WO2017129139A1 - Sulfamic acid ester as indoleamine-2,3-dioxygenase inhibitor, preparation method therefor and use thereof - Google Patents

Abstract

Provided in the present invention are a sulfamic acid ester as an indoleamine-2,3-dioxygenase inhibitor, a preparation method therefor and a use thereof. The structure of the inhibitor in the present invention is shown in general structure I, wherein the definitions of X, R¹, R², R³, R⁴, R⁵ and n are shown in the description and claims. Also disclosed in the present invention is a preparation method for the inhibitor. The compound in general formula I of the present invention can be used as an indoleamine-2,3-dioxygenase inhibitor for the preparation of a drug for preventing and/or treating diseases mediated by indoleamine-2,3-dioxygenase.

Description

Aminosulfonate as a guanamine-2,3-dioxygenase inhibitor, and preparation method and use thereof Technical field

The invention belongs to the technical field of medicinal chemistry, in particular to a sulfamate as a guanamine-2,3-dioxygenase inhibitor, a preparation method and use thereof, and more particularly to a sulfamate containing And IDO inhibitors of 1,2,5-oxadiazole structure, and preparation methods and uses thereof.

Background technique

Indoleamine-2,3-dioxygenase (IDO) is a monomeric enzyme containing heme found in the cell for the first time in 1967 by the Hayaishi group. The cDNA encodes a protein. 403 amino acid composition with a molecular weight of 45 kDa, which is the rate-limiting enzyme of the leucine-kynurenine pathway catabolism and is widely expressed in many mammalian tissues (Hayaishi O. et al. Science, 1969, 164, 389). -396). In tumor cells, IDO often plays an important physiological role in inducing tumor microenvironmental immune tolerance, and its mediated tryptophan (Trp)-kynurenine (Kyn) metabolic pathway is involved in tumors. Immune escape, and IDO also plays an important role as an immune tolerance to induce tumor microenvironment.

Tryptophan is one of the important essential amino acids in mammals. It needs to be ingested in large quantities from food, maintain cell activation and proliferation, and synthesize proteins and some neurotransmitters. Therefore, its lack of dysfunction can lead to some important cells. IDO can catalyze the conversion of tryptophan to N-formyl kynurenine in vivo, degrading the content of tryptophan and causing deficiency of tryptophan in vivo, leading to tumor formation. Immunohistological studies have shown that the kynurenine pathway can lead to an increase in the excitotoxic quinolinic acid, as well as many serious human diseases such as Alzheimer's and other neurological diseases (Guillemin GJet al Neuropathol. and Appl. Neurobiol. 2005, 31, 395–404).

There are two main types of tryptophan rate-limiting enzymes in mammals: tryptophan dioxygenase (TDO) and IDO. In 1937, Kotake et al. purified proteins from rabbit intestines and found that TDO was mainly expressed in mammalian liver. It has not been found to be closely related to the immune system. TDO catalyzes the kynurenine pathway and converts tryptophan to N-formyl kynurenine [Higuchi K. et al J. Biochem. 1937, 25, 71-77; Shimizu T. et al J. Biol. .1978, 253, 4700-4706]. In 1978, the enzyme purified from the intestinal tract of rabbits was identified as a dioxygenase (IDO) containing heme, which is the only one outside the liver that catalyzes the oxidative cleavage of guanidine in the tryptophan molecule. An enzyme of acids and other metabolites. IDO is usually expressed in organs with more mucosa, such as the lungs, small intestine and large intestine, rectum, spleen, kidney, stomach and brain, and is widely distributed (Hayaishi O. et al, Proceedings of the tenth FEBS meeting, 1975, 131– 144). Under certain special or pathological conditions, such as pregnancy, chronic infection, organ transplantation and tumors, IDO expression will increase significantly, and participate in mediating local immunosuppression.

Studies have shown that IDO can inhibit local T cell immune responses in the tumor microenvironment by: tryptophan depletion, toxic metabolism, and induction of regulatory T cell proliferation. In many cases, it is transiently expressed in tumors, thereby consuming local tryptophan and producing a large amount of metabolites such as kynurenine. In fact, in the absence of tryptophan or kynurenine culture conditions, T cells undergo proliferation inhibition, decreased activity, and even apoptosis. And T There is a regulatory point in the cell that is very sensitive to the level of tryptophan. Under the action of IDO, tryptophan can be consumed, which leads to the arrest of T cells in the middle of G1 phase, thereby inhibiting the proliferation of T cells and inhibiting T cells. Immune response. Once T cells stop proliferating, they may not be stimulated any more. This is the mechanism of IDO immunity in vivo (Mellor A. et al Biochem. Biophys. Res. Commun. 2005, 338(1): 20-24) (LeRond S. et al J. Exp. Med. 2002, 196(4): 447-457).

There is still a need in the art to develop novel IDO inhibitors.

Summary of the invention

It is an object of the present invention to provide a novel compound containing a sulfamate and a 1,2,5-oxadiazole structure as a highly potent IDO enzyme inhibitor.

Another object of the present invention is to provide a process for the preparation of the compound.

In a first aspect of the invention, there is provided a compound of formula (I), or a pharmaceutically acceptable salt thereof, a stereoisomer thereof or a tautomer thereof, or a prodrug:

In the formula,

R ¹ , R ² , R ³ and R ⁴ are each independently hydrogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₂ -C ₁₀ alkenyl, substituted or unsubstituted C ₃ -C ₁₀ alkynyl, substituted or unsubstituted C ₆ -C ₂₀ aryl, or substituted or unsubstituted C ₃ -C ₁₄ heteroaryl; R ¹ and R ² , or R ³ and R ⁴ may together form a three- to eight-membered carbocyclic ring or a three- to eight-membered heterocyclic ring (eg, a three-membered ring, a four-membered ring, a five-membered ring, a six-membered ring, a seven-membered ring, and eight-membered ring). Ring) or R ¹ and R ³ may together form a five- to eight-membered carbocyclic ring or a five- to eight-membered heterocyclic ring (such as a five-membered ring, a six-membered ring, a seven-membered ring, or an eight-membered ring), wherein the hetero atom may be sulfur, oxygen, NH or NR ^f;

R ⁵ is C ₆ -C ₂₀ aryl, 5- or 6-membered heteroaryl; R ⁵ may be substituted by one or more groups selected from the group consisting of halogen, C ₁ -C ₆ alkyl, C ₁ - C ₆ alkoxy, hydroxy, amino, nitro, aldehyde, -CF ₃ , -CN, -SF ₅ , NR ^a R ^b , carboxy, -COR ^a , -CO ₂ C ₁ -C ₆ alkyl, - CONR ^a R ^b , -S(O)R ^a , -S(O) ₂ R ^a , -S(O)(NH)R ^a , -S(O)(NR ^d )R ^a , -SO ₂ NR ^a R ^b , -P(O)Me ₂ , -P(O)(OMe) ₂ ; wherein each R ^a and each R ^b are independently hydrogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl, substituted or not Substituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₂ -C ₁₀ alkenyl, substituted or unsubstituted C ₆ -C ₂₀ aryl, or substituted or unsubstituted C ₃ -C ₁₄ heteroaryl And R ^a and R ^b may together form a three to eight membered ring or a four to eight membered heterocyclic ring, wherein the hetero atom may be sulfur, oxygen, NH or NR ^f ;

X is a single bond, O, S, NH or NR ^d ;

R ^d and R ^{f are} independently C ₁ -C ₁₀ alkyl, C ₃ -C ₁₀ cycloalkyl, C ₆ -C ₂₀ aryl, or C ₃ -C ₁₄ heteroaryl;

n is an integer from 2 to 8.

In another preferred embodiment, the "substitution" means having one or more substituents selected from the group consisting of halogen, hydroxy, -NH ₂ , nitro, -CN, C ₁ -C ₄ alkyl, C ₁ -C ₄ haloalkyl, C ₁ -C ₄ alkoxy, C ₃ -C ₆ cycloalkyl, C ₂ -C ₄ alkenyl, C ₂ -C ₄ alkynyl group, a phenyl group, a benzyl group.

In another preferred embodiment, X is NH.

In another preferred embodiment, R ³ and R ⁴ are each independently hydrogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl; R ³ and R ⁴ may together form a three to eight membered ring or a three to eight membered impurity. A ring wherein the hetero atom can be sulfur, oxygen, NH or NR ^f .

In another preferred embodiment, n is an integer from 2 to 6.

In another preferred embodiment, n is an integer from 2 to 6, and each R ^{3 is} independently hydrogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, Substituted or unsubstituted C ₂ -C ₁₀ alkenyl, substituted or unsubstituted C ₃ -C ₁₀ alkynyl, substituted or unsubstituted C ₆ -C ₂₀ aryl, or substituted or unsubstituted C ₃ -C ₁₄ Heteroaryl; each R ^{4 is} independently hydrogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₂ -C ₁₀ alkenyl a substituted or unsubstituted C ₃ -C ₁₀ alkynyl group, a substituted or unsubstituted C ₆ -C ₂₀ aryl group, or a substituted or unsubstituted C ₃ -C ₁₄ heteroaryl group; or R ³ and R ⁴ may be together It may form a three to eight yuan three to eight yuan carbocyclic or heterocyclic ring, wherein the heteroatoms may be sulfur, oxygen, NH or NR ^f.

In another preferred embodiment, R ³ is hydrogen.

In another preferred embodiment, R ⁴ is hydrogen.

In another preferred embodiment, the compound is as shown in the formula (II),

In the formula,

Ar is a benzene ring, and Ar may be substituted by one or more groups selected from the group consisting of halogen, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, hydroxy, amino, nitro, aldehyde, -CF ₃ , -CN, -SF ₅ , NR ^a R ^b , carboxyl group, -COR ^a , -CO ₂ C ₁ -C ₆ alkyl group, -CONR ^a R ^b , -S(O)R ^a , -S( O) ₂ R ^a , -S(O)(NH)R ^a , -S(O)(NR ^d )R ^a , -SO ₂ NR ^a R ^b ;

Wherein R ^a , R ^b , R ^d , R ³ , R ⁴ , R ² and R ¹ are as defined above;

n is an integer of 2-6.

In another preferred embodiment, R ¹ is H, C ₁ -C ₁₀ alkyl, C ₃ -C ₁₀ cycloalkyl, C ₆ -C ₂₀ aryl, or C ₃ -C ₁₀ heteroaryl; R ¹ It may be substituted by one or more halogens.

In another preferred embodiment, R ¹ is C ₁ -C ₁₀ alkyl;

R ² is H, C ₁ -C ₁₀ alkyl;

R ³ and R ⁴ are each independently hydrogen, C ₁ -C ₁₀ alkyl or R ³ and R ⁴ may together form a three to eight membered carbocyclic ring or a three to eight membered heterocyclic ring.

In another preferred embodiment, R ¹ is H, C ₁ -C ₃ alkyl; and/or R ² is H, C ₁ -C ₃ alkyl.

In another preferred embodiment, R ¹ , R ² , R ³ , and R ⁴ are each independently H.

In another preferred embodiment, the prodrug of the compound of formula I is as shown in formula (III),

In the formula,

The definitions of R ¹ , R ² , R ³ , R ⁴ , n and Ar are as described above;

Y is R ⁶ , OR ⁶ , NR ^a R ^b ; R ⁶ is a substituted or unsubstituted C ₆ -C ₂₀ aryl group, a substituted or unsubstituted five- or six-membered heteroaryl group, a substituted or unsubstituted C ₁ a -C ₁₂ alkyl, substituted or unsubstituted C ₃ -C ₁₂ cycloalkyl, substituted or unsubstituted C ₃ -C ₁₂ heterocyclic group, wherein the hetero atom may be sulfur, oxygen, NH or NR ^f ;

R ^a and each R ^b are each independently hydrogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₂ -C ₁₀ alkenyl a substituted or unsubstituted C ₆ -C ₂₀ aryl group, or a substituted or unsubstituted C ₃ -C ₁₄ heteroaryl group; R ^a and R ^b may together form a three to eight membered ring or a four to eight membered heterocyclic ring. Wherein the hetero atom may be sulfur, oxygen, NH or NR ^f ; N ^f is as defined above.

Wherein said "substituted" means having one or more substituents selected from the group consisting of halogen, hydroxy, -NH _2, nitro, -CN, C ₁ -C ₄ alkyl, C ₁ -C ₄ haloalkane A C ₁ -C ₄ alkoxy group, a C ₃ -C ₆ cycloalkyl group, a C ₂ -C ₄ alkenyl group, a C ₂ -C ₄ alkynyl group, a phenyl group, a benzyl group.

In another preferred embodiment, each of groups such as X, R ¹ , R ² , R ³ , R ⁴ , Y and Ar are independently the formulas (I), (II), and (III) prepared in the examples. Corresponding groups in each specific compound.

In another preferred embodiment, the compound is selected from the group consisting of:

In another preferred embodiment, R ¹ , R ² , R ³ , R ⁴ , and R ⁵ in the formula (I) are each independently selected from the corresponding groups of the specific compounds of the above table.

In another preferred embodiment, the compound is a specific compound prepared in the examples.

In another preferred embodiment, the compound is a racemate.

In another preferred embodiment, the compound is an enantiomer.

In another preferred embodiment, the pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, hydrobromide, sulfate, phosphate, methanesulfonate, triflate, benzenesulfonic acid Salt, p-toluenesulfonate (tosylate), 1-naphthalenesulfonate, 2-naphthalenesulfonate, acetate, trifluoroacetate, malate, tartrate, citrate, milk Acid, oxalate, succinate, fumarate, maleate, benzoate, salicylate, phenylacetate, mandelate.

In a second aspect of the invention, there is provided a process for the preparation of a compound of formula (I) according to the first aspect, comprising the steps of:

(a) compound AA is reacted with compound CC to give compound BB;

(b) Compound BB is ring-opened under alkaline hydrolysis conditions (such as aqueous sodium hydroxide) to give the final product of formula I;

In each formula, the definitions of R ¹ , R ² , R ³ , R ⁴ , R ⁵ and n are as defined above.

The invention also provides a preparation method of the second compound of the general formula (I), comprising the following steps:

(a) Compound DD is reacted with compound EE and a reducing agent under acid catalysis to give compound BB;

(b) Compound BB is ring-opened under alkaline hydrolysis conditions (e.g., aqueous sodium hydroxide) to give the final compound of formula (I).

The invention also provides a process for the preparation of a third compound of the general formula (I), comprising the steps of:

(a) Compound EE is reacted with H ₂ O ₂ in the presence of TFA to give compound FF;

(b) compound FF and compound GG are reacted to obtain compound AA;

(c) reacting compound AA with compound CC to obtain compound BB;

(d) Compound BB is ring-opened under alkaline hydrolysis conditions (e.g., aqueous sodium hydroxide) to give the final compound of formula (I).

In a third aspect of the invention, there is provided the use of a compound of formula (I) according to the first aspect, for:

(i) preparing a guanamine-2,3-dioxygenase inhibitor; and/or

(ii) A medicament for the prevention and/or treatment of a guanamine-2,3-dioxygenase mediated disease.

In another preferred embodiment, the guanamine-2,3-dioxygenase mediated disease is a disease characterized by the pathology of an IDO-mediated tryptophan metabolism pathway.

In another preferred embodiment, the guanamine-2,3-dioxygenase mediated disease is cancer, eye disease, heart disorder, depression, anxiety, AIDS, Alzheimer's disease, and/or autoimmune disease.

In another preferred embodiment, the cancer includes, but is not limited to, colon cancer, breast cancer, gastric cancer, lung cancer, colon cancer, pancreatic cancer, ovarian cancer, prostate cancer, kidney cancer, liver cancer, brain cancer, melanoma, multiple Myeloma, chronic myeloid leukemia, hematological tumors, lymphoid tumors, including metastatic lesions in other tissues or organs remote from the primary site of the tumor.

According to a fourth aspect of the invention, a pharmaceutical composition comprising:

a compound of the formula (I), or a pharmaceutically acceptable salt thereof, a stereoisomer thereof or a tautomer thereof, or a prodrug thereof;

A pharmaceutically acceptable carrier.

According to a fifth aspect of the invention, a pharmaceutical composition comprising:

a compound of the formula (I), or a pharmaceutically acceptable salt thereof, a stereoisomer thereof or a tautomer thereof, or a prodrug thereof;

Anti-tumor drugs.

In another preferred embodiment, the anti-tumor drug includes, but is not limited to, an immunotherapeutic drug for cancer: PD-1 antibody, CTLA-4 antibody, PD-L1 antibody, PD-L2 antibody, any other chemotherapeutic drug or target To the treatment of the drug.

According to a sixth aspect of the present invention, a method for preventing and/or treating a guanamine-2,3-dioxygenase-mediated disease comprising administering a compound of the formula (I) according to the first aspect to a patient Or the step of the pharmaceutical composition of the fourth or fifth aspect.

In another preferred embodiment, the indoleamine-2,3-dioxygenase mediated disease is cancer, the method further comprising administering to the patient an additional anticancer agent (also known as an antitumor drug, The steps of the antitumor drug as described above).

The compound of the formula (I) of the present invention has various pharmacological activities such as antitumor, treatment of neurodegenerative diseases (Alzheimer's disease), and anti-inflammatory.

It is to be understood that within the scope of the present invention, the various technical features of the present invention and the various technical features specifically described hereinafter (as in the embodiments) may be combined with each other to constitute a new or preferred technical solution. Due to space limitations, we will not repeat them here.

detailed description

The inventors have extensively and intensively studied, and for the first time, unexpectedly developed a novel compound containing a sulfamate and a 1,2,5-oxadiazole structure, which can be used as a highly potent IDO enzyme inhibitor for Prevention and/or treatment of indoleamine-2,3-dioxygenase mediated diseases can also be used as an anti-inflammatory drug. On the basis of this, the present invention has been completed.

Before the present invention is described, it is to be understood that the invention is not limited to the specific methods and experimental conditions described, as such methods and conditions may vary. It should also be understood that the terms used herein are only intended to describe The specific embodiments are not intended to be limiting, and the scope of the invention is limited only by the appended claims.

All technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs, unless otherwise defined. As used herein, when used in reference to a particular recited value, the term "about" means that the value can vary by no more than 1% from the recited value. For example, as used herein, the expression "about 100" includes all values between 99 and 101 and (eg, 99.1, 99.2, 99.3, 99.4, etc.).

Although any methods and materials similar or equivalent to those described in the present invention can be used in the practice or testing of the present invention, the preferred methods and materials are exemplified herein.

definition

The term "alkyl" refers to a monovalent saturated aliphatic hydrocarbon group having from 1 to 10 carbon atoms, including straight-chain and branched hydrocarbon groups such as methyl (ie, CH ₃ -), ethyl (ie, CH ₃ CH ₂ -). , n-propyl (ie CH ₃ CH ₂ CH ₂ -), isopropyl (ie (CH ₃ ) ₂ CH-), n-butyl (ie CH ₃ CH ₂ CH ₂ CH ₂ -), isobutyl (ie (CH ₃ ) ₂ CHCH ₂ -), sec-butyl (ie (CH ₃ )(CH ₃ CH ₂ )CH-), tert-butyl (ie (CH ₃ ) ₃ C-), n-pentyl (ie CH _{3 )} CH ₂ CH ₂ CH ₂ CH ₂ -), neopentyl (ie (CH ₃ ) ₃ CCH ₂ -). In the present invention, the term includes a substituted or unsubstituted alkyl group.

The term "substituted or unsubstituted" as used herein means that the group may be unsubstituted or that H in the group is one or more (preferably 1-6, more preferably 1- Replaced by three substituents.

As used herein, "substituted" or "substituted" means that the group has one or more (preferably 1-6, more preferably 1-3) substituents selected from the group consisting of: Halogen, hydroxy, -NH ₂ , nitro, -CN, C ₁ -C ₄ alkyl, C ₁ -C ₄ haloalkyl, C ₁ -C ₄ alkoxy, C ₃ -C ₆ cycloalkyl, C ₂ -C ₄ alkenyl, C ₂ -C ₄ alkynyl, phenyl, benzyl.

As used herein, the term "cycloalkyl" refers to a substituted or unsubstituted C ₃ -C ₁₂ cycloalkyl.

As used herein, the term "alkoxy" refers to -O-alkyl, wherein the alkyl group can be saturated or unsaturated, can be branched, straight chain, or cyclic. Preferably, the alkoxy group has 1 to 10 carbon atoms, preferably 1 to 6 carbon atoms. Representative examples include, but are not limited to, methoxy, ethoxy, propoxy.

As used herein, the term "aryl" refers to a monovalent aromatic carbocyclic group of 6 to 20 (preferably 6 to 14) carbon atoms which has a monocyclic (eg phenyl) or fused ring (eg naphthyl) Or sulfhydryl), if the point of attachment is on the aromatic carbon, the fused ring may be non-aromatic (eg 2-benzoxazolone, 2H-1,4-benzoxazine-3(4H)-one-7 - base, etc.). Preferred aryl groups include phenyl and naphthyl. The term includes substituted or unsubstituted forms wherein the substituents are as defined above.

The term "alkenyl" as used herein refers to an alkenyl group having 2 to 10 (eg 2 to 6 or 2 to 4) carbon atoms and having at least 1 (eg 1 to 2) unsaturated ethylenic bonds (>C) =C<). Examples of such groups are vinyl, allyl, but-3-enyl. As used herein, the term "cycloalkyl" refers to a cyclic alkyl group having from 3 to 10 carbon atoms having a single or multiple ring (including fused systems, bridged ring systems, and spiro ring systems). In a fused ring system, one or more of the rings may be a cycloalkyl, heterocyclic, aryl or heteroaryl group as long as the attachment site is a ring through a cycloalkyl group. Examples of suitable cycloalkyl groups include, for example, adamantyl, cyclopropyl, cyclobutyl, cyclopentyl and cyclooctyl.

As used herein, the term "halo" or "halogen" refers to fluoro, chloro, bromo and iodo.

As used herein, the term "heteroaryl" refers to an aromatic group having from 1 to 10 carbon atoms and from 1 to 4 heteroatoms selected from the group consisting of oxygen, nitrogen and sulfur, such heteroaryl groups may be monocyclic Pyridyl Or a furyl group or a fused ring (such as indolizinyl or benzothienyl), wherein the fused ring may be non-aromatic and/or contain a hetero atom as long as the point of attachment is through aromatic heteroaryl The atom of the base. In one embodiment, the ring atom nitrogen and/or sulfur of the heteroaryl group is optionally oxidized to an N-oxide (N-O), a sulfinyl group or a sulfonyl group. Preferred heteroaryl groups include pyridinyl, pyrrolyl, indolyl, thienyl and furanyl. The term includes substituted or unsubstituted heteroaryl.

The term "substituted heteroaryl" as used herein refers to a heteroaryl group substituted with 1 to 5, preferably 1 to 3, more preferably 1 to 2 substituents selected from the group consisting of The same substituent as defined by the aryl group.

As used herein, the term "heterocycle" or "heterocycle" or "heterocycloalkyl" or "heterocyclyl" refers to a saturated, partially saturated or unsaturated group (but not aromatic), Has a single ring or a fused ring (including a bridged ring system and a spiro ring system having from 1 to 10 carbon atoms and from 1 to 4 heteroatoms selected from nitrogen, sulfur or oxygen in the ring, in a fused ring system, one or more The ring may be a cycloalkyl, aryl or heteroaryl group as long as the point of attachment passes through the non-aromatic ring. In one embodiment, the nitrogen and/or sulfur atom of the heterocyclic group is optionally oxidized to provide N-oxide, sulfinyl and sulfonyl moieties.

The term "substituted heterocyclic" or "substituted heterocycloalkyl" or "substituted heterocyclic" as used herein refers to a heterocyclic group substituted by 1 to 5 (eg, 1 to 3) substituents. The substituent is the same as the substituent defined by the substituted cycloalkyl group.

As used herein, the term "stereoisomer" refers to a chiralally different compound of one or more stereocenters. Stereoisomers include enantiomers and diastereomers.

As used herein, the term "tautomer" refers to an alternative form of a compound having a different proton position, such as an enol-ketone and an imine-enamine tautomer, or a tautomeric form of a heteroaryl group. The heteroaryl group comprises a ring atom attached to the -NH- moiety of the ring and the =N- moiety of the ring, such as pyrazole, imidazole, benzimidazole, triazole and tetrazole.

"Prodrug" refers to any derivative of an embodiment compound that, when administered to a subject, is capable of providing, directly or indirectly, a compound of the Examples, or an active metabolite or residue thereof. Particularly preferred derivatives and prodrugs are those which, when administered to a subject, increase the bioavailability of the compound of the example (eg, the compound administered orally is more readily absorbed into the blood) or the parent compound relative to the parent species Derivatives and prodrugs to the bioregional compartment (such as the brain or lymphatic system). Prodrugs include the ester form of the compounds of the invention.

Compound of the invention

As used herein, the term "compound of the invention" refers to a compound of formula (I), formula (II) or formula (III), a deuterated compound thereof, a racemate, a stereoisomer thereof or a mutual variation thereof. A construct, or a prodrug, or a pharmaceutically acceptable salt thereof. The present invention relates to a racemic mixture of these compounds, enriched in a mixture of any one of the enantiomers, and any of the separated enantiomers. For the scope of the invention, it is to be understood that the racemic mixture refers to a mixture of two R and S enantiomers of 50%: 50%. The isolated enantiomer is understood to be the pure enantiomer (ie 100%) or highly enriched in an enantiomer (purity ≥ 98%, ≥ 95%, ≥ 93%, ≥ 90%, ≥ 88%) , ≥85%, ≥80%) of the mixture.

Typically, the invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, a stereoisomer thereof, or a tautomer thereof, or a prodrug thereof,

In the formula,

R ¹ , R ² , R ³ and R ⁴ are each independently hydrogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₂ -C ₁₀ alkenyl group, a substituted or unsubstituted C ₃ -C ₁₀ alkynyl group, a substituted or unsubstituted C ₆ -C ₂₀ aryl group, a substituted or unsubstituted C ₃ -C ₁₄ heteroaryl group; R ¹ and R ² , or R ³ and R ⁴ may together form a three- to eight-membered carbocyclic ring or a three- to eight-membered heterocyclic ring, or that R ¹ and R ³ together may form a five- to eight-membered carbocyclic ring or a five- to eight-membered heterocyclic ring. Wherein the hetero atom may be sulfur, oxygen, NH or NR ^f ;

R ⁵ is C ₆ -C ₂₀ aryl, 5- or 6-membered heteroaryl; R ⁵ may be substituted by one or more groups selected from the group consisting of halogen, C ₁ -C ₆ alkyl, C ₁ - C ₆ alkoxy, hydroxy, amino, nitro, aldehyde, -CF ₃ , -CN, -SF ₅ , NR ^a R ^b , carboxy, -COR ^a , -CO ₂ C ₁ -C ₆ alkyl, - CONR ^a R ^b , -S(O)R ^a , -S(O) ₂ R ^a , -S(O)(NH)R ^a , -S(O)(NR ^d )R ^a , -SO ₂ NR ^a R ^b , -P(O)Me ₂ , -P(O)(OMe) ₂ ; wherein each R ^a and each R ^b are independently hydrogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl, substituted or not Substituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₂ -C ₁₀ alkenyl, substituted or unsubstituted C ₆ -C ₂₀ aryl, or substituted or unsubstituted C ₃ -C ₁₄ heteroaryl And R ^a and R ^b may together form a three to eight membered ring or a four to eight membered heterocyclic ring, wherein the hetero atom may be sulfur, oxygen, NH or NR ^f ;

X is a single bond, O, S, NH or NR ^d ;

R ^d and R ^{f are} independently C ₁ -C ₁₀ alkyl, C ₃ -C ₁₀ cycloalkyl, C ₆ -C ₂₀ aryl, or C ₃ -C ₁₄ heteroaryl;

n is 2 to 8.

In another preferred embodiment, it said "substitution" refers to the group having one or more substituents selected from: halogen, hydroxy, -NH _2, nitro, -CN, C ₁ -C ₄ alkyl, C _1- C ₄ haloalkyl, C ₁ -C ₄ alkoxy, C ₃ -C ₆ cycloalkyl, C ₂ -C ₄ alkenyl, C ₂ -C ₄ alkynyl, phenyl, benzyl.

In another preferred embodiment, X is NH.

In another preferred embodiment, R ³ and R ⁴ are each independently hydrogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl; R ³ and R ⁴ may together form a three to eight membered ring or a three to eight membered impurity. A ring wherein the hetero atom can be sulfur, oxygen, NH or NR ^f .

In another preferred embodiment, n is from 2 to 6.

In another preferred embodiment, R ³ is hydrogen.

In another preferred embodiment, R ⁴ is hydrogen.

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

In the formula,

Ar is a benzene ring, and Ar may be substituted by one or more groups selected from the group consisting of halogen, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, hydroxy, amino, nitro, aldehyde, -CF ₃ , -CN, -SF ₅ , NR ^a R ^b , carboxyl group, -COR ^a , -CO ₂ C ₁ -C ₆ alkyl group, -CONR ^a R ^b , -S(O)R ^a , -S( ^{_{O) 2 R a, -S (}} O) (NH) R a, -S (O) (NR d) R a, -SO 2 NR a R b;

Wherein R ^a , R ^b , R ^d , R ³ , R ⁴ , R ² and R ¹ are as defined above;

n is 2-6.

In another preferred embodiment, R ¹ is H, C ₁ -C ₁₀ alkyl, C ₃ -C ₁₀ cycloalkyl, C ₆ -C ₂₀ aryl, or C ₃ -C ₁₀ heteroaryl; R ¹ It may be substituted by one or more halogens.

In another preferred embodiment, R ¹ is C ₁ -C ₁₀ alkyl;

R ² is H, C ₁ -C ₁₀ alkyl;

R ³ and R ⁴ are each independently hydrogen, C ₁ -C ₁₀ alkyl or R ³ and R ⁴ may together form a three to eight membered carbocyclic ring or a three to eight membered heterocyclic ring.

In another preferred embodiment, R ¹ , R ² , R ³ , and R ⁴ are each independently H.

In another preferred embodiment, the compound of the formula (I) is as shown in the formula (III),

In the formula,

The definitions of R ¹ , R ² , R ³ , R ⁴ , n and Ar are as described above;

Y is R ⁶ , OR ⁶ , NR ^a R ^b ; R ⁶ is a substituted or unsubstituted C ₆ -C ₂₀ aryl group, a substituted or unsubstituted five- or six-membered heteroaryl group, a substituted or unsubstituted C ₁ a -C ₁₂ alkyl, substituted or unsubstituted C ₃ -C ₁₂ cycloalkyl, substituted or unsubstituted C ₃ -C ₁₂ heterocyclic group, wherein the hetero atom may be sulfur, oxygen, NH or NR ^f ;

R ^a and each R ^b are each independently hydrogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₂ -C ₁₀ alkenyl a substituted or unsubstituted C ₆ -C ₂₀ aryl group, or a substituted or unsubstituted C ₃ -C ₁₄ heteroaryl group; R ^a and R ^b may together form a three to eight membered ring or a four to eight membered heterocyclic ring. Wherein the hetero atom may be sulfur, oxygen, NH or NR ^f ; Nf is as defined above.

Wherein said "substituted" means having one or more substituents selected from the group consisting of halogen, hydroxy, -NH _2, nitro, -CN, C ₁ -C ₄ alkyl, C ₁ -C ₄ haloalkane A C ₁ -C ₄ alkoxy group, a C ₃ -C ₆ cycloalkyl group, a C ₂ -C ₄ alkenyl group, a C ₂ -C ₄ alkynyl group, a phenyl group, a benzyl group.

Where a compound of the invention is present as a stereoisomer, the invention includes all stereoisomers of the compound.

In the case where the compounds of the invention are present as tautomers, the invention includes all tautomers of the compounds.

The invention also includes deuterated compounds resulting from the substitution of any one or more of the hydrogen atoms of the compound by its stable isotope oxime.

Unless otherwise specified, among all compounds of the present invention, each chiral carbon atom (chiral center) may optionally be in the R configuration or the S configuration, or a mixture of the R configuration and the S configuration.

Pharmaceutical composition

The invention also provides a pharmaceutical composition comprising an active ingredient in a safe and effective amount, And a pharmaceutically acceptable carrier.

The "active ingredient" as used in the present invention means a compound of the formula (I), or a pharmaceutically acceptable salt thereof, a stereoisomer thereof or a tautomer thereof, or a prodrug thereof.

The "active ingredient" and pharmaceutical compositions described herein are useful as IDO inhibitors. In another preferred embodiment, a medicament for the preparation and prevention or/or treatment of a tumor is prepared. In another preferred embodiment, a medicament for the preparation of a disease preventing and/or treating IDO mediated diseases.

By "safe and effective amount" is meant that the amount of active ingredient is sufficient to significantly improve the condition without causing serious side effects. In general, the pharmaceutical compositions contain from 1 to 2000 mg of active ingredient per dose, more preferably from 10 to 200 mg of active ingredient per dose. Preferably, the "one dose" is a tablet.

"Pharmaceutically acceptable carrier" means: one or more compatible solid or liquid fillers or gel materials which are suitable for human use and which must be of sufficient purity and of sufficiently low toxicity. By "compatibility" it is meant herein that the components of the composition are capable of intermingling with the active ingredients of the present invention and with respect to each other without significantly reducing the efficacy of the active ingredients.

The compounds of the preferred embodiments of the invention may be administered as separate active agents or in combination with one or more other agents useful in the treatment of cancer. The use of the compounds of the preferred embodiments of the invention in combination with known therapeutic agents and anticancer agents is also effective, and combinations of currently known compounds with other anticancer or chemotherapeutic agents are within the scope of the preferred embodiments. Examples of such agents can be found in the Cancer Principles and Practice of Oncology, VT Devita and S. Hellman (editor), 6th edition (February 15, 2001), published by Lippincott Will iams & Wilkins. Society. One of ordinary skill in the art will be able to discern effective combinations of agents based on the particular nature of the drug and the cancer involved. Such anticancer agents include, but are not limited to, HDAC inhibitors, estrogen receptor modulators, androgen receptor modulators, retinol receptor modulators, cytotoxic/cytostatic agents, anti-proliferative Agents, prenyl-protein transferase inhibitors, HMG-CoA reductase inhibitors and other angiogenesis inhibitors, cell proliferation and survival signal inhibitors, apoptosis inducers, and cell cycle checkpoints Reagent, CTLA4 antibody, PD-1 antibody, PD-L1 antibody, and the like. The compounds of the preferred embodiments are also effective when administered concurrently with radiation therapy.

In general, the compounds of the preferred embodiments will be administered in a therapeutically effective amount by any of the accepted modes of administration of agents having similar effects. The actual amount of the compound (i.e., active ingredient) of the preferred embodiment is determined by a number of factors, such as the severity of the condition to be treated, the age and relative health of the patient, the potency of the compound being used, the route and form of administration, and other factors. . The drug can be administered multiple times a day, preferably once or twice a day. All of these factors are within the consideration of the attending physician.

For the purposes of the preferred embodiment, the therapeutically effective dose can generally be the total daily dose administered to the patient in a single or divided dose, for example, from about 0.001 to about 1000 mg/kg body weight per day, preferably from about 1.0 to about daily. 30 mg / kg body weight. A Dosage unit composition can include its dosage factor to form a daily dose. The choice of dosage form will depend on various factors such as the mode of administration and the bioavailability of the drug substance. In general, the compounds of the preferred embodiments can be administered as a pharmaceutical composition by any of the following routes: oral, systemic (e.g., transdermal, intranasal or by suppository), or parenteral (e.g., intramuscular, intravenous). Or subcutaneous). The preferred mode of administration is oral, and a convenient daily dose can be adjusted depending on the degree of bitterness. The compositions may take the form of tablets, pills, capsules, semi-solids, powders, sustained release formulations, solutions, suspensions, elixirs, aerosols or any other suitable compositions. Another preferred mode of administration of the preferred embodiment compounds is inhalation. This is an effective method of delivering a therapeutic agent directly to the respiratory tract (see, e.g., U.S. Patent No. 5,607,915).

Suitable pharmaceutically acceptable carriers or excipients include, for example, treatment and drug delivery modifiers and accelerators such as calcium phosphate, magnesium stearate, talc, monosaccharides, disaccharides, starches, gelatin, cellulose , sodium methylcellulose, carboxymethylcellulose, glucose, hydroxypropyl-B-cyclodextrin, polyvinylpyrrolidone, low melting wax, ion exchange resin, and the like, and combinations of any two or more thereof. The liquid and semi-solid excipients may be selected from the group consisting of glycerin, propylene glycol, water, ethanol, and various oils, including petroleum, animal, vegetable, or synthetic sources such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. Preferred liquid carriers, particularly carriers for injectable solutions, include water, saline, aqueous dextrose and ethylene glycol. Other suitable pharmaceutically acceptable excipients are described in Remington&apos;s Pharmaceutical Sciences, Mack Pub. Co., New Jersey (1991), incorporated herein by reference.

The term "pharmaceutically acceptable salt," as used herein, refers to a non-toxic acid or alkaline earth metal salt of a compound of formula I. These salts can be prepared in situ by final isolation and purification of the compound of formula I, or by separately reacting a suitable organic or inorganic acid or base with a basic or acidic functional group. Representative salts include, but are not limited to, acetate, adipate, alginate, citrate, aspartate, benzoate, besylate, hydrogen sulfate, butyrate , camphorate, camphor sulfonate, digluconate, cyclopentane propionate, lauryl sulfate, ethanesulfonate, glucose heptanoate, glycerol phosphate, hemisulfate, heptanoic acid Salt, hexanoate, fumarate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, nicotinate , 2-naphthyl sulfonate, oxalate, pamoate, pectate, thiocyanate, 3-phenylpropionate, picrate, pivalate, propionate, Succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate and undecanoate. Further, the nitrogen-containing basic group can be quaternized by the following reagents: alkyl halides such as methyl, ethyl, propyl, butyl chloride, bromide and iodide; dialkyl sulfate Such as dimethyl, diethyl, dibutyl and dipentyl sulfate; long chain halides such as sulfhydryl, lauryl, myristyl and stearyl chlorides, bromides and iodides; aralkyl Base halides such as benzyl and phenethyl bromide. A water soluble or oil soluble or dispersible product is thus obtained. Examples of the acid which can be used to form a pharmaceutically acceptable acid addition salt include inorganic acids such as hydrochloric acid, sulfuric acid, phosphoric acid, and organic acids such as oxalic acid, maleic acid, methanesulfonic acid, succinic acid, and citric acid. The base addition salt can be prepared in situ upon final isolation and purification of the compound of formula I, or by separately reacting the carboxylic acid moiety with a suitable base such as a hydroxide, carbonate or carbonate of a pharmaceutically acceptable metal cation. Hydrogen salt) or ammonia, or organic primary, secondary or tertiary amine reaction. Pharmaceutically acceptable salts include, but are not limited to, alkali metal and alkaline earth metal based cations such as sodium, lithium, potassium, calcium, magnesium, aluminum, and the like, as well as non-toxic ammonium, quaternary ammonium, and amine cations, including However, it is not limited to: ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine and the like. Other representative organic amines useful for the formation of base addition salts include diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, and the like.

As used herein, the term "pharmaceutically acceptable prodrug" refers to a prodrug of a compound of those preferred embodiments, which is rapidly converted in vivo to a parent compound of the above formula, for example, hydrolyzed in blood. In "T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, 14 volumes of the ACS15Symposium Series" and "Edward B. Roche, Bioreversible Vectors in Drug Design" A complete discussion is provided in Bioreversible Carriers in Drug Design, The American Pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated herein by reference.

The invention is further illustrated below in conjunction with specific embodiments. It is to be understood that the examples are not intended to limit the scope of the invention. Experimental parties not showing specific conditions in the following examples The method is usually carried out according to the conditions described in conventional conditions such as Sambrook et al., Molecular Cloning: Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or in accordance with the conditions recommended by the manufacturer. Percentages and parts are by weight unless otherwise stated.

The invention is advantageous in that:

(1) providing a novel structure of the compound of the formula (I);

(2) The compound of the present invention can be used as a highly potent IDO enzyme inhibitor;

(3) The synthesis method is mild, the operation is simple and easy, the yield is high, and the derivatization is easy, and it is suitable for industrial scale production;

(4) It has various pharmacological activities such as anti-tumor, neurodegenerative diseases (Alzheimer's disease), and treatment of AIDS.

Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those skilled in the art. In addition, any methods and materials similar or equivalent to those described may be employed in the methods of the invention. The preferred embodiments and materials described herein are for illustrative purposes only.

Preparation of Compound 1 of Example 1

First step: preparation of compound D

Propyl cyanohydrin (32.0 g, 485 mmol) was dissolved in 600 mL of water and heated to complete dissolution. Under ice cooling, sodium nitrite (38.0 g, 533 mmol) and 6N hydrochloric acid (5.5 mL) were added. Under the ice bath After reacting for 0.5 hour, the mixture was heated to room temperature for 2 hours. The reaction solution was cooled in an ice bath, and a 50% aqueous solution of hydroxylamine hydrochloride (99.0 g, 1500 mmol) was added dropwise to the reaction mixture, stirred for half an hour, and then allowed to react to room temperature for 1 hour. The reaction was heated to reflux for 2 hours. After completion of the reaction, the mixture was adjusted to pH 7.0 with 80 mL of 6N hydrochloric acid. The precipitate was filtered, washed once with water and ethyl acetate and dried to yield 69.3 g of white compound.

^{13 C NMR (75MHz, DMSO-} d 6): δ154.4,144.0,140.0.

MS ESI: m/z = 144.0, [M+H] ⁺ .

Step 2: Preparation of Compound E

Compound D (30.0 g, 210 mmol) was dissolved in 213 mL of acetic acid, and then 420 mL of water, 105 mL of hydrochloric acid and sodium chloride (36.7 g, 629 mmol) were added. Under ice cooling, sodium nitrite (14.2 g, 206 mmol) dissolved in 50 mL of water was added dropwise, and the reaction was kept at zero temperature for 1 hour, and then the temperature was raised to room temperature for 5 hours. After the completion of the reaction, the precipitate was filtered, and the filtered cake was washed with water, dried, evaporated, evaporated, evaporated, evaporated.

¹³ C NMR (75 MHz, DMSO-d ₆ ): δ 154.4, 142.3, 126.9.

MS ESI: m/z = 160.9, [M+H] ^- .

The third step: preparation of compound F

Compound E (15.0 g, 92 mmol) was dissolved in ethyl acetate (200 mL), and the mixture was stirred, and then, 2-methoxy-1-ethylamine (7.6 mL, 101 mmol) was added to the reaction mixture. After stirring for ten minutes, triethylamine was added. (14.0, 138 mmol), reacted for 1 hour. The reaction solution was washed once with water, brine and dried over anhydrous sodium sulfate.

^{1 H NMR (300MHz, DMSO-} d 6): δ10.66 (s, 1H), 6.27 (s, 2H), 6.14 (t, 1H), 3.50 (m, 2H), 3.35 (t, 2H), 3.18 (s, 3H).

The fourth step: preparation of compound G

Compound F (15.0 g, 75.0 mmol) was added to 65 mL of water, potassium hydroxide (12.5 g, 224 mmol) and refluxed for 30 hr. The reaction solution was cooled to room temperature and extracted with ethyl acetate three times. The organic layer was dried, and the solvent was evaporated.

^{1 H NMR (300MHz, DMSO-} d 6): δ10.54 (s, 1H), 6.21 (s, 1H), 6.13 (t, 1H), 3.49 (m, 2H), 3.36 (t, 2H), 3.25 (s, 3H).

Step 5: Preparation of Compound H

Compound G (9.0 g, 45 mmol) was dissolved in 50 mL of acetic acid, and then 50 mL of water, 45 mL of hydrochloric acid, and sodium chloride (7.8 g, 135 mmol) were added. Under ice cooling, sodium nitrite (3.2 g, 4.3 mmol) dissolved in 18 mL of water was added dropwise, and the reaction was kept at zero temperature for 1 hour, and then the temperature was raised to room temperature for 5 hours. After completion of the reaction, ethyl acetate was extracted three times, and the organic layer was combined, washed with water and brine, and the solvent was evaporated to give a white solid compound 8.0 g.

¹ H NMR (300 MHz, DMSO-d ₆ ): δ 13.41 (s, 1H), 5.88 (t, 1H) 3.53 (m, 2H), 3.39 (t, 2H), 3.25 (s, 3H).

Step 6: Preparation of Compound I

Compound H (8.0 g, 36 mmol) was dissolved in 50 mL water, heated to 60 ° C, 3-bromo-4-fluoroaniline (6.8 g, 36 mmol) was added and stirred for ten minutes, then aqueous sodium bicarbonate (4.6 g, 54 mmol) ). After reacting for 1 hour, it was cooled to room temperature. After completion of the reaction, ethyl acetate was extracted three times, and the organic layer was combined and evaporated to dryness

^{1 H NMR (300MHz, DMSO-} d 6): δ11.53 (s, 1H), 8.88 (s, 1H), 7.21 (t, 1H), 7.09 (m, 1H), 6.76 (m, 1H), 6.14 (t, 1H), 3.51 (m, 2H), 3.37 (m, 2H), 3.26 (s, 3H).

Step 7: Preparation of Compound J

Compound I (15.0 g, 40 mmol) was dissolved in 150 mL of ethyl acetate, and carbonyldiimidazole (9.8 g, 60 mmol) was added, and the mixture was heated to 60 ° C for 1 hour. The solution was cooled to room temperature, and extracted with EtOAc EtOAc.

^{1 H NMR (300MHz, DMSO-} d 6): δ8.06 (dd, 1H), 7.69 (m, 1H), 7.58 (t, 1H), 6.40 (s, 1H), 3.48 (m, 2H), 3.39 (m, 2H), 3.25 (s, 3H).

Step 8: Preparation of Compound K

Compound J (12.5 g, 31 mmol) was dissolved in 95 mL and cooled to -67 °, boron tribromide (15.7 mL, 62.6 mmol) was slowly added, and after the addition, the reaction mixture was slowly allowed to react to -10 ° for 1 hour. After the completion of the reaction, the pH was adjusted to 7. by a slow-saturated sodium carbonate solution in the mixture, and the mixture was extracted three times with ethyl acetate. The organic layer was combined, dried and evaporated to give a white solid. 91%.

_{^{1 H NMR (300MHz, CDCl 3}} ): δ7.64 (dd, 1H), 7.36 (m, 1H), 7.31 (m, 1H), 5.64 (s, 1H), 3.93 (m, 2H), 3.59 (m , 1H).

Step 9: Preparation of Compound L

Under ice-cooling, chlorosulfonyl isocyanate (139 mg, 0.98 mmol) was dissolved in 2 mL of dichloromethane, and formic acid (45 mg, 0.98 mmol) was slowly added dropwise, and the reaction was kept at zero temperature for 1 hour, and then stirred at room temperature overnight. Compound K (100 mg, 0.26 mmol) was dissolved in 1 mL of DMA under ice-cooling, and was slowly added dropwise to the above reaction mixture, and stirring was continued for 3 hours. Water was added, and ethyl acetate was extracted three times. The organic layer was combined and evaporated to dryness

^{1 H NMR (400MHz, DMSO-} d 6): δ8.11 (dd, 1H), 7.75 (m, 1H), 7.71 (t, 1H), 6.72 (t, 1H), 4.22 (t, 2H), 3.61 (m, 2H).

Step 10: Preparation of Compound 1

The compound L (60 mg, 0.13 mmol) was dissolved in 2 mL of tetrahydrofuran, and 0.5 mL of a 2N sodium hydroxide solution was added thereto in an ice bath, and the mixture was reacted at room temperature for 1 hour, and the pH was adjusted to 4-5 with 2N hydrochloric acid. The mixture was extracted with EtOAc (3 mL).EtOAc.

^{1 H NMR (400MHz, DMSO-} d 6): δ11.56 (s, 1H), 8.94 (s, 1H), 7.56 (s, 2H), 7.16 (t, 1H), 7.13 (m, 1H), 6.79 (m, 1H), 6.35 (t, 1H), 4.21 (t, 2H), 3.57 (m, 2H);

MS ESI: m/z = 439.0, [M+H] ⁺ .

Preparation of Compound 2 of Example 2

According to the preparation method of Example 1, Compound K (50 mg, 0.13 mmol) was dissolved in 1 mL of DMA, and methylaminosulfonyl chloride (34 mg, 0.26 mmol) was added dropwise thereto under ice-cooling, and reacted at room temperature for 3 hours, then the reaction mixture was poured into 3 mL of water. The precipitated solid was collected by filtration to give 60 mg of Intermediate, which was obtained as white solid.

¹ H NMR (400 MHz, CDCl ₃ ): δ 7.46 (s, 1H), 7.24 (dd, 1H), 7.07 (t, 1H), 6.95 (m, 1H), 6.92 (s, 1H), 6.15 (t, 1H), 4.53 (d, 1H), 4.38 (t, 2H), 3.71 (q, 3H), 2.81 (d, 3H);

MS ESI: m / z = 452.6 , [M + H] +.

Preparation of Compound 3 of Example 3

Compound K (50 mg, 0.13 mmol) was reacted with ethylamine sulfonyl chloride according to the procedure of Example 2, and then hydrolyzed to afford Example 3 as a white solid.

¹ H NMR (400 MHz, CDCl ₃ ): δ 7.55 (s, 1H), 7.26 (dd, 1H), 7.03 (t, 1H), 6.93 (m, 1H), 6.92 (s, 1H), 6.14 (t, 1H), 4.58 (t, 1H), 4.38 (t, 2H), 3.72 (m, 2H), 3.21 (m, 2H), 1.20 (t, 3H);

MS ESI: m/z = 466.6, [M+H] ⁺ .

Preparation of Compound 4 of Example 4

Compound K (50 mg, 0.13 mmol) was reacted with isopropylaminosulfonyl chloride according to the procedure of Example 2, and then hydrolyzed to afford Example 4 as a white solid.

_{^{1 HNMR (400MHz, CDCl 3)}} : δ7.46 (s, 1H), 7.24 (dd, 1H), 7.06 (t, 1H), 6.94 (m, 1H), 6.91 (s, 1H), 6.14 (t, 1H), 4.40 (m, 3H), 3.72 (m, 2H), 3.72 (m, 2H), 1.23 (s, 3H), 1.21 (s, 3H);

MS ESI: m/z = 480.7, [M+H] ⁺ .

Preparation of Compound 5 of Example 5

The first step: 2,2-dimethoxyethanol (50 mg, 0.47 mmol) was dissolved in 3 mL of dichloromethane, and ice-cooled, sodium hydrogen (38 mg, 0.94 mmol) was added. After stirring for half an hour, slowly add two. Methylaminosulfonyl chloride (138 mg, 0.94 mmol) was stirred at room temperature for 4 hr, then concentrated, evaporated, evaporated, evaporated, evaporated Product M of a white solid.

¹ H NMR (400 MHz, CDCl ₃ ): δ 7.27 (s, 1H), 4.63 (t, 3H), 4.10 (d, 2H), 3.43 (s, 6H), 2.90 (s, 6H);

MS ESI: m/z = 236.1, [M+Na] ⁺ .

The second step: compound M (53 mg, 0.16 mmol), triethylsilane (125 uL, 0.78 mmol), methanesulfonic acid (80 uL, 1.23 mmol) was dissolved in 5 mL of dichloromethane, then iced, 4-amino-1,2,5-oxadiazol-3-yl)-4-(3-bromo-4-fluorophenyl) 1,2,4-oxazolidine-5(4H)-one (50 mg , 0.23 mmol), slowly rose to room temperature for 9 hours. The aqueous sodium hydrogencarbonate solution was adjusted to pH 7, and then washed with water and a saturated NaCI solution, and the organic phase was concentrated under reduced pressure. The crude product was purified by silica gel column, and the mobile phase was 3:1 to 1:1 n-hexane/ethyl acetate. 50 mg of product N as a white solid.

_{^{1 HNMR (400MHz, CDCl 3)}} : δ7.65 (dd, 1H), 7.35 (m, 1H), 7.33 (m, 1H), 5.66 (t, 1H), 4.38 (t, 2H), 3.76 (m, 2H), 2.90 (s, 6H);

MS ESI: m/z = 492.7, [M+H] ⁺ .

The third step: the compound N (50 mg, 0.10 mmol) was dissolved in 3 mL of tetrahydrofuran, then ice-cold, 0.5 mL of 2N sodium hydroxide solution was added, and the reaction was slowly warmed to room temperature for 3 hours, then the reaction mixture was concentrated under reduced pressure, and ethyl acetate was added. The ester and water were added to the concentrate, and the organic phase was separated by stirring. The organic phase was dried with MgSO ₄ and evaporated. 25 mg of product as a white solid.

_{^{1 HNMR (400MHz, CDCl 3)}} : δ7.43 (s, 1H), 7.23 (dd, 1H), 7.06 (t, 1H), 6.94 (m, 1H), 6.90 (s, 1H), 6.16 (t, 1H), 4.40 (m, 3H), 3.69 (m, 2H), 2.89 (s, 3H), 2.88 (s, 3H);

MS ESI: m/z = 467.0, [M+H] ⁺ .

Preparation of Compound 6 of Example 6

3-(4-Amino-1,2,5-oxadiazol-3-yl)-4-(3-chloro-4-fluorophenyl) 1,2,4-oxazolidine-5(4H) The ketone and the compound M were used as starting materials, and the method described in Example 5 gave Example 6 as a white solid.

^{1 HNMR (400MHz, Acetone-d6} ): δ10.67 (s, 1H), 8.14 (s, 1H), 7.18 (t, 1H), 7.15 (m, 1H), 6.98 (m, 1H), 6.29 (bs , 1H), 4.37 (t, 2H), 3.68 (dt, 2H), 2.87 (s, 6H).

MS ESI: m/z = 423.0, [M+H] ⁺ .

Preparation of Compound 7 of Example 7

3-(4-Amino-1,2,5-oxadiazol-3-yl)-4-(3-trifluoromethyl-4-fluorophenyl) 1,2,4-oxazolidine-5 The (4H)-one and the compound M were used as a starting material.

¹ H NMR (400 MHz, Acetone-d6): δ 10.75 (s, 1H), 8.28 (s, 1H), 7.35 (d, 1H), 7.29 (d, 1H), 7.28 (d, 1H), 6.30 (bs) , 1H), 4.37 (t, 2H), 3.69 (d, 2H), 2.87 (s, 6H).

MS ESI: m/z = 457.0, [M+H] ⁺ .

Preparation of Compound 8 of Example 8

Using the method described in Example 5, the target compound.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ8.32 (s, 1H), 7.47-7.44 (m, 1H), 7.337.29 (m, 2H), 7.21-7.19 (m, 1H), 2.29-6.28 ( m, 1H), 4.39-4.37 (m, 2H), 3.71-3.69 (m, 2H), 2.90-2.89 (m, 6H).

MS ESI: m/z = 438.1, [M+H] ⁺ .

Example 9: Preparation of Compound 9

First step: preparation of compound O

The compound NN (500 mg, 1.46 mmol) was added to the reaction flask, and trifluoroacetic acid (5.0 mL) and aqueous hydrogen peroxide (5 mL) were added at room temperature, and the mixture was heated at 60 ° C for 12 hours, and the starting materials disappeared, and ethyl acetate was added. (20 mL) and water (20 mL), EtOAc (EtOAc)EtOAc. Concentrated by filtration to give crude compound (500 mg, 91% yield)

The second step: preparation of compound P

Compound O (500 mg, 1.34 mmol) was dissolved in tetrahydrofuran (5 mL), and 3-amino 1-propanol (147.0 mg, 2.02 mmol) was added and reacted for 10 minutes at room temperature, and the starting material disappeared completely. Ethyl acetate (20 mL) and water (20 mL) were evaporated and evaporated. The mixture was concentrated by filtration to give a crude compound, which was purified by column chromatography to afford compound P (320 mg, 62% yield).

¹ H NMR (400 MHz, CDCl ₃ ): δ 7.64 - 7.62 (m, 1H), 7.36-7.29 (m, 2H), 5.76-7.74 (m, 1H), 3.84-3.81 (m, 2H), 3.59- 3.54 (m, 2H), 1.98-1.92 (m, 2H).

MS ESI: m/z = 400.1, [M+H] ⁺

The third step: preparation of compound Q

Under ice-cooling, nitrogen-methylsulfonyl chloride (26.0 mg, 0.25 mmol) was added to a DMA solution of the starting material (50.0 mg, 0.13 mmol) of the second step, and reacted for 2 hours. Ethyl acetate (20 mL) and water (20 mL) were added, extracted, and washed with water (5 mL), and purified by column chromatography to give Compound 1 (40.0 mg, 67% yield)

MS ESI: m/z = 493.1, [M+H] ⁺ .

The fourth step: preparation of compound 9

The product of the third step (40 mg, 0.08 mmol) was added to THF (2.0 mL). The TLC monitoring reaction knew that the starting material had disappeared. Ethyl acetate (20 ml) and water (20 ml) were evaporated. The mixture was concentrated by filtration to give the title compound (yield: EtOAc).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ8.39 (s, 1H), 7.24-7.22 (m, 1H), 7.05-7.01 (m, 1H), 6.95-6.91 (m, 1H), 6.86 (s, 1H), 6.64-6.62 (s, 1H), 4.55-4.53 (m, 1H), 4.39-4.37 (m, 2H), 3.56-3.52 (m, 2H), 2.87 (s, 3H), 2.18-2.13 ( m, 2H).

MS ESI: m/z = 467.1, [M+H] ⁺

Example 10: Preparation of Compound 10

According to the production method of Example 9, the title compound (30% yield) was obtained.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ8.40 (s, 1H), 7.24-7.22 (m, 1H), 7.05-7.01 (m, 1H), 6.95-6.92 (m, 1H), 6.85 (s, 1H), 6.66-6.64 (s, 1H), 4.47 (s, 1H), 4.39-4.37 (m, 2H), 3.56-3.51 (m, 2H), 3.27-3.23 (m, 2H), 2.16-2.14 ( m, 2H), 1.26-1.24 (m, 3H).

MS ESI: m/z = 481.1, [M+H] ⁺

Example 11: Preparation of Compound 11

The title compound (34% yield) was obtained according to the preparation method of Example 9.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ8.42 (s, 1H), 7.24-7.22 (m, 1H), 7.05-7.01 (m, 1H), 6.95-6.91 (m, 1H), 6.85 (s, 1H), 6.68-6.65 (m, 1H), 4.38-4.36 (m, 2H), 4.34 - 4.32 (m, 1H), 3.75-3.68 (m, 1H), 3.56-3.51 (m, 2H), 2.18- 2.13 (m, 2H), 1.27 (d, 6H).

MS ESI: m/z = 495.1, [M+H] ⁺

Example 12: Preparation of Compound 12

The title compound (27% yield) was obtained according to the preparation method of Example 9.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ7.22-7.19 (m, 1H), 7.04-7.00 (m, 1H), 6.96 (s, 1H), 6.92-6.89 (m, 1H), 6.36-6.34 ( m, 1H), 4.41-4.38 (m, 2H), 3.54-3.50 (m, 2H), 2.15-2.09 (m, 2H).

MS ESI: m/z = 453.1, [M+H] ⁺

Example 13: Preparation of Compound 13

The title compound (41 mg, 71% yield) was obtained.

1H NMR (400MHz, CDCl ₃ ): δ 9.81 (s, 1H), 7.13 (dd, 1H), 6.92-6.96 (m, 2H), 6.80-6.84 (m, 1H), 6.44 (t, 1H), 5.73 (s, 2H), 3.68 (d, 2H), 1.27 (t, 2H), 0.88 (t, 2H).

MS ESI: m/z = 466.9, [M+H]+.

Example 14: Preparation of Compound 14

According to the production method of Example 9, the title compound (yield: 89%) was obtained.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.19 (s, 1H), 7.14 (dd, 1H), 6.95 (t, 1H), 6.82-6.86 (m, 2H), 6.32 (t, 1H), 5.01 ( s, 1H), 4.28 (s, 1H), 3.68 (d, 2H), 2.70 (d, 3H), 1.24 (t, 2H), 0.92 (t, 2H).

MS ESI: m/z = 479.0, [M+H] ⁺ .

Example 15: Preparation of Compound 15

The title compound (66% yield) was obtained according to the preparation method of Example 9.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.23 (s, 1H), 7.15 (dd, 1H), 6.97 (t, 1H), 6.82-6.88 (m, 2H), 6.35 (t, 1H), 4.97 ( s, 1H), 4.21 (s, 2H), 3.68 (d, 2H), 3.08 (t, 2H), 2.70 (d, 3H), 1.27 (t, 2H), 1.11-1.13 (m, 3H), 0.91 (t, 2H).

MS ESI: m/z = 494.0, [M+H] ⁺ .

Example 16: Preparation of Compound 16

The title compound (54% yield) was obtained according to the preparation method of Example 9.

¹ H NMR (400 MHz, CDCl ₃ ): δ 7.14 (dd, 1H), 6.97 (t, 1H), 6.84-6.88 (m, 2H), 6.32 (t, 1H), 4.49 (d, 1H), 3.67 ( d, 2H), 3.55-3.60 (m, 1H), 1.33 (t, 2H), 1.18 (t, 3H), 1.14 (d, 3H), 0.91 (t, 2H).

MS ESI: m/z = 507.0, [M+H] ⁺ .

Example 17: Preparation of Compound 17

The title compound (76% yield) was obtained according to the preparation method of Example 9.

¹ H NMR (400 MHz, DMSO-d ₆ ): δ 9.01 (s, 1H), 7.69 (s, 2H), 7.26 (t, 1H), 7.20 (s, 1H), 6.85 (s, 1H), 6.60 ( s, 1H), 4.48 (d, 2H), 4.42 (d, 2H), 2.27 (s, 2H), 3.59 (d, 2H).

MS ESI: m/z = 496.9, [M+H] ⁺ .

Example 18: Preparation of Compound 18

According to the production method of Example 9, the title compound (yield: 89%) was obtained.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.49 (s, 1H), 7.19 (d, 1H), 6.97 (t, 1H), 6.87 (s, 1H), 6.82 (s, 1H), 6.55 (s, 1H), 4.77 (s, 1H), 4.55 (d, 2H), 4.49 (s, 2H), 4.42 (d, 2H), 3.75 (d, 2H), 2.81 (d, 3H).

MS ESI: m/z = 509.0, [M+H] ⁺ .

Example 19: Preparation of Compound 19

The title compound (87% yield) was obtained according to the procedure of Example 9.

_{^{1 HNMR (400MHz, CDCl 3)}} : δ8.39 (s, 1H), 7.17 (d, 1H), 6.97 (t, 1H), 6.95-6.99 (m, 1H), 6.87 (s, 1H), 6.58 ( s, 1H), 4.55 (d, 2H), 4.50 (s, 2H), 4.41 (d, 2H), 3.75 (d, 2H), 3.17-3.24 (m, 2H), 1.22 (t, 3H).

MS ESI: m/z =524.1., [M+H] ⁺ .

Example 20: Preparation of Compound 20

The title compound (82% yield) was obtained according to the procedure of Example 9.

¹ H NMR (400 MHz, CDCl ₃ ): δ 7.16 (dd, 1H), 6.96 (t, 1H), 6.84-6.88 (m, 1H), 6.80 (s, 1H), 6.54 (t, 1H), 4.52 ( d, 2H), 4.47 (s, 2H), 4.41 (d, 2H), 3.75 (d, 2H), 3.61-3.67 (m, 1H), 1.23 (s, 3H), 1.21 (s, 3H).

MS ESI: m/z = 535.0, [M+H] ^- .

Example 21: Preparation of Compound 21

The title compound (53% yield) was obtained according to the preparation method of Example 9.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ7.11-7.08 (m, 2H), 6.93-6.91 (m, 1H), 6.34-6.32 (m, 1H), 3.76 (d, 2H), 1.44-1.40 ( m, 2H), 1.00-0.96 (m, 2H).

MS ESI: m/z = 421.1, [M+H] ⁺ .

Example 22: Preparation of Compound 22

The title compound (37% yield) was obtained according to the preparation method of Example 9.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ7.08-7.02 (m, 2H), 6.93 (s, 1H), 6.89-6.86 (m, 1H), 6.42-6.40 (m, 1H), 3.75 (d, 4H), 1.37-1.33 (m, 2H), 1.01-0.97 (m, 2H).

MS ESI: m/z = 435.1, [M+H] ⁺ .

Example 23: Preparation of Compound 23

According to the preparation method of Example 9, the target compound was obtained (38% yield)

_{^{1 H NMR (400MHz, CDCl 3}} ): δ7.65 (s, 1H), 7.09-7.03 (m, 2H), 6.91 (s, 1H), 6.90-6.86 (m, 1H), 6.40-6.37 (m, 1H), 4.70 (s, 1H), 3.75 (d, 4H), 3.19-3.16 (m, 2H) 1.36-1.34 (m, 2H), 1.21-1.17 (m, 3H), 0.99-0.96 (m, 2H) ).

MS ESI: m/z = 449.1, [M+H] ⁺ .

Example 24: Preparation of Compound 24

The title compound (38% yield) was obtained according to the preparation method of Example 9.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ7.43 (s, 1H), 7.09-7.03 (m, 2H), 6.90-6.86 (s, 2H), 6.40-6.37 (m, 1H), 4.54 (d, 1H), 3.75 (d, 2H), 3.67-3.62 (m, 2H), 1.39-1.36 (m, 2H), 1.26-1.21 (m, 6H), 0.99-0.95 (m, 2H).

MS ESI: m/z = 463.1, [M+H] ⁺ .

Example 25: Preparation of Compound 25

The title compound (54% yield) was obtained according to the preparation method of Example 9.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ7.10-7.03 (m, 2H), 6.92 (s, 1H), 6.89-6.88 (m, 1H), 6.42-6.40 (m, 1H), 4.49-4.47 ( m, 2H), 3.56-3.52 (m, 2H), 2.24-2.17 (m, 2H).

MS ESI: m/z = 409.0, [M+H] ⁺

Example 26: Preparation of Compound 26

The title compound (39% yield) was obtained according to the procedure of Example 9.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ7.09-7.03 (m, 2H), 6.90-6.85 (m, 2H), 6.64-6.62 (m, 1H), 4.40-4.37 (m, 2H), 3.56- 3.52 (m, 2H), 2.87 (s, 3H), 2.17-2.15 (m, 2H).

MS ESI: m/z = 423.0, [M+H] ⁺ .

Example 27: Preparation of Compound 27

The title compound (yield: 47%) was obtained according to the procedure of Example 9.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.41 (s, 1H), 7.09-7.03 (m, 2H), 6.90-6.85 (m, 2H), 6.64-6.62 (m, 1H), 4.46-4. m, 2H), 3.56-3.52 (m, 2H), 3.26-3.22 (m, 2H), 2.23-2.12 (m, 2H), 2.282.24 (m, 3H).

MS ESI: m/z = 437.0, [M+H] ⁺ .

Example 28: Preparation of Compound 28

According to the preparation method of Example 9, the target compound was obtained (45% yield)

_{^{1 H NMR (400MHz, CDCl 3}} ): δ8.42 (s, 1H), 7.09-7.03 (m, 2H), 6.90-6.86 (m, 1H), 6.85 (s, 1H), 6.65-6.64 (m, 1H), 4.38-4.369 (m, 2H), 4.32 (d, 1H), 3.73-3.68 (m, 1H), 3.56-3.51 (m, 2H), 2.18-2.12 (m, 2H).

MS ESI: m/z = 451.0, [M+H] ⁺ .

Example 29: Preparation of Compound 29

The title compound (69% yield) was obtained according to the procedure of Example 9.

¹ H NMR (400 MHz, CDCl ₃ ): δ 9.22 (s, 1H), 7.15 (dd, 1H), 6.96 (t, 1H), 6.89 (s, 1H), 6.83-6.87 (m, 1H), 6.49 ( t,1H), 5.42(s,2H), 4.17(s,2H), 3.27(d,2H)0.74(d,2H),0.70(d,2H).

MS ESI: m/z = 479.8, [M+H] ⁺ .

Example 30: Preparation of Compound 30

According to the preparation method of Example 9, the target compound was obtained.

¹ H NMR (400 MHz, CDCl ₃ ): δ 7.24-7.22 (m, 1H), 7.05-7.01 (m, 1H), 6.93-6.89 (m, 1H), 6.35-6.33 (m, 1H), 3.87- 3.85 (m, 2H), 3.25-3.22 (m, 2H), 2.65-2.61 (m, 2H), 2.18-2.13 (m, 2H), 126-1.22 (m, 7H).

MS ESI: m/z = 507.0, [M+H] ^- .

Example 31: Preparation of Compound 31

The title compound (61% yield) was obtained according to the preparation method of Example 9.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.12 (s, 1H), 7.19 (dd, 1H), 6.97 (t, 1H), 6.81-6.86 (m, 2H), 6.27 (t, 1H), 4.74 ( d, 1H), 3.81 (d, 2H), 3.58-3.63 (m, 1H), 2.49-2.57 (m, 2H), 2.19-2.24 (m, 2H), 1.85-1.93 (m, 1H) 1.18 (s , 3H), 1.16 (s, 3H).

MS ESI: m/z = 520.0, [M+H] ^- .

Example 32 Activity Test

(1) Inducible expression and purification method of IDO protein

First, the IDO gene was amplified by PCR, and the amplified PCR product was recovered. Then, the pET28a plasmid (purchased from Shanghai Baoman Biotechnology Co., Ltd.) and the IDO gel recovery product were digested with EcoR I and Xho I restriction enzymes. (37 ° C, digestion 2h), running gelatin, recovery, T4 ligase linkage overnight ligation product added to DH5α competent state, placed on ice for 30min, 42 °C heat shock 90s, shaken plate, pick monoclonal, PCR identification , sequencing identification, all correct, that is, the pET28a-IDO plasmid was successfully constructed.

The constructed BL21 containing the pET28a-IDO plasmid was shaken at 37 ° C to an OD ₆₀₀ of 0.6-0.8, and added to a final concentration of 7 μM hemin and 1 mM IPTG (isopropyl-β-D-thio Galactoside was induced at 28 ° C for 4 h; after induction, the cells were collected by centrifugation at 6000 ° C at 4 ° C, and the collected cells were washed once with 20 mM PBS (pH 6.5), and the cells were collected by centrifugation.

The collected cells were resuspended with lysate (20 mM PBS pH 6.5), sonicated (40% lysis for 20 min, placed on ice), and the lysed bacteria were centrifuged at 13,000 rpm for 15 min, the precipitate was discarded, and the supernatant was retained. The nickel column was equilibrated with lysate (20 mM PBS pH 6.5) for 3 column volumes, and then the lysed supernatant was applied to a nickel column. After loading, it was washed with a rinse solution (20 mM PBS pH 6.5, 20 mM imidazole). 4 column volumes, finally eluted with eluate (20 mM PBS pH 6.5, 250 mM imidazole); the eluted protein solution was dialyzed for 4 h, the dialysis solution was 20 mM PBS pH 6.5, and the protein sample was concentrated after dialysis. Packed, liquid nitrogen frozen, placed at -80 ° C for storage.

(2) IDO enzyme inhibitory activity test method

The compound was firstly diluted 3-fold, and 1 μL of each concentration was added to a 96-well plate; 50 μL of the formulated IDO enzyme solution (final concentration 600 ng/100 μL) was added: 25 μL of the substrate 1 mixed solution was added, and 25 μL of the substrate was added. The mixed solution initiates the reaction. The final OD ₃₂₁ nm reading was 60 min.

(3) Cell activity test method

Hela cells (100 μL) were seeded in 96-well plates in an amount of 5 × 10 ³ per well and grown overnight. On the next day, after the compound was diluted, 1 μL was added to a 96-well plate, and then 100 μL of a medium containing human interferon γ (final concentration: 50 ng/mL) was added to a 96-well plate to give a final volume of 200 μL. After 48 hours of incubation, 140 μL of supernatant from each well was transferred to a new 96-well plate. 10 μL of 6.1 N trichloroacetic acid was added to each well and mixed at 50 ° C for 30 minutes. IDO catalyzed N-formyl canine urea as canine urea. The reaction mixture was centrifuged at 2500 rpm for 10 minutes to remove the precipitate. 100 μL of the supernatant from each well was transferred to a new 96-well plate and mixed with 100 μL of a 2% dimethylaminobenzaldehyde acetic acid solution. After canine urea separation, the values were measured at 480 nm using a SPECTRAmax i3 reader.

The test results of the IDO enzyme inhibitory activity and the cell inhibitory activity of the compound of the present invention are shown in Table 1.

Table 1 IDO enzyme and cytostatic activity test results

The above results indicate that the compound of the present invention has excellent inhibitory activity against IDO enzymes and cells.

Example 33 Pharmacokinetic data of the compound of Example 1 in rats

1. Dosing regimen

Six SD rats, male, weighing 180-200 g, were given the compound of Example 1 by gavage or intravenously (Table 2):

Table 2

The gavage administration was formulated as a suspension in 1% MC and intravenously in DMSO: 30% PEG400 (5:95). Fasting for 12 hours before the test, free to drink water. Uniformly eaten 2 hours after administration.

2. Blood collection time and sample processing:

Administration by intragastric administration: 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0 and 24 h after administration;

Intravenous administration: 5 min after administration, 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0 and 24 h;

At the above set time point, 0.3 ml of venous blood was taken from the posterior venous plexus of the rat eye, placed in an EDTA-2K test tube, centrifuged at 11,000 rpm for 5 min, and the plasma was separated and frozen in a refrigerator at -20 °C.

3. Sample testing and data analysis

The concentration of the compound of Example 1 in rat plasma was determined by LC/MS/MS method.

The pharmacokinetic parameters after administration were calculated using a non-compartmental model of WinNonlin 5.3 software (Pharsight, USA).

The peak concentration C _max and T _max is time to peak found;

The area under the curve of the drug AUC _0-t value: calculated by the trapezoidal method; AUC _0-∞ = AUC _0-t + C _t / k _e , C _t is the blood concentration of the last measurable time point, k _e is Eliminate the rate constant;

Elimination half-life t _1/2 =0.693/k _e ;

Average residence time MRT = AUMC / AUC.

Clearance rate CL=D/AUC _0-∞ ;

Steady state distribution volume V _ss =CL×MRT

Absolute bioavailability F = (AUC _gavage × D _vein ) / (AUC _vein × D _gavage ) × 100%

4. Test results

The pharmacokinetic parameters of the rats administered with 10 mg/kg of the compound of Example 1 and the intravenous injection of 1 mg/kg of the compound of Example 1 were shown in Tables 3 and 4.

Table 3 Pharmacokinetic parameters of rats after intragastric administration of 10 mg/kg of the compound of Example 1

Table 4 Pharmacokinetic parameters of 1 mg/kg of the compound of Example 1 administered intravenously in rats

The results showed that after intravenous administration, the clearance rate (CL) of the compound of Example 1 was 2.85 L/h/kg, and the steady-state volume (Vss) was 6.932 L/kg. The exposure (AUC _0-t ) It was 354 ng·h/mL. After the intragastric administration, the peak time _Tmax of the compound of Example 1 in the rat was 0.33 h, and the exposure (AUC _0-t ) was 1552 ng·h/mL. After the dose was standardized, the absolute bioavailability was 43.8%.

Example 34 Pharmacokinetic data of the compound of Example 1 in mice

1. Experimental purpose

After the compound of Example 1 was orally administered to ICR mice, blood samples were collected at different time points, and the concentration of the compound of Example 1 in the plasma of the mice after the administration of the test substance was determined by LC-MS/MS, and the relevant pharmacokinetic parameters were calculated. 1 Pharmacokinetic properties of the compound in mice.

2. Test method

Same as Example 9.

3. Animal experiment

Experimental design: 3 ICR mice were provided by Suzhou Kangrun New Drug Research and Development Co., Ltd., and experiments were carried out according to Table 5.

table 5

Sample Collection

After each isoflurane anesthesia, each animal took about 25 μL of blood through the eyelids, and EDTAK _{2 was} anticoagulated. The time of collection was: before administration of the test substance (0 hr) and after administration of the test substance for 15 min, 30 min, 1 h, 2 h, 4h, 6h, 8h and 24h. Blood samples were collected and placed on ice, and plasma was centrifuged within 1 hour (centrifugation conditions: 5000 rpm, 10 minutes, 4 ° C). The collected plasma was stored at –20 °C prior to analysis.

4 data processing

The data acquisition and control system software is Analyst 1.5.1 software (Appl ied Biosystem). The peak integration method of the spectrum sample is automatic integration; the ratio of the peak area of the sample to the peak area of the internal standard is used as an index, and the concentration of the sample is used for regression. Regression mode: linear regression, the weight coefficient is 1/X ² . Pharmacokinetic parameters were analyzed by non-compartmental model analysis using WinNonl in Professional v6.3 (Pharsight, USA). C _max is the measured maximum blood concentration, and the area under the blood concentration-time curve AUC _(0→t) is calculated by the trapezoidal method, and T _max is the peak time of the blood drug concentration after administration. The experimental data is expressed as "mean ± standard deviation" (Mean ± SD, n ≥ 3) or "mean" (Mean, n = 2).

5 experimental results

The pharmacokinetic study of the compound of Example 1 after oral administration (dose of 30.0 mg/kg) in ICR mice showed that the average peak time of the blood concentration of the compound of Example 1 in mice was 0.25 hr. The peak concentration was 809.3±254ng/mL, AUC _0→t was 982±151hr*ng/mL, and the average residence time in MRT was 6.76±4.13hr.

The concentration of the compound of Example 1 administered to ICR mice at various time points in plasma (PO, 30 mg/kg) is shown in Table 6.

Table 6

3. Calculate the pharmacokinetic parameters after administration, see pharmacokinetic parameters

According to the plasma concentration data, the pharmacokinetic parameters after administration were calculated using the WinNonlin V6.3 non-compartment model.

Table 7. Pharmacokinetic parameters of the compound of Example 1 in ICR mice (oral: 30 mg/kg)

The results indicate that the compound of Example 1 has good pharmacokinetic parameters.

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

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

Claims (10)

A compound of formula I, or a pharmaceutically acceptable salt thereof, a deuterated compound, a stereoisomer thereof or a tautomer thereof, or a prodrug:

In the formula, R ¹ , R ² , R ³ and R ⁴ are each independently hydrogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₂ -C ₁₀ alkenyl, substituted or unsubstituted C ₃ -C ₁₀ alkynyl, substituted or unsubstituted C ₆ -C ₂₀ aryl, or substituted or unsubstituted C ₃ -C ₁₄ heteroaryl; R ¹ and R ² , or R ³ and R ⁴ may together form a three- to eight-membered carbocyclic ring or a three- to eight-membered heterocyclic ring, or that R ¹ and R ³ may together form a five- to eight-membered carbocyclic ring or a five- to eight-membered heterocyclic ring. Wherein the hetero atom may be sulfur, oxygen, NH or NR ^f ; R ⁵ is C ₆ -C ₂₀ aryl, 5- or 6-membered heteroaryl; R ⁵ may be substituted by one or more groups selected from the group consisting of halogen, C ₁ -C ₆ alkyl, C ₁ - C ₆ alkoxy, hydroxy, amino, nitro, aldehyde, -CF ₃ , -CN, -SF ₅ , NR ^a R ^b , carboxy, -COR ^a , -CO ₂ C ₁ -C ₆ alkyl, - CONR ^a R ^b , -S(O)R ^a , -S(O) ₂ R ^a , -S(O)(NH)R ^a , -S(O)(NR ^d )R ^a , -S(O) ₂ NR ^a R ^b , -P(O)Me ₂ , -P(O)(OMe) ₂ ; wherein each R ^a and each R ^b are independently hydrogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl, Substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₂ -C ₁₀ alkenyl, substituted or unsubstituted C ₆ -C ₂₀ aryl, or substituted or unsubstituted C ₃ -C _C14 heteroaryl group; R ^a and R ^b may together may form a three to eight yuan four to eight yuan or heterocyclic ring, wherein the heteroatoms may be sulfur, oxygen, NH or NR ^f; X is a single bond, O, S, NH or NR ^d ; R ^d and R ^{f are} independently C ₁ -C ₁₀ alkyl, C ₃ -C ₁₀ cycloalkyl, C ₆ -C ₂₀ aryl, or C ₃ -C ₁₄ heteroaryl; n is an integer from 2 to 8. The compound of the formula I according to claim 1, wherein the compound is represented by the formula (II).

In the formula, Ar is a benzene ring, and Ar may be substituted by one or more groups selected from the group consisting of halogen, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, hydroxy, amino, nitro, aldehyde, -CF ₃ , -CN, -SF ₃ , -SF ₅ , NR ^a R ^b , carboxyl group, -COR ^a , -CO ₂ C ₁ -C ₆ alkyl group, -CONR ^a R ^b , -S(O)R ^a , -S(O) ₂ R ^a , -S(O)(NH)R ^a , -S(O)(NR ^d )R ^a , -S(O) ₂ NR ^a R ^b ; Wherein R ^a , R ^b , R ^d , R ³ , R ⁴ , R ² , R ¹ are as defined above; the hydrogen atom in the alkyl group may be substituted by halogen; n is 2-6. The compound of the formula II according to claim 2, wherein R ¹ is H, C ₁ -C ₁₀ alkyl, C ₃ -C ₁₀ cycloalkyl, C ₆ -C ₂₀ aryl, or C ₃ - C ₁₀ heteroaryl; R ¹ may be substituted by one or more halogens; R ² is H, C ₁ -C ₁₀ alkyl; R ¹ and R ² may together form a three to eight membered carbocyclic ring or a three to eight membered heterocyclic ring, wherein the hetero atom may be sulfur, oxygen, NH or NR ^f ; R ³ and R ⁴ are each independently hydrogen, C ₁ -C ₁₀ alkyl or R ³ and R ⁴ may together form a three to eight membered carbocyclic ring or a three to eight membered heterocyclic ring. A prodrug of a compound of formula I according to claim 1 wherein said prodrug is as shown in formula (III).

In the formula, The definitions of R ¹ , R ² , R ³ , R ⁴ , n and Ar are as described above; Y is R ⁶ , OR ⁶ , NR ^a R ^b ; R ⁶ is a substituted or unsubstituted C ₆ -C ₂₀ aryl group, a substituted or unsubstituted five- or six-membered heteroaryl group, a substituted or unsubstituted C ₁ a -C ₁₂ alkyl, substituted or unsubstituted C ₃ -C ₁₂ cycloalkyl, substituted or unsubstituted C ₃ -C ₁₂ heterocyclic group, wherein the hetero atom may be sulfur, oxygen, NH or NR ^f ; R ^a and each R ^b are each independently hydrogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₂ -C ₁₀ alkenyl a substituted or unsubstituted C ₆ -C ₂₀ aryl group, or a substituted or unsubstituted C ₃ -C ₁₄ heteroaryl group; R ^a and R ^b may together form a three to eight membered ring or a four to eight membered heterocyclic ring. Wherein the hetero atom may be sulfur, oxygen, NH or NR ^f ; Nf is as defined above; Wherein said "substituted" means having one or more substituents selected from the group consisting of halogen, hydroxy, -NH _2, nitro, -CN, C ₁ -C ₄ alkyl, C ₁ -C ₄ haloalkane A C ₁ -C ₄ alkoxy group, a C ₃ -C ₆ cycloalkyl group, a C ₂ -C ₄ alkenyl group, a C ₂ -C ₄ alkynyl group, a phenyl group, a benzyl group. A compound of formula I according to claim 1 wherein said compound is selected from the group consisting of:

A compound of formula I according to claim 1 wherein said pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, hydrobromide, sulfate, phosphate, methanesulfonate, Fluoromethanesulfonate, besylate, p-toluenesulfonate (tosylate), 1-naphthalenesulfonate, 2-naphthalenesulfonate, acetate, trifluoroacetate, malate , tartrate, citrate, lactate, oxalate, succinate, fumarate, maleate, benzoate, salicylate, phenylacetate, and mandelate. Use of a compound of formula I according to claim 1 for: (i) preparing a guanamine-2,3-dioxygenase inhibitor; and/or (ii) A medicament for the prevention and/or treatment of a guanamine-2,3-dioxygenase mediated disease. The use according to claim 7, wherein the indoleamine-2,3-dioxygenase-mediated disease is cancer, neurodegenerative disease, eye disease, mental disorder, depression, anxiety, AIDS , Alzheimer's disease and / or autoimmune diseases. A pharmaceutical composition, characterized in that the pharmaceutical composition comprises: A compound of formula I, or a pharmaceutically acceptable salt thereof, a stereoisomer thereof or a tautomer thereof, or a prodrug thereof; A pharmaceutically acceptable carrier and/or an anti-tumor drug. A method for preparing a compound of the formula (I), characterized in that the method comprises the steps of:

(a) compound AA is reacted with compound CC to give compound BB; (b) Compound BB is ring-opened under alkaline hydrolysis conditions (such as aqueous sodium hydroxide) to give the final product of formula (I); In each formula, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , n are as defined above; or The method includes the following steps:

(a) Compound DD is reacted with compound EE and a reducing agent under acid catalysis to give compound BB, (b) Compound BB is ring-opened under alkaline hydrolysis conditions (such as aqueous sodium hydroxide) to give the final product of formula (I); Or (a) Compound EE is reacted with H ₂ O ₂ in the presence of TFA to give compound FF; (b) compound FF and compound GG are reacted to obtain compound AA; (c) reacting compound AA with compound CC to obtain compound BB; (d) Compound BB is ring-opened under alkaline hydrolysis conditions (e.g., aqueous sodium hydroxide) to give the final compound of formula (I).