CN105646389A

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Publication number: CN105646389A
Application number: CN201610059454.5A
Authority: CN
Inventors: 王召印; 朱继东
Original assignee: Shanghai Institute of Organic Chemistry of CAS
Current assignee: Shanghai Institute of Organic Chemistry of CAS
Priority date: 2016-01-28
Filing date: 2016-01-28
Publication date: 2016-06-08
Anticipated expiration: 2036-01-28
Also published as: CN105646389B; WO2017129139A1

Abstract

Translated fromChinese

本发明提供了一种作为吲哚胺-2,3-双加氧酶抑制剂的氨基磺酸脂及其制备方法和用途。本发明的抑制剂，结构如通式I所示，其中，X、R¹、R²、R³、R⁴、R⁵、n的定义如说明书和权利要求书中所示。本发明还公开了抑制剂的制备方法。本发明的通式I化合物，可以作为吲哚胺-2,3-双加氧酶抑制剂，用于制备预防和/或治疗吲哚胺-2,3-双加氧酶介导的疾病的药物。The invention provides a sulfamic acid ester as an indoleamine-2,3-dioxygenase inhibitor, a preparation method and application thereof. The structure of the inhibitor of the present invention is shown in the general formula I, wherein the definitions of X, R¹ , R² , R³ , R⁴ , R⁵ and n are as shown in the specification and claims. The invention also discloses a preparation method of the inhibitor. The compound of 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 drug.

Description

Translated fromChinese

一种作为吲哚胺-2,3-双加氧酶抑制剂的氨基磺酸脂及其制备方法和用途A kind of sulfamate as indoleamine-2,3-dioxygenase inhibitor and its preparation method and application

技术领域technical field

本发明属于药物化学技术领域，具体地涉及一种作为吲哚胺-2,3-双加氧酶抑制剂的氨基磺酸脂及其制备方法和用途，更具体涉及一种含有氨基磺酸脂和1,2,5-噁二唑结构的IDO抑制剂及其制备方法和用途。The invention belongs to the technical field of medicinal chemistry, and in particular relates to a sulfamic acid ester as an indoleamine-2,3-dioxygenase inhibitor and its preparation method and application, more specifically to a sulfamic acid ester containing and IDO inhibitors with 1,2,5-oxadiazole structures, their preparation methods and applications.

背景技术Background technique

吲哚胺-2,3-双加氧酶(Indoleamine-2,3-dioxygenase,IDO)是1967年Hayaishi小组首次在细胞内发现的一种含有亚铁血红素的单体酶，cDNA编码蛋白由403氨基酸组成，分子量为45kDa，它是延着色氨酸-犬尿氨酸途径分解代谢的限速酶，并且在多种哺乳动物的组织中具有广泛的表达(HayaishiO.etalScience,1969,164,389-396)。在肿瘤患者的细胞中，IDO常作为诱导肿瘤微环境免疫耐受产生重要的生理作用，其介导的色氨酸(Tryptophan,Trp)-犬尿氨酸(Kynurenine,Kyn)代谢途径参与了肿瘤免疫逃逸，而IDO作为诱导肿瘤微环境免疫耐受也产生重要的作用。Indoleamine-2,3-dioxygenase (Indoleamine-2,3-dioxygenase, IDO) is a monomeric enzyme containing heme first discovered in cells by the Hayaishi group in 1967. The cDNA-encoded protein is composed of Composed of 403 amino acids, the molecular weight is 45kDa, it is the rate-limiting enzyme of the catabolism of the tryptophan-kynurenine pathway, and it is widely expressed in the tissues of various mammals (HayaishiO.etalScience, 1969,164,389-396 ). In the cells of tumor patients, IDO often plays an important physiological role in inducing immune tolerance in the tumor microenvironment, and its mediated tryptophan (Trp)-kynurenine (Kynurenine, Kyn) metabolic pathway participates in tumor Immune escape, and IDO also plays an important role in inducing immune tolerance in the tumor microenvironment.

色氨酸为哺乳动物体内重要的必须氨基酸之一，需要从食物中大量摄取，维持细胞活化和增殖，以及蛋白质以及一些神经递质的合成。因此，它的缺乏会导致一些重要的细胞的功能失常。IDO在体内能够催化色氨酸转化为N-甲酰犬尿氨酸，降解色氨酸的含量而造成色氨酸体内的不足，导致肿瘤的发生。而免疫组织学研究显示，犬尿氨酸途径能够导致兴奋毒素喹啉酸的增多，还会导致阿兹海默等神经系统疾病等多种严重的人类疾病(GuilleminG.J.etalNeuropathol.andAppl.Neurobiol.2005,31,395–404)。Tryptophan is one of the important essential amino acids in mammals. It needs to be taken in large quantities from food to maintain cell activation and proliferation, as well as the synthesis of proteins and some neurotransmitters. Therefore, its deficiency can lead to malfunction of some important cells. IDO can catalyze the conversion of tryptophan into N-formylkynurenine in the body, degrade the content of tryptophan and cause the deficiency of tryptophan in the body, leading to the occurrence of tumors. Immunohistological studies have shown that the kynurenine pathway can lead to an increase in the excitotoxin quinolinic acid, and can also lead to a variety of serious human diseases such as Alzheimer's and other neurological diseases (GuilleminG.J.etalNeuropathol.andAppl.Neurobiol .2005, 31, 395–404).

哺乳动物体内色氨酸限速酶主要有两种：色氨酸双加氧酶(TDO)和IDO。1937年，Kotake等从兔子肠中纯化出蛋白，并首次发现TDO主要在哺乳动物肝脏中表达，目前尚未发现他与免疫系统有密切联系。TDO能够催化犬尿氨酸途径，使色氨酸转变为N-甲酰犬尿氨酸[HiguchiK.etalJ.Biochem.1937,25,71-77；ShimizuT.etalJ.Biol.Chem.1978,253,4700-4706]。1978年，从兔子肠道中纯化的酶，被鉴定是含有亚铁血红素的双加氧酶(IDO)，IDO是肝脏以外唯一可以催化色氨酸分子中的吲哚氧化裂解，产生犬尿氨酸及其它代谢物的酶。IDO通常在粘膜较多的器官中表达，如肺，小肠和大肠，直肠，脾，肾，胃和脑等，分布比较广泛(HayaishiO.etal,ProceedingsofthetenthFEBSmeeting,1975,131–144)。在某些特殊或病理条件下，如妊娠，慢性感染，器官移植和肿瘤等，IDO表达会明显增加，参与介导局部的免疫抑制。There are two main tryptophan rate-limiting enzymes in mammals: tryptophan dioxygenase (TDO) and IDO. In 1937, Kotake et al. purified the protein from rabbit intestines and discovered for the first time that TDO is mainly expressed in the liver of mammals. It has not been found that it is closely related to the immune system. TDO can catalyze the kynurenine pathway to convert tryptophan into N-formylkynurenine [HiguchiK.etalJ.Biochem.1937,25,71-77; ShimizuT.etalJ.Biol.Chem.1978,253, 4700-4706]. In 1978, the enzyme purified from the rabbit intestine was identified as a heme-containing dioxygenase (IDO). IDO is the only one other than the liver that can catalyze the oxidative cleavage of indole in the tryptophan molecule to produce kynurenine Acid and other metabolite enzymes. IDO is usually expressed in organs with more mucous membranes, such as the lung, small and large intestine, rectum, spleen, kidney, stomach and brain, and is widely distributed (HayaishiO.etal, ProceedingsthetenthFEBSmeeting, 1975, 131-144). In some special or pathological conditions, such as pregnancy, chronic infection, organ transplantation and tumors, the expression of IDO will increase significantly, participating in mediating local immunosuppression.

研究表明，IDO在肿瘤微环境中可以通过以下几种方式来抑制局部T细胞免疫反应：色氨酸耗竭、毒性代谢和诱导调节性T细胞增殖。很多情况是在肿瘤中过渡表达，从而消耗局部的色氨酸，产生大量的犬尿氨酸等代谢产物。事实上，在无色氨酸或犬尿氨酸的培养条件下，T细胞会发生增殖抑制、活性降低甚至凋亡。而T细胞中存在一个对色氨酸水平非常敏感的调节点，IDO的作用下，能够使色氨酸消耗，从而导致T细胞停滞于G1期中期，从而抑制了T细胞的增殖，也抑制了T细胞的免疫应答。而T细胞一旦停止增殖，可能就不会再被刺激作用，这是IDO在体内免疫作用机制(MellorA.etalBiochem.Biophys.Res.Commun.2005,338(1):20-24)(LeRondS.etalJ.Exp.Med.2002,196(4):447-457)。Studies have shown that IDO can suppress local T cell immune responses in several ways in the tumor microenvironment: tryptophan depletion, toxic metabolism, and induction of regulatory T cell proliferation. In many cases, it is over-expressed in tumors, thereby consuming local tryptophan and producing a large amount of metabolites such as kynurenine. In fact, under culture conditions without tryptophan or kynurenine, T cells will undergo proliferation inhibition, decreased activity and even apoptosis. However, there is a regulatory point in T cells that is very sensitive to the level of tryptophan. Under the action of IDO, tryptophan can be consumed, thereby causing T cells to stagnate in the middle of G1 phase, thereby inhibiting the proliferation of T cells and inhibiting the growth rate of T cells. T cell immune response. Once T cells stop proliferating, they may not be stimulated again. This is the mechanism of IDO's immune function in vivo (MellorA.etalBiochem.Biophys.Res.Commun.2005,338(1):20-24) (LeRondS.etalJ . Exp. Med. 2002, 196(4): 447-457).

本领域尚需研发新型的IDO抑制剂。There is still a need to develop new IDO inhibitors in this field.

发明内容Contents of the invention

本发明的目的在于提供一种新型的含有氨基磺酸脂和1,2,5-噁二唑结构的化合物作为高效的IDO酶抑制剂。The purpose of the present invention is to provide a novel compound containing sulfamic acid ester and 1,2,5-oxadiazole structure as a high-efficiency IDO enzyme inhibitor.

本发明的另一目的在于提供该化合物的制备方法。Another object of the present invention is to provide a preparation method of the compound.

本发明的第一方面，提供一种通式(I)化合物或其药学上可以接受的盐、其立体异构体或其互变异构体、或前药：The first aspect of the present invention provides a compound of general formula (I) or a pharmaceutically acceptable salt thereof, a stereoisomer or a tautomer thereof, or a prodrug:

式中，In the formula,

R¹,R²,R³和R⁴各自独立为氢、取代或未取代的C₁-C₁₀烷基、取代或未取代的C₃-C₁₀环烷基、取代或未取代的C₂-C₁₀烯基、取代或未取代的C₃-C₁₀炔基、取代或未取代的C₆-C₂₀芳基、或取代或未取代的C₃-C₁₄杂芳基；R¹和R²，R¹和R³或R³和R⁴可以一起可以形成三至八元碳环或三至八元杂环，其中杂原子可以是硫、氧、NH或NR^f；R¹ , R² , R³ and R⁴ are each independently hydrogen, substituted or unsubstituted C₁ -C₁₀ alkyl, substituted or unsubstituted C₃ -C₁₀ cycloalkyl, substituted or unsubstituted C₂ -C₁₀ alkenyl, substituted or unsubstituted C₃ -C₁₀ alkynyl, substituted or unsubstituted C₆ -C₂₀ aryl, or substituted or unsubstituted C₃ -C₁₄ heteroaryl; R¹ and R² , R¹ and R³ or R³ and R⁴ can together form a three- to eight-membered carbocyclic ring or a three- to eight-membered heterocyclic ring, wherein the heteroatom can be sulfur, oxygen, NH or NR^f ;

R⁵为C₆-C₂₀芳基、五元或六元杂芳基；R⁵可以被一个或多个选自下组的基团取代：卤素、C₁-C₆烷基、C₁-C₆烷氧基、羟基、氨基、硝基、醛基、-CF₃、-CN、-SF₅、NR^aR^b、羧基、-COR^a、-CO₂C₁-C₆烷基、-CONR^aR^b、-S(O)R^a,-S(O)₂R^a、-S(O)(NH)R^a、-S(O)(NR^d)R^a、-SO₂NR^aR^b、-P(O)Me₂、-P(O)(OMe)₂；其中各R^a和各R^b各自独立为氢、取代或未取代的C₁-C₁₀烷基、取代或未取代的C₃-C₁₀环烷基、取代或未取代的C₂-C₁₀烯基、取代或未取代的C₆-C₂₀芳基、或取代或未取代的C₃-C₁₄杂芳基；R^a和R^b可以一起可以形成三至八元环或四至八元杂环，其中杂原子可以是硫、氧、NH或NR^f；R⁵ is C₆ -C₂₀ aryl, five-membered or six-membered heteroaryl; R⁵ can be substituted by one or more groups selected from the group: halogen, C₁ -C₆ alkyl, C₁ - C₆ alkoxy, hydroxyl, amino, nitro, aldehyde, -CF₃ , -CN, -SF₅ , NR^a R^b , carboxyl, -COR^a , -CO₂ C₁ -C₆ 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 , -SO₂ NR^a R^b , -P(O)Me₂ , -P(O)(OMe)₂ ; wherein each R^a and each R^b is independently hydrogen, substituted or unsubstituted C₁ -C₁₀ alkyl, substituted or unsubstituted Substituted C₃ -C₁₀ cycloalkyl, substituted or unsubstituted C₂ -C₁₀ alkenyl, substituted or unsubstituted C₆ -C₂₀ aryl, or substituted or unsubstituted C₃ -C₁₄ heteroaryl R^a and R^b together can form three to eight membered rings or four to eight membered heterocyclic rings, wherein the heteroatoms can be sulfur, oxygen, NH or NR^f ;

X为一个单键、O，S、NH或NR^d；X is a single bond, O, S, NH or NR^d ;

R^d和R^f独立地为C₁-C₁₀烷基、C₃-C₁₀环烷基、C₆-C₂₀芳基、或C₃-C₁₄杂芳基；R^d and R^f are independently C₁ -C₁₀ alkyl, C₃ -C₁₀ cycloalkyl, C₆ -C₂₀ aryl, or C₃ -C₁₄ heteroaryl;

n为2至8的整数。n is an integer of 2 to 8.

在另一优选例中，所述的“取代”指具有一个或多个选自下组的取代基：卤素、羟基、-NH₂、硝基、-CN、C₁-C₄烷基、C₁-C₄卤代烷基、C₁-C₄烷氧基、C₃-C₆环烷基、C₂-C₄链烯基、C₂-C₄炔基、苯基、苄基。In another preferred example, the "substitution" refers to having one or more substituents selected from the following group: halogen, hydroxyl, -NH₂ , nitro, -CN, C₁ -C₄ alkyl, C₁ -C₄ haloalkyl, C₁ -C₄ alkoxy, C₃ -C₆ cycloalkyl, C₂ -C₄ alkenyl, C₂ -C₄ alkynyl, phenyl, benzyl.

在另一优选例中，X为NH。In another preferred example, X is NH.

在另一优选例中，R³和R⁴各自独立为氢、取代或未取代的C₁-C₁₀烷基；R³和R⁴可以一起可以形成三至八元环或三至八元杂环，其中杂原子可以是硫、氧、NH或NR^f。In another preferred example, R³ and R⁴ are each independently hydrogen, substituted or unsubstituted C₁ -C₁₀ alkyl; R³ and R⁴ can together form a three- to eight-membered ring or a three- to eight-membered hetero Ring, where the heteroatom can be sulfur, oxygen, NH or NR^f .

在另一优选例中，n为2至6的整数。In another preferred example, n is an integer of 2-6.

在另一优选例中，R³为氢。In another preferred embodiment,^R3 is hydrogen.

在另一优选例中，R⁴为氢。^In another preferred embodiment, R4 is hydrogen.

在另一优选例中，所述化合物如通式(II)所示，In another preferred example, the compound is represented by general formula (II),

式中，In the formula,

Ar为苯环，Ar可以被一个或多个选自下组的基团取代：卤素、C₁-C₆烷基、C₁-C₆烷氧基、羟基、氨基、硝基、醛基、-CF₃、-CN、-SF₅、NR^aR^b、羧基、-COR^a、-CO₂C₁-C₆烷基、-CONR^aR^b、-S(O)R^a,-S(O)₂R^a、-S(O)(NH)R^a、-S(O)(NR^d)R^a、-SO₂NR^aR^b；Ar is a benzene ring, and Ar can be substituted by one or more groups selected from the group consisting of: halogen, C₁ -C₆ alkyl, C₁ -C₆ alkoxy, hydroxyl, amino, nitro, aldehyde, -CF₃ , -CN, -SF₅ , NR^a R^b , carboxyl, -COR^a , -CO₂ C₁ -C₆ 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 , -SO₂ NR^a R^b ;

其中，R^a、R^b、R^d、R³、R⁴、R²、R¹的定义如上所述；Wherein, R^a , R^b , R^d , R³ , R⁴ , R² , R¹ are as defined above;

n为2-6的整数。n is an integer of 2-6.

在另一优先选例中，R¹为H、C₁-C₁₀烷基、C₃-C₁₀环烷基、C₆-C₂₀芳基、或C₃-C₁₀杂芳基；R¹可以被一个或多个卤素取代。In another preferred embodiment, R¹ is H, C₁ -C₁₀ alkyl, C₃ -C₁₀ cycloalkyl, C₆ -C₂₀ aryl, or C₃ -C₁₀ heteroaryl; R¹ Can be substituted by one or more halogens.

在另一优选例中，R¹为C₁-C₁₀烷基；In another preferred embodiment, R¹ is C₁ -C₁₀ alkyl;

R²为H、C₁-C₁₀烷基；R² is H, C₁ -C₁₀ alkyl;

R³和R⁴各自独立为氢、C₁-C₁₀烷基；R³ and R⁴ are each independently hydrogen, C₁ -C₁₀ alkyl;

在另一优选例中，R¹、R²、R³、R⁴为各自独立为H。In another preferred example, R¹ , R² , R³ , and R⁴ are each independently H.

在另一优选例中，所述化合物如通式(III)所示，In another preferred example, the compound is represented by the general formula (III),

式中，In the formula,

R¹、R²、R³、R⁴、n和Ar的定义如上所述；R¹ , R² , R³ , R⁴ , n and Ar are as defined above;

Y为R⁶、OR⁶、NR^aR^b；R⁶为取代或未取代的C₆-C₂₀芳基、取代或未取代的五元或六元杂芳基、取代或未取代的C₁-C₁₂烷基、取代或未取代的C₃-C₁₂环烷基、取代或未取代的C₃-C₁₂杂环基，其中杂原子可以是硫、氧、NH或NR^f；Y is R⁶ , OR⁶ , NR^a R^b ; R⁶ is substituted or unsubstituted C₆ -C₂₀ aryl, substituted or unsubstituted five-membered or six-membered heteroaryl, substituted or unsubstituted C₁ -C₁₂ alkyl, substituted or unsubstituted C₃ -C₁₂ cycloalkyl, substituted or unsubstituted C₃ -C₁₂ heterocyclyl, wherein the heteroatom can be sulfur, oxygen, NH or NR^f ;

R^a和各R^b各自独立为氢、取代或未取代的C₁-C₁₀烷基、取代或未取代的C₃-C₁₀环烷基、取代或未取代的C₂-C₁₀烯基、取代或未取代的C₆-C₂₀芳基、或取代或未取代的C₃-C₁₄杂芳基；R^a和R^b可以一起可以形成三至八元环或四至八元杂环，其中杂原子可以是硫、氧、NH或NR^f；Nf定义如上所述。R^a and each R^b are each independently hydrogen, substituted or unsubstituted C₁ -C₁₀ alkyl, substituted or unsubstituted C₃ -C₁₀ cycloalkyl, substituted or unsubstituted C₂ -C₁₀ alkenyl , a substituted or unsubstituted C₆ -C₂₀ aryl group, or a substituted or unsubstituted C₃ -C₁₄ heteroaryl group; R^a and R^b can together form a three to eight-membered ring or a four to eight-membered heterocyclic ring, Wherein the heteroatom may be sulfur, oxygen, NH or NR^f ; Nf is as defined above.

其中，所述的“取代”指具有一个或多个选自下组的取代基：卤素、羟基、-NH_2、硝基、-CN、C₁-C₄烷基、C₁-C₄卤代烷基、C₁-C₄烷氧基、C₃-C₆环烷基、C₂-C₄链烯基、C₂-C₄炔基、苯基、苄基。Wherein, the "substitution" refers to having one or more substituents selected from the following group: halogen, hydroxyl, -NH_{2 ,} nitro, -CN, C₁ -C₄ alkyl, C₁ -C₄ haloalkane C₁ -C₄ alkoxy, C₃ -C₆ cycloalkyl, C₂ -C₄ alkenyl, C₂ -C₄ alkynyl, phenyl, benzyl.

在另一优选例中，X、R¹、R²、R³、R⁴、Y及Ar等各基团分别独立地为实施例中制备的式(I)、(II)、和(III)的各具体化合物中的相应基团。In another preferred example, each group of X, R¹ , R² , R³ , R⁴ , Y and Ar is independently the formulas (I), (II) and (III) prepared in the examples The corresponding groups in each specific compound.

在另一优选例中，所述化合物选自下组：In another preferred embodiment, the compound is selected from the following group:

(Z)-2-((4-(氮-(3-溴-4-氟苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基磺酰氨；(Z)-2-((4-(nitrogen-(3-bromo-4-fluorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxy Sulfonamide;

(Z)-2-((4-(氮-(3-溴-4-氟苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基甲基磺酰氨；(Z)-2-((4-(nitrogen-(3-bromo-4-fluorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxy Methylsulfonamide;

(Z)-2-((4-(氮-(3-溴-4-氟苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基乙基磺酰氨；(Z)-2-((4-(nitrogen-(3-bromo-4-fluorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxy Ethylsulfonamide;

(Z)-2-((4-(氮-(3-溴-4-氟苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基异丙基磺酰氨；(Z)-2-((4-(nitrogen-(3-bromo-4-fluorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxy Isopropylsulfonamide;

(Z)-2-((4-(氮-(3-溴-4-氟苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基二甲基磺酰氨；(Z)-2-((4-(nitrogen-(3-bromo-4-fluorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxy Dimethylsulfonamide;

(Z)-2-((4-(氮-(3-氯苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基磺酰氨；(Z)-2-((4-(nitrogen-(3-chlorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxysulfonamide;

(Z)-2-((4-(氮-(3-氯苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基甲基磺酰氨；(Z)-2-((4-(nitrogen-(3-chlorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxymethylsulfonyl ammonia;

(Z)-2-((4-(氮-(3-氯苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基乙基磺酰氨；(Z)-2-((4-(nitrogen-(3-chlorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxyethylsulfonyl ammonia;

(Z)-2-((4-(氮-(3-氯苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基异丙基磺酰氨；(Z)-2-((4-(nitrogen-(3-chlorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxyisopropylsulfonate Amide;

(Z)-2-((4-(氮-(3-氯苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基二甲基磺酰氨；(Z)-2-((4-(nitrogen-(3-chlorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxydimethylsulfonate Amide;

(Z)-2-((4-(氮-(4-氯苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基磺酰氨；(Z)-2-((4-(nitrogen-(4-chlorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxysulfonamide;

(Z)-2-((4-(氮-(4-氯苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基甲基磺酰氨；(Z)-2-((4-(nitrogen-(4-chlorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxymethylsulfonyl ammonia;

(Z)-2-((4-(氮-(4-氯苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基乙基磺酰氨；(Z)-2-((4-(nitrogen-(4-chlorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxyethylsulfonyl ammonia;

(Z)-2-((4-(氮-(4-氯苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基异丙基磺酰氨；(Z)-2-((4-(nitrogen-(4-chlorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxyisopropylsulfonate Amide;

(Z)-2-((4-(氮-(4-氯苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基二甲基磺酰氨；(Z)-2-((4-(nitrogen-(4-chlorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxydimethylsulfonate Amide;

(Z)-2-((4-(氮-(3-氯-4-氟苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基磺酰氨；(Z)-2-((4-(nitrogen-(3-chloro-4-fluorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxy Sulfonamide;

(Z)-2-((4-(氮-(3-氯-4-氟苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基甲基磺酰氨；(Z)-2-((4-(nitrogen-(3-chloro-4-fluorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxy Methylsulfonamide;

(Z)-2-((4-(氮-(3-氯-4-氟苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基乙基磺酰氨；(Z)-2-((4-(nitrogen-(3-chloro-4-fluorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxy Ethylsulfonamide;

(Z)-2-((4-(氮-(3-氯-4-氟苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基异丙基磺酰氨；(Z)-2-((4-(nitrogen-(3-chloro-4-fluorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxy Isopropylsulfonamide;

(Z)-2-((4-(氮-(3-氯-4-氟苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基二甲基磺酰氨；(Z)-2-((4-(nitrogen-(3-chloro-4-fluorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxy Dimethylsulfonamide;

(Z)-2-((4-(氮-(3-三氟甲基苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基磺酰氨；(Z)-2-((4-(nitrogen-(3-trifluoromethylphenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxysulfonium Amide;

(Z)-2-((4-(氮-(3-三氟甲基苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基甲基磺酰氨；(Z)-2-((4-(nitrogen-(3-trifluoromethylphenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxymethyl Sulfonamide;

(Z)-2-((4-(氮-(3-三氟甲基苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基乙基磺酰氨；(Z)-2-((4-(nitrogen-(3-trifluoromethylphenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxyethyl Sulfonamide;

(Z)-2-((4-(氮-(3-三氟甲基苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基异丙基磺酰氨；(Z)-2-((4-(nitrogen-(3-trifluoromethylphenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxyiso Propylsulfonamide;

(Z)-2-((4-(氮-(3-三氟甲基苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基二甲基磺酰氨；(Z)-2-((4-(nitrogen-(3-trifluoromethylphenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxydi Methylsulfonamide;

(Z)-2-((4-(氮-(4-三氟甲基苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基磺酰氨；(Z)-2-((4-(nitrogen-(4-trifluoromethylphenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxysulfonium Amide;

(Z)-2-((4-(氮-(4-三氟甲基苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基甲基磺酰氨；(Z)-2-((4-(nitrogen-(4-trifluoromethylphenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxymethyl Sulfonamide;

(Z)-2-((4-(氮-(4-三氟甲基苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基乙基磺酰氨；(Z)-2-((4-(nitrogen-(4-trifluoromethylphenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxyethyl Sulfonamide;

(Z)-2-((4-(氮-(4-三氟甲基苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基异丙基磺酰氨；(Z)-2-((4-(nitrogen-(4-trifluoromethylphenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxyiso Propylsulfonamide;

(Z)-2-((4-(氮-(4-三氟甲基苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基二甲基磺酰氨；(Z)-2-((4-(nitrogen-(4-trifluoromethylphenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxydi Methylsulfonamide;

(Z)-2-((4-(氮-(3-氟-4-氯苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基磺酰氨；(Z)-2-((4-(nitrogen-(3-fluoro-4-chlorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxy Sulfonamide;

(Z)-2-((4-(氮-(3-氟-4-氯苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基甲基磺酰氨；(Z)-2-((4-(nitrogen-(3-fluoro-4-chlorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxy Methylsulfonamide;

(Z)-2-((4-(氮-(3-氟-4-氯苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基乙基磺酰氨；(Z)-2-((4-(nitrogen-(3-fluoro-4-chlorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxy Ethylsulfonamide;

(Z)-2-((4-(氮-(3-氟-4-氯苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基异丙基磺酰氨；(Z)-2-((4-(nitrogen-(3-fluoro-4-chlorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxy Isopropylsulfonamide;

(Z)-2-((4-(氮-(3-氟-4-氯苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基二甲基磺酰氨；(Z)-2-((4-(nitrogen-(3-fluoro-4-chlorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino)ethoxy Dimethylsulfonamide;

(Z)-2-((4-(氮-(3-三氟甲基-4-氟苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基磺酰氨；(Z)-2-((4-(nitrogen-(3-trifluoromethyl-4-fluorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino) Ethoxysulfonamide;

(Z)-2-((4-(氮-(3-三氟甲基-4-氟苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基甲基磺酰氨；(Z)-2-((4-(nitrogen-(3-trifluoromethyl-4-fluorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino) Ethoxymethylsulfonamide;

(Z)-2-((4-(氮-(3-三氟甲基-4-氟苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基乙基磺酰氨；(Z)-2-((4-(nitrogen-(3-trifluoromethyl-4-fluorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino) Ethoxyethylsulfonamide;

(Z)-2-((4-(氮-(3-三氟甲基-4-氟苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基异丙基磺酰氨；(Z)-2-((4-(nitrogen-(3-trifluoromethyl-4-fluorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino) Ethoxyisopropylsulfonamide;

(Z)-2-((4-(氮-(3-三氟甲基-4-氟苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基二甲基磺酰氨；(Z)-2-((4-(nitrogen-(3-trifluoromethyl-4-fluorophenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino) Ethoxydimethylsulfonamide;

(Z)-2-((4-(氮-(3-氟-4-三氟甲基苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基磺酰氨；(Z)-2-((4-(nitrogen-(3-fluoro-4-trifluoromethylphenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino) Ethoxysulfonamide;

(Z)-2-((4-(氮-(3-氟-4-三氟甲基苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基甲基磺酰氨；(Z)-2-((4-(nitrogen-(3-fluoro-4-trifluoromethylphenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino) Ethoxymethylsulfonamide;

(Z)-2-((4-(氮-(3-氟-4-三氟甲基苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基乙基磺酰氨；(Z)-2-((4-(nitrogen-(3-fluoro-4-trifluoromethylphenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino) Ethoxyethylsulfonamide;

(Z)-2-((4-(氮-(3-氟-4-三氟甲基苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基异丙基磺酰氨；(Z)-2-((4-(nitrogen-(3-fluoro-4-trifluoromethylphenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino) Ethoxyisopropylsulfonamide;

(Z)-2-((4-(氮-(3-氟-4-三氟甲基苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基二甲基磺酰氨；(Z)-2-((4-(nitrogen-(3-fluoro-4-trifluoromethylphenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino) Ethoxydimethylsulfonamide;

(Z)-2-((4-(氮-(3，5-二氟-4甲氧基苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基磺酰氨；(Z)-2-((4-(nitrogen-(3,5-difluoro-4methoxyphenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino ) ethoxysulfonamide;

(Z)-2-((4-(氮-(3，5-二氟-4甲氧基苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基甲基磺酰氨；(Z)-2-((4-(nitrogen-(3,5-difluoro-4methoxyphenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino ) ethoxymethylsulfonamide;

(Z)-2-((4-(氮-(3，5-二氟-4甲氧基苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基乙基磺酰氨；(Z)-2-((4-(nitrogen-(3,5-difluoro-4methoxyphenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino ) ethoxyethylsulfonamide;

(Z)-2-((4-(氮-(3，5-二氟-4甲氧基苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基异丙基磺酰氨；和(Z)-2-((4-(nitrogen-(3,5-difluoro-4methoxyphenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino ) ethoxyisopropylsulfonamide; and

(Z)-2-((4-(氮-(3，5-二氟-4甲氧基苯基)-N'-羟基亚胺-1,2,5-噁二唑-3)胺基)乙氧基二甲基磺酰氨。(Z)-2-((4-(nitrogen-(3,5-difluoro-4methoxyphenyl)-N'-hydroxyimine-1,2,5-oxadiazole-3)amino ) Ethoxydimethylsulfonamide.

在另一优选例中，所述的化合物是实施例中所制备的化合物1-5。In another preferred embodiment, the compound is compound 1-5 prepared in the examples.

在另一优选例中，所述的化合物是外消旋体。In another preferred example, the compound is a racemate.

在另一优选例中，所述的化合物是对映异构体。In another preferred embodiment, the compounds are enantiomers.

在另一优选例中，所述药学上可接受的盐选自下组：盐酸盐、氢溴酸盐、硫酸盐、磷酸盐、甲磺酸盐、三氟甲磺酸盐、苯磺酸盐、对甲苯磺酸盐(甲苯磺酸盐)、1-萘磺酸盐、2-萘磺酸盐、乙酸盐、三氟乙酸盐、苹果酸盐、酒石酸盐、柠檬酸盐、乳酸盐、草酸盐、琥珀酸盐、富马酸盐、马来酸盐、苯甲酸盐、水杨酸盐、苯乙酸盐、扁桃酸盐。In another preferred embodiment, the pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, hydrobromide, sulfate, phosphate, methanesulfonate, trifluoromethanesulfonate, benzenesulfonic acid Salt, p-toluenesulfonate (toluenesulfonate), 1-naphthalenesulfonate, 2-naphthalenesulfonate, acetate, trifluoroacetate, malate, tartrate, citrate, milk Salicylate, Oxalate, Succinate, Fumarate, Maleate, Benzoate, Salicylate, Phenylacetate, Mandelate.

本发明的第二方面，提供第一方面所述的通式(I)化合物的制备方法，包括以下步骤：The second aspect of the present invention provides the preparation method of the compound of general formula (I) described in the first aspect, comprising the following steps:

(a)化合物A与化合物C反应，得到化合物B；(a) compound A reacts with compound C to obtain compound B;

(b)化合物B在碱水解条件下(如氢氧化钠水溶液)开环，得到终产物通式I化合物；(b) Compound B is ring-opened under alkaline hydrolysis conditions (such as aqueous sodium hydroxide solution) to obtain the final product compound of formula I;

各式中，R¹、R²、R³、R⁴、R⁵、n的定义如上所示。In each formula, R¹ , R² , R³ , R⁴ , R⁵ , and n are as defined above.

本发明也提供了第二种通式(I)化合物的制备方法，包括以下步骤：The present invention also provides a second preparation method of the compound of general formula (I), comprising the following steps:

(a)化合物D在酸的催化下与化合物E和一个还原剂反应，得到化合物F；(a) compound D reacts with compound E and a reducing agent under the catalysis of acid to obtain compound F;

(b)化合物F在碱水解条件下(如氢氧化钠水溶液)开环，得到终产物通式(I)化合物。(b) Compound F is ring-opened under alkaline hydrolysis conditions (such as aqueous sodium hydroxide solution) to obtain the compound of general formula (I) as the final product.

本发明的第三方面，提供第一方面所述的通式(I)化合物的用途，用于：The third aspect of the present invention provides the purposes of the compound of general formula (I) described in the first aspect, for:

(i)制备吲哚胺-2,3-双加氧酶抑制剂；(i) preparing an indoleamine-2,3-dioxygenase inhibitor;

(ii)制备预防和/或治疗吲哚胺-2,3-双加氧酶介导的疾病的药物；或(ii) preparing a medicament for the prevention and/or treatment of diseases mediated by indoleamine-2,3-dioxygenase; or

(iii)制备抗炎药物。(iii) Preparation of anti-inflammatory drugs.

在另一优选例中，所述吲哚胺-2,3-双加氧酶介导的疾病为IDO介导的色氨酸代谢途径的病理学特征的疾病。In another preferred example, the disease mediated by indoleamine-2,3-dioxygenase is a disease characterized by the pathology of IDO-mediated tryptophan metabolic pathway.

在另一优选例中，所述吲哚胺-2,3-双加氧酶介导的疾病为癌症、眼疾、心里障碍、抑郁症、焦虑症、老年痴呆症和/或自身免疫性疾病。In another preferred example, the disease mediated by indoleamine-2,3-dioxygenase is cancer, eye disease, heart disorder, depression, anxiety, Alzheimer's disease and/or autoimmune disease.

在另一优选例中，所述癌症包括但不限于：结肠癌、乳腺癌、胃癌、肺癌、大肠癌、胰腺癌、卵巢癌、前列腺癌、肾癌、肝癌、脑癌、黑色素瘤、多发性骨髓瘤、慢性粒细胞性白血病、血液肿瘤、淋巴肿瘤，包括在其他远离肿瘤原发部位的组织或器官的转移病变。In another preferred example, the cancers include, but are not limited to: colon cancer, breast cancer, gastric cancer, lung cancer, colorectal cancer, pancreatic cancer, ovarian cancer, prostate cancer, kidney cancer, liver cancer, brain cancer, melanoma, multiple Myeloma, chronic myelogenous leukemia, hematological neoplasms, lymphoid neoplasms, including metastatic lesions in other tissues or organs distant from the primary site of the tumor.

本发明的第四方面，提供一种药物组合物，所述药物组合物包含：A fourth aspect of the present invention provides a pharmaceutical composition comprising:

第一方面的通式(I)化合物或其药学上可以接受的盐、其立体异构体或其互变异构体、或其前药；以及The compound of general formula (I) of the first aspect or a pharmaceutically acceptable salt thereof, a stereoisomer or a tautomer thereof, or a prodrug thereof; and

药学上可接受的载体。pharmaceutically acceptable carrier.

本发明的第五方面，提供一种药物组合物，所述药物组合物包含：In a fifth aspect of the present invention, a pharmaceutical composition is provided, which comprises:

第一方面所述的通式(I)化合物或其药学上可以接受的盐、其立体异构体或其互变异构体、或其前药；以及The compound of general formula (I) described in the first aspect or its pharmaceutically acceptable salt, its stereoisomer or its tautomer, or its prodrug; and

抗肿瘤药物。antineoplastic drugs.

在另一优选例中，所述抗肿瘤药物包括但不限于癌症的免疫治疗药物：PD-1抗体，CTLA-4抗体，PD-L1抗体，PD-L2抗体，任何一种其它化疗药物或靶向治疗药物。In another preferred example, the anti-tumor drugs include but not limited to cancer immunotherapy drugs: PD-1 antibody, CTLA-4 antibody, PD-L1 antibody, PD-L2 antibody, any other chemotherapy drugs or target to therapeutic drugs.

本发明的第六方面，提供一种预防和/或治疗吲哚胺-2,3-双加氧酶介导的疾病的方法，包括对患者给予第一方面所述的通式(I)化合物或第四或第五方面所述的药物组合物的步骤。The sixth aspect of the present invention provides a method for preventing and/or treating diseases mediated by indoleamine-2,3-dioxygenase, comprising administering the compound of general formula (I) described in the first aspect to the patient Or the step of the pharmaceutical composition described in the fourth or fifth aspect.

在另一优选例中，所述吲哚胺-2,3-双加氧酶介导的疾病为癌症，所述方法进一步包含对患者施用额外的抗癌剂(也称为抗肿瘤药物，所述抗肿瘤药物如上所述)的步骤。In another preferred example, the disease mediated by indoleamine-2,3-dioxygenase is cancer, and the method further comprises administering additional anticancer agents (also known as antitumor drugs, so The steps of antineoplastic drugs as described above).

本发明的通式(I)化合物，具有抗肿瘤、治疗神经退行疾病(阿尔茨海默病)、抗炎等多种药理活性。The compound of the general formula (I) of the present invention has multiple pharmacological activities such as anti-tumor, treatment of neurodegenerative diseases (Alzheimer's disease), and anti-inflammation.

应理解，在本发明范围内中，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。限于篇幅，在此不再一一赘述。It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described in the following (such as embodiments) can be combined with each other to form new or preferred technical solutions. Due to the limited space, I won't go into details here.

具体实施方式detailed description

本发明人经过广泛而深入地研究，首次意外研发出一种新型的包含氨基磺酸脂和1,2,5-噁二唑结构的化合物，该化合物可作为高效的IDO酶抑制剂，用于预防和/或治疗吲哚胺-2,3-双加氧酶介导的疾病，也可作为抗炎药物使用。在此基础上，完成了本发明。After extensive and in-depth research, the inventors have accidentally developed a novel compound containing sulfamic acid ester and 1,2,5-oxadiazole structure for the first time, which can be used as a highly efficient IDO enzyme inhibitor for It can prevent and/or treat diseases mediated by indoleamine-2,3-dioxygenase, and can also be used as an anti-inflammatory drug. On this basis, the present invention has been accomplished.

在描述本发明之前，应当理解本发明不限于所述的具体方法和实验条件，因为这类方法和条件可以变动。还应当理解本文所用的术语其目的仅在于描述具体实施方案，并且不意图是限制性的，本发明的范围将仅由所附的权利要求书限制。Before the present invention is described, it is to be understood that this invention is not limited to the particular methods and experimental conditions described, as such methods and conditions may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, the scope of the present invention being limited only by the appended claims.

除非另外定义，否则本文中所用的全部技术与科学术语均具有如本发明所属领域的普通技术人员通常理解的相同含义。如本文所用，在提到具体列举的数值中使用时，术语“约”意指该值可以从列举的值变动不多于1％。例如，如本文所用，表述“约100”包括99和101和之间的全部值(例如，99.1、99.2、99.3、99.4等)。Unless defined otherwise, 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. As used herein, the term "about" when used in reference to a specifically recited value means that the value may 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 in between (eg, 99.1, 99.2, 99.3, 99.4, etc.).

虽然在本发明的实施或测试中可以使用与本发明中所述相似或等价的任何方法和材料，本文在此处例举优选的方法和材料。Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are exemplified herein.

定义definition

术语“烷基”是指一价饱和脂族烃基，具有1至10个碳原子，包括直链和支链烃基，如甲基(即CH₃-)、乙基(即CH₃CH₂-)、正丙基(即CH₃CH₂CH₂-)、异丙基(即(CH₃)₂CH-)、正丁基(即CH₃CH₂CH₂CH₂-)、异丁基(即(CH₃)₂CHCH₂-)、仲丁基(即(CH₃)(CH₃CH₂)CH-)、叔丁基(即(CH₃)₃C-)、正戊基(即CH₃CH₂CH₂CH₂CH₂-)、新戊基(即(CH₃)₃CCH₂-)。在本发明中，该术语包括取代或未取代的烷基。The term "alkyl" refers to a monovalent saturated aliphatic hydrocarbon group having 1 to 10 carbon atoms, including straight and branched chain hydrocarbon groups, such as methyl (ie CH₃ -), ethyl (ie CH₃ CH₂ -) , n-propyl (ie CH₃ CH₂ CH₂ -), isopropyl (ie (CH₃ )₂ CH-), n-butyl (ie CH₃ CH₂ CH₂ CH₂ -), isobutyl (ie (CH₃ )₂ CHCH₂ -), sec-butyl (ie (CH₃ )(CH₃ CH₂ )CH-), tert-butyl (ie (CH₃ )₃ C-), n-pentyl (ie CH₃_{CH2CH2CH2CH2-}₎ , neopentyl₍ ie (_CH3₎_3CCH2-₎ . In the present invention, the term includes substituted or unsubstituted alkyl groups.

如本文所用，术语“取代或未取代的”指所述基团可以是未取代的，或者所述基团中的H被一个或多个(较佳地1-6个，更佳地1-3个)取代基所取代。As used herein, the term "substituted or unsubstituted" means that the group can be unsubstituted, or that H in the group is replaced by one or more (preferably 1-6, more preferably 1- 3) Substituents are substituted.

如本文所用，所述的“取代”或“取代的”指所述基团具有一个或多个(较佳地1-6个，更佳地1-3个)选自下组的取代基：卤素、羟基、-NH₂、硝基、-CN、C1-C4烷基、C1-C4卤代烷基、C1-C4烷氧基、C3-C6环烷基、C2-C4链烯基、C2-C4炔基、苯基、苄基。As used herein, the "substituted" or "substituted" means that the group has one or more (preferably 1-6, more preferably 1-3) substituents selected from the following group: Halogen, hydroxyl, -NH₂ , nitro, -CN, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkoxy, C3-C6 cycloalkyl, C2-C4 alkenyl, C2-C4 Alkynyl, phenyl, benzyl.

如本文所用，术语“环烷基”指取代或未取代的C3-C12环烷基。As used herein, the term "cycloalkyl" refers to a substituted or unsubstituted C3-C12 cycloalkyl group.

如本文所用，术语“烷氧基”指-O-烷基，其中所述烷基可以是饱和或不饱和的、可以是支链、直链的、或环状的。优选地，烷氧基具有1-10个碳原子，较佳地1-6个碳原子。代表性的例子包括(但并不限于)：甲氧基、乙氧基、丙氧基。As used herein, the term "alkoxy" refers to an -O-alkyl group, wherein the alkyl group may be saturated or unsaturated, branched, straight chained, or cyclic. Preferably, the alkoxy group has 1-10 carbon atoms, preferably 1-6 carbon atoms. Representative examples include, but are not limited to: methoxy, ethoxy, propoxy.

如本文所用，术语“芳基”是指6至20个(较佳6-14个)碳原子的单价芳香族碳环基团，它具有单环(如苯基)或稠环(如萘基或蒽基)，如果连接点在芳香碳原上，稠环可能是非芳香性的(如2-苯并噁唑酮，2H-1,4-苯并噁嗪-3(4H)-酮-7-基等)。优选的芳基包括苯基和萘基。该术语包括取代或未取代的形式，其中取代基的定义如上。As used herein, the term "aryl" refers to a monovalent aromatic carbocyclic group of 6 to 20 (preferably 6-14) carbon atoms, which has a single ring (such as phenyl) or condensed rings (such as naphthyl or anthracenyl), if the point of attachment is on an aromatic carbon, the fused ring may be non-aromatic (such as 2-benzoxazolone, 2H-1,4-benzoxazin-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.

如本文所用，术语“烯基”是指具有2至10(如2至6或2至4)个碳原子的烯基，且具有至少1(如1至2)个不饱和烯键(>C＝C<)。这类基团的例如有乙烯基、烯丙基、丁-3-烯基。如本文所用，术语“环烷基”是指具有3至10个碳原子的、具有单环或多环(包括稠合体系，桥环体系和螺环体系)的环状烷基。在稠环体系中，一个或多个环可以是环烷基、杂环的、芳基或杂芳基，只要连接位点是通过环烷基的环。合适的环烷基的例子包括：例如，金刚烷基、环丙基、环丁基、环戊基和环辛基。As used herein, the term "alkenyl" refers to an alkenyl group having 2 to 10 (such as 2 to 6 or 2 to 4) carbon atoms and having at least 1 (such as 1 to 2) ethylenically unsaturated 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 3 to 10 carbon atoms, having a single ring or multiple rings (including fused systems, bridged ring systems and spiro ring systems). In fused ring systems, one or more rings can be cycloalkyl, heterocyclic, aryl, or heteroaryl as long as the point of attachment is through the cycloalkyl ring. Examples of suitable cycloalkyl groups include, for example, adamantyl, cyclopropyl, cyclobutyl, cyclopentyl and cyclooctyl.

如本文所用，术语“卤代”或“卤素”是指氟、氯、溴和碘。As used herein, the term "halo" or "halogen" refers to fluorine, chlorine, bromine and iodine.

如本文所用，术语“杂芳基”是指环内具有1至10个碳原子和1至4个选自氧、氮和硫的杂原子的芳香基团，这样的杂芳基可以是单环的(如吡啶基或呋喃基)或稠环(如吲嗪基(indolizinyl)或苯并噻吩基)，其中，所述稠环可以是非芳香性的和/或含有一个杂原子，只要连接点是通过芳香性杂芳基的原子。在一实施例中，杂芳基的环原子氮和/或硫任选地被氧化为N-氧化物(N-O)，亚磺酰基或磺酰基。优选地杂芳基包括吡啶基、吡咯基、吲哚基、噻吩基和呋喃基。该术语包括取代或未取代的杂芳基。As used herein, the term "heteroaryl" refers to an aromatic group having 1 to 10 carbon atoms and 1 to 4 heteroatoms selected from oxygen, nitrogen and sulfur within the ring, such heteroaryl groups may be monocyclic (such as pyridyl or furyl) or fused ring (such as indolizinyl (indolizinyl) or benzothienyl), wherein the fused ring can be non-aromatic and/or contain a heteroatom as long as the point of attachment is through An atom of an aromatic heteroaryl group. In one embodiment, the ring atom nitrogen and/or sulfur of the heteroaryl is optionally oxidized to N-oxide (N-O), sulfinyl or sulfonyl. Preferred heteroaryl groups include pyridyl, pyrrolyl, indolyl, thienyl and furyl. The term includes substituted or unsubstituted heteroaryl groups.

如本文所用，术语“取代的杂芳基”是指被1至5个、优选1至3个、更优选1至2个的取代基所取代的杂芳基，所述取代基选自与取代的芳基所定义的相同取代基。As used herein, the term "substituted heteroaryl" refers to a heteroaryl group substituted by 1 to 5, preferably 1 to 3, more preferably 1 to 2, substituents selected from and substituted The same substituents defined for the aryl group.

如本文所用，术语“杂环”或“杂环的”或“杂环烷基”或“杂环基”是指饱和的、部分饱和的或不饱和的基团(但不是芳香性的)，具有单环或稠环(包括桥环体系和螺环体系，环内具有1至10个碳原子和1至4个选自氮、硫或氧的杂原子，在稠环体系中，一个或多个环可以是环烷基、芳基或杂芳基，只要连接点通过非芳香性环。在一实施例中，杂环基团的氮原子和/或硫原子任选地被氧化，以提供N-氧化物、亚磺酰基和磺酰基部分。As used herein, the term "heterocycle" or "heterocyclic" or "heterocycloalkyl" or "heterocyclyl" refers to a saturated, partially saturated or unsaturated group (but not aromatic), Has a single ring or condensed ring (including bridged ring system and spiro ring system, with 1 to 10 carbon atoms and 1 to 4 heteroatoms selected from nitrogen, sulfur or oxygen in the ring, in the condensed ring system, one or more Each ring can be a cycloalkyl, aryl or heteroaryl as long as the point of attachment is through a non-aromatic ring. In one embodiment, the nitrogen and/or sulfur atoms of the heterocyclic group are optionally oxidized to provide N-oxide, sulfinyl and sulfonyl moieties.

如本文所用，术语“取代的杂环的”或“取代的杂环烷基”或“取代的杂环基”是指被1到5(如1至3)个取代基所取代的杂环基团，所述取代基与取代的环烷基所定义的取代基相同。As used herein, the term "substituted heterocyclic" or "substituted heterocycloalkyl" or "substituted heterocyclyl" refers to a heterocyclyl group substituted with 1 to 5 (eg 1 to 3) substituents group, the substituents are the same as defined for substituted cycloalkyl groups.

如本文所用，术语“立体异构体”是指一个或多个立体中心的手性不同的化合物。立体异构体包括对映异构体和非对映异构体。As used herein, the term "stereoisomer" refers to compounds that differ in chirality at one or more stereocenters. Stereoisomers include enantiomers and diastereomers.

如本文所用，术语“互变异构体”是指质子位置不同的化合物的替代形式，如烯醇-酮和亚胺-烯胺互变异构体，或杂芳基的互变异构形式，所述杂芳基包含与环的-NH-部分和环的＝N-部分连接的环原子，如吡唑、咪唑、苯并咪唑、三唑和四唑。As used herein, the term "tautomer" refers to alternative forms of compounds that differ in the position of the proton, such as enol-keto and imine-enamine tautomers, or tautomeric forms of heteroaryl groups , the heteroaryl group contains ring atoms attached to the -NH- portion of the ring and the =N- portion of the ring, such as pyrazole, imidazole, benzimidazole, triazole and tetrazole.

“前药”是指实施例化合物的任何衍生物，当被施用于受试者时，其能够直接或间接地提供实施例的化合物或其活性代谢物或残余物。特别优选的衍生物和前药是那些，当被施用于受试者时，提高实施例化合物的生物利用度(如口服给药的化合物更容易被吸收进入血液)或相对于母体种类提高母体化合物到生物区室(如脑或淋巴系统)的运送的衍生物和前药。前药包括本发明化合物的酯类形式。"Prodrug" refers to any derivative of a compound of the Examples 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 compounds of the Examples (e.g., an orally administered compound is more readily absorbed into the bloodstream) or increase the parent compound relative to the parent species. Delivery of derivatives and prodrugs to biological compartments such as the brain or lymphatic system. Prodrugs include ester forms of the compounds of the invention.

本发明化合物Compounds of the invention

如本文所用，术语“本发明化合物”指通式(I)、通式(II)或通式(III)化合物、其氘代化合物、外消旋体、其立体异构体或其互变异构体、或前药、或其药学上可以接受的盐。本发明涉及：这些化合物的外消旋混合物，富集任一种对映体的混合物，以及任一种分离的对映体。对于本发明的范围，应当理解为，所述外消旋混合物指两种R和S对映体50％:50％的混合物。所述分离的对映体应理解为纯的对映体(即100％)或者高度富集某种对映体(纯度≥98％、≥95％、≥93％、≥90％、≥88％、≥85％、≥80％)的混合物。As used herein, the term "compound of the present invention" refers to a compound of general formula (I), general formula (II) or general formula (III), its deuterated compound, racemate, its stereoisomer or its tautomer Constructs, or prodrugs, or pharmaceutically acceptable salts thereof. The present invention relates to: racemic mixtures of these compounds, mixtures enriched in either enantiomer, and either isolated enantiomer. For the scope of the present invention, it is understood that said racemic mixture refers to a 50%:50% mixture of the two R and S enantiomers. The separated enantiomers should be understood as pure enantiomers (i.e. 100%) or highly enriched enantiomers (purity ≥ 98%, ≥ 95%, ≥ 93%, ≥ 90%, ≥ 88% , ≥85%, ≥80%) mixture.

典型地，本发明提供通式(I)化合物或其药学上可以接受的盐、其立体异构体或其互变异构体、或前药，Typically, the present invention provides a compound of general formula (I) or a pharmaceutically acceptable salt thereof, a stereoisomer or a tautomer thereof, or a prodrug,

式中，In the formula,

X为一个单键、O，S、NH或NR^d；X is a single bond, O, S, NH or NR^d ;

n为2至8；n is 2 to 8;

在另一优选例中，X为NH。In another preferred example, X is NH.

在另一优选例中，n为2至6。In another preferred example, n is 2-6.

在另一优选例中，所述通式(I)化合物为：In another preferred example, the compound of general formula (I) is:

式中，In the formula,

n为2-6。n is 2-6.

R²为H、C₁-C₁₀烷基；R² is H, C₁ -C₁₀ alkyl;

在另一优选例中，所述通式(I)化合物如通式(III)所示，In another preferred example, the compound of general formula (I) is represented by general formula (III),

式中，In the formula,

在本发明所述的化合物有立体异构体存在的情况下，本发明包括化合物的所有立体异构体。Where a compound described herein exists as a stereoisomer, the present invention includes all stereoisomers of the compound.

在本发明所述的化合物有互变异构体存在的情况下，本发明包括化合物的所有互变异构体。Where compounds described herein exist as tautomers, the present invention includes all tautomers of the compounds.

本发明还包括所述化合物中的任何一个或多个氢原子被其稳定同位素氘取代而产生的氘代化合物。The present invention also includes deuterated compounds in which any one or more hydrogen atoms in the compounds are replaced by its stable isotope deuterium.

除特别说明之处，本发明的所有化合物之中，各手性碳原子(手性中心)可以任选地为R构型或S构型，或R构型和S构型的混合物。Unless otherwise specified, in all compounds of the present invention, each chiral carbon atom (chiral center) may optionally be in R configuration or S configuration, or a mixture of R configuration and S configuration.

药物组合物pharmaceutical composition

本发明还提供了一种药物组合物，它包含安全有效量范围内的活性成分，以及药学上可接受的载体。The present invention also provides a pharmaceutical composition, which contains active ingredients in a safe and effective dose range, and a pharmaceutically acceptable carrier.

本发明所述的“活性成分”是指本发明所述的通式(I)化合物或其药学上可以接受的盐、其立体异构体或其互变异构体、或其前药。The "active ingredient" in the present invention refers to the compound of general formula (I) or its pharmaceutically acceptable salt, its stereoisomer or its tautomer, or its prodrug in this invention.

本发明所述的“活性成分”和药物组合物可用作IDO抑制剂。在另一优选例中，用于制备预防和/或治疗肿瘤的药物。在另一优选例中，用于制备预防和/或治疗IDO介导的疾病的药物。The "active ingredients" and pharmaceutical compositions described in the present invention can be used as IDO inhibitors. In another preferred embodiment, it is used for preparing drugs for preventing and/or treating tumors. In another preferred embodiment, it is used for preparing medicines for preventing and/or treating diseases mediated by IDO.

“安全有效量”指的是：活性成分的量足以明显改善病情，而不至于产生严重的副作用。通常，药物组合物含有1-2000mg活性成分/剂，更佳地，含有10-200mg活性成分/剂。较佳地，所述的“一剂”为一个药片。"Safe and effective amount" means: the amount of the active ingredient is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 1-2000 mg active ingredient/dose, more preferably 10-200 mg active ingredient/dose. Preferably, the "one dose" is a tablet.

“药学上可接受的载体”指的是：一种或多种相容性固体或液体填料或凝胶物质，它们适合于人使用，而且必须有足够的纯度和足够低的毒性。“相容性”在此指的是组合物中各组份能和本发明的活性成分以及它们之间相互掺和，而不明显降低活性成分的药效。"Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use, and must have sufficient purity and low toxicity. "Compatibility" here means that each component in the composition can be blended with the active ingredient of the present invention and with each other without significantly reducing the efficacy of the active ingredient.

本发明优选实施例的化合物可以作为单独活性药剂给药，也可以与一个或多个其它用于治疗癌症的试剂组合使用。本发明优选实施例的化合物与已知的治疗剂和抗癌剂组合使用也是有效的，目前已知的化合物和其它抗癌剂或化疗剂的组合在优选实施例范围之内。这类药剂的例子可参见《癌症原理和实践肿瘤学》(CancerPrinciplesandPracticeofOncology)，V.T.Devita和S.Hellman(编者)，第6版(2001年2月15日)，LippincottWilliams&Wilkins出版社。基于药物的特殊性质和所涉及的癌症，本领域普通技术人员能够辨别有效的药剂组合。这种抗癌剂包括(但不限于)如下：HDAC抑制剂、雌激素受体调节剂、雄激素受体调节剂、视黄醇类受体调节剂、细胞毒性/细胞生长抑制剂、抗增殖剂、异戊烯基蛋白转移酶抑制剂、HMG-CoA还原酶抑制剂和其他血管生成抑制剂、细胞增殖和生存信号抑制剂、凋亡诱导剂和干扰细胞周期检查点(cellcyclecheckpoint)的试剂、CTLA4抗体、PD-1抗体、PD-L1抗体等。优选实施例的化合物与放射治疗同时施用时也有效。The compounds of the preferred embodiments of the present invention may be administered as the sole active agent or in combination with one or more other agents useful in the treatment of cancer. Compounds of the preferred embodiments of this invention are also effective in combination with known therapeutic and anticancer agents, 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 Cancer Principles and Practice of Oncology, V.T. Devita and S. Hellman (Eds.), 6th Edition (February 15, 2001), Lippincott Williams & Wilkins Press. One of ordinary skill in the art will be able to discern effective combinations of agents based on the particular properties of the drugs and the cancer involved. Such anticancer agents include, but are not limited to, the following: HDAC inhibitors, estrogen receptor modulators, androgen receptor modulators, retinoid receptor modulators, cytotoxic/cytostatic, antiproliferative agents, prenyl protein transferase inhibitors, HMG-CoA reductase inhibitors and other angiogenesis inhibitors, cell proliferation and survival signaling inhibitors, apoptosis inducers and reagents that interfere with cell cycle checkpoints, CTLA4 antibody, PD-1 antibody, PD-L1 antibody, etc. The compounds of the preferred embodiments are also effective when administered concurrently with radiation therapy.

通常，优选实施例的化合物将以治疗有效量、通过具有类似作用的药剂的任意一种可接受的模式施用。优选实施例的化合物(即活性成分)的实际用量根据多个因素确定，如待治疗疾病的严重程度、患者的年龄和相对健康程度、被使用化合物的效力、施用的路径和形式，以及其他因素。该药物可一天施用多次，优选地，每天一次或两次。所有这些因素都在主治医生的考虑范围内。In general, the compounds of the preferred embodiments will be administered in a therapeutically effective amount by any of the accepted modes of agents having similar effects. The actual amount of the compound (i.e., active ingredient) of the preferred embodiments to be used will depend on a number of factors, such as the severity of the disease being 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.

优选实施例的目的，治疗有效剂量通常可以是对患者一次性施用或分次施用的每日总剂量，例如，每日约0.001至约1000毫克/公斤体重，优选地，每日约1.0至约30毫克/千克体重。单位剂量组合物(Dosageunitcomposition)可包含其剂量因数以形成每日剂量。剂型的选择取决于各种因素，例如给药模式和药物物质的生物利用度。通常，优选实施例的化合物可作为药物组合物通过以下任意一种路线给药：口服、全身给药(如透皮、鼻内或通过栓剂)、或肠外给药(如肌内、静脉内或皮下)。优选的给药方式为口服，可根据苦的程度调节方便的日剂量。组合物可采取的形式为片剂、丸剂、胶囊、半固体、粉剂、缓释制剂、溶液、悬浮液、酏剂、气雾剂或任何其他适当的组合物。另一种优选的施用优选实施例化合物的方式为吸入。这是一种将治疗剂直接运送给呼吸道的有效方法(参见，如美国专利号5,607,915)。For the purposes of preferred embodiments, the therapeutically effective dose can generally be a total daily dose administered to the patient once or in divided doses, for example, about 0.001 to about 1000 mg/kg body weight per day, preferably about 1.0 to about 30 mg/kg body weight. Dosage unit compositions may contain dosage factors thereof to give a daily dose. The choice of dosage form depends 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 pharmaceutical compositions by any of the following routes: oral, systemic (such as transdermal, intranasal, or via suppository), or parenteral (such as intramuscular, intravenous or subcutaneous). The preferred mode of administration is oral administration, and the convenient daily dose can be adjusted according to 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 composition. Another preferred mode of administering the compounds of the preferred embodiments is by inhalation. This is an effective method of delivering therapeutic agents directly to the respiratory tract (see, eg, US Patent No. 5,607,915).

合适的药学上可接受的载体或赋形剂包括：如处理剂和药物运送改性剂和促进剂，诸如磷酸钙、硬脂酸镁、滑石、单糖、二糖、淀粉、明胶、纤维素、甲基纤维素钠、羧甲基纤维素、葡萄糖、羟丙基-B-环糊精、聚乙烯吡咯烷酮、低熔点蜡、离子交换树脂等，及其任意两种或多种的组合。液体和半固体的赋形剂可以选自甘油、丙二醇、水、乙醇和各种油，包括石油、动物油、植物油或合成来源，如花生油、豆油、矿物油、芝麻油等。优选的液体载体，特别是用于可注射溶液的载体，包括水、盐水、葡萄糖水性溶液和乙二醇。其它适宜的药学上可接受的赋形剂在《雷明顿药物科学》(Remington’sPharmaceuticalSciences),MackPub.Co.,新泽西(1991)有描述，通过引用纳入本文。Suitable pharmaceutically acceptable carriers or excipients include, for example, treating agents and drug delivery modifiers and enhancers such as calcium phosphate, magnesium stearate, talc, monosaccharides, disaccharides, starch, gelatin, cellulose , sodium methylcellulose, carboxymethylcellulose, glucose, hydroxypropyl-B-cyclodextrin, polyvinylpyrrolidone, low melting point wax, ion exchange resin, etc., and any combination of two or more thereof. Liquid and semisolid excipients can be selected from glycerol, propylene glycol, water, ethanol and various oils, including petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. Preferred liquid carriers, especially for injectable solutions, include water, saline, aqueous dextrose and glycol. Other suitable pharmaceutically acceptable excipients are described in Remington's Pharmaceutical Sciences, Mack Pub. Co., New Jersey (1991), incorporated herein by reference.

如本文所用，术语“药学上可接受的盐”是指通式I化合物的非毒性酸或碱土金属盐。这些盐可在最终分离和纯化通式I化合物时原位制得、或分别将合适的有机或无机酸或碱与碱性或酸性官能团反应制得。代表性的盐包括，但不限于：乙酸盐、己二酸盐、藻酸盐、柠檬酸盐、天冬氨酸盐、苯甲酸盐、苯磺酸盐、硫酸氢盐、丁酸盐、樟脑酸盐、樟脑磺酸盐、二葡糖酸盐、环戊烷丙酸盐、十二烷基硫酸盐、乙磺酸盐、葡萄糖庚酸盐、甘油磷酸盐、半硫酸盐、庚酸盐、己酸盐、富马酸盐、盐酸盐、氢溴酸盐、氢碘酸盐、2-羟基乙磺酸盐、乳酸盐、马来酸盐、甲磺酸盐、烟酸盐、2-萘基磺酸盐、草酸盐、双羟萘酸盐、果胶酸盐、硫氰酸盐、3-苯基丙酸盐、苦味酸盐、新戊酸盐、丙酸盐、琥珀酸盐、硫酸盐、酒石酸盐、硫氰酸盐、对甲苯磺酸盐和十一烷酸盐。此外，含氮的碱性基团可被如下试剂季铵盐化：烷基卤化物，如甲基、乙基、丙基、丁基的氯化物、溴化物和碘化物；二烷基硫酸盐，如二甲基、二乙基、二丁基和二戊基硫酸酯；长链卤化物如癸基、月桂基、肉豆蔻基和硬脂基的氯化物、溴化物和碘化物；芳烷基卤化物如苄基和苯乙基溴化物等。由此得到水溶性或油溶性或可分散产品。可被用于形成药学上可接受的酸加成盐的酸的例子包括如盐酸、硫酸、磷酸的无机酸，和如草酸、马来酸、甲磺酸、琥珀酸、柠檬酸的有机酸。碱加成盐可在最终分离和纯化通式I的化合物时原位制得、或使羧酸部分分别与合适的碱(如药学上可接受的金属阳离子的氢氧化物，碳酸盐或碳酸氢盐)或氨、或有机伯、仲或叔胺反应制得。药学上可接受的盐包括，但不限于，基于碱金属和碱土金属的阳离子，如钠、锂、钾、钙、镁、铝的盐等，以及无毒的铵、季铵和胺阳离子，包括，但不限于：铵、四甲基铵、四乙基铵、甲胺、二甲胺、三甲胺、三乙胺、乙胺等。其它代表性的用于形成碱加成盐的有机胺包括二乙胺、乙二胺、乙醇胺、二乙醇胺、哌嗪等。As used herein, the term "pharmaceutically acceptable salt" refers to non-toxic acid or alkaline earth metal salts of compounds of general formula I. These salts can be prepared in situ during the final isolation and purification of the compounds of general formula I, or by reacting suitable organic or inorganic acids or bases with basic or acidic functional groups, respectively. Representative salts include, but are not limited to: acetate, adipate, alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate , camphorate, camphorsulfonate, digluconate, cyclopentanepropionate, lauryl sulfate, ethanesulfonate, glucose heptanoate, glycerophosphate, hemisulfate, heptanoic acid Salt, Caproate, Fumarate, Hydrochloride, Hydrobromide, Hydroiodide, 2-Hydroxyethanesulfonate, Lactate, Maleate, Methanesulfonate, Nicotinate , 2-naphthylsulfonate, oxalate, pamoate, pectate, thiocyanate, 3-phenylpropionate, picrate, pivalate, propionate, Succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate and undecanoate. In addition, nitrogen-containing basic groups can be quaternized with the following reagents: alkyl halides, such as chlorides, bromides, and iodides of methyl, ethyl, propyl, and butyl groups; dialkyl sulfates , such as dimethyl, diethyl, dibutyl, and dipentyl sulfate; long-chain halides such as decyl, lauryl, myristyl, and stearyl chlorides, bromides, and iodides; arane Halides such as benzyl and phenethyl bromide, etc. Water-soluble or oil-soluble or dispersible products are thus obtained. Examples of acids which can be used to form pharmaceutically acceptable acid addition salts include inorganic acids such as hydrochloric acid, sulfuric acid, phosphoric acid, and organic acids such as oxalic acid, maleic acid, methanesulfonic acid, succinic acid, citric acid. Base addition salts can be prepared in situ during the final isolation and purification of the compounds of general formula I, or by separately reacting the carboxylic acid moiety with a suitable base such as a pharmaceutically acceptable metal cation hydroxide, carbonate or carbonic acid 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 salts, etc., as well as non-toxic ammonium, quaternary ammonium and amine cations, including , but not limited to: ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, etc. Other representative organic amines for the formation of base addition salts include diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, and the like.

如本文所用，术语“药学上可接受的前药”是指那些优选实施例的化合物的前药，在体内迅速转化为上述通式所示的母体化合物的化合物，例如在血液中水解。在“T.Higuchi和V.Stella，作为新型运送系统的前药(Pro-drugsasNovelDeliverySystems)，A.C.S.15SymposiumSeries的14卷”和“EdwardB.Roche编，药物设计中的生物可逆载体(BioreversibleCarriersinDrugDesign)，美国药学协会和Pergamon出版社，1987年”中提供了完整的讨论，这两者都引入本文作为参考。As used herein, the term "pharmaceutically acceptable prodrug" refers to those prodrugs of the compounds of the preferred embodiments, which are rapidly converted into the parent compound of the above general formula in vivo, eg, by hydrolysis in blood. In "T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems (Pro-drugs as Novel Delivery Systems), Volume 14 of A.C.S. 15 Symposium Series" and "Edward B. Roche, eds., Bioreversible Carriers in Drug Design (Bioreversible Carriers in Drug Design), American Pharmaceutical Association and Pergamon Press, 1987", both of which are incorporated herein by reference.

下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件如Sambrook等人，分子克隆：实验室手册(NewYork:ColdSpringHarborLaboratoryPress,1989)中所述的条件，或按照制造厂商所建议的条件。除非另外说明，否则百分比和份数按重量计算。Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. For the experimental methods without specific conditions indicated in the following examples, generally follow conventional conditions such as the conditions described in Sambrook et al., Molecular Cloning: Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the conditions suggested by the manufacturer. Percentages and parts are by weight unless otherwise indicated.

本发明的有益之处在于：The benefits of the present invention are:

(1)提供一种结构新颖的通式(I)化合物；(1) provide a compound of general formula (I) with a novel structure;

(2)本发明的化合物可以作为高效的IDO酶抑制剂；(2) The compounds of the present invention can be used as efficient IDO enzyme inhibitors;

(3)合成方法温和，操作简单易行，收率较高，易于衍生化，适合工业放大量生产；(3) The synthesis method is mild, the operation is simple and easy, the yield is high, and it is easy to derivatize, and is suitable for large-scale industrial production;

(4)具有抗肿瘤、神经退行性疾病(阿尔茨海默病)、抗炎等多种药理活性。(4) It has various pharmacological activities such as anti-tumor, neurodegenerative diseases (Alzheimer's disease), and anti-inflammation.

除非另行定义，文中所使用的所有专业与科学用语与本领域熟练人员所熟悉的意义相同。此外，任何与所记载内容相似或均等的方法及材料皆可应用于本发明方法中。文中所述的较佳实施方法与材料仅作示范之用。Unless otherwise defined, all professional and scientific terms used herein have the same meanings as commonly understood by those skilled in the art. In addition, any methods and materials similar or equivalent to those described can be applied to the method of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

实施例1Example 1

第一步：4-氨基-氮’-羟基-1,2,5-噁二唑-3-甲脒(化合物D)的制备The first step: the preparation of 4-amino-nitrogen'-hydroxyl-1,2,5-oxadiazole-3-carboxamidine (compound D)

将丙二氰(32.0g,485mmol)溶于600mL水中，加热至完全溶解。冰水浴冷却下，加入亚硝酸钠(38.0g,533mmol)和6N盐酸(5.5mL)。在冰浴下反应0.5小时后，升温至室温反应2小时。将反应液继续冰浴冷却，将50％的盐酸羟胺水溶液(99.0g,1500mmol)滴加入反应液，搅拌半小时后，升至室温反应1小时。加热回流反应2小时，反应完全后，冰浴下，用80mL6N盐酸调节pH至7.0。过滤沉淀，用水和乙酸乙酯各洗一次，干燥，得到白色化合物69.3g,收率92％。Dissolve malondicyanide (32.0g, 485mmol) in 600mL water and heat until completely dissolved. Under cooling in an ice-water bath, sodium nitrite (38.0 g, 533 mmol) and 6N hydrochloric acid (5.5 mL) were added. After reacting in an ice bath for 0.5 hours, the temperature was raised to room temperature and reacted for 2 hours. The reaction solution was continued to be cooled in an ice bath, and 50% aqueous solution of hydroxylamine hydrochloride (99.0 g, 1500 mmol) was added dropwise to the reaction solution. After stirring for half an hour, it was raised to room temperature for 1 hour of reaction. Heat to reflux for 2 hours. After the reaction is complete, adjust the pH to 7.0 with 80 mL of 6N hydrochloric acid in an ice bath. The precipitate was filtered, washed once with water and ethyl acetate, and dried to obtain 69.3 g of a white compound with a yield of 92%.

¹³CNMR(75MHz,DMSO-d₆):δ154.4,144.0,140.0。¹³ CNMR (75MHz, DMSO-d₆ ): δ 154.4, 144.0, 140.0.

MSESI:m/z＝144.0,[M+H]⁺.MSESI: m/z＝144.0,[M+H]⁺ .

第二步：4-氨基-氮-羟基-1,2,5-噁二唑-3-甲脒氯(化合物E)的制备The second step: the preparation of 4-amino-nitrogen-hydroxyl-1,2,5-oxadiazole-3-carboxamidine chloride (compound E)

将化合物D(30.0g,210mmol)溶于213mL乙酸，分别加入水420mL，盐酸105mL和氯化钠(36.7g,629mmol)。冰浴下，滴加入溶于50mL水的亚硝酸钠(14.2g,206mmol)，保持零度反应1小时，后升温至室温反应5小时。反应结束后，过滤沉淀，滤饼用水洗，抽干后，用乙酸乙酯溶解，无水硫酸钠干燥，过滤，浓缩绿叶，得到类白色固体17.1g，收率51％。Compound D (30.0 g, 210 mmol) was dissolved in 213 mL of acetic acid, and 420 mL of water, 105 mL of hydrochloric acid and sodium chloride (36.7 g, 629 mmol) were added respectively. Under ice-cooling, sodium nitrite (14.2 g, 206 mmol) dissolved in 50 mL of water was added dropwise, kept at zero temperature for 1 hour, and then heated to room temperature for 5 hours. After the reaction, the precipitate was filtered, the filter cake was washed with water, drained, dissolved in ethyl acetate, dried over anhydrous sodium sulfate, filtered, and the green leaves were concentrated to obtain 17.1 g of an off-white solid with a yield of 51%.

¹³CNMR(75MHz,DMSO-d₆):δ154.4,142.3,126.9。¹³ CNMR (75MHz, DMSO-d₆ ): δ 154.4, 142.3, 126.9.

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

第三步：4-氨基-氮’-羟基-N-((2-甲氧)乙基))-1,2,5-噁二唑-3-甲脒(化合物F)的制备The third step: the preparation of 4-amino-nitrogen'-hydroxyl-N-((2-methoxy)ethyl))-1,2,5-oxadiazole-3-carboxamidine (compound F)

将化合物E(15.0g,92mmol)溶于200mL乙酸乙酯，冰浴下，2-甲氧基-1-乙胺(7.6mL,101mmol)加入反应液中，搅拌十分钟后，加入三乙胺(14.0,138mmol)，反应1小时。将反应液用水洗一次，盐水洗一次，无水硫酸钠干燥，得棕色油状物15.0g，收率81％。Compound E (15.0g, 92mmol) was dissolved in 200mL ethyl acetate, under ice cooling, 2-methoxy-1-ethylamine (7.6mL, 101mmol) was added to the reaction solution, after stirring for ten minutes, triethylamine was added (14.0, 138mmol), reacted for 1 hour. The reaction solution was washed once with water and once with brine, and dried over anhydrous sodium sulfate to obtain 15.0 g of a brown oil with a yield of 81%.

¹HNMR(300MHz,DMSO-d₆)：δ10.66(s,1H),6.27(s,2H),6.14(t,1H),3.50(m,2H),3.35(t,2H),3.18(s,3H)。¹ HNMR (300MHz, DMSO-d₆ ): δ10.66(s,1H),6.27(s,2H),6.14(t,1H),3.50(m,2H),3.35(t,2H),3.18( s, 3H).

第四步：氮’-羟基-4-(((2-甲氧)乙基)氨基)-1,2,5-噁二唑-3-甲脒(化合物G)的制备The fourth step: the preparation of nitrogen'-hydroxyl-4-(((2-methoxy)ethyl)amino)-1,2,5-oxadiazole-3-carboxamidine (compound G)

将化合物F(15.0g,75.0mmol)加入65mL水中，氢氧化钾(12.5g,224mmol)，回流反应30小时。将反应液冷却到室温，乙酸乙酯萃取3次，合并有机层，干燥，旋干溶剂，得白色固体粗品9.0g，收率60％。Compound F (15.0 g, 75.0 mmol) was added into 65 mL of water, potassium hydroxide (12.5 g, 224 mmol), and refluxed for 30 hours. The reaction solution was cooled to room temperature, extracted three times with ethyl acetate, the organic layers were combined, dried, and the solvent was spin-dried to obtain 9.0 g of a white solid crude product with a yield of 60%.

¹HNMR(300MHz,DMSO-d₆)：δ10.54(s,1H),6.21(s,1H),6.13(t,1H),3.49(m,2H),3.36(t,2H),3.25(s,3H)。¹ HNMR (300MHz, DMSO-d₆ ): δ10.54(s,1H),6.21(s,1H),6.13(t,1H),3.49(m,2H),3.36(t,2H),3.25( s, 3H).

第五步：氮-羟基-4-(((2-甲氧)乙基)氨基)1,2,5-噁二唑-3-甲脒氯(化合物H)的制备Step 5: Preparation of nitrogen-hydroxyl-4-(((2-methoxy)ethyl)amino)1,2,5-oxadiazole-3-carboxamidine chloride (compound H)

将化合物G(9.0g,45mmol)溶于50mL乙酸，分别加入水50mL，盐酸45mL，氯化钠(7.8g,135mmol)。冰浴下，滴加入溶于18mL水的亚硝酸钠(3.2g,4.3mmol)，保持零度反应1小时，后升温至室温反应5小时。反应结束后，乙酸乙酯萃取3次，合并有机层，水和盐水各洗一次，旋干溶剂，干燥，得白色固体化合物8.0g,收率81％。Compound G (9.0 g, 45 mmol) was dissolved in 50 mL of acetic acid, and 50 mL of water, 45 mL of hydrochloric acid and sodium chloride (7.8 g, 135 mmol) were added respectively. Under ice-cooling, sodium nitrite (3.2 g, 4.3 mmol) dissolved in 18 mL of water was added dropwise, kept at zero temperature for 1 hour, and then heated to room temperature for 5 hours. After the reaction was completed, extracted three times with ethyl acetate, combined the organic layers, washed with water and brine each time, spin-dried the solvent, and dried to obtain 8.0 g of white solid compound with a yield of 81%.

¹HNMR(300MHz,DMSO-d₆):δ13.41(s,1H),5.88(t,1H)3.53(m,2H),3.39(t,2H),3.25(s,3H)。¹ H NMR (300 MHz, DMSO-d₆ ): δ13.41 (s, 1H), 5.88 (t, 1H) 3.53 (m, 2H), 3.39 (t, 2H), 3.25 (s, 3H).

第六步：氮-(3-溴-4-氟苯基)-N’-羟基-4-(((2-甲氧)乙基)氨基)-1,2,5-噁二唑-3-甲脒(化合物I)的制备Step 6: Nitrogen-(3-bromo-4-fluorophenyl)-N'-hydroxy-4-(((2-methoxy)ethyl)amino)-1,2,5-oxadiazole-3 - Preparation of formamidine (compound I)

将化合物H(8.0g,36mmol)溶于50mL水，加热到60℃，加入3-溴-4-氟苯胺(6.8g,36mmol)，搅拌十分钟后，加入碳酸氢钠水溶液(4.6g,54mmol)。反应1小时后，冷却至室温。反应结束后，乙酸乙酯萃取3次，合并有机层，旋干溶剂，干燥，得黄色固体15.0g，收率111％。Dissolve compound H (8.0g, 36mmol) in 50mL of water, heat to 60°C, add 3-bromo-4-fluoroaniline (6.8g, 36mmol), stir for ten minutes, add aqueous sodium bicarbonate (4.6g, 54mmol ). After reacting for 1 hour, it was cooled to room temperature. After the reaction, ethyl acetate was extracted three times, the organic layers were combined, the solvent was spin-dried, and dried to obtain 15.0 g of a yellow solid with a yield of 111%.

¹HNMR(300MHz,DMSO-d₆):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)。¹ HNMR (300MHz, DMSO-d₆ ): 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).

第七步：4-(3-溴-4-氟苯基)-3-4-(((2-甲氧)乙基)氨基)-1,2,5-噁二唑-3-基)-1,2-4-噁二唑酮(化合物J)的制备The seventh step: 4-(3-bromo-4-fluorophenyl)-3-4-(((2-methoxy)ethyl)amino)-1,2,5-oxadiazol-3-yl) Preparation of -1,2-4-oxadiazolone (compound J)

将化合物I(15.0g,40mmol)溶于150mL乙酸乙酯，加入羰基二咪唑(9.8g,60mmol)，加热到60℃反应1小时。溶液冷却至室温，乙酸乙酯萃取3次，合并有机层，旋干溶剂，干燥，得黄色固体12.7g，收率79％。Compound I (15.0 g, 40 mmol) was dissolved in 150 mL of ethyl acetate, carbonyldiimidazole (9.8 g, 60 mmol) was added, and heated to 60° C. for 1 hour. The solution was cooled to room temperature, extracted three times with ethyl acetate, the organic layers were combined, the solvent was spin-dried, and dried to obtain 12.7 g of a yellow solid with a yield of 79%.

¹HNMR(300MHz,DMSO-d₆):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)。¹ HNMR (300MHz, DMSO-d₆ ): 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).

第八步：4-(3-溴-4-氟苯基)3-4-(((2-羟基乙氧基)-1,2,5-噁二唑-3-基)-1,2,4-噁二唑酮(化合物K)的制备The eighth step: 4-(3-bromo-4-fluorophenyl) 3-4-(((2-hydroxyethoxy)-1,2,5-oxadiazol-3-yl)-1,2 , Preparation of 4-oxadiazolone (compound K)

将化合物J(12.5g,31mmol)溶于95mL并降温至-67°，缓慢加入三溴化硼，(15.7mL,62.6mmol)，加完后将反应混合物缓慢生至-10°反应1小时。反应结束后，冰浴下在混合液中缓慢低价饱和碳酸钠溶液调节pH至7.用乙酸乙酯萃取3次，合并有机层，干燥，旋干溶剂，得白色固体11.0g,收率91％。Compound J (12.5g, 31mmol) was dissolved in 95mL and cooled to -67°, boron tribromide (15.7mL, 62.6mmol) was slowly added, and after the addition was completed, the reaction mixture was slowly brought to -10° for 1 hour. After the reaction was over, slowly adjust the pH to 7 with a low-priced saturated sodium carbonate solution in the mixed solution under an ice bath. Extracted 3 times with ethyl acetate, combined the organic layers, dried, and spin-dried the solvent to obtain 11.0 g of a white solid with a yield of 91 %.

¹HNMR(300MHz,CDCl3-d₆):δ7.64(dd,1H),7.36(m,1H),7.31(m,1H),5.64(s,1H),3.93(m,2H),3.59(m,1H)。¹ HNMR (300MHz, CDCl3-d₆ ): δ7.64(dd,1H),7.36(m,1H),7.31(m,1H),5.64(s,1H),3.93(m,2H),3.59( m, 1H).

第九步：2-((4-(4-(3-溴-4-氟苯基)-5-酮-4,5-二羟-1,2,4-噁二唑-3-基)-1,2,5-噁二唑-3-基)胺)乙基)磺酰胺(化合物L)的制备The ninth step: 2-((4-(4-(3-bromo-4-fluorophenyl)-5-keto-4,5-dihydroxy-1,2,4-oxadiazol-3-yl) Preparation of -1,2,5-oxadiazol-3-yl)amine)ethyl)sulfonamide (compound L)

冰浴下，将氯磺酰异氰酸酯(139mg,0.98mmol)溶于2mL二氯甲烷溶液中，缓慢滴加入甲酸(45mg,0.98mmol)，保持零度反应1小时，然后室温搅拌过夜。冰浴下，将化合物K(100mg,0.26mmol)溶于1mLDMA,缓慢滴加入上述反应液中，继续搅拌3小时。加水，乙酸乙酯萃取3次，合并有机层，旋干溶剂，干燥，得粗品60mg，收率50％。Under ice-cooling, chlorosulfonyl isocyanate (139mg, 0.98mmol) was dissolved in 2mL of dichloromethane solution, formic acid (45mg, 0.98mmol) was slowly added dropwise, kept at zero temperature for 1 hour, then stirred overnight at room temperature. Under ice-cooling, compound K (100 mg, 0.26 mmol) was dissolved in 1 mL DMA, slowly added dropwise to the above reaction solution, and stirring was continued for 3 hours. Add water, extract 3 times with ethyl acetate, combine the organic layers, spin to dry the solvent, and dry to obtain 60 mg of crude product with a yield of 50%.

¹HNMR(400MHz,DMSO-d₆):δ8.11(dd,1H),7.75(m,1H),7.71(t,1H),6.72(t,1H),4.22(t,2H),3.61(m,2H)。¹ HNMR (400MHz, DMSO-d₆ ): δ8.11(dd,1H),7.75(m,1H),7.71(t,1H),6.72(t,1H),4.22(t,2H),3.61( m,2H).

第十步：(Z)-2-((4-(N-(3-溴-4-氟苯基)-N’-羟基亚胺基)-1,2,5-噁二唑-3-基)胺)乙基)磺酰胺(化合物1)的制备The tenth step: (Z)-2-((4-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyimino)-1,2,5-oxadiazole-3- base) amine) ethyl) sulfonamide (compound 1) preparation

将化合物L(60mg,0.13mmol)，溶于入2mL四氢呋喃，冰浴下加入2N氢氧化钠溶液0.5mL，室温反应1小时，用2N盐酸调节pH至4-5。乙酸乙酯萃取3次，合并有机层，干燥，过滤，旋干溶剂，柱层析(二氯甲烷：甲醇＝20:1)，得白色固体50mg，收率88％。Compound L (60mg, 0.13mmol) was dissolved in 2mL of tetrahydrofuran, 0.5mL of 2N sodium hydroxide solution was added under ice-cooling, and reacted at room temperature for 1 hour, and the pH was adjusted to 4-5 with 2N hydrochloric acid. Extracted three times with ethyl acetate, combined the organic layers, dried, filtered, spin-dried the solvent, and column chromatography (dichloromethane:methanol=20:1) to obtain 50 mg of white solid with a yield of 88%.

¹HNMR(400MHz,DMSO-d₆):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)；¹ HNMR (400MHz, DMSO-d₆ ): 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);

MSESI:m/z＝439.0,[M+H]⁺。MSESI: m/z = 439.0, [M+H]⁺ .

实施例2Example 2

根据实施例1制备方法，将化合物K(50mg,0.13mmol)溶于1mLDMA，冰浴下滴加甲胺基磺酰氯(34mg,0.26mmol)，室温反应3小时，然后将反应混合物倒入3mL水中，将析出固体过滤收集得到60mg呈白色固体的中间体，然后与实例1的第十步相同的条件处理此中间体得到20mg呈白色固体的实例2。According to the preparation method of Example 1, compound K (50mg, 0.13mmol) was dissolved in 1mLDMA, methylaminosulfonyl chloride (34mg, 0.26mmol) was added dropwise under ice cooling, and reacted at room temperature for 3 hours, and then the reaction mixture was poured into 3mL of water , the precipitated solid was collected by filtration to obtain 60 mg of an intermediate as a white solid, and then the intermediate was treated under the same conditions as in the tenth step of Example 1 to obtain 20 mg of Example 2 as a white solid.

¹HNMR(400MHz,CDCl₃):δ7.46(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)；¹ HNMR (400MHz, CDCl₃ ): δ7.46(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);

MSESI:m/z＝452.6,[M+H]⁺。MSESI: m/z = 452.6, [M+H]⁺ .

实施例3Example 3

根据实施例2制备方法，将化合物K(50mg,0.13mmol)与乙胺基磺酰氯反应，然后水解得到呈白色固体的实例3。According to the preparation method of Example 2, compound K (50 mg, 0.13 mmol) was reacted with ethylsulfonyl chloride, and then hydrolyzed to obtain Example 3 as a white solid.

¹HNMR(400MHz,CDCl₃):δ7.55(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)；¹ HNMR (400MHz, CDCl₃ ): δ7.55(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);

MSESI:m/z＝466.6,[M+H]⁺。MSESI: m/z = 466.6, [M+H]⁺ .

实施例4Example 4

根据实施例2制备方法，将化合物K(50mg,0.13mmol)与异丙胺基磺酰氯反应，然后水解得到呈白色固体的实例4。According to the preparation method of Example 2, compound K (50 mg, 0.13 mmol) was reacted with isopropylaminosulfonyl chloride, and then hydrolyzed to obtain Example 4 as a white solid.

¹HNMR(400MHz,CDCl₃):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)；¹ HNMR (400MHz, CDCl₃ ): 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);

MSESI:m/z＝480.7,[M+H]⁺。MSESI: m/z = 480.7, [M+H]⁺ .

实施例5Example 5

第一步：将2，2-二甲氧基乙醇(50mg,0.47mmol)溶于3mL二氯甲烷，冰浴下，加入钠氢(38mg,0.94mmol).搅拌半小时后，缓慢滴加入二甲胺基磺酰氯(138mg,0.94mmol)，室温搅拌4小时，减压浓缩，15mL乙酸乙酯加入到反应浓缩物，然后加热回流2小时，此混合物冷却至室温、过滤、减压浓缩得到70mg白色固体的产物M。Step 1: Dissolve 2,2-dimethoxyethanol (50mg, 0.47mmol) in 3mL of dichloromethane, add sodium hydrogen (38mg, 0.94mmol) under ice-cooling. After stirring for half an hour, slowly add two Methylaminosulfonyl chloride (138 mg, 0.94 mmol), stirred at room temperature for 4 hours, concentrated under reduced pressure, 15 mL of ethyl acetate was added to the reaction concentrate, then heated to reflux for 2 hours, the mixture was cooled to room temperature, filtered, and concentrated under reduced pressure to obtain 70 mg Product M as white solid.

¹HNMR(400MHz,CDCl₃):δ7.27(s,1H),4.63(t,3H),4.10(d,2H),3.43(s,6H),2.90(s,6H)；¹ HNMR (400MHz, CDCl₃ ): δ7.27(s, 1H), 4.63(t, 3H), 4.10(d, 2H), 3.43(s, 6H), 2.90(s, 6H);

MSESI:m/z＝236.1,[M+Na]⁺。MSESI: m/z = 236.1, [M+Na]⁺ .

第二步：将化合物M(53mg,0.16mmol)，三乙基硅烷(125uL，0.78mmol),甲磺酸(80uL，1.23mmol)溶于5mL二氯甲烷，然后冰浴下，加入3-(4-氨基-1，2，5-噁二唑-3-基)-4-(3-溴-4-氟苯基)1，2，4-噁唑烷-5(4H)-酮(50mg,0.23mmol),缓慢升至室温反应9小时。碳酸氢钠水溶液调节pH至7，然后分别用水和饱和NaCl溶液洗，减压浓缩有机相，粗产物用硅胶柱纯化，流动相是3:1至1:1的正己烷/乙酸乙酯，得到50mg呈白色固体的产物N。The second step: Dissolve compound M (53mg, 0.16mmol), triethylsilane (125uL, 0.78mmol), methanesulfonic acid (80uL, 1.23mmol) in 5mL dichloromethane, then add 3-( 4-Amino-1,2,5-oxadiazol-3-yl)-4-(3-bromo-4-fluorophenyl)1,2,4-oxazolidin-5(4H)-one (50mg , 0.23mmol), slowly rose to room temperature for 9 hours. Adjust the pH to 7 with aqueous sodium bicarbonate solution, then wash with water and saturated NaCl solution respectively, and concentrate the organic phase under reduced pressure. The crude product is purified with a silica gel column, and the mobile phase is n-hexane/ethyl acetate from 3:1 to 1:1 to obtain 50 mg of product N as a white solid.

¹HNMR(400MHz,CDCl₃):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)；¹ HNMR (400MHz, CDCl₃ ): 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);

MSESI:m/z＝492.7,[M+H]⁺。MSESI: m/z = 492.7, [M+H]⁺ .

第三步：将化合物N(50mg,0.10mmol)溶于3mL四氢呋喃，然后冰浴下，加入2N氢氧化钠溶液0.5mL,缓慢升至室温反应3小时，然后减压浓缩反应混合物，加入乙酸乙酯和水至浓缩物，混合摇动后分出有机相，用MgSO₄干燥后减压浓缩，粗产物用硅胶柱纯化，流动相是3:1至1:1的正己烷/乙酸乙酯，得到25mg呈白色固体的产物。Step 3: Dissolve compound N (50mg, 0.10mmol) in 3mL tetrahydrofuran, then add 0.5mL 2N sodium hydroxide solution under ice bath, slowly rise to room temperature and react for 3 hours, then concentrate the reaction mixture under reduced pressure, add ethyl acetate Ester and water to the concentrate, mixed and shaken, the organic phase was separated, dried with MgSO₄ and concentrated under reduced pressure, the crude product was purified with a silica gel column, and the mobile phase was n-hexane/ethyl acetate from 3:1 to 1:1 to obtain 25 mg of product as a white solid.

¹HNMR(400MHz,CDCl₃):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)；¹ HNMR (400MHz, CDCl₃ ): 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);

MSESI:m/z＝467.0,[M+H]⁺。MSESI: m/z = 467.0, [M+H]⁺ .

实施例6Example 6

以3-(4-氨基-1，2，5-噁二唑-3-基)-4-(3-氯-4-氟苯基)1，2，4-噁唑烷-5(4H)-酮和化合物M为原料，用实例5描述的方法，得到白色固体的实例6。With 3-(4-amino-1,2,5-oxadiazol-3-yl)-4-(3-chloro-4-fluorophenyl)1,2,4-oxazolidine-5(4H) - Ketone and compound M as starting materials, using the method described in Example 5, to obtain Example 6 as a white solid.

¹HNMR(400MHz,Acetone-d₃):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).¹ HNMR (400MHz, Acetone-d₃ ): 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).

MSESI:m/z＝423.0,[M+H]⁺。MSESI: m/z = 423.0, [M+H]⁺ .

实施例7Example 7

以3-(4-氨基-1，2，5-噁二唑-3-基)-4-(3-三氟甲基-4-氟苯基)1，2，4-噁唑烷-5(4H)-酮和化合物M为原料，用实例5描述的方法，得到白色固体的实例7。With 3-(4-amino-1,2,5-oxadiazol-3-yl)-4-(3-trifluoromethyl-4-fluorophenyl)1,2,4-oxazolidine-5 Starting from (4H)-ketone and compound M, using the method described in Example 5, Example 7 was obtained as a white solid.

¹HNMR(400MHz,Acetone-d₃):δ10.75(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(dt,2H),2.87(s,6H).¹ HNMR (400MHz, Acetone-d₃ ): δ10.75(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(dt,2H), 2.87(s,6H).

MSESI:m/z＝457.0,[M+H]⁺。MSESI: m/z = 457.0, [M+H]⁺ .

实施例8Example 8

活性测试activity test

(1)IDO蛋白的诱导表达及纯化方法(1) Induced expression and purification method of IDO protein

首先PCR扩增IDO基因，扩增的PCR产物回收，然后将pET28a质粒(购自上海宝曼生物科技有限公司)和IDO胶回收产物用EcoRI和XhoI两种限制性内切酶进行酶切(37℃，酶切2h)，跑胶，回收，T4连接酶链接过夜连接产物加入到DH5α感受态，冰上放置30min，42℃热击90s，摇菌涂板，挑取单克隆，PCR鉴定，测序鉴定，全部正确，即pET28a-IDO质粒构建成功。First, the IDO gene was amplified by PCR, and the amplified PCR product was recovered, and then the pET28a plasmid (purchased from Shanghai Baoman Biotechnology Co., Ltd.) and the product recovered from IDO gel were digested with two restriction enzymes EcoRI and XhoI (37 ℃, enzyme digestion 2h), running gel, recovery, T4 ligase linked overnight ligation product was added to DH5α competent, placed on ice for 30min, heat shock at 42℃ for 90s, shake the plate, pick a single clone, PCR identification, sequencing The identification was all correct, that is, the pET28a-IDO plasmid was constructed successfully.

将构建好的含有pET28a-IDO质粒的BL21，37℃大摇至OD600为0.6-0.8，加入至终浓度为7μM的氯高铁血红素和1mM的IPTG(异丙基-β-D-硫代半乳糖苷)，28℃诱导4h；诱导后，4℃，6000rpm离心收集菌体，收集的菌体用20mMPBS(pH6.5)清洗一次，再离心收集菌体。The constructed BL21 containing the pET28a-IDO plasmid was shaken at 37°C until the OD600 was 0.6-0.8, and the final concentration was 7 μM hemin and 1 mM IPTG (isopropyl-β-D-thiosemi Lactose), induced at 28° C. for 4 h; after induction, the cells were collected by centrifugation at 4° C. at 6000 rpm, washed once with 20 mMPBS (pH 6.5), and collected by centrifugation again.

将收集的菌体用裂解液(20mMPBSpH6.5)重新悬起，超声裂解(功率40％裂解20min，冰上放置)，将裂解后的细菌，13000rpm离心15min，弃去沉淀，保留上清；将镍柱用裂解液(20mMPBSpH6.5)平衡3个柱体积，然后将裂解上清上样到镍柱上，上样之后，用漂洗液(20mMPBSpH6.5，20mM咪唑)清洗4个柱体积，最后用洗脱液(20mMPBSpH6.5，250mM咪唑)洗脱蛋白；将洗脱的蛋白溶液进行透析4h，透析溶液为20mMPBSpH6.5，透析之后蛋白样品浓缩，分装，液氮速冻，放入-80℃保存备用。The collected bacteria were resuspended with lysate (20mMPBSpH6.5), ultrasonically lysed (power 40% lysed for 20min, placed on ice), the lysed bacteria were centrifuged at 13000rpm for 15min, the precipitate was discarded, and the supernatant was retained; The nickel column was equilibrated for 3 column volumes with lysate (20mMPBSpH6.5), and then the lysed supernatant was loaded onto the nickel column. After loading, 4 column volumes were washed with rinse solution (20mMPBSpH6.5, 20mM imidazole), and finally Elute the protein with the eluent (20mMPBSpH6.5, 250mM imidazole); dialyze the eluted protein solution for 4h, the dialysis solution is 20mMPBSpH6.5, after dialysis, the protein sample is concentrated, subpackaged, quick-frozen in liquid nitrogen, and put Store at ℃ for later use.

(2)IDO酶抑制活性测试方法(2) Test method for IDO enzyme inhibitory activity

首先将化合物进行3倍梯度稀释，各个浓度取1μL加入到96孔板中；加入50μL配好的IDO酶溶液(终浓度600ng/100μL)：加入25μL底物1混合溶液，加入25μL的底物2混合溶液起始反应。最后OD₃₂₁nm读数60min。Firstly, the compound was serially diluted 3 times, and 1 μL of each concentration was added to a 96-well plate; 50 μL of prepared IDO enzyme solution (final concentration 600ng/100 μL) was added: 25 μL of substrate 1 mixed solution was added, and 25 μL of substrate 2 was added. The mixed solution initiates the reaction. The final OD₃₂₁ nm reading was 60 min.

(3)细胞活性测试方法(3) Cell Viability Test Method

Hela细胞(100μL)接种在96孔板上，接种量为每个孔5×10³，生长过夜。第二天，化合物稀释后，取1μL加入到96孔板中，然后将含有人的干扰素γ(终浓度50ng/mL)的培养基100μL加入到96孔板中，使最终体积为200μL。48小时孵化后，每个孔取140μL上清液转移到一个新的96孔板上。10μL6.1N三氯乙酸加入每个孔混合，50℃孵化30分钟，IDO催化N甲酰犬尿素为犬尿素。反应混合物2500转离心10分钟去掉沉淀物。每个孔100μL上清液转移到一个新的96孔板与100μL2％二甲氨基苯甲醛乙酸溶液混合。犬尿素分离后，用SPECTRAmaxi3reader在480nm测定数值。Hela cells (100 μL) were inoculated on a 96-well plate at an inoculum size of 5×10³ 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 the 96-well plate to make the final volume 200 μL. After 48 hours of incubation, transfer 140 μL of supernatant from each well to a new 96-well plate. Add 10 μL of 6.1N trichloroacetic acid to each well to mix and incubate at 50°C for 30 minutes. IDO catalyzes N-formyl kynurene into kynurene. The reaction mixture was centrifuged at 2500 rpm for 10 minutes to remove the precipitate. Transfer 100 μL supernatant from each well to a new 96-well plate and mix with 100 μL 2% dimethylaminobenzaldehyde acetic acid solution. After separation of kynurene, the value was measured at 480nm with SPECTRAmaxi3reader.

本发明化合物的IDO酶抑制活性和细胞抑制活性的测试结果如表1所示。Table 1 shows the test results of IDO enzyme inhibitory activity and cell inhibitory activity of the compounds of the present invention.

表1IDO酶和细胞抑制活性测试结果Table 1 IDO enzyme and cytostatic activity test result

上述结果表明，本发明化合物具有优异的针对IDO酶和细胞的抑制活性。The above results show that the compounds of the present invention have excellent inhibitory activity against IDO enzymes and cells.

实施例9化合物1在大鼠中的药代动力学数据Pharmacokinetic data of embodiment 9 compound 1 in rats

1.给药方案1. Dosing regimen

SD大鼠6只，雄性，体重180-200g，灌胃或静脉给予化合物1(表2)：6 SD rats, male, with a body weight of 180-200g, were given compound 1 by intragastric or intravenous administration (Table 2):

表2Table 2

灌胃给药以1％MC配制为悬浮液，静脉注射以DMSO:30％PEG400(5:95)配制。试验前禁食12h，自由饮水。给药后2h统一进食。It is prepared as a suspension with 1% MC for intragastric administration, and as a suspension with DMSO:30% PEG400 (5:95) for intravenous injection. Fasting 12h before the test, free to drink water. Eat uniformly 2 hours after administration.

2.采血时间点及样品处理：2. Blood collection time point and sample processing:

灌胃给药：给药后0.25，0.5，1.0，2.0，4.0，6.0，8.0和24h；Oral administration: 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0 and 24 hours after administration;

静脉给药：给药后5min，0.25，0.5，1.0，2.0，4.0，6.0，8.0和24h；Intravenous administration: 5min, 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0 and 24h after administration;

在以上设定时间点经大鼠眼球后静脉丛取静脉血0.3ml，置EDTA-2K试管中，11000rpm离心5min，分离血浆，于–20℃冰箱中冷冻。At the above set time points, 0.3ml of venous blood was collected through the retroocular venous plexus of rats, placed in EDTA-2K test tubes, centrifuged at 11000rpm for 5min, separated from plasma, and frozen in a -20°C refrigerator.

3.样品测试和数据分析3. Sample testing and data analysis

采用LC/MS/MS法测定大鼠血浆中化合物1的浓度。The concentration of compound 1 in rat plasma was determined by LC/MS/MS method.

采用WinNonlin5.3软件(美国Pharsight公司)的非房室模型计算给药后的药代动力学参数。The non-compartmental model of WinNonlin5.3 software (Pharsight, USA) was used to calculate the pharmacokinetic parameters after administration.

达峰浓度C_max和达峰时间T_max为实测值；The peak concentration C_max and the peak time T_max are measured values;

药时曲线下面积AUC_0-t值：采用梯形法计算；AUC_0-∞＝AUC_0-t+C_t/k_e，C_t为最后一个可测得时间点的血药浓度，k_e为消除速率常数；AUC_0-t value of the area under the drug-time curve: calculated by trapezoidal method; AUC_0-∞ = AUC_0-t + C_t /k_e , C_t is the blood drug concentration at the last measurable time point, k_e is Elimination rate constant;

消除半衰期t_1/2＝0.693/k_e；Elimination half-life t_1/2 = 0.693/_ke ;

平均滞留时间MRT＝AUMC/AUC。Mean residence time MRT=AUMC/AUC.

清除率CL＝D/AUC_0-∞；Clearance rate CL=D/AUC_0-∞ ;

稳态分布容积V_ss＝CL×MRTSteady-state volume of distribution V_ss =CL×MRT

绝对生物利用度F＝(AUC_灌胃×D_静脉)/(AUC_静脉×D_灌胃)×100％Absolute bioavailability F = (AUC_gavage × D_vein ) / (AUC_intravenous × D_gavage ) × 100%

4.试验结果4. Test results

大鼠灌胃给予10mg/kg实例1、静脉注射1mg/kg化合物1后的药动学参数见表3和表4。See Table 3 and Table 4 for the pharmacokinetic parameters after intragastric administration of 10 mg/kg of Example 1 and intravenous injection of 1 mg/kg of Compound 1 in rats.

表3大鼠灌胃10mg/kg实例1后的药动学参数Pharmacokinetic parameters after table 3 rat gavage 10mg/kg example 1

表4大鼠静脉胃给药1mg/kg实例1后的药动学参数Pharmacokinetic parameters after table 4 rat intravenous gastric administration 1mg/kg example 1

结果表明，静脉注射给药后，实例1在大鼠体内清除率(CL)2.85L/h/kg，稳态分布容积(Vss)为6.932L/kg，暴露量(AUC_0-t)为354ng·h/mL。灌胃给药后实例1在大鼠体内血浆达峰时间T_max为0.33h，暴露量(AUC_0-t)为1552ng·h/mL，剂量标准化后，绝对生物利用度为43.8％。The results showed that after intravenous administration, the clearance rate (CL) of Example 1 in rats was 2.85L/h/kg, the steady state volume of distribution (Vss) was 6.932L/kg, and the exposure (AUC_0-t ) was 354ng h/mL. After intragastric administration, the peak plasma time T_max of Example 1 in rats was 0.33 h, the exposure (AUC_0-t ) was 1552 ng·h/mL, and the absolute bioavailability was 43.8% after dose standardization.

实施例10实例1在小鼠中的药代动力学数据The pharmacokinetic data of embodiment 10 example 1 in mice

1.实验目的1. Purpose of the experiment

实例1口服给药ICR小鼠后，于不同时间点采集血样，LC-MS/MS测定给予受试物后小鼠血浆中实例1的浓度并计算相关药代参数，考察实例1在小鼠体内的药代属性。Example 1 After oral administration to ICR mice, blood samples were collected at different time points, and LC-MS/MS was used to measure the concentration of Example 1 in the mouse plasma after administration of the test substance and calculate relevant pharmacokinetic parameters, and investigate Example 1 in mice. pharmacokinetic properties.

2.测试方法2. Test method

与实例9相同。Same as Example 9.

3.动物实验3. Animal experiments

实验设计：ICR小鼠3只，由苏州康润新药研发有限公司提供，按表6进行实验。Experimental design: 3 ICR mice were provided by Suzhou Kangrun New Drug Research and Development Co., Ltd., and the experiment was carried out according to Table 6.

表6Table 6

样品采集Sample Collection

每只动物每次异氟烷麻醉后通过眼眶取血约25μL血液，EDTAK₂抗凝，采集时间点为：给予受试物前(0hr)和给予受试物后15min，30min，1h，2h，4h，6h，8h和24h。血液样本采集后置于冰上，并于1小时之内离心分离血浆(离心条件：5000转/分钟，10分钟，4℃)。收集的血浆分析前存放于–20℃。Each animal was anesthetized with isoflurane each time and about 25 μL of blood was collected through the orbit, anticoagulated with EDTAK₂ , and the collection time points were: before the administration of the test substance (0hr) and 15min, 30min, 1h, 2h after the administration of the test substance, 4h, 6h, 8h and 24h. Blood samples were placed on ice after collection, and the plasma was centrifuged within 1 hour (centrifugation conditions: 5000 rpm, 10 minutes, 4°C). The collected plasma was stored at –20°C until analysis.

4数据处理4 Data processing

数据采集及控制系统软件为Analyst1.5.1软件(AppliedBiosystem)。图谱样品峰积分方式为自动积分；采用样品峰面积和内标峰面积的比值作为指标，和样品的浓度进行回归。回归方式：线性回归，权重系数为1/X²。药代动力学参数用WinNonlinProfessionalv6.3(Pharsight,USA)用非房室模型分析处理。C_max为实测的最大血药浓度，血药浓度-时间曲线下面积AUC_(0→t)由梯形法计算得到，T_max为给药后血药浓度达峰时间。实验数据用“均数±标准差”(Mean±SD,n≥3)或“均数”(Mean,n＝2)表示。Data acquisition and control system software is Analyst1.5.1 software (AppliedBiosystem). 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 the index, and the concentration of the sample is used for regression. Regression method: linear regression, the weight coefficient is 1/X² . Pharmacokinetic parameters were analyzed using WinNonlin Professional v6.3 (Pharsight, USA) with non-compartmental model. C_max is the measured maximum blood drug concentration, the area under the blood drug concentration-time curve AUC_(0→t) is calculated by the trapezoidal method, and T_max is the peak time of blood drug concentration after administration. The experimental data are expressed by "mean ± standard deviation" (Mean ± SD, n≥3) or "mean" (Mean, n=2).

5实验结果5 Experimental results

实例1的口服给药后(剂量为30.0mg/kg)在ICR小鼠体内药代动力学研究结果表明，实例1在小鼠体内血药浓度平均达峰时间为0.25hr，达峰浓度为809.3±254ng/mL，AUC_0→t为982±151hr*ng/mL，体内平均滞留时间MRT为6.76±4.13hr.After the oral administration of example 1 (dosage is 30.0mg/kg) in vivo pharmacokinetics research result in ICR mice shows, example 1 is 0.25hr on average in the blood drug concentration peak time in mouse body, and peak concentration is 809.3 ±254ng/mL, AUC_0→t is 982±151hr*ng/mL, mean residence time MRT in vivo is 6.76±4.13hr.

ICR小鼠给药实例1在血浆中不同时间点的浓度(PO,30mg/kg)如表7所示。Table 7 shows the concentrations (PO, 30 mg/kg) of Example 1 in plasma at different time points in ICR mice.

表7Table 7

3.计算给药后的药代参数，见药代动力学参数3. Calculate the pharmacokinetic parameters after administration, see pharmacokinetic parameters

根据血药浓度数据，使用WinNonlinV6.3非房室模型计算给药后的药代参数见表8。According to the blood drug concentration data, the pharmacokinetic parameters after administration were calculated using the WinNonlinV6.3 non-compartmental model, as shown in Table 8.

表8.实例1在ICR小鼠中的药代动力学参数(口服:30mg/kg)Table 8. Pharmacokinetic parameters of example 1 in ICR mice (oral: 30mg/kg)

结果表明，实例1具有良好的药代动力学参数。The results show that Example 1 has good pharmacokinetic parameters.

在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。All documents mentioned in this application are incorporated by reference in this application as if each were individually incorporated by reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.