophan catabolism Science 1998 281 (5380): 1191-3). In addition, some diseases caused by abnormal immune responses, such as transplant rejection and autoimmune diseases, are also closely related to indolamine 2,3-dioxygenase.

Despite the fact that significant progress has been made in the treatment of tumors in recent years, clinical efficacy is still unsatisfactory. Escape from the immune response is one of the main biological mechanisms of oncogenesis and tumor metastasis, and has become an important factor influencing the therapeutic effect in relation to tumors. Indolamine 2,3-dioxygenase, as an immune regulatory enzyme, can effectively inhibit T cell function, enhance Treg cell function, and induce NK cell dysfunction, while tumor cells can use these innate immunoregulatory mechanisms in the body to elude identification. and destruction by the immune system (Jia Yunlong, Wang Yu, Chinese Journal of Cancer Biotherapy, 2004, 21 (6): 693-7). In order to ensure optimal treatment benefit for patients with tumors, it is necessary to rationally adjust the treatment strategy in relation to the escape of tumors from the immune response. The indolamine-2,3-dioxygenase inhibitor of the present invention can effectively regulate the patient's immune system, block the escape of tumor cells from the immune response, and have a good therapeutic effect on most spontaneous tumors. Based on the regulation of the immune system, in addition to treating tumors, the indolamine-2,3-dioxygenase inhibitor of the present invention can also treat other immune-related diseases such as chronic infection and AIDS.

Indolamine 2,3-dioxygenase is also closely associated with neurological diseases. It can lower 5-hydroxytryptamine levels and cause mental illnesses such as depression and anxiety. It can also cause accumulation in the brain of neurotoxic metabolites such as quinolinic acid, which is closely associated with the onset of neurodegenerative diseases such as Alzheimer's disease. Indolamine 2,3-dioxygenase may affect brain function through at least two mechanisms: 1) in an inflammatory response, tryptophan catabolism may lead to a decrease in circulating tryptophan concentrations, thereby leading to a decrease in 5-hydroxytryptamine levels, leading to depression ; 2) indolamine 2,3-dioxygenase catabolizes tryptophan to products that are involved in the kynurenine pathway, leading to accumulation of kynurenine and neurotoxic quinoline acid (Kong Linglei, Kuang Chunxiang, Yang Qing, Chinese Journal of Medicinal Chemistry, 2009, 19(2 ): 147-154).

Content of the present invention

The present invention relates to a compound or a pharmaceutically acceptable salt thereof, a composition containing the compound or a pharmaceutically acceptable salt thereof, and a method for inhibiting indolamine-2,3-dioxygenase (IDO) activity by using the present compound or a pharmaceutically acceptable salt thereof, or a method for treating a disease characterized by an abnormal tryptophan metabolism pathway mediated by indolamine 2,3-dioxygenase by using the compound or a pharmaceutically acceptable salt thereof, and to using the compound or a pharmaceutically acceptable salt for the manufacture of a medicament for inhibiting the activity of indolamine 2,3-dioxygenase or for treatment of a disease characterized by an abnormal tryptophan metabolism pathway mediated by indolamine-2,3-dioxygenase.

The compound, or a pharmaceutically acceptable salt thereof, has excellent indolamine-2,3-dioxygenase inhibitory activity, and this activity is substantially higher than other IDO inhibitors. In addition, by measuring the body weight of mice before and after administration of the compound or a pharmaceutically acceptable salt thereof, it was found that, compared with other IDO inhibitors, the compound of the present invention or a pharmaceutically acceptable salt thereof can significantly improve the quality of life of mice during tumor treatment and significantly reduce side effects. In clinical practice, the compound of the present invention, or a pharmaceutically acceptable salt thereof, will not only improve the patient's quality of life, but also significantly improve the patient's drug compliance and drug efficacy. The compound of the present invention, or a pharmaceutically acceptable salt thereof, can significantly improve learning and memory impairments in animals and enhance knowledge acquisition and spatial memory ability, have a positive therapeutic value for neurodegenerative diseases such as Alzheimer's syndrome, and are superior to other IDO inhibitors. The compound of the present invention or a pharmaceutically acceptable salt thereof can enhance the function of DC to stimulate T cell proliferation, whereby it can be used for the treatment of neoplastic diseases, autoimmune diseases, transplant rejection and infectious diseases, and is superior to other IDO inhibitors.

DETAILED DESCRIPTION OF THE PRESENT INVENTION

In some embodiments, the invention relates to a compound represented by formula I ₀ , or a pharmaceutically acceptable salt thereof:

where each of R ₁ and R ₂ is independently selected from the group consisting of: H, substituted or unsubstituted C _1-10 alkyl, aldehyde group, substituted or unsubstituted carbonyl, cyano, CF ₃ , substituted or unsubstituted C _1-10 alkoxy, substituted or unsubstituted sulfonyl, substituted or unsubstituted C _3-10 cycloalkyl, substituted or unsubstituted C _2-10 alkenyl, substituted or unsubstituted C _6-20 aryl, substituted or unsubstituted C _3-14 heteroaryl;

each of R ₃ and R ₄ independently represents a mono substituent selected from the group consisting of: H, substituted or unsubstituted C _1-10 alkyl, substituted or unsubstituted C _3-10 cycloalkyl, cyano, substituted or unsubstituted C _1-10 alkoxy, substituted or unsubstituted sulfonyl, substituted or unsubstituted C _6-20 aryl, substituted or unsubstituted C _3-14 heteroaryl; or

,

или

, где C представляет собой атом C в a- или b-положении, m равен целому числу, выбранному из 0-6, такому как 0 или 1, или 2, или 3, или 4, или 5, или 6; кроме того, группа, сформированная R₃ и R₄ вместе с атомом C в a- или b-положении, представляет собой C=CH₂,

,

,

,

или

;each of R ₃ and R ₄ is independently selected from disubstituents thereby forming together with the C atom in the a- or b-position the following groups:

,

or

where C is a C atom at the a or b position, m is an integer selected from 0-6 such as 0 or 1 or 2 or 3 or 4 or 5 or 6; in addition, the group formed by R ₃ and R ₄ together with a C atom in the a- or b-position is C=CH ₂ ,

,

,

,

or

;

n is an integer selected from 0-6 such as 0, 1, 2, 3, 4, 5, or 6; preferably, n is 0, 1, 2 or 3.

In some embodiments, the present invention relates to a compound represented by formula I ₀ as described above, or a pharmaceutically acceptable salt thereof, wherein R ₁ and R ₂ are each independently selected from the group consisting of: C _1-6 alkyl, carbonyl, C _{1 -6} alkoxy, sulfonyl, amidino, sulfinyl which is optionally substituted with one or more substituents selected from the group consisting of: halogen, hydroxy, carboxy, carbonyl, aldehyde group, cyano, amino, aryl, heteroaryl, C _1-6 alkyl, C _3-12 cycloalkyl, C _2-6 alkenyl and C _3-12 cycloalkenyl, where carboxy, carbonyl, aldehyde group, cyano, amino, aryl, heteroaryl, C _3-12 cycloalkyl, C _2-6 alkenyl, C _3-12 cycloalkenyl as a substituent C _1-6 alkyl, carbonyl, C _1-6 alkoxy, sulfonyl, amidino or sulfinyl is optionally substituted with one or more substituents selected from the group consisting of: H, halogen, C _1-6 alkyl, carbonyl, C _1-6 alkoxy, sulfinyl and sulfonyl, and halogen is chosen come from the group consisting of F, Cl, Br and I.

In some embodiments, the present invention relates to a compound represented by formula I ₀ as described above, or a pharmaceutically acceptable salt thereof, wherein R ₁ and R ₂ are each independently selected from the group consisting of: H, methyl, ethyl, propyl, isopropyl, R ₅ C(O)-, R ₅ S(O) _x -; R ₅ is selected from the group consisting of C _1-10 alkyl, C _3-12 cycloalkyl, substituted C _1-10 alkyl and substituted C _3-12 cycloalkyl, where substituted C _1-10 alkyl or substituted C _3-12 cycloalkyl is substituted with hydroxy , cyano, C _1-6 alkyl, C _3-12 cycloalkyl, C _1-6 alkoxy, aryl or heteroaryl, where x is 1 or 2;

each of R ₃ and R ₄ independently represents a mono substituent selected from the group consisting of: H, substituted or unsubstituted C _1-10 alkyl, substituted or unsubstituted C _3-10 cycloalkyl, cyano, substituted or unsubstituted C _1-10 alkoxy, substituted or unsubstituted sulfonyl, substituted or unsubstituted C _6-20 aryl, substituted or unsubstituted C _3-14 heteroaryl; or

,

, C=CH₂,

,

,

или

.each of R ₃ and R ₄ is independently selected from disubstituents thereby forming together with the C atom in the a- or b-position the following groups:

,

, C=CH ₂ ,

,

,

or

.

In some embodiments, the present invention relates to a compound represented by formula I ₀ as described above, or a pharmaceutically acceptable salt thereof, wherein R ₁ and R ₂ are each independently selected from the group consisting of: H, methyl, ethyl, propyl, isopropyl, R ₅ C(O)-, R ₅ S(O) _x -; R ₅ is selected from the group consisting of C _1-10 alkyl, C _3-12 cycloalkyl, substituted C _1-6 alkyl and substituted C _3-8 cycloalkyl, wherein substituted C _1-6 alkyl or substituted C _3-8 cycloalkyl is substituted by hydroxy , cyano, C _1-6 alkyl, C _3-12 cycloalkyl, C _1-6 alkoxy, aryl, or heteroaryl; x is 2; R ₃ and R ₄ are both H.

According to some embodiments, the present invention relates to a compound represented by formula I ₀ as described above, or a pharmaceutically acceptable salt thereof, where if R ₂ , R ₃ and R ₄ in formula I ₀ respectively represent H, then the compound is represented by formula I:

,

,

where R ₁ is selected from the group consisting of: H, amino, sulfonyl, nitro, carbonyl, amidino, C _1-6 alkyl, substituted amidino, substituted C _1-6 alkyl, substituted C _1-6 alkoxy, substituted carbonyl, substituted sulfonyl and a substituted sulfinyl, wherein the substituted amidino, substituted C _1-6 alkyl, substituted C _1-6 alkoxy, substituted carbonyl, substituted sulfonyl, or substituted sulfinyl is substituted with halogen, hydroxy, carboxy, carbonyl, aldehyde, cyano, amino, aryl, heteroaryl, C _3-12 cycloalkyl, C _2-6 alkenyl, C _3-12 cycloalkenyl; n is an integer selected from 0-6 such as 0, 1, 2, 3, 4, 5, or 6; preferably, n is 1 or 2.

In some embodiments, the present invention relates to a compound represented by formula I ₀ as described above, or a pharmaceutically acceptable salt thereof, wherein if R ₁ in formula I is H, then the compound is represented by formula II:

,

,

where n is an integer selected from 0-6 such as 0, 1, 2, 3, 4, 5, or 6.

In some embodiments, the present invention relates to a compound represented by formula I ₀ as described above, or a pharmaceutically acceptable salt thereof, wherein if R ₁ in formula I is an R ₀ -substituted carbonyl, then the compound is represented by formula III:

,

,

where R ₀ is selected from the group consisting of H, C _1-6 alkyl, substituted C _1-6 alkyl and substituted C _1-6 alkoxy, where substituted C _1-6 alkyl or substituted C _1-6 alkoxy is substituted by halogen, hydroxy, carboxy, carbonyl, aldehyde group, cyano, amino, aryl, heteroaryl, C _3-12 cycloalkyl, C _2-6 alkenyl, C _3-12 cycloalkenyl, n is an integer selected from 0-6, such as 0, 1, 2, 3, 4, 5 or 6.

In addition, R ₀ is selected from the group consisting of C _1-6 alkyl, substituted C _1-6 alkyl and substituted C _1-6 alkoxy, where substituted C _1-6 alkyl or substituted C _1-6 alkoxy is substituted with halogen, hydroxy, carboxy, carbonyl, aldehyde group, cyano, amino, aryl, heteroaryl, C _3-12 cycloalkyl, C _2-6 alkenyl, C _3-12 cycloalkenyl.

In some embodiments, the present invention relates to a compound represented by formula I ₀ as described above, or a pharmaceutically acceptable salt thereof, wherein if R ₁ in formula I ₀ is H, then the compound is represented by formula IV:

,

,

where R ₂ is selected from the group consisting of H, methyl, ethyl, propyl, isopropyl, R ₅ C(O)-, R ₅ S(O) _m -; R ₅ is selected from the group consisting of H, C _1-10 alkyl, C _3-12 cycloalkyl, substituted C _1-10 alkyl and substituted C _3-12 cycloalkyl, where substituted C _1-10 alkyl and substituted C _3-12 cycloalkyl substituted with hydroxy, cyano, CF ₃ , C _1-6 alkyl, C _3-10 cycloalkyl, alkoxy, aryl, or heteroaryl; m is selected from 1 or 2;

each of R ₃ and R ₄ is independently selected from the group consisting of H, C _1-10 alkyl, C _3-12 cycloalkyl, substituted C _1-10 alkyl, substituted C _3-12 cycloalkyl, substituted C _1-10 alkoxy, substituted sulfonyl and substituted C _3-14 heteroaryl, where substituted C _1-10 alkyl, substituted C _3-12 cycloalkyl, substituted C _1-10 alkoxy, substituted sulfonyl or substituted C _3-14 heteroaryl is substituted with hydroxy, cyano, halogen, C _{1- 6} alkyl, C _3-10 cycloalkyl, alkoxy, aryl or heteroaryl;

where n is an integer selected from 0-6, such as 0, 1, 2, 3, 4, 5, or 6;

in addition, R ₂ is selected from the group consisting of H, R ₅ C(O)-, R ₅ S(O) _m -; R ₅ is selected from the group consisting of H, C _1-10 alkyl, C _3-12 cycloalkyl, substituted C _1-10 alkyl and substituted C _3-12 cycloalkyl, where substituted C _1-10 alkyl or substituted C _3-12 cycloalkyl substituted with hydroxy, cyano, CF ₃ , C _1-6 alkyl, C _3-10 cycloalkyl, C _1-6 alkoxy, aryl, or heteroaryl; m is selected from 1 or 2;

in addition, R ₂ is selected from the group consisting of R ₅ C(O)- and R ₅ S(O) _x -; R ₅ is selected from the group consisting of C _1-6 alkyl, C _3-8 cycloalkyl, substituted C _1-6 alkyl and substituted C _3-8 cycloalkyl, where substituted C _1-6 alkyl or substituted C _3-8 cycloalkyl is substituted with hydroxy , cyano, C _1-6 alkyl, C _3-8 cycloalkyl, C _1-6 alkoxy, aryl, or heteroaryl; x is 2; R ₃ and R ₄ are both H.

In some embodiments, the invention relates to the following compound, or a pharmaceutically acceptable salt thereof:

,

,

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.

Based on the general formula or the synthesis method of the general formula according to the present invention, compounds not limited to these specific compounds can be obtained, and taking into account the methodological principles for obtaining the general formula or the synthesis method of the general formula according to the present invention, all specific compounds that can be obtained specialists in the art without the need for creative effort, fall within the scope of the present invention.

According to some embodiments, the present invention relates to a method for the synthesis of the above-described compound represented by formula I ₀ :

,

,

those. to the general synthetic route I, which is characterized by the following steps:

1) Oxidation of compound 1a to obtain compound 2a:

,

,

wherein the oxidizing agent used above includes, without limitation, at least one of hydrogen peroxide, ozone, peroxyacetic acid;

2) Reaction of a compound represented by formula 1 with a compound represented by formula 2a under alkaline conditions to obtain a compound represented by formula 2:

,

,

wherein the alkali used above includes, without limitation, alkali metal hydroxides, preferably sodium hydroxide, potassium hydroxide, barium hydroxide;

3) Reaction of a compound represented by formula 2 with a compound represented by formula 3a to obtain a compound represented by formula I ₀ :

where each of R ₁ and R ₂ is independently selected from the group consisting of: H, substituted or unsubstituted C _1-10 alkyl, aldehyde group, substituted or unsubstituted carbonyl, cyano, CF ₃ , substituted or unsubstituted C _1-10 alkoxy, substituted or unsubstituted sulfonyl, substituted or unsubstituted C _3-10 cycloalkyl, substituted or unsubstituted C _2-10 alkenyl, substituted or unsubstituted C _6-20 aryl, substituted or unsubstituted C _3-14 heteroaryl;

each of R ₃ and R ₄ independently represents a mono substituent selected from the group consisting of: H, substituted or unsubstituted C _1-10 alkyl, substituted or unsubstituted C _3-10 cycloalkyl, cyano, substituted or unsubstituted C _1-10 alkoxy, substituted or unsubstituted sulfonyl, substituted or unsubstituted C _6-20 aryl, and substituted or unsubstituted C _3-14 heteroaryl;

,

или

, где C представляет собой атом C в a- или b-положении, m равен целому числу, выбранному из 0-6;or each of R ₃ and R ₄ is independently selected from disubstituents thereby forming together with the C atom in the a- or b-position the following groups:

,

or

, where C represents a C atom in the a - or b-position, m is an integer selected from 0-6;

n is an integer selected from 0-6.

According to some embodiments, the present invention relates to a method for synthesizing the above-described compound represented by Formula I:

,

,

those. to the general method for the synthesis of IA, which is characterized by the following steps:

1) Oxidation of compound 1a to obtain compound 2a:

;

;

2) Reaction of the compound represented by formula 1 with compound 2a under alkaline conditions to obtain the compound represented by formula 2:

;

;

3) The interaction of the compound represented by formula 2 with the compound represented by formula 3a to obtain the compound represented by formula I:

,

,

where R ₁ is selected from the group consisting of: H, amino, sulfonyl, nitro, carbonyl, amidino, C _1-6 alkyl, substituted amidino, substituted C _1-6 alkyl, substituted C _1-6 alkoxy, substituted carbonyl, substituted sulfonyl and a substituted sulfinyl, wherein the substituted amidino or substituted C _1-6 alkyl, substituted C _1-6 alkoxy, substituted carbonyl, substituted sulfonyl or substituted sulfinyl is substituted with halogen, hydroxy, carboxy, carbonyl, aldehyde group, cyano, amino, aryl, heteroaryl, C _3-12 cycloalkyl, C _2-6 alkenyl, C _3-12 cycloalkenyl; n is an integer selected from 0-6;

the oxidizing agent used above is preferably, without limitation, at least one of hydrogen peroxide, ozone, or peroxyacetic acid;

the alkali is preferably selected without limitation from alkali metal hydroxides, preferably sodium hydroxide, potassium hydroxide or barium hydroxide.

According to some embodiments, the present invention relates to a method for synthesizing the above-described compound represented by formula II:

,

,

those. to the general synthetic route II, which is characterized by the following steps:

1) Oxidation of the compound represented by formula 2 with compound 3a' to obtain the compound represented by formula IIa:

;

;

2) Deprotection of the compound represented by formula IIa under acidic conditions followed by reaction under basic conditions to obtain the compound represented by formula II:

,

,

where n is 0, 1, 2, 3 or 4,

the alkali is chosen without limitation from alkali metal and alkaline earth metal hydroxides, preferably sodium hydroxide, potassium hydroxide, barium hydroxide.

According to some embodiments, the present invention relates to a method for the synthesis of the above-described compound represented by formula II':

,

,

those. to the general synthetic route III, which is characterized by the following steps:

,

,

n is 0, 1, 2, 3 or 4,

the compound represented by formula IIa reacts under acidic conditions to obtain the compound represented by formula II'.

According to some embodiments, the present invention relates to a method for the synthesis of the above-described compound represented by formula III:

,

,

those. to the general synthesis route IV, which is characterized by the following steps:

1) Reacting a compound represented by formula II with a compound represented by formula 4a to obtain a compound represented by formula IIIa:

;

;

where R ₃ is selected from the group consisting of H, OH, CN, CH _3-m X _m , nitro, C _1-9 alkyl, C _1-9 alkoxy, C _3-9 cycloalkoxy, C _3-12 cycloalkyl, C _{1 -6} heteroalkyl, 3-12-membered heterocycloalkyl, aryl, heteroaryl, substituted C _1-6 alkyl, substituted C _1-9 alkoxy, substituted aryl, substituted heteroaryl and substituted carbonyl, where substituted C _1-6 alkyl, substituted C _{1- 9} alkoxy, substituted aryl, substituted heteroaryl or substituted carbonyl substituted with halogen, hydroxy, carboxy, carbonyl, aldehyde group, cyano, amino, sulfonyl, aryl, heteroaryl, C _3-12 cycloalkyl, C _3-12 cycloalkenyl, m is 1, 2 or 3; preferably m is 2 or 3;

2) Reaction of a compound represented by formula IIIa under alkaline conditions to obtain a compound represented by formula III:

.

.

More specifically, R ₃ is selected from the group consisting of H, OH, CN, CH _3-m X _m , nitro, C _1-9 alkyl, C _3-9 cycloalkoxy, C _3-12 cycloalkyl, C _1-6 heteroalkyl, 3-12-membered heterocycloalkyl, aryl, heteroaryl, substituted C _1-6 alkyl, substituted C _1-9 alkoxy, substituted aryl and substituted heteroaryl, where substituted C _1-6 alkyl, substituted C _1-9 alkoxy, substituted aryl or substituted heteroaryl is substituted with halogen, hydroxy, carboxy, carbonyl, aldehyde group, cyano, amino, sulfonyl, aryl, heteroaryl, C _3-12 cycloalkyl, C _3-12 cycloalkenyl. In addition, R ₃ is selected from the group consisting of H, OH, CN, CF ₃ , CHCl ₂ , CH ₂ Cl, nitro, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl , 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 1,2-dimethylpropyl, 1-ethylpropyl, hexyl, pentylmethyl, pentylethyl, pentylpropyl, pentylbutyl, hexylmethyl, hexylethyl, hexylpropyl , cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropylmethyl, cyclopropylethyl, cyclopropylpropyl, cyclobutylmethyl, cyclobutylethyl, cyclobutylpropyl, cyclopentylmethyl, cyclopentylethyl, cyclopentylpropyl, cyclohexylmethyl, cyclohexylethyl, cyclohexylpropyl, p-methoxybenzyl (PMB), benzyl (Bn); or

R ₃ is any one selected from the group consisting of substituted furan, pyrrole, thiophene, pyrazole, imidazole, oxazole, thiophene, isoxazole, isothiazole, pyridine, pyran, thiopyran, pyridazine, pyridine, pyrazine and piperazine, or is a disubstituent on a benzene ring, i.e. forms benzofuran, benzopyrrole, benzopiperazine together with the benzene ring; or

в формуле 4a выбирают из группы, состоящей из карбазола, акридина, феназина или фенотиазина.group

in formula 4a is selected from the group consisting of carbazole, acridine, phenazine or phenothiazine.

The synthesis method according to the present invention is only one way to obtain each synthesized target compound or its intermediate, in which each step and number, such as 1a, 2a, 3a, 4a, 1, 2, 3, etc., are independent , and their production is not limited to the method according to the present invention.

Unless otherwise indicated, the solvent used in each step of the reactions of the present invention, described above and hereinafter, is a solvent generally accepted in the art, and the principle of selection is that the solvent is capable of dissolving the chemicals, but did not interact, extract the product, or allow the corresponding product to crystallize in solution so as to be separated from impurities. Examples of the solvent include water, halogenated alkanes, alkylamines, aliphatic hydrocarbons, esters, alcohols, aromatic hydrocarbons, ethers, heterocyclic solvents. More specifically, the solvent is selected without limitation from the following: methanol, ethanol, propanol, isopropyl, diethyl ether, ethyl acetate, acetic acid, cyclohexane, dichloromethane, chloroform, tetrahydrofuran, pyridine, diethylamine, triethylamine, dimethylformamide, toluene, and mixtures of at least two of them.

Unless otherwise indicated, in each of the reactions according to the present invention described above and hereinafter, if the reagent is present in excess, then the completion of the reaction can be carried out by adding a substance that interacts with the excess reagent.

Unless otherwise indicated, in each of the reactions according to the present invention described above and hereinafter, the method of purification of the product at each stage of the reactions is selected from the group consisting of extraction, crystallization, solvent removal, column chromatography, and all manipulations in their carrying out are generally accepted techniques in the art, and a person skilled in the art can manipulate them in accordance with certain conditions.

The numerals used in the general formula of the present invention are used to conveniently describe the general formula, and in specific embodiments, they may be modified to other numerals such as 1, 2, 3, etc., for convenience of description, and represent are expressions of the general formula and general reaction equations that do not affect the essence of the structural formula and the reaction equations with its participation.

For the compounds covered by Formulas I-III and their specific compound examples, chiral or cis-trans isomers and mixtures of said compound isomers in any ratio are also within the scope of the compounds covered by Formulas I-III and their specific compound examples.

According to some embodiments, the present invention relates to a pharmaceutical composition containing the compounds described above, i. compounds covered by formulas I-III, and the specific compounds described above, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable adjuvants.

As used herein, the term "pharmaceutically acceptable salt" refers to addition salts formed with a pharmaceutically acceptable acid or alkali, or a solvate thereof. Such pharmaceutically acceptable salts include acid salts, where the acids include hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrobromic acid, hydroiodic acid, formic acid, acetic acid, p-toluenesulfonic acid, sulfinic acid, methanesulfonic acid, benzoic acid, fumaric acid, citric acid, tartaric acid, maleic acid, fatty acid. Non-toxic pharmaceutically acceptable alkali addition salts include alkali salts, and such alkalis include sodium, potassium, calcium, magnesium, aluminum, ammonium alkalis.

In some embodiments, the present invention relates to a compound as described above, or a pharmaceutically acceptable salt thereof, that is capable of inhibiting the activity of indolamine-2,3-dioxygenase and is useful in the manufacture of a medicament for the treatment of a disease characterized by an abnormal indoleamine-mediated metabolism of tryptophan. -2,3-dioxygenase; the drug is used to treat cancer, infectious disease, neurodegenerative disease, depression, anxiety, or age-related cataract;

wherein the cancer is selected from the group consisting of lung cancer, liver cancer, colon cancer, pancreatic cancer, breast cancer, prostate cancer, brain cancer, ovarian cancer, cervical cancer , testicular cancer, kidney cancer, head and neck cancer, lymphoma, melanoma, or leukemia;

the neurodegenerative disease refers to Alzheimer's disease;

infectious disease refers to an infection caused by a bacterium, fungus, virus, or parasite.

In some embodiments, the present invention relates to a method for inhibiting indolamine 2,3-dioxygenase activity by using a compound described above or a pharmaceutically acceptable salt thereof, or for treating a disease characterized by an abnormal tryptophan metabolism pathway mediated by indolamine 2,3-dioxygenase, wherein the method includes administering a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof. The disease is selected from the group consisting of cancer, infectious disease, neurodegenerative disease, depression, anxiety, or age-related cataract; wherein the cancer is selected from the group consisting of lung cancer, liver cancer, colon cancer, pancreatic cancer, breast cancer, prostate cancer, brain cancer, ovarian cancer, cervical cancer , testicular cancer, kidney cancer, head and neck cancer, lymphoma, melanoma, or leukemia; the neurodegenerative disease refers to Alzheimer's disease; infectious disease refers to an infection caused by a bacterium, fungus, virus, or parasite.

According to the examples according to the present invention, the results of the activity test demonstrate that the compounds obtained by the present invention have an excellent inhibitory activity against indolamine 2,3-dioxygenase, and this activity is significantly higher than that of the compound INCB024360. The results of in vivo tests demonstrate that the compounds of the present invention have a high degree of inhibition against tumors, and the therapeutic effect against tumors is significantly better than that of the INCB024360 compound and other IDO inhibitors. In addition, by measuring the body weight of mice before and after administration, it was found that, compared with other IDO inhibitors, the indolamine-2,3-dioxygenase inhibitors of the present invention can significantly reduce side effects during tumor treatment, significantly improve the quality of life of mice. , and in clinical practice, not only improve the patient's quality of life, but also significantly improve the patient's drug compliance and drug efficacy.

The compounds of the present invention can significantly improve learning and memory impairments, enhance the ability to acquire knowledge and the ability to spatial memory, and have a positive therapeutic value in neurodegenerative diseases such as Alzheimer's syndrome, and their effect is superior to other IDO inhibitors.

In the T cell proliferation reaction experiment, it was found that the compound of the present invention can enhance the function of DC to promote T cell proliferation, whereby it can be used for the treatment of tumor diseases, autoimmune diseases, transplant rejection and infectious diseases, and certainly superior to other IDO inhibitors.

When the indolamine 2,3-dioxygenase inhibitor of the present invention is used for the manufacture of a medicament for the treatment of a disease characterized by an abnormal tryptophan metabolism pathway mediated by indolamine 2,3-dioxygenase, it exhibits the following technical advantages:

(1) The antitumor effect is remarkable. The compound of the present invention has a significant inhibitory activity against indolamine 2,3-dioxygenase, and in vivo test shows that the degree of tumor inhibition of the compound of the present invention is significantly higher than that of cyclophosphamide as a positive control drug and compound INCB024360.

(2) The side effect is reduced. The compound of the present invention is an indolamine 2,3-dioxygenase inhibitor that reverses the inhibition of T cell proliferation and regulates the immune function of the body by inhibiting the activity of indolamine 2,3-dioxygenase, thereby performing the functions of monitoring and eliminating tumor cells by the immune system. person. Based on such a special mechanism of action, this compound does not negatively affect the growth of normal cells in the human body, while inhibiting the growth of tumor cells, thereby significantly reducing side effects. In addition, the compound has a significant therapeutic effect against autoimmune diseases, transplant rejection and infectious diseases associated with T cell proliferation.

(3) The treatment of Alzheimer's disease and other neurodegenerative diseases is significantly effective, learning and memory impairments in animals can be significantly improved, and learning ability and spatial memory ability can be significantly enhanced.

Specific models for carrying out the invention

The present invention is described in more detail below with respect to specific embodiments, but the present invention is not limited to them. In addition, taking into account the methodological principles for obtaining the general formula or the synthesis method of the general formula (general synthesis method I, general synthesis method IA, general synthesis method II, general synthesis method III, general synthesis method IV) and specific embodiments of the present invention, all specific compounds obtained by those skilled in the art without the need for creative effort fall within the scope of the present invention.

Example 1

Reaction 1

Compound 1 (903 mg, 10 mmol) was dissolved in acetone (10 ml), then potassium carbonate (2.76 g, 20 mmol) was added at room temperature, stirred at room temperature for 0.5 h, benzenesulfonyl chloride was added dropwise, stirred at room temperature overnight, quenched by adding water, the resulting substance was extracted with ethyl acetate, dried over anhydrous sodium sulfate and concentrated to obtain 900 mg of a white powder.

Reaction 2

Compound 2 (230 mg, 1 mmol) was dissolved in acetonitrile (5 ml), stirred at room temperature for 0.5 h, then compound 2a (320 mg, 2 mmol) was added, heated at 60°C for 24 h, the resulting material was immediately evaporated to dryness under reduced pressure and purified by preparative HPLC to give 210 mg of the expected product.

Reaction 3

Compound 3 (171 mg, 0.5 mmol) was dissolved in dichloromethane (10 ml), stirred at room temperature for 5 min, trifluoroacetic acid (5 ml) was added, reacted at room temperature for 2 h, and then the resulting substance was immediately evaporated to dryness under reduced pressure to give the desired product 4, which was immediately used as the crude product in the next step.

Reaction 4

Compound 4 (120 mg, 0.5 mmol) was dissolved in tetrahydrofuran, compound 4a (120 mg, 0.5 mmol) was added, the mixture was stirred at room temperature for 10 min, 1N sodium hydroxide (1 ml) was added dropwise, and the mixture was stirred at room temperature for 10 minutes. The reaction solution was concentrated and purified by preparative HPLC to obtain 5 mg of the desired product.

¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.45 (s, 1H), 8.89 (s, 1H), 7.76-7.77 (m, 2H ), 7.48-7.52 (m, 3H), 7.10-7.20 (m, 3H), 6.75-6.77 (m, 2H), 6.27 (s, 1H), 3.73-3.77 (m, 2H), 2.52-2.69 (m, 2H).

Purity according to HPLC: @214 nm 99.5%, @254 nm 100%.

LC-MS: m/z 541 [M+1].

Based on the general synthesis method I and Example 1, the following compounds were synthesized.

Example No. Deputy Substance No. MS Example 2

, n равен 1, R₂, R₃, R₄ представляют собой H.R ₁ is

, n is 1, R ₂ , R ₃ , R ₄ are H. XSD3-059 574.0 Example 3 R ₁ is

, n is 1, R ₂ , R ₃ , R ₄ are H. XSD3-060 558.0 Example 4 R ₁ is

, n is 1, R ₂ , R ₃ , R ₄ are H. XSD3-061 546.1 Example 5 R ₁ is

, n is 1, R ₂ , R ₃ , R ₄ are H. XSD3-062 478.0 Example 6 R ₁ is

(methylcarbonyl), n is 2, R ₂ , R ₃ , R ₄ are H. 3050 456.1 Example 7 R ₁ is

, n is 1, R ₂ , R ₃ , R ₄ are H. 3051 600.2 Example 8 R ₁ is ethyl carbonyl, n is 1, R ₂ , R ₃ , R ₄ is H. XSD3-052 456.1 Example 9 R ₁ is

(methylcarbonyl), n is 1, R ₂ , R ₃ , R ₄ are H. XSD3-056 442.1

Example 10

Reaction 1

Compound 1a (682 mg, 2 mmol) was dissolved in trifluoroacetic acid (13 ml), then 30% H ₂ O ₂ was added dropwise at room temperature, the mixture was reacted overnight at 50° C., and the reaction solution changed from cloudy to transparent yellow. After completion of the reaction, the reaction mixture was quenched by adding saturated sodium sulfate solution. After the KI-starch indicator paper turned colorless, the resulting substance was extracted with ethyl acetate (50 ml × 2), and the organic phase was dried over anhydrous sodium sulfate, concentrated, and then subjected to purification by column chromatography (petroleum ether:ethyl acetate = 1:1 ) to give a pale yellow solid compound 2a (500 mg, 67% yield).

Reaction 2

To a solution of compound 2a (170 mg, 0.5 mmol) in tetrahydrofuran (10 ml) was added ethylenediamine 1 (30 mg, 0.5 mmol), then 1N NaOH (0.4 ml) was added, the reaction solution was stirred at room temperature in for 0.5 h, and immediately the reaction solution was used to obtain compound 2 (120 mg).

Reaction 3

A mixed solution of compound 2 (120 mg, 0.33 mmol) and compound 3a (100 mg, 0.33 mmol) in methanol (10 ml) was stirred at room temperature overnight, the reaction solution was concentrated and purified by column chromatography (petroleum ether:ethyl acetate = 1:1) to give compound 3 (42 mg, 23%).

Reaction 4

To a solution of compound 3 (31 mg, 0.05 mmol) in dichloromethane (3 ml) was added trifluoroacetic acid (0.6 ml) and stirred at room temperature overnight. The reaction solution was concentrated and purified by preparative HPLC to give compound XSD3-047 (9 mg, 68% yield). For target compound: ¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.4 (s, 1H), 8.88 (s, 1H), 7.22-7.11 (m, 2H), 6.71-6.5 (m, 3H), 6.38-6.21 (m, 2H), 3.37 (m, 3H), 3.01 (m, 2H). MS: m/z 401.2 [M+1].

Example 11

Reaction 1

To a solution of compound 1 (170 mg, 0.5 mmol, the method of synthesis of which is described for the synthesis of compound 2a in the summary report for XSD3-047) in tetrahydrofuran (10 ml) was added 1,3-diaminopropane (35 mg, 0.5 mmol) , the reaction solution was stirred at room temperature for 0.5 h, and the reaction solution was immediately used to obtain 130 mg of the desired product.

Reaction 2

A mixed solution of compound 2 (130 mg, 0.33 mmol) and compound 1a (100 mg, 0.33 mmol) in methanol (10 ml) was stirred at room temperature overnight, the reaction solution was concentrated and purified by column chromatography (petroleum ether:ethyl acetate = 1:1) to give compound 3 (78 mg, 45%).

Reaction 3

To a solution of compound 3 (31 mg, 0.05 mmol) in dichloromethane (3 ml) was added trifluoroacetic acid (0.6 ml), stirred at room temperature overnight, then adjusted to pH=12 by adding 1N NaOH solution, stirred continuously for 20 min, and followed the development of the reaction by LCMS. After completion of the reaction, the reaction solution was adjusted to neutral by adding 0.5 N HCl, the reaction solution was concentrated and purified by preparative HPLC to obtain the desired product (9 mg, 68% yield). For desired product: ¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.4 (s, 0.6 H), 8.88 (s, 1H), 7.6 ( s, 1H), 7.4-6.7 (m, 6H), 6.7 (s, 1H), 6.24 (s, 1H), 3.19-3.14 (m, 4H), 1 .75 (m, 2H). MS: m/z 415.2 [M+1].

Example 12

Reaction 1

A mixed solution of compound 1 (384 mg, 1 mmol, the method of which is described in the preparation example of compound 3047) and compound 1a (100 mg, 1.1 mmol) in tetrahydrofuran (10 ml) was stirred at room temperature for 48 h, and the reaction the solution was immediately used to obtain the target product 2 (89 mg, 20% yield).

Reaction 2

Compound 2 (89 mg, 0.2 mmol) was dissolved in THF (10 ml), then adjusted to pH=12 with 1N NaOH, and stirred continuously for 20 minutes. The progress of the reaction was monitored by LCMS. After completion of the reaction, the reaction solution was adjusted to neutral by adding 0.5 N HCl, the reaction solution was concentrated and purified by preparative HPLC to obtain the desired product (13 mg, 15% yield). For target compound: ¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.47 (s, 0.6 H), 8.92 (s, 1H), 7.50- 7.09 (m, 5H), 6.70 (s, 1H), 6.28 (s, 1H), 2.72-2.50 (m, 4H), 2.50 (m, 3H). MS: m/z 416.2 [M+1].

Example 13

Reaction 1

Compound 1 (102 mg, 0.016 mmol, synthetic method described in the summary report for XSD3-047) was dissolved in THF (10 ml), then 1N NaOH solution (0.1 ml) was added at room temperature, stirred at room temperature for 0.2 h, and monitored the progress of the reaction by LCMS. After completion of the reaction, the pH was adjusted to neutral values by adding 0.5 N HCl solution, to obtain 52 mg of the target compound. For target compound: ¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.45 (s, 1H), 8.89 (s, 1H), 8.24 (m, 1H ), 7.76-7.77 (m, 2H), 7.14-7.20 (m, 2H), 7.18 (s, 1H), 6.31 (s, 1H), 3.33- 3.67 (m, 4H), 1.44-1.48 (m, 18H). MS: m/z 601.2 [M+1].

Example 14

Compound 1 (52 mg, 0.1 mmol, synthetic method described in the summary report for XSD3-048) was dissolved in THF (10 ml), then TFA (0.5 ml) was added at room temperature, stirred at room temperature for 12 h, and followed the development of the reaction by LCMS. After purification by preparative HPLC, 15 mg of the title compound was obtained. For target compound: ¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.44 (s, 1H), 10.85 (s, 1H), 8.91 (s, 1H ), 8.49 (s, 1H), 7.18-7.09 (m, 3H), 6.78-6.75 (m, 1H), 6.28-6.25 (m, 1H), 3.33-3.19 (m, 4H), 1.85-1.78 (m, 2H), 1.5 (s, 3H). Characterization result by LC-MS: m/z 515 [M+1].

Example 15

Reaction 1

Compound 1 (100 mg, 0.23 mmol) was dissolved in acetone (10 ml), then potassium carbonate (0.276 g, 2.0 mmol) was added, stirred at room temperature for 0.5 h, benzenesulfonyl chloride was added dropwise, stirred at room temperature overnight. The reaction mixture was quenched by adding water, the resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate and concentrated to give a crude product which was immediately used in the next step.

Reaction 2

Compound 2 (120 mg, 0.5 mmol) was dissolved in tetrahydrofuran/water, 1N sodium hydroxide (1 ml) was added dropwise, and stirred at room temperature for 10 minutes. The reaction solution was concentrated and purified by preparative HPLC to obtain 20 mg of the desired product.

¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.45 (s, 1H), 8.90 (s, 1H), 7.75-7.77 (m, 2H ), 7.46-7.52 (m, 3H), 7.12-7.20 (m, 1H), 7.10-7.11 (m, 1H), 6.75-6.78 (m , 2H), 6.19 (s, 1H), 4.25 (m, 2H), 3.11-3.19 (m, 4H), 1.67-1.70 (m, 2H).

Purity according to HPLC: @214 nm 99.2%, @254 nm 99.3%.

LC-MS: m/z 555 [M+1].

Example 16

Reaction 1

Compound 1 (106 mg, 0.25 mmol) was dissolved in acetone (10 ml), potassium carbonate (138 mg, 1 mmol) was added, compound 1a (49 mg, 0.25 mmol) was added dropwise, stirred at room temperature in overnight, quenched by adding water, the resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate and concentrated to obtain 100 mg of the crude product.

Reaction 2

Compound 2 (100 mg) was dissolved in methanol, stirred at room temperature for 10 min, 1N sodium hydroxide (1 ml) was added dropwise, stirred at room temperature for 10 min. The reaction solution was concentrated and purified by preparative HPLC to obtain 12 mg of the desired product.

¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.45 (s, 1H), 8.90 (s, 1H), 7.10-7.21 (m, 2H ), 6.76-7.17(m, 4H), 6.28(m, 1H), 3.33-3.40(m, 4H), 2.76(m, 2H), 0.98- 1.86 (m, 11H).

Purity according to HPLC: @214 nm 93.7%, @254 nm 97.6%.

LC-MS: m/z 563 [M+1].

Example 17

Reaction 1

Compound 1 (110 mg, 0.25 mmol) was dissolved in acetone (10 ml), potassium carbonate (138 mg, 1 mmol) was added, compound 1a (47.5 mg, 0.25 mmol) was added dropwise, stirred at room temperature temperature overnight, quenched by adding water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated to give 100 mg of a crude product.

Reaction 2

Compound 2 (100 mg) was dissolved in methanol, stirred at room temperature for 10 min, 1N sodium hydroxide (1 ml) was added dropwise, stirred at room temperature for 10 min, the reaction solution was concentrated and purified by preparative HPLC to obtain 15 mg of the target product.

¹ H-NMR (400 MHz, CDCl ₃ ): δ (ppm): 9.96 (s, 1H), 7.77-7.79 (m, 2H), 6.18-7.32 ( m, 9H), 3.31-3.42 (m, 4H), 2.40 (s, 3H), 1.76 (m, 2H).

Purity according to HPLC: @214 nm 98.6%, @254 nm 98.9%.

LC-MS: m/z 571 [M+1].

Example 18

Reaction 1

Compound 1 (96 mg, 10 mmol) and compound 1a (48 mg, 0.25 mmol) were dissolved in acetonitrile (10 ml), stirred at room temperature overnight, quenched by adding water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 100 mg of a crude product.

Reaction 2

Compound 2 (100 mg) was dissolved in methanol, stirred at room temperature for 10 min, 1N sodium hydroxide (1 ml) was added dropwise, stirred at room temperature for 10 min. The reaction solution was concentrated and purified by preparative HPLC to obtain 15 mg of the desired product.

¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.76 (s, 1H), 11.45 (s, 1H), 8.68-9.04 (m, 3H ), 7.09-7.20 (m, 2H), 6.73-6.77 (m, 1H), 6.33-6.36 (m, 1H), 3.53-3.54 (m , 2H), 3.44-3.46(m, 2H), 1.18(m, 1H), 1.00-1.02(m, 2H), 0.90-0.92(m, 2H ).

Purity according to HPLC: @214 nm 98.8%, @254 nm 99.8%.

LC-MS: m/z 471 [M+1].

Example 19

Reaction 1

Compound 1 (96 mg, 0.25 mmol) and compound 1a (48 mg, 0.25 mmol) were dissolved in acetonitrile (10 ml), stirred at room temperature overnight, quenched by adding water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 100 mg of a crude product.

Reaction 2

Compound 2 (100 mg) was dissolved in methanol, stirred at room temperature for 10 min, 1N sodium hydroxide (1 ml) was added dropwise, stirred at room temperature for 10 min. The reaction solution was concentrated and purified by preparative HPLC to obtain 12 mg of the desired product.

¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.48 (s, 1H), 11.28 (s, 1H), 8.75-8.92 (m, 3H ), 7.09-7.20 (m, 2H), 6.73-6.77 (m, 1H), 6.35-6.38 (m, 1H), 3.53-3.54 (m , 2H), 3.44-3.46 (m, 2H), 3.24-3.28 (m, 2H), 2.13-2.20 (m, 3H), 1.77-1.98 (m, 2H).

Purity according to HPLC: @214 nm 97.9%, @254 nm 98.6%.

LC-MS: m/z 485 [M+1].

Example 20

Reaction 1

Compound 1 (500 mg, 5.5 mmol) was dissolved in 3 ml of dichloromethane, O-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (128 mg, 0.336 mmol), cyclohexanecarboxylic acid (800 mg, 6.25 mmol) and diisopropylethylamine (1.16 g, 896 mmol) in that order, reacted at room temperature overnight under a protective layer of nitrogen, quenched by adding water, the resulting substance was extracted with ethyl acetate (50 ml × 5), the liquid extracts were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure to give 40 mg of product, which was immediately used in the next step.

Reaction 2

Compound 2 (530 mg, 2.65 mmol) was dissolved in acetonitrile (5 ml), compound 2a (850 mg, 5.3 mmol) was added, stirred at room temperature for 24 h and immediately evaporated to dryness under reduced pressure to give 300 mg of the product, which was immediately used in the next step.

Reaction 3

Compound 3 (300 mg, 0.99 mmol) was dissolved in dichloromethane (10 ml), stirred at room temperature for 5 min, trifluoroacetic acid (5 ml) was added, reacted at room temperature for 2 h, and the resulting to dryness under reduced pressure to give 200 mg of expected product 4, which was a crude product and was used immediately in the next step.

Reaction 4

Compound 4 (200 mg, 0.99 mmol) was dissolved in tetrahydrofuran, compound 4a (250 mg, 1.0 mmol) was added, stirred at room temperature for 10 min, 1N sodium hydroxide (1 ml) was added dropwise, stirred at room temperature for 10 min. The reaction solution was concentrated and purified by preparative HPLC to obtain 3 mg of the desired product.

¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.90 (s, 1H), 11.55 (s, 1H), 8.67-9.12 (m, 3H ), 7.10-7.20 (m, 2H), 6.73-6.77 (m, 1H), 6.37 (s, 1H), 3.33-3.54 (m, 4H), 2.16-2.21 (m, 1H), 1.56-1.81 (m, 6H), 1.15-1.36 (m, 1H).

Purity according to HPLC: @214 nm 97.7%, @254 nm 98.0%.

LC-MS: m/z 511 [M+1].

Example 21

Reaction 1:

Compound 1 (35 mg, 0.2 mmol) and compound 2 (90 mg, 0.22 mmol) were dissolved in 10 ml of acetonitrile and stirred at 90°C for 16 hours. The reaction solution was immediately evaporated to dryness under reduced pressure to obtain compound 3 which was a crude product and which was used immediately in the next step (100 mg).

Reaction 2:

Compound 3 (100 mg, 0.1 mmol) was dissolved in 1.5N NaOH (2 ml) in THF (5 ml), stirred at room temperature for 0.5 hour, and purified by preparative HPLC to give target compound (30 mg, yield: 30%).

¹ H-NMR (DMSO, 400M): 11.40 (s, 1H), 11.33 (s, 1H), 9.09 (s, 1H), 8.93 (s, 1H), 8..7 (s, 1H), 7.09-7.20 (m, 2H), 6.73-6.77 (m, 1H), 6.36 (s, 1H), 3.45-3.55 (m , 4H), 2.31-2.33 (m, 2H), 0.95-0.99 (m, 1H), 0.47-0.51 (m, 2H), 0.16-0.20 (m, 2H).

Purity according to HPLC: @214 nm 98.8%, @254 nm 98.9%.

LC-MS: m/z 483.1 [M+1].

Example 22

Reaction 1:

Compound 1 (44 mg, 0.2 mmol) and compound 2 (90 mg, 0.22 mmol) were dissolved in 10 ml of acetonitrile and stirred at 90°C for 16 hours. The reaction solution was immediately evaporated to dryness under reduced pressure to obtain compound 3 which was a crude product and which was used immediately in the next step (100 mg).

Reaction 2:

Compound 3 (100 mg, 0.1 mmol) was dissolved in 1N NaOH (4 ml) in tetrahydrofuran (5 ml), stirred at room temperature for 1 hour, and purified by preparative HPLC to give the title compound (35 mg, yield : 35%).

¹ H-NMR (DMSO, 400M): 11.40 (s, 1H), 11.33 (s, 1H), 9.18 (s, 1H), 9.02 (s, 1H), 8.78 ( s, 1H, 7.10-7.17(m, 2H), 6.73-6.76(m, 1H), 6.35-6.38(m, 1H), 3.46-3.65 (m, 4H), 2.26-2.38 (m, 2H), 1.47-1.69 (m, 5H), 0.80-1.22 (m, 6H),

Purity according to HPLC: @214 nm 96.5%, @254 nm 96.5%.

LC-MS: m/z 527.2 [M+1].

Example 23

Reaction 1:

Compound 1 (35 mg, 0.26 mmol) and compound 2 (90 mg, 0.22 mmol) were dissolved in 10 ml of acetonitrile and stirred at 90°C for 20 hours. The reaction solution was immediately evaporated to dryness under reduced pressure to obtain compound 4, which was a crude product and which was used immediately in the next step (110 mg).

Reaction 2:

Compound 3 (100 mg, 0.2 mmol) was dissolved in 1N NaOH (1.5 mL) in tetrahydrofuran (5 mL), stirred at room temperature for 0.5 hour, and purified by preparative HPLC to give target compound (15 mg, yield: 15%).

¹ H-NMR (DMSO, 400M): 12.50 (s, 1H), 11.50 (broad, 1H), 9.19 (s, 1H), 8.65-8.91 (m, 2H) , 7.12-7.21(m, 2H), 6.73-6.77(m, 1H), 6.36(s, 1H), 3.24-3.36(m, 4H), 1 .77-1.84 (m, 3H), 0.90-1.02 (m, 4H).

Purity according to HPLC: @214 nm 98.6%, @254 nm 98.3%.

LC-MS: m/z 481.1 [M+1].

Example 24

Reaction 1:

Compound 1 (42 mg, 0.2 mmol) and compound 2 (90 mg, 0.22 mmol) were dissolved in 10 ml of acetonitrile and stirred at 90°C for 20 hours. The reaction solution was immediately evaporated to dryness under reduced pressure to obtain compound 4, which was a crude product and which was used immediately in the next step (120 mg).

Reaction 2:

Compound 3 (110 mg, 0.2 mmol) was dissolved in 1N NaOH (5 mL) in tetrahydrofuran (5 mL), stirred at room temperature for 1 hour, and purified by preparative HPLC to give the title compound (35 mg, yield: 35%).

¹ H-NMR (DMSO, 400M): 11.44 (s, 1H), 11.35 (s, 1H), 9.06 (s, 1H), 8.92 (s, 1H), 8.64 ( s, 3H), 7.16-7.21 (t, 1H), 7.09-7.11 (q, 1H), 6.74-6.78 (m, 1H), 6.26-6, 29 (m, 1H), 3.23-3.34 (m, 4H), 2.49-2.50 (m, 1H), 1.61-1.81 (m, 7H), 1.17- 1.32 (m, 5H).

Purity according to HPLC: @214 nm 99.4%, @254 nm 99.8%.

LC-MS: m/z 527.2 [M+1].

Example 25

Reaction 1

When run simultaneously, in two vials, compound 1 (500 mg, 8.77 mmol) was dissolved in DCM, compound 1a (3.3 g, 21.93 mmol), HATU (4.01 g, 5.52 mmol) was added and DIEA (3.71 g, 56.5 mmol), stirred at room temperature overnight, quenched with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate and purified by column chromatography to give 128 mg of product.

Reaction 2

Compound 2 (128 mg, 0.743 mmol) was dissolved in ACN, compound 2a (300 mg, 0.752 mmol) was added, stirred at 90° C. overnight. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 300 mg of a crude product.

Reaction 3

Compound 3 (300 mg) was dissolved in THF/H ₂ O, stirred at room temperature for 10 min, 1N sodium hydroxide (1 ml) was added dropwise, and stirred at room temperature for 10 min. After hydrolysis was completed according to LCMS, 1M HCl was added dropwise until neutral, the reaction solution was extracted with ethyl acetate, the extract was concentrated and purified by preparative HPLC (acidic method) to obtain 25 mg of the desired product.

¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.341 (s, 2H), 8.649-8.923 (m, 4H), 6.274-7.188 (m, 4H), 3.255-3.324 ( m, 4H), 1.694-2.067 (m, 9H).

Purity according to HPLC: @214 nm 99.05%, @254 nm 98.1%.

LC-MS: m/z 497[M+1].

Example 26

Reaction 1:

Compound 1 (1.0 g, 10 mmol) and compound 2 (2.7 mg, 30 mmol) were dissolved in 20 ml DMF and stirred at room temperature, then HATU (3.8 g, 10 mmol), DIEA (6 .5 g, 50 mmol) and stirred for 10 h. The reaction solution was added to 60 ml of water, extracted with ethyl acetate, the extract was evaporated to dryness under reduced pressure and subjected to purification by column chromatography to obtain compound 3 (0.4 g, yield : 24%).

Reaction 2:

Compound 3 (35 mg, 0.2 mmol) and compound 4 (90 mg, 0.22 mmol) were dissolved in 10 ml of acetonitrile and stirred at 90°C for 20 h, the reaction solution was immediately evaporated to dryness under reduced pressure to obtain compound 5 which was a crude product and which was used immediately in the next step (120 mg).

Reaction 3:

Compound 3 (110 mg, 0.2 mmol) was dissolved in 1N NaOH (4 mL) in tetrahydrofuran (5 mL), stirred at room temperature for 1 hour, and purified by preparative HPLC to give the title compound (15 mg, yield: 15%).

¹ H-NMR (DMSO, 400M): 11.43 (s, 1H), 11.27 (s, 1H), 9.06 (s, 1H), 8.92 (s, 1H), 8.64 ( s, 3H), 7.16-7.21 (t, 1H), 7.09-7.11 (q, 1H), 6.74-6.78 (m, 1H), 6.26-6, 29 (m, 1H), 3.23-3.35 (m, 4H), 2.32-2.34 (m, 2H), 1.80-1.84 (m, 2H), 0.95- 1.01 (m, 1H), 0.49-0.51 (m, 5H), 0.18-0.19 (m, 2H).

Purity according to HPLC: @214 nm 96.5%, @254 nm 97.9%.

LC-MS: m/z 497.2 [M+1].

Example 27

Reaction 1:

Compound 1 (45 mg, 0.2 mmol) and compound 2 (90 mg, 0.22 mmol) were dissolved in 10 ml of acetonitrile and stirred at 90°C for 20 h, the reaction solution was immediately evaporated to dryness under reduced pressure to obtain compound 4, which was a crude product and was used immediately in the next step (120 mg, crude, >99% yield).

Reaction 2:

Compound 3 (110 mg, 0.2 mmol) was dissolved in 1N NaOH (4 ml) in tetrahydrofuran (5 ml) and stirred at room temperature for 1 hour, the resulting substance was purified by preparative HPLC to obtain the target compound (55 mg , yield = 50%).

¹ H-NMR (DMSO, 400M): 11.53 (s, 1H), 11.44 (s, 1H), 9.13 (s, 1H), 8.92 (s, 1H), 8.67 ( s, 1H), 7.16-7.21 (t, 1H), 7.09-7.11 (q, 1H), 6.74-6.78 (m, 1H), 6.26-6, 29 (m, 1H), 3.23-3.34 (m, 4H), 2.27-2.28 (m, 2H), 1.66-1.84 (m, 8H), 0.91- 1.17 (m, 5H).

LC-MS: m/z 583.2 [M+1].

Example 28

Reaction 1

Compound 1 (106 mg, 0.25 mmol) was dissolved in acetone (10 ml), potassium carbonate (138 mg, 1 mmol) was added, compound 1a (43.5 mg, 0.25 mmol) was added dropwise, stirred at room temperature temperature overnight, quenched by adding water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 100 mg of a crude product.

Reaction 2

Compound 2 (100 mg) was dissolved in methanol, stirred at room temperature for 10 min, 1N sodium hydroxide (1 ml) was added dropwise, stirred at room temperature for 10 min, the reaction solution was concentrated and purified by preparative HPLC to obtain 9 mg of the target product.

¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.48 (s, 1H), 11.44 (s, 1H), 8.92-8.95 (m, 3H ), 7.27-7.34 (m, 5H), 7.09-7.17 (m, 2H), 6.74-6.76 (m, 1H), 6.32-6.35 (m , 1H), 3.75-3.77 (m, 2H), 3.50-3.56 (m, 2H), 3.44-3.46 (m, 2H).

Purity according to HPLC: @214 nm 97.9%, @254 nm 99.2%.

LC-MS: m/z 521 [M+1].

Example 29

Reaction 1

Compound 1 (110 mg, 0.25 mmol) was dissolved in acetone (10 ml), potassium carbonate (138 mg, 1 mmol) was added, compound 1a (43.5 mg, 0.25 mmol) was added dropwise, stirred at room temperature temperature overnight, quenched by adding water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 100 mg of a crude product.

Reaction 2

Compound 2 (100 mg) was dissolved in methanol, stirred at room temperature for 10 min, 1N sodium hydroxide (1 ml) was added dropwise, stirred at room temperature for 10 min, the reaction solution was concentrated and purified by preparative HPLC to obtain 9 mg of the target product.

¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.64 (s, 1H), 11.42 (s, 1H), 9.03-9.07 (m, 3H ), 7.27-7.34 (m, 5H), 7.09-7.17 (m, 2H), 6.74-6.76 (m, 1H), 6.32-6.35 (m , 1H), 3.75-3.77 (m, 2H), 3.14-3.34 (m, 4H), 1.80-1.84 (m, 2H).

Purity according to HPLC: @214 nm 96.0%, @254 nm 96.0%.

LC-MS: m/z 535 [M+1].

Example 30

Reaction 1

Compound 1 (330 mg, 0.75 mmol) was dissolved in acetone (30 ml), potassium carbonate (414 mg, 3 mmol) was added, compound 1a (130.5 mg, 0.75 mmol) was added dropwise, stirred at room temperature temperature overnight, quenched by adding water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 300 mg of a crude product.

Reaction 2

Compound 2 (300 mg) was dissolved in methanol, stirred at room temperature for 10 min, 1N sodium hydroxide (1 ml) was added dropwise, stirred at room temperature for 10 min, the reaction solution was concentrated and purified by preparative HPLC to obtain 59 mg of the target product.

¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.64 (s, 1H), 11.42 (s, 1H), 9.03-9.07 (m, 4H ), 7.32-7.35 (m, 2H), 7.15-7.32 (m, 3H), 7.08-7.11 (m, 1H) 6.74-6.77 (m, 1H), 6.25-6.28 (m, 1H), 3.75-3.77 (m, 2H), 3.14-3.34 (m, 4H), 1.80-1.84 ( m, 2H).

Purity according to HPLC: @214 nm 99.90%, @254 nm 99.70%.

LC-MS: m/z 569[M+1].

Example 31

Reaction 1

Compound 1 (330 mg, 0.75 mmol) was dissolved in acetone (30 ml), potassium carbonate (414 mg, 3 mmol) was added, compound 1a (130.5 mg, 0.75 mmol) was added dropwise, stirred at room temperature temperature overnight, quenched by adding water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 300 mg of a crude product.

Reaction 2

Compound 2 (300 mg) was dissolved in methanol, stirred at room temperature for 10 min, 1N sodium hydroxide (1 ml) was added dropwise, stirred at room temperature for 10 min, the reaction solution was concentrated and purified by preparative HPLC to obtain 35 mg of the target product.

¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.64 (s, 1H), 11.42 (s, 1H), 9.03-9.07 (m, 4H ), 7.32-7.35 (m, 2H), 7.15-7.32 (m, 3H), 7.08-7.11 (m, 1H), 6.74-6.77 (m , 1H), 6.25-6.28 (m, 1H), 3.75-3.77 (m, 2H), 3.14-3.34 (m, 4H), 1.80-1.84 (m, 2H).

Purity according to HPLC: @214 nm 99.40%, @254 nm 99.25%.

LC-MS: m/z 553[M+1].

Example 32

Reaction 1

Compound 1 (330 mg, 0.75 mmol) was dissolved in tetrahydrofuran (10 ml), potassium carbonate (414 mg, 3 mmol) was added, stirred at room temperature for 20 min, a solution of compound 1a (184 mg, 1 mmol) in tetrahydrofuran (1 ml), stirred at room temperature for 2 h, quenched by adding water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 500 mg of a crude product.

Reaction 2

Compound 2 (500 mg) was dissolved in tetrahydrofuran (20 ml), stirred at room temperature for 5 min, 1N sodium hydroxide (5 ml) added dropwise, stirred at room temperature for 30 min. The reaction solution was extracted with water and ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give a crude product. The crude product was dissolved in acetonitrile and purified by preparative HPLC to give 29 mg of the desired product.

¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.91 (s, 1H), 11.43 (s, 1H), 9.04 (s, 1H), 8, 90 (s, 1H), 8.70 (s, 2H), 7.25-7.23 (m, 2H), 7.20-7.16 (m, 1H), 7.11-7.09 ( m, 1H), 6.91-6.88 (m, 2H), 6.78-6.75 (m, 1H), 6.25 (m, 1H), 3.73 (s, 3H), 3 .68 (s, 2H), 3.28-3.23 (m, 4H), 1.83-1.80 (m, 2H).

Purity according to HPLC: @214 nm 99.2%, @254 nm 99.7%.

LC-MS: m/z 563.2[M+1].

Example 33

Reaction 1

Compound 1 (150 mg, 0.47 mmol) was dissolved in DCM, Et ₃ N was added, stirred at room temperature for 1 h, quenched with water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 100 mg of a crude product.

Reaction 2

Compound 2 (100 mg, 1.08 mmol) was dissolved in ACN, compound 2a (120 mg, 1.06 mmol) was added, stirred at 90° C. overnight. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 100 mg of a crude product.

Reaction 3

Compound 3 (100 mg) was dissolved in THF/H ₂ O, stirred at room temperature for 10 min, 1N sodium hydroxide (1 ml) was added dropwise, stirred at room temperature for 10 min, the reaction solution was concentrated and subjected to purification by the method preparative HPLC to obtain 8 mg of the target product.

¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.579 (s, 1H), 11.480 (s, 1H), 8.718-8.917 (m, 3H), 6.286-7.187 (m, 4H), 3.255-3.324 (m, 4H), 2.499-2.507 (m, 1H), 1.570-1.843 (m, 8H).

Purity according to HPLC: @214 nm 97.3%, @254 nm 98.1%.

LC-MS: m/z 497[M+1].

Example 34

Reaction 1

Compound 1 (500 mg, 8.77 mmol) was dissolved in DCM, compound 1a (3.3 g, 21.93 mmol), HATU (4.01 g, 5.52 mmol) and DIEA (3.71 g, 56.5 mmol), stirred at room temperature overnight, quenched with water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain 120 mg of the desired product.

Reaction 2

Compound 2 (120 mg, 1.08 mmol) was dissolved in ACN, compound 2a (100 mg, 1.06 mmol) was added, stirred at 90° C. overnight. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 100 mg of a crude product.

Reaction 3

Compound 3 (100 mg) was dissolved in THF/H ₂ O, stirred at room temperature for 10 min, 1N sodium hydroxide (1 ml) was added dropwise, stirred at room temperature for 10 min, the reaction solution was concentrated and subjected to purification by the method preparative HPLC to obtain 12 mg of the target product.

¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 12.041 (s, 1H), 8.649-8.923 (m, 3H), 6.274-7.188 (m, 4H), 3.255-3.324 ( m, 4H), 2.509-2.683 (m, 2H), 1.594-2.067 (m, 6H).

Purity according to HPLC: @214 nm 95.9%, @254 nm 95.4%.

LC-MS: m/z 497[M+1].

Example 35

Reaction 1

Compound 1 (1 g, 8.77 mmol) was dissolved in DCM, compound 1a (6.3 g, 21.93 mmol), HATU (4.01 g, 10.52 mmol) and DIEA (5.71 g, 56.5 mmol), stirred at room temperature overnight, quenched with water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain 200 mg of product.

Reaction 2

Compound 2 (200 mg, 1.08 mmol) was dissolved in ACN, compound 2a (420 mg, 1.06 mmol) was added, stirred at 90° C. overnight. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 100 mg of a crude product.

Reaction 3

Compound 3 (100 mg) was dissolved in THF/H ₂ O, stirred at room temperature for 10 min, 1N sodium hydroxide (1 ml) was added dropwise, stirred at room temperature for 10 min, the reaction solution was concentrated and subjected to purification by the method preparative HPLC to obtain 12 mg of the target product.

¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.579 (s, 1H), 11.480 (s, 1H), 8.718-8.917 (m, 3H), 6.286-7.187 (m, 4H), 3.255-3.324 (m, 4H), 2.499-2.507 (m, 1H), 1.570-1.843 (m, 10H).

Purity according to HPLC: @214 nm 91.2%, @254 nm 96.8%.

LC-MS: m/z 511[M+1].

Example 36

Reaction 1

Compound 1 (500 mg, 8.77 mmol) was dissolved in DCM, compound 1a (3.3 g, 21.93 mmol), HATU (4.01 g, 10.52 mmol) and DIEA (5.71 g, 56.5 mmol), stirred at room temperature overnight, quenched with water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain 120 mg of the desired product.

Reaction 2

Compound 2 (120 mg, 1.08 mmol) was dissolved in ACN, compound 2a (100 mg, 1.06 mmol) was added, stirred at 90° C. overnight. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 100 mg of a crude product.

Reaction 3

Compound 3 (100 mg) was dissolved in THF/H ₂ O, stirred at room temperature for 10 min, 1N sodium hydroxide (1 ml) was added dropwise, stirred at room temperature for 10 min, the reaction solution was concentrated and subjected to purification by the method preparative HPLC to obtain 12 mg of the target product.

¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.441 (s, 2H), 8.649-8.923 (m, 3H), 6.274-7.188 (m, 4H), 3.255-3.324 ( m, 4H), 1.694-2.067 (m, 11H).

Purity according to HPLC: @214 nm 99.1%, @254 nm 99.3%.

LC-MS: m/z 511[M+1].

Example 37

Reaction 1

Compound 1 (110 mg, 0.25 mmol) was dissolved in acetone (10 ml), potassium carbonate (138 mg, 1 mmol) was added, compound 1a (113 mg, 1 mmol) was added dropwise, stirred at room temperature overnight , quenched by adding water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 100 mg of a crude product.

Reaction 2

Compounds 2 and 2a (100 mg) were dissolved in methanol, stirred at room temperature for 10 min, 1N sodium hydroxide (1 ml) was added dropwise, stirred at room temperature for 10 min. The reaction solution was concentrated and purified by preparative HPLC to give the desired products XSD3-087 (15 mg) and XSD3-087-01 (17 mg).

XSD3-087 ¹ H-NMR (400 MHz, DMSO): δ (ppm): 11.41 (s, 1H), 8.90 (m, 1H), 6.18-7.32 (s, 1H), 7.10-7.21 (m, 2H), 7.10-7.12 (m, 2H), 6.76-6.79 (m, 1H), 6.23 (m, 1H) , 3.12-3.23(m, 4H), 6.76-6.79(m, 1H), 2.67(s, 3H), 1.72-1.75(m, 2H).

Purity according to HPLC: @214 nm 93.5%, @254 nm 94.6%.

LC-MS: m/z 492 [M+1].

XSD3-087-01 ¹ H-NMR (400 MHz, DMSO): δ (ppm): 11.43 (s, 1H), 8.90 (m, 1H), 8.01 (m, 2H) , 7.10-7.20 (s, 2H), 6.85 (m, 1H), 6.23 (m, 1H), 3.72-3.76 (m, 2H), 3.40 (s , 3H), 3.24-3.26(m, 2H), 2.93(s, 3H), 1.87-1.91(m, 2H).

Purity according to HPLC: @214 nm 98%, @254 nm 98.6%.

LC-MS: m/z 570 [M+3].

Example 38

Reaction 1

Compound 1 (100 mg, 0.21 mmol) was dissolved in acetone (10 ml), potassium carbonate (78 mg, 0.55 mmol) was added, compound 1a (41 mg, 0.23 mmol) was added dropwise, stirred at room temperature. temperature overnight, quenched by adding water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 100 mg of a crude product.

Reaction 2

Compound 2 (100 mg) was dissolved in THF/H ₂ O, stirred at room temperature for 10 min, 1N sodium hydroxide (1 ml) was added dropwise, stirred at room temperature for 10 min. The reaction solution was concentrated and subjected to preparative HPLC to obtain 5 mg of the desired product.

¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.421 (s, 1H), 8.897 (s, 1H), 7.153-7.197 (m, 2H), 7.105-7.120 (m, 1H), 6.746-7.098 (m, 1H), 6.220 (s, 1H), 4.078 (s, 2H), 3.104-3.129 (m, 4H), 2.735-2.761 (m, 2H), 1.535-1.853 (m, 7H), 0.976-1.202 (m, 6H).

Purity according to HPLC: @214 nm 99.8%, @254 nm 99.3%.

LC-MS: m/z 575[M+1].

Example 39

Reaction 1

Compound 1 (1.0 g, 3.6 mmol) was dissolved in acetone (16 ml), potassium carbonate (993 mg, 7.2 mmol) was added, compound 1a (254 mg, 2.7 mmol) was added dropwise, stirred at room temperature overnight, quenched by adding water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 300 mg of a crude product.

Reaction 2

Compound 2a (100 mg) was dissolved in acetonitrile, compound 2 (40 mg, 0.27 mmol) was added, stirred at 90°C for 12 h, quenched by adding water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 80 mg of a crude product.

Reaction 3

Compound 3 (80 mg) was dissolved in DMF, stirred at room temperature for 10 minutes, potassium carbonate (132 mg, 0.96 mmol) was added, stirred at room temperature for 12 hours, quenched with water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated to give 60 mg of a crude product, which was purified by preparative HPLC to give 14.2 mg of the desired product.

¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.53 (s, 1H), 8.91 (s, 1H), 8.58-8.27 (m, 3H ), 7.38-7.12(m, 2H), 6.78(s, 1H), 6.28(s, 1H), 3.65(s, 3H), 3.28-3.24( m, 4H), 1.79 (s, 2H).

Purity according to HPLC: @214 nm 94.5%, @254 nm 95.4%.

LC-MS: m/z 473 [M+1].

Example 40

Reaction 1

Compound 1 (1.0 g, 3.6 mmol) was dissolved in acetone (16 ml), potassium carbonate (993 mg, 7.2 mmol) was added, compound 1a (294 mg, 2.7 mmol) was added dropwise, stirred at room temperature overnight, quenched by adding water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 400 mg of a crude product.

Reaction 2

Compound 2a (100 mg) was dissolved in acetonitrile, compound 2 (42 mg, 0.27 mmol) was added, stirred at 90°C for 12 h, quenched by adding water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 66 mg of a crude product.

Reaction 3

Compound 3 (66 mg) was dissolved in DMF, stirred at room temperature for 10 min, potassium carbonate (108 mg, 0.78 mmol) was added, stirred at room temperature for 12 h, quenched by adding water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 50 mg of a crude product, which was purified by preparative HPLC to give 26.5 mg of the desired product.

¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.47 (s, 1H), 11.24 (s, 1H), 8.92 (s, 1H), 8, 62 (s, 2H), 7.21-7.09 (m, 2H), 6.79-6.75 (m, 1H), 6.29-6.25 (m, 1H), 4.25- 4.20 (m, 2H), 3.32-3.24 (m, 4H), 1.85-1.80 (m, 2H), 1.28-1.24 (m, 3H).

Purity according to HPLC: @214 nm 97.8%, @254 nm 98.6%.

LC-MS: m/z 487 [M+1].

Example 41

Reaction 1

Compound 1 (1.0 g, 3.6 mmol) was dissolved in acetone (16 ml), potassium carbonate (993 mg, 7.2 mmol) was added, compound 1a (732 mg, 6.0 mmol) was added dropwise, stirred at room temperature overnight, quenched by adding water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 360 mg of a crude product.

Reaction 2

Compound 2a (100 mg) was dissolved in acetonitrile, compound 2 (45 mg, 0.27 mmol) was added, stirred at 90° C. for 12 h, quenched by addition of water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 51 mg of a crude product.

Reaction 3

Compound 3 (51 mg) was dissolved in DMF, stirred at room temperature for 10 min, potassium carbonate (108 mg, 0.78 mmol) was added, stirred at room temperature for 12 h, quenched with water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 50 mg of a crude product, which was purified by preparative HPLC to give 10.6 mg of the desired product.

¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.48 (s, 1H), 11.18 (s, 1H), 8.92 (s, 1H), 8, 60 (s, 2H), 7.21-7.09 (m, 2H), 6.79-6.75 (m, 1H), 6.30-6.25 (m, 1H), 4.98- 4.91(m, 1H), 3.32-3.24(m, 4H), 1.83-1.78(m, 2H), 1.28-1.27(m, 6H).

Purity according to HPLC: @214 nm 95.2%, @254 nm 98.0%.

LC-MS: m/z 501 [M+1].

Example 42

Reaction 1

Compound 1 (1.0 g, 3.6 mmol) was dissolved in acetone (16 ml), potassium carbonate (993 mg, 7.2 mmol) was added, compound 1a (540 mg, 3.6 mmol) was added dropwise, stirred at room temperature overnight, quenched by adding water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 330 mg of a crude product.

Reaction 2

Compound 2a (100 mg) was dissolved in acetonitrile, compound 2 (50 mg, 0.27 mmol) was added, stirred at 90° C. for 12 h, quenched by addition of water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 90 mg of a crude product.

Reaction 3

Compound 3 (90 mg) was dissolved in DMF, stirred at room temperature for 10 minutes, potassium carbonate (132 mg, 0.96 mmol) was added, stirred at room temperature for 12 hours, quenched with water. The resulting material was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give 70 mg of a crude product, which was purified by preparative HPLC to give 32.5 mg of the desired product.

¹ H-NMR (400 MHz, DMSO-d6): δ (ppm): 11.51 (s, 1H), 11.24 (s, 1H), 8.91 (s, 1H), 8, 55-8.27(m, 3H), 7.21-7.09(m, 2H), 6.79-6.75(m, 1H), 6.29-6.25(m, 1H), 4.25-4.20 (m, 2H), 3.32-3.24 (m, 4H), 1.80 (s, 2H), 1.57 (s, 2H), 1.28 (s, 4H), 0.88-0.85 (m, 3H).

¹ H-NMR (400 MHz, CDCl3): δ (ppm): 7.23-7.20 (m, 1H), 7.10 (s, 1H), 7.04-7.00 (m , 1H), 6.92-6.88 (m, 1H), 5.85 (s, 1H), 4.09-4.06 (m, 2H), 3.58-3.53 (m, 2H ), 3.38-3.35 (m, 2H), 1.98-1.97 (m, 2H), 1.70-1.63 (m, 2H), 1.38-1.35 (m , 4H), 0.91-0.88 (m, 3H).

Purity according to HPLC: @214 nm 98.1%, @254 nm 97.8%.

LC-MS: m/z 529[M+1]

Based on Synthesis Method I and the Examples described above, the following compounds were synthesized.

Example No. Deputy Substance No. MS Example 43

, n равен 2, R₁, R₃, R₄ представляют собой H.R ₂ is

, n is 2, R ₁ , R ₃ , R ₄ are H. XSD3-075 560.3 Example 44 R ₂ is ethyl carbonyl, n is 1, R ₁ , R ₃ , R ₄ are H. XSD3-063 456.3 Example 45 R ₂ is isopropylcarbonyl, n is 1, R ₁ , R ₃ , R ₄ are H. XSD3-064 470.2 Example 46 R ₂ is methyl carbonyl, n is 2, R ₁ , R ₃ , R ₄ are H. XSD3-086 456.1 Example 47 R ₂ is cyano, n is 1, R ₁ , R ₃ , R ₄ are H. XSD3-073 425.1 Example 48 R ₂ is cyano, n is 2, R ₁ , R ₃ , R ₄ are H. XSD3-085 439.1 Example 49 R ₂ is

, n is 1, R ₁ , R ₃ , R ₄ are H. XSD3-071 510.2 Example 50 R ₂ is

, n is 2, R ₁ , R ₃ , R ₄ are H. XSD3-077 469.1 Example 51 R ₂ is

, n is 2, R ₁ , R ₃ , R ₄ are H. XSD3-083 524.2 Example 52 R ₃ is 1,2-ethanediyl, n is 1, R ₁ , R ₂ , R ₄ are H. XSD3-090 426.1 Example 53 R ₃ is 1,2-ethanediyl, n is 2, R ₁ , R ₂ , R ₄ are H. XSD3-091 440.1 Example 54 R ₂ is

, n is 1, R ₁ , R ₃ , R ₄ are H. XSD3-099 554.1

Example 55: Determination of inhibitory activity against indolamine-2,3-dioxygenase and IC ₅₀

The construction of a plasmid containing the human indolamine 2,3-dioxygenase gene, its expression in E. coli, extraction and purification were performed according to the method described in Littlejohn et al. (Takikawa O, Kuroiwa T, Yamazaki F, et al. J. Biol. Chem. 1988, 263, 2041-2048). In a 96-well plate, 50 mM potassium phosphate buffer (pH 6.5), 20 mM ascorbate, 20 μM methylene blue, and purified human indolamine 2,3-dioxygenase protein were mixed, and 200 μM L-tryptophan and inhibitor were added to the mixture. . The reaction was carried out at 37°C for 60 minutes, then the reaction was stopped by adding 30% trichloroacetic acid, the mixture was incubated at 65°C for 15 minutes to hydrolyze N-formylkynurenine to kynurenine, and centrifuged at 3400 rpm for 5 minutes to remove the precipitated protein. The supernatant was placed in a new 96-well plate, a solution of 2% (w/v) para-dimethylaminobenzaldehyde in acetic acid was added, reacted by incubation at 25°C for 10 minutes, and a spectrophotometer was read at 480 nm. Control wells did not contain an indolamine 2,3-dioxygenase inhibitor or indolamine 2,3-dioxygenase itself and were used as the necessary non-linear regression parameters to determine the IC ₅₀ for each compound. Non-linear regression and determination of IC ₅₀ values were performed using GraphPad Prism 4 software. Those compounds with IC ₅₀ values less than 10 μM were considered effective inhibitors in this assay method. The compounds of Examples according to the present invention had excellent inhibitory activity against indolamine-2,3-dioxygenase.

Table 1: IC ₅₀ values of compounds

Structural formula No. IC ₅₀ (nM) 3047 37 3048 twenty 3049 22 3051 23 3053 16 INCB024360 102

Note: INCB024360 was an indolamine-2,3-dioxygenase inhibitor with the following structure:

.

.

The IC ₅₀ values for the following compounds were determined by the method described in Example 55 and the specific results are presented in Table 2:

Table 2: IC ₅₀ values of compounds

Structural formula No. IC ₅₀ (nM) XSD3-058 57 XSD3-079 95 XSD3-093 44 XSD3-093-1 56 XSD3-093-2 61 XSD3-093-3 76 XSD3-074 63 XSD3-088 72 XSD3-089 67 XSD3-076 102 XSD3-081 89 XSD3-069 345 XSD3-052 343 XSD3-065 117.9 XSD3-066 131.5 XSD3-067 142 XSD3-068 539 XSD3-070 345 XSD3-072 502 XSD3-078 145 XSD3-080 87 XSD3-082 101 XSD3-084 160 XSD3-092 399

Example 56 In Vivo Test for Antitumor Activity of Indolamine 2,3-Dioxygenase Inhibitors

1. Division of animals into groups and testing method

LLC cells in the logarithmic growth phase were taken, cell viability was determined by the trypan blue staining method, the concentration of viable cells was adjusted to 1×10 ⁷ cells/ml, and administered to homologous C57BL6 mice by subcutaneous injection at a dose of 0.2 ml/mouse. After tumor formation, mice were randomly divided into a control group, a cyclophosphamide (CTX) group, an INCB024360 compound group, a 3047 compound group, according to their tumor weight and body weight; there were 10 mice in each group, and the CTX group was administered by intraperitoneal injection at a dose of 150 mg/kg, the INCB024360 compound group and the 3047 compound group were intragastrically administered, the control group received the same volume of normal saline in the same time, and the frequency of administration for each group was once a day. The test was terminated after 21 days of administration.

24 hours after the last injection, the animals were weighed and sacrificed, the tumors were removed and weighed, and the average tumor inhibition rate (I) was calculated according to the following formula: I=(1 - average tumor mass in the drug administration group/average tumor mass in the control group) × 100%

2. Statistics and processing method

Experimental data were analyzed by spss16.0 and one-way analysis of variance, and the difference with p<0.05 was considered statistically significant.

3. Test results and discussion

Table 3: Results of Compound Inhibition of LLC Tumors in Mice

Group Number of animals (mouse) Tumor weight (g) Degree of tumor inhibition (%) Control group ten 1.514±0.312 / Group with the introduction of CTX ten 0.701±0.314 ^＃＃ 53.7 Group with introduction INCB024360 ten 0.653±0.178 ^＃＃ 56.9 Group with the introduction of compound 3047 ten 0.370±0.209 ^＃＃ 75.6

Compared with the control group, ^＃＃ P<0.01;

Compared with the CTX administration group and the INCB024360 administration group, ^＃ P<0.05;

As can be seen from Table 3, tumor weight in each drug group was significantly different from that in the control group (P<0.01); the compound 3047 group was significantly different from the cyclophosphamide group and the INCB024360 group (P<0.05). This result indicates that the compound of the present invention is superior to the existing chemotherapeutic drugs cyclophosphamide and compound INCB024360 in therapeutic effect on tumors.

Table 4: Effects of Compounds on Body Weight in Mice

Group Average body weight before testing (g) Average body weight after testing (g) Control group 21±4.2 28±3.2 Group with the introduction of CTX 22±3.1 19±3.3 Group with the introduction of compound 3047 22±3.4 29±5.2 ^＃＃

Compared with the cyclophosphamide group, ^＃＃ P<0.01

As can be seen from Table 4, the 3047 group did not show a significant difference in body weight compared to the control group, but showed a significant difference compared to the CTX group. This result indicates that the compound of the present invention increases body weight in mice while controlling tumor growth, that is, the side effect was reduced and the quality of life of mice was significantly improved. Clinically, this means that the quality of life of patients can be improved, and the patient's drug compliance and drug efficacy can be significantly enhanced.

In addition, we also tested Colon26 cells of mouse colon cancer, Hepa1-6 cells of mouse liver cancer, 4T1 cells of mouse breast cancer and other cell lines, and the results showed that the compounds of the present invention have significant inhibitory effects against indicated tumors.

The in vivo antitumor activity of the following compounds was determined by the method of Example 56 and the specific results are shown in the following table:

Table 5: Results of Compound Inhibition of LCC Tumors in Mice

Group Number of animals (mouse) Tumor weight (g) Degree of tumor inhibition (%) Control group ten 1.445±0.217 / Group with the introduction of the connection XSD3-058 ten 0.400±0.112 ^＃＃ 72.3 Group with the introduction of the connection XSD3-079 ten 0.384±0.092 ^＃＃ 73.4

Compared with the control group, ^＃＃ P<0.01.

As can be seen from Table 5, the tumor weight in each drug group was significantly different from that of the control group (P<0.01). This result indicates that the compound of the present invention has a significant therapeutic effect on the tumor.

The effect of each compound on mouse body weight was tested. No significant difference in body weight was found between the XSD3-058 compound administration group and the XSD3-079 compound administration group compared to the control group. This result indicates that the compounds of the present invention can increase body weight in mice by regulating tumor growth, reduce drug side effects, and significantly improve mouse quality of life. Clinically, the compounds of the present invention can improve the quality of life of patients and significantly enhance patient drug compliance and drug efficacy.

Example 57 Determination of Behavioral Changes in the Morris Water Maze Test in Mice with Alzheimer's Disease

1. Division of animals into groups and methods of testing

According to the present invention, in order to generate AD models according to the method of Richardson et al. with a single injection of aggregated Aβ1-42 in the bilateral CA3 region of the hippocampus, mice were selected at the age of 9 months, then they were divided into a control group, a group with the introduction of the compound INCB024360, a group with the introduction of the compound 3047; a total of 10 mice per group, five males and five females. Mouse behavior analysis was performed using a Morris water maze (Ethovision XT monitoring and analysis software, Morris water maze system manufactured by Noldus, Netherlands). The water maze test procedure was divided into two parts, the hidden platform detection test on the 5th consecutive day, and the spatial test on the 6th day, with each test administered to mice according to test groups and doses designed. Mice were trained 4 times a day, each time immersing mice in different areas, the water maze was divided into areas 1, 2, 3 and 4 according to south, east, north and west, and the platform was area 5 located in area 4 Each swimming time was 60 s, the interval for each training was approximately 1 hour, and if the mouse did not find the platform, then the latency period was considered to be 60 s. The hidden platform test was used to determine the learning ability of the mice, while the spatial test was used to determine the mice's spatial memory ability.

2. Statistics and processing method

The rescue latency in the hidden platform detection test was analyzed by multi-measurement analysis of variance using SPSS16.0 statistical analysis software; and analyzed the swimming time in each quadrant and the number of target crossings in the spatial test by one-way analysis of variance. Data were expressed as mean ± standard deviation and the significant level of difference in the two-tailed test was given as P=0.05.

3. Test results and discussion

Table 6: Latent period(s) of each animal group for platform seeking in the hidden platform test

Group 1st day 2nd day 3rd day 4th day 5th day Control group 54.0±5.9 52.8±1.9 43.5±6.7 39.8±6.9 38.4±5.6 Group with introduction INCB024360 53.2±2.3 51.4±4.8 38.6±5.5 28.7±3.9 ^＃ 19.9±6.7 ^＃＃ Group with the introduction of compound 3047 52.3±5.2 44.0±4.4 ^＃ 22.1±4.5 ^＃＃ 10.5±5.7 ^＃＃ 5.4±6.0 ^＃＃

Compared with the control group, ^＃ P<0.05, ^＃＃ P<0.01,

Compared with INCB024360 administration group, ^＃ P<0.05, ^＃＃ P<0.01.

Table 7: Time and number of stops in the area of the platform for each group of animals

Group Stop time (s) Number of stops (times) 4th region 5th region 4th region 5th region Control group 14.9±3.8 1.9±0.8 4.8±0.7 1.0±0.3 Group with introduction INCB024360 16.2±2.4 3.9±0.7 ^＃ 8.4±1.0 ^＃ 2.1±0.4 ^＃＃ Group with the introduction of compound 3047 25.9±5.2 ^＃＃ 5.9±0.7 ^＃＃ 13.0±1.5 ^＃＃ 3.8±0.6 ^＃＃

Compared with the control group, ^＃ P<0.05, ^＃＃ P<0.01,

Compared with INCB024360 injection group, ^＃ P<0.05.

As can be seen from Tables 6 and 7, compound 3047 can significantly improve learning and memory impairment in animals, significantly improve knowledge acquisition ability and spatial memory ability, and is superior to INCB024360 compound, indicating that the development of the compound according to the present invention has of great importance in the treatment of Alzheimer's syndrome.

The effect of the following compounds on the behavior of Alzheimer's mice was tested by the method of Example 57. Specific results are shown in the following table:

Table 8: Latent period(s) for finding the platform in the hidden platform test for each group of animals

Group 1st day 2nd day 3rd day 4th day 5th day Control group 59.7±4.6 59.8±1.8 50.6±5.5 42.8±4.9 38.7±5.2 Group with the introduction of the connection XSD3-058 45.3±3.7 ^＃ 41.3±3.2 ^＃ 25.4±4.6 ^＃＃ 15.5±4.9 ^＃＃ 7.3±6.1 ^＃＃ Group with the introduction of the connection XSD3-079 40.5±5.5 ^＃ 39.3±44 ^＃ 20.3±4.2 ^＃＃ 17.7±5.2 ^＃＃ 9.9±5.2 ^＃＃

Compared with the control group, ^＃P <0.05, ^＃＃ P<0.01.

Table 9: Time and number of stops in the area of the platform for each group of animals

Group Stop time (s) Number of stops (times) 4th region 5th region 4th region 5th region Control group 15.8±3.1 2.0±0.9 4.1±0.9 1.2±0.4 Connection group XSD3-058 25.2±4.2 ^＃＃ 5.9±0.6 ^＃＃ 13.3±1.8 ^＃＃ 3.5±0.9 ^＃＃ Connection group XSD3-079 26.7±3.3 ^＃＃ 5.8±0.7 ^＃＃ 14.4±1.3 ^＃＃ 3.9±0.5 ^＃＃

Compared with the control group, ^＃＃ P<0.01.

As can be seen from Tables 8 and 9, the compounds according to the present invention can significantly improve learning and memory impairment, and significantly improve the ability to acquire knowledge and the ability to spatial memory, and the results indicate that the development of the compound according to the present invention is of great importance. in the treatment of Alzheimer's syndrome.

Example 58 DC Stimulated T Cell Proliferative Response After Drug Treatment

The dendritic cell (DC) is the most potent antigen presenting cell (APC) that can effectively activate intact T cells to proliferate, which is the most important difference between DC and other APCs. DC is the initiator of the immune response. DC has become one of the hottest applications of modern immunological research due to its key role in the CD4 ⁺ , CD8 ⁺ T-cell immune response. To date, DC research has mainly focused on the prevention and treatment of neoplastic diseases, autoimmune diseases, transplant rejection, and infection control.

1. Isolation and cultivation of human peripheral blood dendritic cells

The human peripheral blood buffy coat was removed and diluted with the same volume of 0.01 mol/l PBS. PBMC were isolated in a standard manner using lymphocytic separation medium, and the cell concentration was adjusted to 3×10 ⁶ ml ^-1 in RPMI1640 complete medium; cells were introduced into a 6-well plate at 3 ml per well were cultured in an incubator at 5% CO ₂ , 37°C for 2 hours, washed 3 times with PBS to remove non-adherent cells, then culture medium containing IL-4 (100 u /ml), GM-CSF (150 ng/ml) and TNF-α (500 U/ml), and then cultivated in a standard way, renewing the medium half every day. After 8 days of cultivation, the cells were used for identification and experiment.

2. Preparation of T cells

The human PBMC layer was separated by the method described in step 1. Macrophages were removed by an adhesive method, B cells were removed with a nylon cotton syringe, and the obtained T cells were adjusted to a cell concentration of 1×10 ⁶ cells/ml.

3. Cooking DC

Mature DC with a purity of 99% were centrifuged, RPMI1640 was added to obtain cell concentrations of 1×10 ⁵ , 4×10 ⁴ , 2×10 ⁴ /ml, and added to a 96-well plate in the amount of 100 μl per well, two wells per every concentration. Separately, the INCB024360 compound and the 3047 compound were added and cultured for 2 days.

4. T cell proliferation assay (MLR) method

In each of the above drug groups, T cells were added to the DC at 100 μl per well, cultured in an incubator at 5% CO ₂ , 37° C. for 72 hours; 6 hours before the end of cultivation, 100 μl of the culture solution was carefully removed from each well, 10 μl of MTT (5 mg/ml) was added, then cultivated in an incubator for 6 hours, then 100 μl of 0.01 mol/l HCl-10% was added SDS, left overnight at 37°C, and determined the optical density (A _{570 nm} ) in a microplate to demonstrate the level of T cell proliferation.

5. Experimental results and analysis

Table 10: Effect of compound 3047 on DC-stimulated T cell proliferation

Group T:DC (optical density) 10:1 25:1 50:1 Control group 0.801±0.003 0.782±0.002 0.755±0.005 Group with introduction INCB024360 0.830±0.002 ^＃ 0.804±0.004 ^＃ 0.799±0.003 ^＃ Group with the introduction of compound 3047 0.864±0.006 ^＃＃ 0.834±0.004 ^＃＃ 0.825±0.007 ^＃＃

Compared with the control group, ^＃ P<0.05, ^＃＃ P<0.01,

Compared with INCB024360 group, ^＃ P<0.05.

As can be seen from Table 10, compared with the control group, the number of T cells in the compound 3047 group and the INCB024360 group were significantly increased, showing significant differences ( ^＃P <0.05, ^＃＃ P<0, 01); and compared with the INCB024360 compound group, the 3047 compound group had a more significant effect on T cell proliferation, showing a significant difference ( ^＃P <0.05). This indicates that the compound of the present invention has a significant effect of enhancing DC-stimulated T cell proliferation, and this effect is significantly better than that of the compound INCB024360, and therefore can be used for the treatment of neoplastic diseases, autoimmune diseases, transplant rejection, and infectious diseases.

DC-stimulated T cell proliferative responses after treatment with the following compounds were determined by the method of Example 58, and the specific results are shown in the following table:

Table 11: Effect of each compound on DC-stimulated T cell proliferation

Group T:DC (optical density) 10:1 25:1 50:1 Control group 0.810±0.004 0.793±0.004 0.760±0.002 Group with the introduction of the connection XSD3-058 0.880±0.003 ^＃＃ 0.849±0.002 ^＃＃ 0.831±0.002 ^＃＃ Group with the introduction of the connection XSD3-079 0.876±0.004 ^＃＃ 0.850±0.003 ^＃＃ 0.837±0.004 ^＃＃

Compared with the control group, ^＃＃ P<0.01.

As can be seen from Table 11, compared with the control group, the number of T cells in each compound administration group was significantly increased, showing a significant difference ( ^＃＃ P<0.01), indicating that the compound of the present invention can significantly enhance DC-stimulated T cell proliferation, and therefore can be used to treat diseases associated with IDO, such as tumor diseases, autoimmune diseases, transplant rejection, and infectious diseases.

Claims (31)

1. The compound represented by formula I, or a pharmaceutically acceptable salt thereof

, where R ₁ is selected from the group consisting of H, C _1-6 alkyl and C _1-6 alkyl-C(O)-; n is an integer selected from 0-6. 2. A compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein R ₁ is selected from the group consisting of H and C _1-6 alkyl. 3. A compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that R ₁ is selected from the group consisting of H, methyl, ethyl, propyl, isopropyl, butyl. 4. A compound or pharmaceutically acceptable salt thereof according to claim 1, characterized in that n is selected from 0, 1, 2, 3 and 4. 5. A compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that if R ₁ in formula I is H, then the compound is represented by formula II

. 6. A compound or pharmaceutically acceptable salt thereof according to claim 5, characterized in that n is 0, 1, 2 or 3. 7. A compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that if R ₁ in formula I is R ₀ -substituted carbonyl, then the compound is represented by formula III

, where R ₀ is C _1-6 alkyl. 8. A compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:

,

,

,

,

,

. 9. Synthetic method for obtaining the compound represented by the following formula

(formula II), where n is selected from 0, 1, 2, 3 and 4; characterized in that it includes the following stages: 1) interaction of the compound represented by formula 2 with compound 3a' to obtain the compound represented by formula IIa

; 2) carrying out the deprotection of the compound represented by formula IIa under acidic conditions, followed by carrying out the reaction under alkaline conditions to obtain the compound represented by formula IIa

. 10. Synthetic method for obtaining the compound represented by the following formula

(formula II'), characterized in that it includes the following stage:

, where n is selected from 0, 1, 2, 3 or 4; the reaction of the compound represented by formula IIa is carried out under acidic conditions to obtain the compound represented by formula II'. 11. Pharmaceutical composition for inhibition of indolamine-2,3-dioxygenase, containing a compound or a pharmaceutically acceptable salt thereof according to any one of paragraphs. 1-8 and one or more pharmaceutically acceptable adjuvants. 12. A method of treating a disease, wherein the method comprises administering to a patient in need of such treatment a therapeutically effective amount of at least one of the compound or a pharmaceutically acceptable salt thereof according to any one of paragraphs. 1-8, or its pharmaceutical composition according to claim 11, wherein the disease is a disease characterized by an abnormal tryptophan metabolism pathway mediated by indolamine-2,3-dioxygenase, and the disease is a malignant tumor or Alzheimer's disease. 13. The method of claim 12, wherein the cancer is selected from the group consisting of lung cancer, liver cancer, colon cancer, pancreatic cancer, breast cancer, prostate cancer, brain cancer, ovarian cancer, cervical cancer, testicular cancer, kidney cancer, head and neck cancer, lymphoma, melanoma, and leukemia.