WO2019007140A1

WO2019007140A1 - 4-aminopyrimidine derivative used as adenosine a2a receptor antagonist and application thereof

Info

Publication number: WO2019007140A1
Application number: PCT/CN2018/085021
Authority: WO
Inventors: 钟燕; 曹西蓉; 王永临
Original assignee: 上海肇钰医药科技有限公司
Priority date: 2017-07-05
Filing date: 2018-04-28
Publication date: 2019-01-10
Also published as: CN107286146B; CN107286146A

Abstract

Disclosed is a 4-aminopyrimidine derivative used as an adenosine A2A receptor antagonist, having general structural formula (I). R1 is selected from halogen, cyano group, or trifluoromethyl; R2 is selected from pyrazolyl, pyrrolidyl, or pyrazolyl or pyrrolidyl substituted by one or more halogens or C1-3 alkyl groups; R3 is selected from oxazolyl, oxadiazole group, triazole group, or oxazolyl substituted by one or more halogens or C1-3 alkyl groups. The 4-aminopyrimidine derivative provided by the present invention has an obvious antagonistic action on a human-derived adenosine A2A receptor, and can be applied to compositions or combination products for treating diseases or pathological symptoms having responses to the A2A antagonistic action, in particular for treating neurodegenerative disease, extrapyramidal syndrome, depression, hyperkinetic syndrome, sleep disorder, anxiety disorder, diabetes, tumor and other diseases.

Description

作为腺苷A 2A受体拮抗剂的4-氨基嘧啶衍生物及其用途 Adenosine A 4-aminopyrimidine derivative of 2A receptor antagonist and use thereof

技术领域Technical field

本发明涉及药物和有机化学领域，具体涉及一种作为腺苷A _2A受体拮抗剂的4-氨基嘧啶衍生物及其用途。 The present invention relates to the field of pharmaceuticals and organic chemistry, and in particular to a 4-aminopyrimidine derivative as an adenosine A _2A receptor antagonist and uses thereof.

背景技术Background technique

作为中枢(CNS)和外周神经***中多数生理功能的内源性调节剂，腺苷广泛分布于神经***、心血管***、消化***、呼吸***等多组织多器官中，起到调节各种重要生理过程的作用。它通过一类膜特异性受体来发挥其生物作用，这些受体属于G蛋白偶联受体超家族，目前已经确定了4种腺苷受体亚型，它们分别是：A ₁、A _2A、A _2B和A ₃。其中A ₁和A _2A是高表达受体，在生理状态下低水平腺苷浓度时就可产生作用，而A _2B和A ₃的表达量较低，仅在病理情况下，腺苷大量增加时才可活化，产生一系列病理效应。A ₁和A ₃受体通过它们与抑制腺苷酸环化酶的G蛋白偶联来向下调节细胞的水平；相反，A _2A和A _2B受体与活化腺苷酸环化酶的Gs蛋白偶联，并提高细胞内的水平。通过这些受体的作用，腺苷能够实现广泛的生理功能调节。 As an endogenous regulator of most physiological functions in the central (CNS) and peripheral nervous system, adenosine is widely distributed in multiple tissues and organs such as the nervous system, cardiovascular system, digestive system, and respiratory system. The role of physiological processes. It exerts its biological effects through a class of membrane-specific receptors belonging to the G-protein coupled receptor superfamily. Four adenosine receptor subtypes have been identified, respectively: A ₁ , A _2A , A _2B and A ₃ . Among them, A ₁ and A _2A are high expression receptors, which can produce a low level of adenosine concentration under physiological conditions, while A _2B and A ₃ have low expression levels, and only a large amount of adenosine increases under pathological conditions. It can be activated and produces a series of pathological effects. The A ₁ and A ₃ receptors regulate the level of cells down by their coupling to the G protein that inhibits adenylate cyclase; in contrast, the A _2A and A _2B receptors and the Gs protein that activates adenylate cyclase Coupling and increasing intracellular levels. Through the action of these receptors, adenosine can achieve a wide range of physiological function regulation.

A _2A受体大多出现在富含多巴胺的区域，例如基底的神经节元件、各种哺乳动物包括人类的纹状体和苍白球。基底神经节与纹状体作为中心元件，参与皮质、丘脑和边缘叶的信息整合以产生运动行为。在纹状体中已经发现A _2A受体和多巴胺D2受体紧密地共同集中在纹状体苍白球γ-氨基丁酸能神经元，自纹状体形成所谓的间接输出途径，其与运动性抑制有关。A _2A受体通过多种途径调节γ-氨基丁酸、多巴胺、乙酰胆碱和谷氨酸盐的神经传递促进运动行为的控制。通常，A _2A通过和D2受体的相互作用，尤其是A _2A作为拮抗剂的作用对治疗帕金森病有很大益处，其可以导致多巴胺水平的降低。A _2A受体紧密地并且拮抗地与D2受体相互作用，引起D2受体受刺激时对多巴胺亲和力的降低。因此，A _2A受体拮抗剂可以增强内源性多巴胺以及临床应用的多巴胺受体激动剂的作用并且增加多巴胺能的药物响应时限。 A _2A receptors are mostly found in areas rich in dopamine, such as basal ganglion elements, various mammals including human striatum and globus pallidus. The basal ganglia and striatum act as central components involved in the integration of information in the cortex, thalamus and marginal lobes to produce motor behavior. It has been found in the striatum that the A _2A receptor and the dopamine D2 receptor are closely concentrated in the striatum globus gamma-aminobutyric acid neuron, forming a so-called indirect export pathway from the striatum, which is motility Inhibition related. The A _2A receptor regulates the neurotransmission of gamma-aminobutyric acid, dopamine, acetylcholine, and glutamate through various pathways to promote the control of motor behavior. In general, the interaction of _A2A with the D2 receptor, particularly _A2A as an antagonist, is of great benefit in the treatment of Parkinson's disease, which can result in a decrease in dopamine levels. The _A2A receptor interacts closely and antagonistically with the D2 receptor, causing a decrease in dopamine affinity when the D2 receptor is stimulated. Thus, _A2A receptor antagonists can potentiate the effects of endogenous dopamine and clinically applied dopamine receptor agonists and increase the drug response time of dopaminergic agents.

选择性A _2A受体激动剂和拮抗剂在啮齿类和非人类的灵长类的药理学、行为和神经保护的实验中已有广泛描述。在D2受体拮抗剂和A _2A受体激动剂诱导的僵住症模型中能够清楚地例证D2和A _2A受体在僵住症模型中的紧密相互作用，其被A _2A受体拮抗剂和D2受体激动剂反作用。目前，许多研究者已经报道了A _2A受体拮抗剂抗震颤***潜力。例如，腺苷A _2A受体拮抗剂SCH58261和KW-6002，均增强在单侧6-羟基多巴胺(6-OHDA)损伤的小鼠和大鼠中由亚域剂量左旋多巴引起的双侧旋转。此外，已经普遍报道了腺苷A _2A受体拮抗剂KW-6002显著改善在非人类灵长类中因用多巴胺受体激动剂左旋多巴长期治疗由1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)诱导的未引起运动障碍的运动损伤。因此，由于其不仅逆转运动损伤并且能够通过延长细胞寿命减慢或停止疾病进程，A _2A受体拮抗剂作为帕金森氏病患者长期用药的未来药物显示出很大潜力。 Selective _A2A receptor agonists and antagonists have been extensively described in the pharmacological, behavioral, and neuroprotective experiments of rodent and non-human primates. In catalepsy model D2 receptor antagonist and agonist-induced A _2A receptor can be clearly exemplified in close D2 and A _2A receptor interaction catalepsy model, which is A _2A receptor antagonist and D2 receptor agonist reaction. At present, many researchers have reported the potential of A _2A receptor antagonists to resist tremor paralysis. For example, the adenosine A _2A receptor antagonists SCH58261 and KW-6002 both enhanced bilateral rotation caused by subdomain dose of levodopa in unilateral 6-hydroxydopamine (6-OHDA) injured mice and rats. . In addition, it has been widely reported that adenosine A _2A receptor antagonist KW-6002 significantly improves long-term treatment with non-human primates by dopamine receptor agonist levodopa from 1-methyl-4-phenyl-1 , 2,3,6-tetrahydropyridine (MPTP)-induced motor injury that does not cause dyskinesia. Therefore, A _2A receptor antagonists have shown great potential as future drugs for long-term use in patients with Parkinson's disease, as they not only reverse motor damage and can slow or stop disease progression by prolonging cell life.

数个临床前研究表明，腺苷A _2A受体拮抗剂对于治疗神经退行性疾病，如帕金森病、亨廷顿病或阿尔茨海默病具有有效性。且已有报道A _2A受体拮抗剂在不同神经退行性疾病的体内和体外模型中具有神经保护作用。总而言之，A _2A受体拮抗剂能够有效地保护来自各种形式的损伤诱导的神经退行性病变的不同的神经元。 Several preclinical studies have shown that adenosine _A2A receptor antagonists are effective in treating neurodegenerative diseases such as Parkinson's disease, Huntington's disease or Alzheimer's disease. It has been reported that _A2A receptor antagonists have neuroprotective effects in both in vivo and in vitro models of different neurodegenerative diseases. In summary, _A2A receptor antagonists are effective in protecting different neurons from various forms of damage-induced neurodegenerative diseases.

有研究发现A _2A受体敲除的小鼠对“抑制药”攻击比它们的野生型同类敏感性更低。与此研究一致，在小鼠尾巴悬浮实验中A _2A受体拮抗剂SCH58261和KW6002能减少总的不动时间。也发现当对为了有高不动时间，事先筛选的小鼠给药时，拮抗剂SCH8261和ZM241385减少了不动性，而在这个模型中SCH58261减少了为了它们的“无助”选择性饲养的小鼠的不动性。用A _2A敲除的小鼠研究表明这些动物对精神***，例如安非他明和***，反应迟钝。因此现有证据可以表明，腺苷A _2A受体拮抗剂通过调节中纹状体或中肾上腺皮质的多巴胺能的途径，可能具有抗抑郁和/或抗精神病功能。A _2A受体激活可以有助于改善一定范围的神经精神性疾病和障碍，例如抑郁、白天睡眠过多、腿多动综合征(RLS),注意缺陷伴多动障碍和认知疲劳。 Studies have found that A _2A receptor knockout mice are less sensitive to "inhibitory drugs" than their wild type counterparts. Consistent with this study, the A _2A receptor antagonists SCH58261 and KW6002 reduced the total immobility time in the mouse tail suspension assay. It was also found that the antagonists SCH8261 and ZM241385 reduced immobility when administered to mice that had been screened for high immobility time, whereas in this model SCH58261 reduced their selective selection for "helpless" feeding. The immobility of the mouse. Mouse studies with A _2A knockouts showed that these animals were unresponsive to psychostimulants such as amphetamines and ***e. Therefore, available evidence may indicate that adenosine A _2A receptor antagonists may have antidepressant and/or antipsychotic functions by regulating the dopaminergic pathway of the striatum or the middle adrenocortical. A _2A receptor activation can help to improve a range of neuropsychiatric disorders and disorders such as depression, excessive daytime sleep, leg hyperactivity syndrome (RLS), attention deficit hyperactivity disorder, and cognitive fatigue.

锥体外综合征(EPS)是一系列与使用抗精神病药物有关的不利神经反应的通称。有6类不同的EPS-有关神经综合征，其中4种，即张力失常、静坐不能、假性帕金森病(帕金森综合征)及延发性运动障碍在服用抗精神病药物治疗的患者中特别普遍。张力失常是肌肉群，特别是颈部、下颚、背部、咽头及喉头的疼痛痉挛。其最常见于用抗精神病药物治疗的年轻男性，但是也可与使用***、三环类抗抑郁剂、锂盐抗惊厥剂(例如苯英妥和卡马西平)有关。假性帕金森病自身表现为运动不能(强直、僵硬和缓慢随意运动、驼背、曳步行走)和颤抖，以及在疗法开始后数周或数月内发生的这些症状。静坐不能自身表现为运动多动、主观内感觉苦恼或不舒服，常误解为激动或焦虑，此常见的综合征常不能诊断并且对治疗有最小的反应。延发性运动障碍为后期出现的、与长期使用精神安定药物有关的综合征。其更常发生于年长患者，并且特性是脸部、眼睑、口部、舌头、四肢和躯体的刻板、反复、不随意、快速的舞蹈症样运动。Extrapyramidal syndrome (EPS) is a generic term for a series of adverse neurological reactions associated with the use of antipsychotics. There are 6 different types of EPS-related neurological syndromes, 4 of which are dystonia, sedation, pseudo Parkinson's disease (Parkinson's syndrome) and delayed dyskinesia in patients taking antipsychotic medications. universal. Tension disorder is the pain of the muscles, especially the neck, jaw, back, pharynx and throat. It is most commonly found in young men treated with antipsychotics, but can also be associated with the use of ***e, tricyclic antidepressants, lithium salt anticonvulsants (such as phenytoin and carbamazepine). Pseudo Parkinson's disease manifests itself as an inability to exercise (tonic, stiff and slow voluntary movement, hunchback, walking) and tremors, as well as these symptoms that occur within weeks or months after the start of therapy. Meditation can not manifest itself as hyperactivity, subjective feelings of distress or discomfort, often misunderstood as agitation or anxiety, this common syndrome is often not diagnosed and has minimal response to treatment. Delayed dyskinesia is a late-onset syndrome associated with long-term use of neuroleptic drugs. It occurs more often in older patients and is characterized by stereotyped, repetitive, involuntary, and rapid dance-like movements of the face, eyelids, mouth, tongue, limbs, and body.

静坐不能也是RLS和PLMS(睡眠期周期性肢体运动)以及PLMD(周期性腿(或肢体)运动障碍)的特征。RLS为一般障碍，其导致患者对于移动其腿有不能抵抗和不愉快的渴望；其常在静止时和/或夜间显现，并且可能会扰乱睡眠。不具典型RLS症状，但会表现间歇性腿移动而对睡眠带来不利影响的患者被诊断为PLMS。RLS和PLMS的治疗已包括左旋多巴/卡比多巴、左旋多巴/苄丝肼、多巴胺激动剂(如普拉克索和罗匹尼罗)，苯并二氮杂

类药物，阿片类药物，抗惊厥剂及铁(硫酸亚铁)。 Sedation can also be a feature of RLS and PLMS (periodic limb movement during sleep) and PLMD (periodic leg (or limb) dyskinesia). RLS is a general disorder that causes the patient to have an unbearable and unpleasant desire to move their legs; it often appears at rest and/or at night and may disturb sleep. Patients who do not have typical RLS symptoms but who exhibit intermittent leg movements that adversely affect sleep are diagnosed with PLMS. Treatment of RLS and PLMS has included levodopa/carbidopa, levodopa/benserazide, dopamine agonists (such as pramipexole and ropinirole), benzodiazepines

Drugs, opioids, anticonvulsants and iron (ferrous sulfate).

在CNS中，数据显示A _2A受体以高密度存在于基础神经节中，其在控制良好运动中很重要。此外，A _2A受体的选择性拮抗剂在药理上很重要，因为其表现出降低运动损伤的效力，因而改善了神经退行性疾病，例如帕金森病及相关运动障碍(例如亨廷顿舞蹈病)中的功能。与带来治疗指数提高的当前多巴胺能治疗相比，A _2A拮抗剂似乎表现出降低副作用的倾向(例如无运动障碍)。A _2A拮抗剂还具有抗抑郁性质和刺激认知功能。 In the CNS, the data show that the _A2A receptor is present in the basal ganglia at high density, which is important in controlling good exercise. In addition, selective antagonists of the A _2A receptor are pharmacologically important because they exhibit reduced efficacy for motor impairment, thereby improving neurodegenerative diseases such as Parkinson's disease and related dyskinesias (eg Huntington's disease). The function. A _2A antagonists appear to exhibit a tendency to reduce side effects (eg, no dyskinesia) as compared to current dopaminergic treatments that result in an increased therapeutic index. A _2A antagonists also have antidepressant properties and stimulate cognitive function.

因此，人们对发现新型高效且具选择性的腺苷A _2A受体拮抗剂的兴趣逐渐增加。制药公司发现的一些有效的腺苷A _2A拮抗剂已经进入到临床试验并显示出阳性结果，表明A _2A受体拮抗剂不仅有望治疗神经退行性疾病，例如帕金森病、亨廷顿病或阿尔茨海默病，而且也可以用于治疗其他CNS相关的疾病，例如抑郁、多动综合征、睡眠障碍和焦虑症。 Therefore, there is an increasing interest in discovering novel and highly effective and selective adenosine A _2A receptor antagonists. Some effective adenosine A _2A antagonists discovered by pharmaceutical companies have entered clinical trials and showed positive results, suggesting that A _2A receptor antagonists are not only expected to treat neurodegenerative diseases such as Parkinson's disease, Huntington's disease or Alzheim It is also a disease, and can also be used to treat other CNS-related diseases such as depression, hyperactivity syndrome, sleep disorders and anxiety.

此外，腺苷A _2A受体也与免疫调节紧密相关。免疫调节是机体保持内环境稳定、抵御外来有害刺激的重要手段。腺苷作为机体的一种重要递质及调质，在代谢障碍及细胞损伤时会大幅升高,激活腺苷受体而发挥生物学效应，参与机体的免疫调节。近年来的研究表明，在缺血低氧、炎症、创伤、移植等诸多病理过程中，腺苷A _2A受体的激活可以发挥重要的免疫调节作用，这可能与A _2A受体在T细胞、B细胞、单核巨噬细胞、中性粒细胞等多种免疫细胞上表达水平较高有关。 In addition, the adenosine A _2A receptor is also closely related to immune regulation. Immune regulation is an important means for the body to maintain a stable internal environment and resist external harmful stimuli. As an important transmitter and temperament of the body, adenosine is greatly increased in metabolic disorders and cell damage, activates adenosine receptors and exerts biological effects, and participates in the immune regulation of the body. Recent studies have shown that activation of adenosine A _2A receptors may play an important immunomodulatory role in many pathological processes such as ischemia, hypoxia, inflammation, trauma, transplantation, etc., which may be related to A _2A receptors in T cells, B cells, mononuclear macrophages, neutrophils and other immune cells have higher expression levels.

A _2A受体与肿瘤密切相关。正常情况下，机体可以依赖完整的免疫机制来有效地监视和排斥癌变细胞，如：在细胞免疫方面，T淋巴细胞、抗体依赖性细胞毒细胞(K细胞)、NK细胞和巨噬细胞对肿瘤细胞均具杀伤作用。但如果癌变细胞本身或上述免疫细胞功能发生改变，则可能逃脱机体免疫***的清除，恶性增生形成肿瘤。研究已证明，A _2A受体的活化可以促使机体产生免疫耐受,密切参与了肿瘤细胞“免疫逃逸”或“免疫抑制”的形成，为肿瘤的发生发展创造了有利条件。A _2A受体活化可以通过抑制血管内皮细胞血小板反应蛋白的表达而呈剂量依赖性地促进血管生成，为血管依赖性肿瘤的生长创造有利环境；A _2A受体的活化也可以通过升高cAMP，激活PKA抑制自然杀伤细胞对肿瘤细胞的杀伤；可以促进黑色素瘤A375细胞、成纤维瘤NIH3T3细胞及嗜铬细胞瘤PC12细胞等肿瘤细胞的增殖。已有研究表明A _2A受体拮抗剂可应用于治疗各种肿瘤，如肺癌，尤其是非小细胞肺癌。 A _2A receptors are closely related to tumors. Under normal circumstances, the body can rely on the complete immune mechanism to effectively monitor and reject cancerous cells, such as: in cellular immunity, T lymphocytes, antibody-dependent cytotoxic cells (K cells), NK cells and macrophages to tumors The cells have a killing effect. However, if the cancerous cells themselves or the functions of the above-mentioned immune cells are changed, the immune system of the off-line is cleared, and the malignant hyperplasia forms a tumor. Studies have shown that the activation of A _2A receptor can promote the body to produce immune tolerance, and closely participate in the formation of "immune escape" or "immunosuppression" of tumor cells, which creates favorable conditions for the occurrence and development of tumors. A _2A receptor activation can promote angiogenesis in a dose-dependent manner by inhibiting the expression of thrombospondin in vascular endothelial cells, creating a favorable environment for the growth of vascular-dependent tumors; activation of A _2A receptors can also be achieved by increasing cAMP. Activation of PKA inhibits the killing of tumor cells by natural killer cells; it can promote the proliferation of tumor cells such as melanoma A375 cells, fibroblast NIH3T3 cells and pheochromocytoma PC12 cells. Studies have shown that A _2A receptor antagonists can be used to treat various tumors such as lung cancer, especially non-small cell lung cancer.

适合A _2A受体拮抗剂的候选产品，需要强效的与A _2A受体结合但不能与其他腺苷受体强效结合，即需要有较高的A _2A受体亚型选择性，这样有助于减少潜在副作用。现已有多个小分子腺苷A _2A受体拮抗剂进入到临床I期研究，用于***。如Corvus公司的CPI-444，显示结合A _2A受体的亲和力(Ki)为3.5nm，A ₁受体的亲和力(Ki)为192nm，对A ₁受体亚型的选择性达到54倍。基于这些结果，研究者认为在目前预期的剂量水平，CPI-444在合理的剂量范围内具有足够的安全性和潜在疗效。CPI-444在3个生长不同肿瘤的小鼠模型，即EL-4淋巴瘤模型、MC38结肠肿瘤模型和CT26结肠肿瘤模型，中进行了测试,结果显示或癌细胞处***数量显著性减少，或原发部位肿瘤体积明显降低，或肿瘤体积稳定或消退等良好反应，甚至有些小鼠显示已完全治愈。Heptares公司的AZD4635也进入了临床I期治疗晚期实体瘤。这些结果均显示高选择性的A _2A受体拮抗剂可应用于***。 Candidates for A _2A receptor antagonists need to bind strongly to the A _2A receptor but not to other adenosine receptors, ie, require a higher A _2A receptor subtype selectivity, so Helps reduce potential side effects. A number of small-molecule adenosine A _2A receptor antagonists have been introduced into clinical phase I studies for the treatment of tumors. Corvus companies such as CPI-444, show a binding affinity A _2A receptor (Ki) of 3.5nm, A ₁ receptor affinity (Ki) of 192nm, the selectivity to the A ₁ receptor subtype of 54 times. Based on these results, the investigators believe that CPI-444 has sufficient safety and potential efficacy in a reasonable dose range at the current expected dose level. CPI-444 was tested in three mouse models of different tumors, namely the EL-4 lymphoma model, the MC38 colon tumor model, and the CT26 colon tumor model, and the results showed that the number of lymph nodes at the cancer cells was significantly reduced, or The tumor volume at the primary site was significantly reduced, or the tumor volume was stable or resolved, and even some mice showed complete cure. Heptares' AZD4635 also entered the clinical phase I treatment of advanced solid tumors. These results show that highly selective _A2A receptor antagonists can be used to treat tumors.

此外，A _2A受体在哮喘、动脉粥样硬化等慢性炎性疾病中的免疫调节作用也逐渐被人们认识和受到重视，A _2A受体与伤口愈合或心房颤动也有着密切的关系。很多研究已表明，A _2A受体拮抗剂还可应用于治疗糖尿病。 In addition, the immunomodulatory effects of A _2A receptors in chronic inflammatory diseases such as asthma and atherosclerosis have gradually been recognized and valued. A _2A receptors are also closely related to wound healing or atrial fibrillation. Many studies have shown that A _2A receptor antagonists can also be used to treat diabetes.

中国发明专利(CN102892761)提供了一种作为腺苷A _2A受体拮抗剂的4-氨基嘧啶衍生物： The Chinese invention patent (CN102892761) provides a 4-aminopyrimidine derivative as an adenosine A _2A receptor antagonist:

其中R ¹代表任选被一个或两个卤原子取代的或被一个或两个甲基或三氟甲基取代的吡唑、噻唑或***环。该化合物对于腺苷A _2A受体具有较佳的拮抗作用，表现为其对A _2A的亲和力较强，但是其对于A ₁的选择性也很低，导致在A ₁受体存在下，其对A _2A受体的选择性较低。同时，该发明所述的吡唑类化合物的药代动力学特征较差，血浆清除率较高，在大鼠体内的半衰期也很短。 Wherein R ¹ represents a pyrazole, thiazole or triazole ring optionally substituted by one or two halogen atoms or substituted by one or two methyl or trifluoromethyl groups. This compound has a better antagonistic effect on the adenosine A _2A receptor, showing its affinity for A _2A , but its selectivity for A ₁ is also low, resulting in the presence of the A ₁ receptor. The selectivity of the A _2A receptor is low. At the same time, the pyrazole compounds of the invention have poor pharmacokinetic characteristics, high plasma clearance, and a short half-life in rats.

发明的公开Disclosure of invention

本发明的目的是提供作为腺苷A _2A受体拮抗剂的4-氨基嘧啶衍生物，并将其用于治疗对A _2A拮抗作用有反应的疾病或病症。 It is an object of the present invention to provide a 4-aminopyrimidine derivative as an adenosine A _2A receptor antagonist and to use it for the treatment of a disease or condition responsive to A _2A antagonism.

为达到上述目的，本发明提供了一种作为腺苷A _2A受体拮抗剂的4-氨基嘧啶衍生物，其结构通式(Ⅰ)如下： In order to achieve the above object, the present invention provides a 4-aminopyrimidine derivative as an adenosine A _2A receptor antagonist, which has the following structural formula (I):

其中：among them:

R ₁选自卤素、氰基或三氟甲基； R _{1 is} selected from halogen, cyano or trifluoromethyl;

R ₂选自吡唑基、吡咯烷基，或经一个或多个卤素或C _1-3烷基取代的吡唑基或吡咯烷基； R _{2 is} selected from pyrazolyl, pyrrolidinyl, or pyrazolyl or pyrrolidinyl substituted by one or more halogen or C _1-3 alkyl;

R ₃选自噁唑基、噁二唑基、三氮唑基，或经一个或多个卤素或C _1-3烷基取代的噁唑基。 R _{3 is} selected from oxazolyl, oxadiazolyl, triazolyl, or oxazolyl substituted with one or more halogen or C _1-3 alkyl.

优选地，上述的作为腺苷A _2A受体拮抗剂的4-氨基嘧啶衍生物，其中，R ₂选自吡唑基或吡咯烷基；R ₃选自噁唑基、噁二唑基、三氮唑基，或经一个C _1-3烷基取代的噁唑基。 Preferably, the above 4-aminopyrimidine derivative as an adenosine A _2A receptor antagonist, wherein R _{2 is} selected from pyrazolyl or pyrrolidinyl; and R _{3 is} selected from oxazolyl, oxadiazolyl, and tri A oxazolyl group or an oxazolyl group substituted with a C _1-3 alkyl group.

优选地，上述的作为腺苷A _2A受体拮抗剂的4-氨基嘧啶衍生物，其中，该4-氨基嘧啶衍生物选自下述化合物： Preferably, the above 4-aminopyrimidine derivative as an adenosine A _2A receptor antagonist, wherein the 4-aminopyrimidine derivative is selected from the group consisting of:

优选地，该4-氨基嘧啶衍生物选自下述化合物：(1)、(5)、(8)、(10)、(13)、(14)、(17)或(18)。Preferably, the 4-aminopyrimidine derivative is selected from the group consisting of (1), (5), (8), (10), (13), (14), (17) or (18).

本发明还提供了一种上述的4-氨基嘧啶衍生物在制备用于治疗对A _2A拮抗作用有反应的疾病或病症的药物中的用途。 The invention also provides the use of a 4-aminopyrimidine derivative as described above for the manufacture of a medicament for the treatment of a disease or condition responsive to _A2A antagonism.

进一步地，所述疾病或病症包含神经退行性疾病、锥体外综合征、抑郁、多动综合征、睡眠障碍、焦虑症、糖尿病或肿瘤中的任意一种或多种。Further, the disease or condition comprises any one or more of a neurodegenerative disease, an extrapyramidal syndrome, depression, hyperactivity syndrome, sleep disorder, anxiety, diabetes or a tumor.

进一步地，所述肿瘤为肺癌、卵巢癌、胰腺癌、胃癌、乳腺癌、胶质母细胞瘤、黑色素瘤、肾细胞癌、三阴性乳腺癌、结直肠癌、头颈部癌、膀胱癌、***癌、肝细胞肝癌或胆管癌。Further, the tumor is lung cancer, ovarian cancer, pancreatic cancer, gastric cancer, breast cancer, glioblastoma, melanoma, renal cell carcinoma, triple negative breast cancer, colorectal cancer, head and neck cancer, bladder cancer, Prostate cancer, hepatocellular carcinoma or cholangiocarcinoma.

进一步地，所述肺癌为非小细胞肺癌。Further, the lung cancer is non-small cell lung cancer.

本发明还提供了一种药物组合物，其中，其包含上述的作为腺苷A _2A受体拮抗剂的4-氨基嘧啶衍生物，及其药学上可接受的赋形剂。 The present invention also provides a pharmaceutical composition comprising the above-described 4-aminopyrimidine derivative as an adenosine A _2A receptor antagonist, and a pharmaceutically acceptable excipient thereof.

本发明还提供了一种复方药物，其中，包含上述的4-氨基嘧啶衍生物以及能够与其联用的药物。The present invention also provides a compound drug comprising the above 4-aminopyrimidine derivative and a drug capable of being used therewith.

进一步地，所述联用的药物为用于治疗以下疾病或病症的化合物：神经退行性疾病、锥体外综合征、抑郁、多动综合征、睡眠障碍、焦虑症、糖尿病或肿瘤。Further, the combination drug is a compound for treating a disease or a condition: a neurodegenerative disease, an extrapyramidal syndrome, depression, hyperactivity syndrome, sleep disorder, anxiety, diabetes, or tumor.

本发明具有以下有益效果：本发明所提供的4-氨基嘧啶衍生物对腺苷A _2A受体具有明显的选择性拮抗作用，具有较好的药代动力学特征，可应用于治疗对A _2A拮抗作用有反应的疾病或病症的组合物或组合产品中，尤其是用于治疗神经退行性疾病、锥体外综合征、抑郁、多动综合征、睡眠障碍、焦虑症、糖尿病或肿瘤等疾病或病症。 The invention has the following beneficial effects: the 4-aminopyrimidine derivative provided by the invention has obvious selective antagonism to the adenosine A _2A receptor, has good pharmacokinetic characteristics, and can be applied to the treatment of A _2A A composition or combination of diseases or conditions in which a response is responsive, particularly for the treatment of diseases such as neurodegenerative diseases, extrapyramidal syndrome, depression, hyperactivity syndrome, sleep disorders, anxiety disorders, diabetes or tumors or Illness.

实现本发明的最佳方式The best way to implement the invention

以下通过具体实施例对本发明作进一步的描述，这些实施例仅用于说明本发明，并不是对本发明保护范围的限制。The invention is further described by the following examples, which are intended to illustrate the invention and not to limit the scope of the invention.

本发明提供了一种作为腺苷A _2A受体拮抗剂的4-氨基嘧啶衍生物，其结构通式(Ⅰ)如下： The present invention provides a 4-aminopyrimidine derivative as an adenosine A _2A receptor antagonist, which has the following structural formula (I):

其中：among them:

本发明所提供的作为腺苷A _2A受体拮抗剂的4-氨基嘧啶衍生物可以通过以下合成路线之一合成： The 4-aminopyrimidine derivatives provided by the present invention as adenosine A _2A receptor antagonists can be synthesized by one of the following synthetic routes:

合成路线1：Synthetic route 1:

试剂和条件：(a)、乙酸酐，回流；(b)、吡唑，碳酸铯，二甲基甲酰胺(DMF)，80℃；(c)、反-β-苯乙烯硼酸，碳酸钠，二氧六环，室温；四(三苯基膦)钯(Pd(PPh ₃) ₄)，90℃；(d)、臭氧，甲醇(MeOH)/二氯甲烷(CH ₂Cl ₂)，-78℃；(e)、对甲基苯磺酰甲基异腈(TOSMIC)，碳酸钾(K ₂CO ₃)，MeOH，80℃；(f)、N-溴代丁二酰亚胺，DMF。 Reagents and conditions: (a), acetic anhydride, reflux; (b), pyrazole, cesium carbonate, dimethylformamide (DMF), 80 ° C; (c), trans-β-styrene boronic acid, sodium carbonate, Dioxane, room temperature; tetrakis(triphenylphosphine)palladium (Pd(PPh ₃ ) ₄ ), 90 ° C; (d), ozone, methanol (MeOH) / dichloromethane (CH ₂ Cl ₂ ), -78 °C; (e), p-toluenesulfonylmethyl isocyanide (TOSMIC), potassium carbonate (K ₂ CO ₃ ), MeOH, 80 ° C; (f), N-bromosuccinimide, DMF.

化合物(1)-(3)，(6)-(8)，(11)-(13)，(15)-(17)的制备按合成路线1实施。具体实施方法参见实施例1。其中嘧啶5位氟取代的化合物也可从含氟原子的起始原料开始制备。The preparation of the compounds (1) to (3), (6) to (8), (11) to (13), and (15) to (17) was carried out in the same manner as in Scheme 1. For specific implementation methods, see Example 1. Compounds in which the pyrimidine 5-position fluorine is substituted may also be prepared starting from a fluorine atom-containing starting material.

合成路线2：Synthetic route 2:

试剂和条件：(a)、间-氯过苯甲酸，二氯甲烷，室温；(b)、N-溴代琥珀酰亚胺(NBS)，DMF，室温；(c)、1.噁唑，丁基锂(n-BuLi)，四氢呋喃(THF)， -78℃～-20℃，2.Pd(PPh ₃) ₄，80℃；(d)、1H-吡唑，碳酸铯，DMF，90℃；(e)、吡咯烷，碳酸铯，DMF，90℃。 Reagents and conditions: (a), m-chloroperbenzoic acid, dichloromethane, room temperature; (b), N-bromosuccinimide (NBS), DMF, room temperature; (c), 1. oxazole, Butyllithium (n-BuLi), tetrahydrofuran (THF), -78 ° C ~ -20 ° C, 2. Pd (PPh ₃ ) ₄ , 80 ° C; (d), 1H-pyrazole, cesium carbonate, DMF, 90 ° C (e), pyrrolidine, cesium carbonate, DMF, 90 ° C.

使用DMF作为溶剂，在碱(例如碳酸铯或丁基锂)存在下，式(G)的亚砜与不同的商业销售的五元杂环(例如，吡唑、噁唑、噁二唑或三氮唑)衍生物在室温或低温下反应，生成2位被五元杂环取代的衍生物。例如，式(G)的中间体与噁唑在这些条件下反应，则得到式(H)的衍生物。The use of DMF as a solvent in the presence of a base such as cesium carbonate or butyl lithium, a sulfoxide of formula (G) with a different commercially available five-membered heterocyclic ring (eg, pyrazole, oxazole, oxadiazole or tri The azole derivative is reacted at room temperature or at a low temperature to form two derivatives substituted with a five-membered heterocyclic ring. For example, an intermediate of formula (G) is reacted with an oxazole under these conditions to provide a derivative of formula (H).

进一步在碱(例如碳酸铯或甲醇钠)存在下使用DMF作为溶剂，通过五元杂环衍生物(例如，吡唑或吡咯烷)来取代嘧啶衍生物的6位氯原子。例如，衍生物(H)与吡唑或吡咯烷在这些条件下反应可制得化合物(19)和(23)。The 6-position chlorine atom of the pyrimidine derivative is further substituted by a five-membered heterocyclic derivative (for example, pyrazole or pyrrolidine) using DMF as a solvent in the presence of a base such as cesium carbonate or sodium methoxide. For example, the compound (19) and (23) can be obtained by reacting the derivative (H) with a pyrazole or pyrrolidine under these conditions.

化合物(19)，(20)，(23-25)的制备按合成路线2实施。具体实施方法参见实施例15。The preparation of the compounds (19), (20), (23-25) was carried out in accordance with Scheme 2. For specific implementation methods, see Example 15.

合成路线3：Synthetic route 3:

试剂和条件：(a)、氰化亚铜，吡啶，微波(MW)，250℃下反应进行20分钟。(b)、氟化钾(KF)，双(二亚苄基丙酮)钯(Pd(dba) ₂)，2-(二环己基磷)-3,6-二甲氧基-2',4',6'-三异丙基-1,1'-联苯，三甲基(三氟甲基)硅烷，二氧六环，20h，140℃。 Reagents and conditions: (a), cuprous cyanide, pyridine, microwave (MW), and the reaction was carried out at 250 ° C for 20 minutes. (b) potassium fluoride (KF), bis(dibenzylideneacetone)palladium (Pd(dba) ₂ ), 2-(dicyclohexylphosphino)-3,6-dimethoxy-2',4 ',6'-Triisopropyl-1,1'-biphenyl, trimethyl(trifluoromethyl)silane, dioxane, 20 h, 140 °C.

为了合成如上述定义的其中嘧啶5-位取代基R ₁是氰基或三氟甲基的嘧啶衍生物，可以采用如化合物(1)所示的类似前体化合物，使用合成路线3中所述的方法分别制备。 In order to synthesize a pyrimidine derivative wherein the pyrimidine 5-position substituent R ₁ is a cyano group or a trifluoromethyl group as defined above, a similar precursor compound as shown in the compound (1) may be employed, as described in the synthesis scheme 3. The methods were prepared separately.

化合物(4)，(5)，(9)，(10)，(14)，(18)，(21)，(22)的制备按合成路线3实施。具体实施方法参见实施例4和实施例5。The preparation of the compounds (4), (5), (9), (10), (14), (18), (21), (22) was carried out in accordance with Scheme 3. For details, see Example 4 and Example 5.

实施例1：Example 1:

化合物(1)的制备，其结构式如下：The preparation of the compound (1) has the following structural formula:

化合物(1)的制备按上述合成路线1实施：The preparation of the compound (1) is carried out in accordance with the above Scheme 1:

第一步，制备中间体(A)First step, preparation of intermediate (A)

将4-氨基-2，6-二氯嘧啶(DCAP，4g，24.4mmol)悬浮于乙酸酐(80mL，860mmol)中，搅拌下加热回流4小时。反应液冷却后，真空浓缩，残余的乙酸酐加入甲苯后蒸馏除去。残渣溶于乙酸乙酯和水中，加入10％的NaHCO ₃溶液至溶液PH为7。有机层用饱和食盐水洗，回收溶剂后残渣溶于乙酸酐(40mL)，0-5℃下搅拌2小时，过滤收集沉淀，40℃下真空干燥得到中间体(A)。MS m/z(ESI):206.0[M+1] ⁺。 4-Amino-2,6-dichloropyrimidine (DCAP, 4 g, 24.4 mmol) was suspended in acetic anhydride (80 mL, 860 mmol) and heated under reflux for 4 hours. After the reaction mixture was cooled, it was concentrated in vacuo, and the residual acetic acid was added to toluene and then evaporated. The residue was dissolved in ethyl acetate and water, addition of 10% NaHCO ₃ solution until the solution PH 7. The organic layer was washed with saturated brine, and the solvent was evaporated. The residue was dissolved in acetic acid (40 mL), and the mixture was stirred at 0 to 5 ° C for 2 hours, and the precipitate was collected by filtration and dried under vacuum at 40 ° C to afford intermediate (A). MS m/z (ESI): 206.0 [M+1] ⁺ .

第二步，制备中间体(B)The second step, the preparation of intermediate (B)

将中间体(A)(1g，5mmol)溶于无水DMF(15mL)中，再加入吡唑(340mg，5mmol)和碳酸铯(1.6g，5mmol)。混和物于80℃搅拌2小时后倒入水中，用乙酸乙酯提取。有机层依次用水洗、饱和食盐水洗涤后，无水硫酸钠干燥，减压除去溶剂。残余物经硅胶柱层析(3％甲醇：二氯甲烷)分离纯化得产物(B)。MS m/z(ESI):238.0[M+1] ⁺。 Intermediate (A) (1 g, 5 mmol) was dissolved in anhydrous DMF (15 mL)EtOAc. The mixture was stirred at 80 ° C for 2 hours, poured into water and extracted with ethyl acetate. The organic layer was washed with water and brine, and dried over anhydrous sodium sulfate. The residue was purified by silica gel column chromatography (yield MS m/z (ESI): 238.0 [M+1] ⁺ .

第三步，制备中间体(C)The third step is to prepare the intermediate (C)

将中间体(B)(0.4g，1.68mmol)、反-β-苯乙烯硼酸(0.5g，3.36mmol)和碳酸钠(1.08g，10.1mmol)加入到二氧六环/水溶液里，通入氮气约30min后，加入Pd(PPh ₃) ₄(0.2g，0.16mmol)，混和物加热到90℃搅拌20小时后倒入到水里，乙酸乙酯提取。有机层依次用水洗、饱和食盐水洗涤，无水硫酸钠干燥，减压除去溶剂。残余物硅胶柱层析(3％甲醇：二氯甲烷)分离纯化得产物(C)。MS m/z(ESI):306.1[M+1] ⁺。 Intermediate (B) (0.4 g, 1.68 mmol), trans-β-styrene boronic acid (0.5 g, 3.36 mmol) and sodium carbonate (1.08 g, 10.1 mmol) were added to the dioxane/water solution. After about 30 min of nitrogen, Pd(PPh ₃ ) ₄ (0.2 g, 0.16 mmol) was added, and the mixture was heated to 90 ° C for 20 hours, poured into water, and extracted with ethyl acetate. The organic layer was washed with water and brine, dried over anhydrous sodium sulfate The residue was purified by silica gel column chromatography (yield MS m/z (ESI): 306.1 [M+1] ⁺ .

第四步，制备中间体(D)The fourth step, the preparation of intermediate (D)

将中间体(C)(0.4g，1.6mmol)溶于甲醇/二氯甲烷(4/1，20ml)混和溶液中，冷却到-78℃，通入臭氧10分钟。反应完毕后通入氮气20分钟，加入甲醚，反应液升温到室温后再通入氮气使溶剂挥发完全后得粗品(D)。MS m/z(ESI):232.1[M+1] ⁺。 The intermediate (C) (0.4 g, 1.6 mmol) was dissolved in a mixture of methanol/dichloromethane (4/1, 20 ml), cooled to -78 ° C After completion of the reaction, nitrogen gas was introduced for 20 minutes, methyl ether was added, and the reaction liquid was heated to room temperature, and then nitrogen gas was introduced to completely evaporate the solvent to obtain a crude product (D). MS m/z (ESI): 2321. [M+1] ⁺ .

第五步，制备中间体(E)The fifth step, preparation of intermediate (E)

中间体(D)(460mg，2mmol)，TOSMIC(对甲苯磺酰甲基异氰，50mg，4mmol),碳酸钾(860mg，6mmol)混和物加入到甲醇中，加热到80℃，反应16小时后，回收甲醇，残余物倒入水中，乙酸乙酯提取。有机层水洗、饱和食盐水洗涤，无水硫酸钠干燥，减压除去溶剂。残余物硅胶柱层析(5％甲醇：二氯甲烷)，得产物(E)。MS m/z(ESI):229.1[M+1] ⁺。 Intermediate (D) (460 mg, 2 mmol), TOSMIC (p-toluenesulfonylmethylisocyanide, 50 mg, 4 mmol), potassium carbonate (860 mg, 6 mmol) mixture was added to methanol and heated to 80 ° C for 16 hours. Methanol was recovered, and the residue was poured into water and extracted with ethyl acetate. The organic layer was washed with water and brine, dried over anhydrous sodium sulfate The residue was subjected to silica gel column chromatography (yield MS m/z (ESI): 229.1 [M+1] ⁺ .

第五步，制备化合物(1)The fifth step, the preparation of the compound (1)

将0.2g(1.25mmol)NBS(N-溴代丁二酰亚胺)缓慢加入到悬浮有0.2g中间体(E)的冷DMF溶液中。室温下搅拌1小时后，减压除去溶剂。残余物倒入水中，乙酸乙酯提取。有机层水洗、饱和食盐水洗涤，无水硫酸钠干燥，减压除去溶剂。残余物用硅胶柱层析(5％甲醇：二氯甲烷)纯化，制得化合物(1)。MS m/z(ESI):307.1[M+1] ⁺, ¹HNMR(400MHz,DMSO-d6)δ8.55(d,1H),8.32(s,1H),7.84(d,1H),7.53(s,1H),7.58(s,2H),6.58(dd,1H)。 0.2 g (1.25 mmol) of NBS (N-bromosuccinimide) was slowly added to a cold DMF solution in which 0.2 g of the intermediate (E) was suspended. After stirring at room temperature for 1 hour, the solvent was removed under reduced pressure. The residue was poured into water and extracted with ethyl acetate. The organic layer was washed with water and brine, dried over anhydrous sodium sulfate The residue was purified by silica gel column chromatography (yield MS m/z (ESI): 307.1 [M+1] ⁺ , ¹ H NMR (400 MHz, DMSO-d6) δ 8.55 (d, 1H), 8.32 (s, 1H), 7.84 (d, 1H), 7. s, 1H), 7.58 (s, 2H), 6.58 (dd, 1H).

实施例2：Example 2:

化合物(2)的制备，其结构式如下：The preparation of the compound (2) has the following structural formula:

化合物(2)的制备按合成路线1实施，具体方法参考实施例1。第6步反应时用NCS(N-氯代丁二酰亚胺)替换NBS(N-溴代丁二酰亚胺)进行氯代得到化合物(2)。MS m/z(ESI):263.0[M+1] ⁺, ¹HNMR(400MHz,DMSO-d6)δ8.53(d,1H),8.26(s,1H),7.82(d,1H),7.49(s,1H),7.54(s,2H),6.55(dd,1H)。 The preparation of the compound (2) is carried out in accordance with the synthesis scheme 1, and the specific method is referred to in the embodiment 1. In the sixth step, NBS (N-chlorosuccinimide) was replaced with NCS (N-chlorosuccinimide) for chlorination to give compound (2). MS m/z (ESI): 263.0 [M+1] ⁺ , ¹ H NMR (400 MHz, DMSO-d6) δ 8.53 (d, 1H), 8.26 (s, 1H), 7.82 (d, 1H), 7.49 ( s, 1H), 7.54 (s, 2H), 6.55 (dd, 1H).

实施例3Example 3

化合物(3)的制备，其结构式如下：The preparation of the compound (3) has the following structural formula:

按合成路线1实施制得化合物(3)。MS m/z(ESI):247.0[M+1] ⁺, ¹HNMR(400MHz,DMSO-d6)δ8.51(d,1H),8.27(s,1H),7.83(d,1H),7.48(s,1H),7.55(s,2H),6.54(dd,1H)。 Compound (3) was obtained in the same manner as in Scheme 1. MS m/z (ESI): 247.0 [M+1] ⁺ , ¹ H NMR (400 MHz, DMSO-d6) δ 8.51 (d, 1H), 8.27 (s, 1H), 7.83 (d, 1H), 7.48 ( s, 1H), 7.55 (s, 2H), 6.54 (dd, 1H).

实施例4：Example 4:

化合物(4)的制备，其结构式如下：The preparation of the compound (4) has the following structural formula:

把含11.3mg(20μmol)Pd(dba) ₂和15.8mg(29.4μmol)2-(二环己基磷)-3,6-二甲氧基-2',4',6'-三异丙基-1,1'-联苯的混和溶液加入到3ml二氧六环中，然后将混和溶液加入到含0.1g(0.33mmol)化合物(1)(实施例1)，0.04g(0.65mmol)氟化钾的混和物中，加入0.093g(0.65mmol)的三甲基(三氟甲基)硅烷，反应液在140℃下搅拌反应20小时后用硅藻土过滤，滤液真空浓缩。残渣用硅胶柱层析(二氯甲烷：甲醇)纯化得到化合物(4)。MS m/z(ESI):297.1[M+1] ⁺, ¹HNMR(400MHz,DMSO-d6)δ8.51(d,1H),8.31(s,1H),7.85(d,1H),7.52(s,1H),7.55(s,2H),6.58(dd,1H)。 11.3 mg (20 μmol) of Pd(dba) ₂ and 15.8 mg (29.4 μmol) of 2-(dicyclohexylphosphino)-3,6-dimethoxy-2',4',6'-triisopropyl A mixed solution of -1,1'-biphenyl was added to 3 ml of dioxane, and then the mixed solution was added to 0.1 g (0.33 mmol) of the compound (1) (Example 1), 0.04 g (0.65 mmol) of fluorine. To the mixture of potassium, 0.093 g (0.65 mmol) of trimethyl(trifluoromethyl)silane was added, and the reaction mixture was stirred at 140 ° C for 20 hours, then filtered over Celite, and evaporated. The residue was purified by silica gel column chromatography (dichlorohexane: methanol) to afford Compound (4). MS m/z (ESI): 297.1 [M+1] ⁺ , ¹ H NMR (400 MHz, DMSO-d6) δ 8.51 (d, 1H), 8.31 (s, 1H), 7.85 (d, 1H), 7.52 ( s, 1H), 7.55 (s, 2H), 6.58 (dd, 1H).

实施例5：Example 5:

化合物(5)的制备，其结构式如下：The preparation of the compound (5) has the following structural formula:

按合成路线3制备。将化合物(1)和0.06g(0.72mmol)的氰化亚铜加入到吡啶中，混合物置于微波反应器中250℃反应20分钟。薄层色谱(TLC) 监测反应完全后，加入乙酸乙酯后用硅藻土过滤，依次用饱和碳酸氢钠溶液和饱和食盐水洗涤，有机层用无水硫酸镁干燥，浓缩得产物(5)。MS m/z(ESI):254.1[M+1] ⁺, ¹HNMR(400MHz,DMSO-d6)δ8.52(d,1H),8.29(s,1H),7.83(d,1H),7.51(s,1H),7.57(s,2H),6.59(dd,1H)。 Prepared according to Synthetic Route 3. Compound (1) and 0.06 g (0.72 mmol) of cuprous cyanide were added to pyridine, and the mixture was placed in a microwave reactor at 250 ° C for 20 minutes. After the reaction was completed by TLC, ethyl acetate was added, and the mixture was filtered over Celite, washed with saturated sodium hydrogen carbonate and brine, and then evaporated. . MS m/z (ESI): 254.1 [M+1] ⁺ , ¹ H NMR (400 MHz, DMSO-d6) δ 8.52 (d, 1H), 8.29 (s, 1H), 7.83 (d, 1H), 7.51 ( s, 1H), 7.57 (s, 2H), 6.59 (dd, 1H).

实施例6:Example 6

化合物(6)的制备，其结构式如下：The preparation of the compound (6) has the following structural formula:

按合成路线1制备得到化合物(6)，其中第5步反应时用甲基-对甲苯磺酰甲基异氰(Me-TOSMIC)替换对甲苯磺酰甲基异氰(TOSMIC)制备得到化合物(E)的类似物。MS m/z(ESI):261.1[M+1] ⁺, ¹HNMR(400MHz,DMSO-d6)δ8.55(d,1H),8.36(s,1H),7.84(d,1H),7.57(s,2H),6.61(dd,1H),2.16(s,3H)。 Compound (6) is obtained according to the synthesis scheme 1, wherein the compound obtained by the step 5 is prepared by replacing methyl p-toluenesulfonylmethyl isocyanide (Me-TOSMIC) with p-toluenesulfonylmethyl isocyanide (TOSMIC) to obtain a compound ( An analog of E). MS m/z (ESI): 261.1 [M+1] ⁺ , ¹ H NMR (400 MHz, DMSO-d6) δ 8.55 (d, 1H), 8.36 (s, 1H), 7.84 (d, 1H), 7.57 ( s, 2H), 6.61 (dd, 1H), 2.16 (s, 3H).

实施例7:Example 7

化合物(7)的制备，其结构式如下：The preparation of the compound (7) has the following structural formula:

按合成路线1制备。第5步反应时用Me-TOSMIC替换TOSMIC制备化合物(E)的类似物，然后用NCS(N-氯代丁二酰亚胺)替换NBS(N-溴代丁二酰亚胺)进行氯代得到化合物(7)。MS m/z(ESI):277.1[M+1] ⁺, ¹HNMR(400MHz,DMSO-d6)δ8.52(d,1H),8.33(s,1H),7.82(d,1H),7.55(s,2H),6.60(dd,1H),2.14(s,3H)。 Prepared according to Synthetic Route 1. In the fifth step, the analog of the compound (E) is prepared by replacing the TOSMIC with Me-TOSMIC, and then replacing the NBS (N-bromosuccinimide) with NCS (N-chlorosuccinimide) for chlorination. Compound (7) was obtained. MS m/z (ESI): 277.1 [M+1] ⁺ , ¹ H NMR (400 MHz, DMSO-d6) δ 8.52 (d, 1H), 8.33 (s, 1H), 7.82 (d, 1H), 7.55 ( s, 2H), 6.60 (dd, 1H), 2.14 (s, 3H).

实施例8:Example 8

化合物(8)的制备，其结构式如下：The preparation of the compound (8) has the following structural formula:

按合成路线1制备。第5步反应时用Me-TOSMIC替换TOSMIC制备化合物(E)的类似物，然后用NBS(N-溴代丁二酰亚胺)溴代得到化合物(8)。MS m/z(ESI):321.0[M+1] ⁺, ¹HNMR(400MHz,DMSO-d6)δ8.53(d,1H),8.34(s,1H),7.82(d,1H),7.56(s,2H),6.59(dd,1H),2.12(s,3H)。 Prepared according to Synthetic Route 1. In the fifth step, an analog of the compound (E) was prepared by substituting Me-TOSMIC for the TOSMIC, followed by bromination with NBS (N-bromosuccinimide) to obtain the compound (8). MS m/z (ESI): 321.0 [M+1] ⁺ , ¹ H NMR (400 MHz, DMSO-d6) δ 8.53 (d, 1H), 8.34 (s, 1H), 7.82 (d, 1H), 7.56 ( s, 2H), 6.59 (dd, 1H), 2.12 (s, 3H).

实施例9：Example 9

化合物(9)的制备，其结构式如下：The preparation of the compound (9) has the following structural formula:

用化合物(8)替换化合物(1)，按实施例4的方法制备化合物(9)。MS m/z(ESI):311.1[M+1] ⁺, ¹HNMR(400MHz,DMSO-d6)δ8.50(d,1H),8.31(s,1H),7.79(d,1H),7.55(s,2H),6.57(dd,1H),2.13(s,3H)。 Compound (9) was prepared as in Example 4 by substituting Compound (8) for Compound (1). MS m/z (ESI): 311.1 [M+1] ⁺ , ¹ H NMR (400 MHz, DMSO-d6) δ 8.50 (d, 1H), 8.31 (s, 1H), 7.79 (d, 1H), 7.55 ( s, 2H), 6.57 (dd, 1H), 2.13 (s, 3H).

实施例10：Example 10:

化合物(10)的制备，其结构式如下：The preparation of the compound (10) has the following structural formula:

用化合物(8)替换化合物(1)，按实施例5的方法制备化合物(10)。MS m/z(ESI):268.1[M+1] ⁺, ¹HNMR(400MHz,DMSO-d6)δ8.51(d,1H),8.30(s,1H),7.81(d,1H),7.53(s,2H),6.58(dd,1H),2.16(s,3H)。 Compound (10) was prepared as in Example 5 by substituting compound (8) for compound (1). MS m/z (ESI): 268.1 [M+1] ⁺ , ¹ H NMR (400 MHz, DMSO-d6) δ 8.51 (d, 1H), 8.30 (s, 1H), 7.81 (d, 1H), 7. s, 2H), 6.58 (dd, 1H), 2.16 (s, 3H).

实施例11：Example 11

化合物(13)的制备，其结构式如下：The preparation of the compound (13) has the following structural formula:

按合成路线1制备。第二步反应时用吡咯烷替换吡唑得到中间体(B)的类似物，再经偶联、环合、溴代等反应制得化合物(13)。MS m/z(ESI):310.0[M+1] ⁺, ¹HNMR(400MHz,DMSO-d6)δ8.51(d,1H),7.82(d,1H),6.90(s,2H),3.51-3.74(m,4H),1.82-1.85(m,4H)。 Prepared according to Synthetic Route 1. In the second step, the pyrazole is replaced with pyrrolidine to obtain an analog of the intermediate (B), which is then subjected to a coupling, cyclization, bromination or the like to obtain the compound (13). MS m/z (ESI): 310.0 [M+1] ⁺ , ¹ H NMR (400 MHz, DMSO-d6) δ 8.51 (d, 1H), 7.82 (d, 1H), 6.90 (s, 2H), 3.51 3.74 (m, 4H), 1.82-1.85 (m, 4H).

实施例12：Example 12

化合物(14)的制备，其结构式如下：The preparation of the compound (14) has the following structural formula:

用化合物(13)替换化合物(1)，按实施例5的方法制得化合物(14)。MS m/z(ESI):257.1[M+1] ⁺, ¹HNMR(400MHz,DMSO-d6)δ8.50(d,1H),7.81(d,1H),6.94(s,2H),3.53-3.75(m,4H),1.83-1.87(m,4H)。 Compound (14) was obtained by the procedure of Example 5, substituting compound (13) for compound (1). MS m/z (ESI): 257.1 [M+1] ⁺ , ¹ H NMR (400 MHz, DMSO-d6) δ 8.50 (d, 1H), 7.81 (d, 1H), 6.94 (s, 2H), 3.53- 3.75 (m, 4H), 1.83-1.87 (m, 4H).

实施例13：Example 13

化合物(17)的制备，其结构式如下：The preparation of the compound (17) has the following structural formula:

按合成路线1制备。第2步反应用吡咯烷替换吡唑制备中间体(B)的类似物，第5步反应时用Me-TOSMIC替换TOSMIC制备中间体(E)的类似物，然后用NBS(N-溴代丁二酰亚胺)溴代得到化合物(17)。MS m/z(ESI):324.0[M+1] ⁺, ¹HNMR(400MHz,DMSO-d6)δ8.50(s,1H),7.70(s,2H),3.52-3.76(m,4H),2.16(s,3H),1.82-1.86(m,4H)。 Prepared according to Synthetic Route 1. The second step of the reaction is to replace the pyrazole with pyrrolidine to prepare the analog of the intermediate (B). In the fifth step, the analog of the intermediate (E) is prepared by replacing the TOSMIC with Me-TOSMIC, and then NBS (N-brominated) The diimide) is brominated to give the compound (17). MS m/z (ESI): 324.0 [M+1] ⁺ , ¹ H NMR (400 MHz, DMSO-d6) δ 8.50 (s, 1H), 7.70 (s, 2H), 3.52-3.76 (m, 4H), 2.16 (s, 3H), 1.82-1.86 (m, 4H).

实施例14：Example 14

化合物(18)的制备，其结构式如下：The preparation of the compound (18) has the following structural formula:

化合物(17)替换化合物(1)，按实施例5的方法制得化合物(18)。MS m/z(ESI):271.1[M+1] ⁺, ¹HNMR(400MHz,DMSO-d6)δ8.52(d,1H),7.69(s,2H),3.53-3.77(m,4H),1.82-1.86(m,4H)。 Compound (17) was replaced with Compound (1), and Compound (18) was obtained by the method of Example 5. MS m/z (ESI): 271.1 [M+1] ⁺ , ¹ H NMR (400 MHz, DMSO-d6) δ 8.52 (d, 1H), 7.69 (s, 2H), 3.53-3.77 (m, 4H), 1.82-1.86 (m, 4H).

实施例15：Example 15

化合物(19)的制备，其结构式如下：The preparation of the compound (19) has the following structural formula:

按合成路线2制备。Prepared according to Synthetic Route 2.

第一步：制备中间体6-氯-2-(甲基亚磺酰基)嘧啶-4-胺(F)First step: preparation of the intermediate 6-chloro-2-(methylsulfinyl)pyrimidine-4-amine (F)

在30分钟里向溶有1.0g(5.7mmol)6-氯-2-(甲硫基)嘧啶-4-胺的50ml二氯甲烷溶液中加入溶于30ml二氯甲烷的1.5g(6.9mmol)间-氯过苯甲酸(77％)溶液。将该反应混合物在室温下搅拌4小时。过滤生成的白色沉淀，用二氯甲烷洗涤数次，干燥，得到1.0g中间体(F)。MS m/z(ESI):192.0[M+1] ⁺。 To a solution of 1.0 g (5.7 mmol) of 6-chloro-2-(methylthio)pyrimidine-4-amine in 50 ml of dichloromethane over 30 min, 1.5 g (6.9 mmol) dissolved in dichloromethane A solution of m-chloroperbenzoic acid (77%). The reaction mixture was stirred at room temperature for 4 hours. The resulting white precipitate was filtered, washed with dichloromethane several times and dried to give &lt MS m/z (ESI): 192.0 [M+1] ⁺ .

第二步：制备中间体5-溴-6-氯-2-(甲基亚磺酰基)嘧啶-4-胺(G)：将1.12g(6.3mmol的N-溴代丁二酰亚胺(NBS)缓慢加入到1.0g(5.3mmol)6-氯-2-(甲基亚磺酰基)嘧啶-4-胺的30ml DMF冷却的混悬液中。室温下搅拌50分钟后，过滤沉淀，用冷DMF洗涤，用冷水洗涤数次，真空干燥得到(G)。MS m/z(ESI):271.9[M+1] ⁺。 Second step: Preparation of intermediate 5-bromo-6-chloro-2-(methylsulfinyl)pyrimidine-4-amine (G): 1.12 g (6.3 mmol of N-bromosuccinimide ( NBS) was slowly added to a suspension of 1.0 g (5.3 mmol) of 6-chloro-2-(methylsulfinyl)pyrimidine-4-amine in 30 ml of DMF. After stirring at room temperature for 50 minutes, the precipitate was filtered. washed with cold with DMF, washed several times with cold water, and dried in vacuo to give (G) .MS m / z ( ESI): 271.9 [m + 1] +.

第三步：制备中间体5-溴-6-氯-2-(噁唑-2-基〕嘧啶-4-胺(H)：The third step: preparation of the intermediate 5-bromo-6-chloro-2-(oxazol-2-yl)pyrimidine-4-amine (H):

-78℃下把噁唑(260mg，3.7mmol)溶于无水THF，加入nBuLi(1.6M，溶于正己烷)搅拌15分钟，自然升温至-20℃。将1g(3.7mmol)中间体(G)和Pd(PPh ₃) ₄加入到无水THF中。再加入到上述噁唑溶液中。混和液在80℃下反应2小时后，将溶液倒至1N的盐酸中，乙酸乙酯提取，有机层水洗，饱和食盐水洗，无水硫酸钠干燥，硅胶柱层析(5％甲醇的二氯甲烷溶液)得所需产物。MS m/z(ESI):276.9[M+1] ⁺。 The oxazole (260 mg, 3.7 mmol) was dissolved in anhydrous THF at -78 ° C, and nBuLi (1.6M, dissolved in n-hexane) was added and stirred for 15 minutes, and the temperature was naturally raised to -20 °C. 1 g (3.7 mmol) of intermediate (G) and Pd(PPh ₃ ) _{4 were} added to anhydrous THF. Further added to the above oxazole solution. After the reaction mixture was reacted at 80 ° C for 2 hours, the solution was poured into 1 N hydrochloric acid, extracted with ethyl acetate, and the organic layer was washed with water, washed with brine, dried over anhydrous sodium sulfate, silica gel column chromatography The methane solution) gave the desired product. MS m / z (ESI): 276.9 [M + 1] +.

第四步：制备化合物(19)Step 4: Preparation of Compound (19)

向溶有0.15g中间体(H)的3ml DMF溶液中加入0.11g(1.64mmol)的1H-吡唑和0.18g(0.55mmol)的碳酸铯。将该混合物在90℃下搅拌24小时。减压浓缩溶剂DMF。用水洗涤粗品，干燥得化合物(19)。MS m/z(ESI):307.1[M+1] ⁺。 To a solution of 0.15 g of the intermediate (H) in 3 ml of DMF was added 0.11 g (1.64 mmol) of 1H-pyrazole and 0.18 g (0.55 mmol) of cesium carbonate. The mixture was stirred at 90 ° C for 24 hours. The solvent DMF was concentrated under reduced pressure. The crude product was washed with water and dried to give Compound (19). MS m/z (ESI): 307.1 [M+1] ⁺ .

实施例16：Example 16:

化合物(25)的制备，其结构式如下：The preparation of the compound (25) has the following structural formula:

按上述合成路线2制备，具体方法参见实施例15。第三步反应中，中间体(G)和1H-三氮唑改为在碳酸铯作用下，于DMF溶液中室温搅拌反应数小时制备相应中间体(H)类似物。再用实施例15的类似方法用吡唑取代嘧啶6位氯得化合物(25)。MS m/z(ESI):307.1[M+1] ⁺, ¹HNMR(400MHz,DMSO-d6)δ9.14(s,1H),8.60(s,1H),8.45(d,1H),7.64(d,1H),6.55(dd,1H)。 Prepared according to the above synthetic route 2, and the specific method is shown in Example 15. In the third step of the reaction, the intermediate (G) and 1H-triazole are replaced by a reaction of cesium carbonate in a DMF solution at room temperature for several hours to prepare a corresponding intermediate (H) analog. The compound (25) was obtained by substituting a pyrazole at the 6-position of the pyrimidine with a similar method of Example 15. MS m/z (ESI): 307.1 [M+1] ⁺ , ¹ H NMR (400 MHz, DMSO-d6) δ 9.14 (s, 1H), 8.60 (s, 1H), 8.45 (d, 1H), 7.64 ( d, 1H), 6.55 (dd, 1H).

药理学活性：Pharmacological activity:

(1)对腺苷A _2A受体结合亲和力分析： (1) Analysis of adenosine A _2A receptor binding affinity:

使用标准技术，通过确定人源腺苷A _2A受体选择性放射配体[ ³H]CGS-21680的转换来测定本发明化合物在体外结合人源腺苷A _2A受体的结合亲和力，结果总结在表1中。 The binding affinity of the compounds of the invention to the human adenosine A _2A receptor in vitro was determined by determining the conversion of the human adenosine A _2A receptor selective radioligand [ ³ H]CGS-21680 using standard techniques. In Table 1.

用编码人源腺苷A _2A受体的质粒在HEK-293细胞上稳定转染，使用该细胞在三(羟甲基)氨基甲烷(Tris-HCl)缓冲液(pH＝7.4)中采用标准技术制备膜。约15μg的膜蛋白和50nM放射性配体[ ³H]CGS-21680、10μM待测化合物混和并在25℃下共孵育90分钟。添加50μM NECA(腺苷-5’-N-乙基羧基酰胺)测定非特异性结合。将膜过滤并洗涤3次以除去未结合的放射性配体。过滤器用闪烁计数器测定结合的配体。通过分析多个不同的浓度来确定浓度-响应结合竞争性曲线。使用非线性拟合程序计算IC ₅₀值。通过Cheng-Prusoff等式(Ⅱ)来计算化合物的抑制常数(Ki)。 The plasmid encoding the human adenosine A _2A receptor was stably transfected on HEK-293 cells using standard techniques in tris-HCl buffer (pH = 7.4). A film was prepared. About 15 μg of membrane protein was mixed with 50 nM radioligand [ ³ H]CGS-21680, 10 μM test compound and co-incubated for 90 minutes at 25 °C. Non-specific binding was determined by the addition of 50 μM NECA (adenosine-5'-N-ethylcarboxyamide). The membrane was filtered and washed 3 times to remove unbound radioligand. The filter was assayed for the bound ligand using a scintillation counter. The concentration-response binding competition curve was determined by analyzing a plurality of different concentrations. IC ₅₀ values _were calculated using a nonlinear fitting program. The inhibition constant (Ki) of the compound was calculated by Cheng-Prusoff equation (II).

Ki＝IC ₅₀/(1+[L]/KD) (Ⅱ) Ki=IC ₅₀ /(1+[L]/KD) (II)

其中IC ₅₀是转换50％的放射性配体结合时的化合物浓度，[L]是放射性配体的游离浓度，KD是放射性配体的解离常数。IC ₅₀值是通过Prism软件用非线性回归拟合该数据而得到。Ki数值越小说明化合物对人源腺苷A _2A受体的拮抗作用就越明显。 Wherein IC ₅₀ is the concentration of the compound when 50% of the radioligand is converted, [L] is the free concentration of the radioligand, and KD is the dissociation constant of the radioligand. The IC ₅₀ value was obtained by fitting the data by nonlinear regression using Prism software. The smaller the Ki value, the more obvious the antagonism of the compound to the human adenosine A _2A receptor.

(2)对腺苷A ₁受体结合亲和力分析： (2) Analysis of adenosine A ₁ receptor binding affinity:

化合物DPCPX(1,3-二丙基-环戊黄嘌呤)是已知的高活性的腺苷A ₁受体拮抗剂(文献报道其Ki＝0.45nM)。使用标准技术，通过确定人源腺苷A ₁受体选择性放射配体[ ³H]DPCPX的转换来测定本发明化合物在体外结合人源腺苷A ₁受体的结合亲和力，以判断本发明化合物对A _2A受体的选择性强度。结果总结在表1中。 The compound DPCPX (1,3-dipropyl-cyclopentanthine) is a known highly active adenosine A ₁ receptor antagonist (Ki = 0.45 nM reported in the literature). The binding affinity of a compound of the invention to a human adenosine A ₁ receptor in vitro is determined by determining the conversion of the human adenosine A ₁ receptor selective radioligand [ ³ H]DPCPX using standard techniques to determine the present invention. The selective strength of the compound to the A _2A receptor. The results are summarized in Table 1.

用编码人源腺苷A ₁受体的质粒在CHO细胞上稳定转染，使用该细胞在改良的HEPES缓冲液(pH＝7.4)中采用标准技术制备膜。约10μg的膜蛋白和1nM放射性配体[ ³H]DPCPX、10μM待测化合物混和并在25℃下共孵育90分钟。添加100μM R(-)-PIA(R-苯异丙基腺苷)测定非特异性结合。将膜过滤并洗涤3次以除去未结合的放射性配体。过滤器用闪烁计数器测定结合的配体。通过分析多个不同的浓度来确定浓度-响应结合竞争性曲线。按前述方法计算化合物的抑制常数(Ki)。 Encoding human adenosine A ₁ receptor stably transfected with a plasmid in CHO cells, the cells were used in modified HEPES buffer (pH = 7.4) films prepared using standard techniques. About 10 μg of membrane protein was mixed with 1 nM radioligand [ ³ H]DPCPX, 10 μM of the test compound and co-incubated for 90 minutes at 25 °C. Non-specific binding was determined by the addition of 100 μM R(-)-PIA (R-phenylisopropyladenosine). The membrane was filtered and washed 3 times to remove unbound radioligand. The filter was assayed for the bound ligand using a scintillation counter. The concentration-response binding competition curve was determined by analyzing a plurality of different concentrations. The inhibition constant (Ki) of the compound was calculated as described above.

(3)对人源腺苷A _2A受体抗血小板聚集活性分析： (3) Analysis of anti-platelet aggregation activity of human adenosine A _2A receptor:

CGS-21680是腺苷A _2A受体的高活性激动剂，能作用于腺苷A _2A受体促使血小板聚集。通过测试化合物抑制CGS-21680诱导的血小板聚集可获得本发明化合物在细胞水平上对腺苷A _2A受体的拮抗活性。结果总结在表1中。 CGS-21680 is a high adenosine A _2A receptor agonist activity, can function to promote platelet aggregation adenosine A _2A receptor. The antagonistic activity of the compounds of the invention on the adenosine A _2A receptor at the cellular level can be obtained by testing the compound to inhibit CGS-21680-induced platelet aggregation. The results are summarized in Table 1.

37℃下，富含人血小板的血浆在血栓素A ₂受体激动剂U-46619(10μM)作用下，上层富含血小板的血浆(6×10 ⁸血小板/毫升)产生聚集，以光学凝集计检测。以1μM CGS-21680作为对照药，如果5分钟内测试化合物(30μM)体系中血小板聚集达到50％或更多(≥50％)，表明测试物质可能具有腺苷A _2A受体激动剂活性。 At 37 ° C, human platelet-rich plasma under the action of thromboxane A ₂ receptor agonist U-46619 (10 μM), the upper layer of platelet-rich plasma (6 × 10 ⁸ platelets / ml) produced aggregation, with optical agglutination Detection. With 1 μM CGS-21680 as a control, if the platelet aggregation in the test compound (30 μM) system reached 50% or more (≥50%) within 5 minutes, it was indicated that the test substance may have adenosine A _2A receptor agonist activity.

当在某一测试物浓度下未观察到明显的激动活性时，其降低CGS-21680(1μM)诱导的抑制反应达到50％或更多(≥50％)，则表明测试化合物具有腺苷A _2A受体拮抗活性。按前述方法计算化合物的抑制常数(Ki)，Ki数值越小说明化合物对人源腺苷A _2A受体的拮抗作用就越明显。 When no significant agonistic activity was observed at a certain test substance concentration, which reduced CGS-21680 (1 μM)-induced inhibition by 50% or more (≥50%), the test compound had adenosine A _2A Receptor antagonistic activity. The inhibition constant (Ki) of the compound was calculated as described above, and the smaller the Ki value, the more obvious the antagonistic effect of the compound on the human adenosine A _2A receptor.

表1.化合物(1)-(25)在亲和力分析及对血小板聚集分析中得到的对人源腺苷A _2A和A ₁受体的抑制常数(“/”表示未测试) Table 1. Inhibition constants for human adenosine A _2A and A ₁ receptors obtained by affinity analysis and platelet aggregation analysis of compounds (1)-(25) ("/" indicates untested)

由表1可知，本发明所制备的化合物在nM浓度级别上即可对人源腺苷A _2A受体具有明显的拮抗作用。且当R ₁基团的吸电性增强，如卤素原子由溴变为氯，或由卤素原子变为氰基时，本发明化合物对人源腺苷A _2A受体的拮抗活性均有明显提高。 As can be seen from Table 1, the compound prepared by the present invention has a significant antagonistic effect on the human adenosine A _2A receptor at the nM concentration level. And when the hydrogen absorption property of the R ₁ group is enhanced, such as a halogen atom changing from bromine to chlorine, or from a halogen atom to a cyano group, the antagonistic activity of the compound of the present invention on the human adenosine A _2A receptor is markedly improved. .

此外，上述化合物VI是CN102892761中实施例1化合物，其为吡唑类化合物，结构式如下：Further, the above compound VI is a compound of Example 1 in CN102892761, which is a pyrazole compound, and has the following structural formula:

现将其在本发明中作为阳性对照物。在专利CN102892761中，报道了该化合物VI对人源A _2A受体的亲和力Ki值为12nM，在CHO-A _2A细胞上对cAMP的Ki值为25nM。本发明采用了上述的测试方法，所测化合物VI的拮抗活性比CN102892761专利报道的结果较差(见表1)。 It is now used as a positive control in the present invention. In the patent CN102892761, it is reported that the affinity of the compound VI for the human _A2A receptor is 12 nM, and the Ki value for cAMP on CHO-A _2A cells is 25 nM. The above test method is adopted in the present invention, and the antagonistic activity of the tested compound VI is inferior to that reported in the CN102892761 patent (see Table 1).

在腺苷的4种受体亚型中，由于A ₁和A _2A是高表达受体，在生理状态下低水平腺苷浓度时就可产生作用，而A _2B和A ₃的表达量较低，仅在病理情况下，腺苷大量增加时才可活化产生病理效应。本发明所述噁唑类化合物相对于A ₁受体，对A _2A受体具有明显的高选择性(选择性系数为表1中化合物对人源腺苷A ₁受体亲和力的抑制常数和对人源腺苷A _2A受体亲和力的抑制常数的比值)，且部分化合物的选择性系数3-4倍于CN102892761中报道化合物VI的选择性系数，在本技术领域内即可被认为具有显著的进步。 Among the four receptor subtypes of adenosine, since A ₁ and A _2A are highly expressed receptors, low levels of adenosine concentration can be produced under physiological conditions, while A _2B and A ₃ are expressed at lower levels. Only in the pathological situation, a large increase in adenosine can activate the pathological effect. The oxazole compound of the present invention has a markedly high selectivity to the A _2A receptor relative to the A ₁ receptor (the selectivity coefficient is the inhibition constant and the pair of compounds in Table 1 for the affinity of the human adenosine A ₁ receptor) The ratio of the inhibition constant of human adenosine A _2A receptor affinity), and the selectivity coefficient of some compounds 3-4 times the selectivity coefficient of the reported compound VI in CN102892761, can be considered significant in the art. progress.

通过本发明化合物对人源腺苷A _2A受体抗血小板聚集活性的分析，验证了它们在细胞水平上的拮抗A _2A受体的功能作用。大多数测试的本发明化合物表现出比CN102892761中所述的吡唑类化合物VI更强的细胞活性，如化合物(5)的细胞活性比VI强约4.5倍。 Analysis of the anti-platelet aggregation activity of the human adenosine A _2A receptor by the compounds of the present invention demonstrated their antagonism of the functional role of the A _2A receptor at the cellular level. Most of the compounds of the invention tested exhibited greater cell viability than the pyrazoles VI described in CN102892761, such as the compound (5) having a cell activity about 4.5 times stronger than VI.

因而，相比于CN102892761中所述的吡唑类化合物VI，本发明所述的噁唑类化合物对人源腺苷A _2A受体的拮抗活性和功能活性相当或更优，且对人源腺苷A ₁受体的亲和力明显降低，表现出对A _2A受体的高度选择性。 Thus, the antagonistic and functional activity of the oxazole compound of the present invention against the human adenosine A _2A receptor is comparable or superior to that of the pyrazole compound VI described in CN102892761, and is directed to the human gland. The affinity of the glycoside A ₁ receptor is significantly reduced, exhibiting a high selectivity for the A _2A receptor.

药代动力学评价：Pharmacokinetic evaluation:

测试本发明化合物(1)、(5)、(8)、(10)、(13)的药代动力学。The pharmacokinetics of the compounds (1), (5), (8), (10), (13) of the present invention were tested.

按常规方法，以SD大鼠为受试动物，应用LC/MS/MS法测定大鼠静脉注射给予本发明化合物(1)、(5)、(8)、(10)、(13)后不同时刻血浆中的药物浓度，研究本发明化合物在大鼠体内的药代动力学行为，评价其药动学特征。现将专利CN102892761报道的化合物VI作为阳性对照物，采用上述方法同时测试。其结果总结于表2。SD rats were used as test animals according to a conventional method, and the rats were intravenously administered with the compounds of the present invention (1), (5), (8), (10), and (13) by LC/MS/MS method. The drug concentration in the plasma at the time of the study, the pharmacokinetic behavior of the compound of the present invention in rats was investigated, and its pharmacokinetic characteristics were evaluated. Compound VI reported in the patent CN102892761 is now used as a positive control and tested simultaneously by the above method. The results are summarized in Table 2.

表2.本发明化合物(1)、(5)、(8)、(10)、(13)的药代动力学参数(SD大鼠静脉注射1.0mg/kg)Table 2. Pharmacokinetic parameters of the compounds (1), (5), (8), (10), (13) of the present invention (SD rats intravenously 1.0 mg/kg)

由表2结果可见，本发明所述的噁唑类化合物具有更好的药代动力学特征，比CN102892761中所述的吡唑类化合物VI具有明显更低的血浆清除率，从而可以维持较高的血药浓度和较长的有效时间；本发明所述的噁唑类化合物在大鼠体内的半衰期是化合物VI的半衰期的3倍以上，而在本技术领域内半衰期延长20％-40％即可认为更优，延长1倍以上即可认为具有显著的进步；因而本发明化合物相较于CN102892761中所述的吡唑类化合物VI在维持更高的血药浓度和更长的有效治疗时间上具有显著的进步。As can be seen from the results of Table 2, the oxazole compound of the present invention has better pharmacokinetic characteristics and has a significantly lower plasma clearance than the pyrazole compound VI described in CN102892761, thereby maintaining high The blood concentration and the longer effective time; the half-life of the oxazole compound of the present invention in rats is more than three times the half life of the compound VI, and the half-life is extended by 20% to 40% in the technical field. It can be considered that it is better, and it is considered to have a significant improvement by more than one time; therefore, the compound of the present invention maintains a higher blood concentration and a longer effective treatment time than the pyrazole compound VI described in CN102892761. Significant progress.

综上所述，本发明所提供的4-氨基嘧啶衍生物对人源腺苷A _2A受体具有明显的拮抗作用，并显示出高度的A _2A受体选择性，同时在大鼠体内的代谢吸收良好，可应用于治疗对A _2A拮抗作用有反应的疾病或病症的组合物或组合产品中，尤其是用于治疗神经退行性疾病、锥体外综合征、抑郁、多动综合征、睡眠障碍、焦虑症、糖尿病或肿瘤等疾病或病症。 In summary, the 4-aminopyrimidine derivative provided by the present invention has a significant antagonistic effect on the human adenosine A _2A receptor and exhibits high A _2A receptor selectivity and metabolism in rats. Good absorption, can be applied to compositions or combination products for treating diseases or conditions responsive to A _2A antagonism, especially for treating neurodegenerative diseases, extrapyramidal syndrome, depression, hyperactivity syndrome, sleep disorders A disease or condition such as anxiety, diabetes or cancer.

尽管本发明的内容已经通过上述优选实施例作了详细介绍，但应当认识到上述的描述不应被认为是对本发明的限制。在本领域技术人员阅读了上述内容后，对于本发明的多种修改和替代都将是显而易见的。因此，本发明的保护范围应由所附的权利要求来限定。Although the present invention has been described in detail by the preferred embodiments thereof, it should be understood that the foregoing description should not be construed as limiting. Various modifications and alterations of the present invention will be apparent to those skilled in the art. Therefore, the scope of the invention should be defined by the appended claims.

Claims

一种作为腺苷A _2A受体拮抗剂的4-氨基嘧啶衍生物，其特征在于，结构通式(Ⅰ)如下： A 4-aminopyrimidine derivative as an adenosine A _2A receptor antagonist, characterized in that the structural formula (I) is as follows:

其中：among them:

R ₁选自卤素、氰基或三氟甲基； R _{1 is} selected from halogen, cyano or trifluoromethyl;

R ₂选自吡唑基、吡咯烷基，或经一个或多个卤素或C _1-3烷基取代的吡唑基或吡咯烷基； R _{2 is} selected from pyrazolyl, pyrrolidinyl, or pyrazolyl or pyrrolidinyl substituted by one or more halogen or C _1-3 alkyl;

R ₃选自噁唑基、噁二唑基、三氮唑基，或经一个或多个卤素或C _1-3烷基取代的噁唑基。 R _{3 is} selected from oxazolyl, oxadiazolyl, triazolyl, or oxazolyl substituted with one or more halogen or C _1-3 alkyl.
如权利要求1所述的作为腺苷A _2A受体拮抗剂的4-氨基嘧啶衍生物，其特征在于，R ₂选自吡唑基或吡咯烷基；R ₃选自噁唑基、噁二唑基、三氮唑基，或经一个C _1-3烷基取代的噁唑基。 The 4-aminopyrimidine derivative as an adenosine A _2A receptor antagonist according to claim 1, wherein R _{2 is} selected from pyrazolyl or pyrrolidinyl; and R _{3 is} selected from oxazolyl, oxazide An oxazolyl group, a triazolyl group, or an oxazolyl group substituted with a C _1-3 alkyl group.
如权利要求1所述的作为腺苷A _2A受体拮抗剂的4-氨基嘧啶衍生物，其特征在于，该4-氨基嘧啶衍生物选自下述化合物： The 4-aminopyrimidine derivative as an adenosine A _2A receptor antagonist according to claim 1, wherein the 4-aminopyrimidine derivative is selected from the group consisting of the following compounds:
一种如权利要求1-3中任意一项所述的4-氨基嘧啶衍生物在制备用于治疗对A _2A拮抗作用有反应的疾病或病症的药物中的用途。 Use of a 4-aminopyrimidine derivative according to any one of claims 1 to 3 for the manufacture of a medicament for the treatment of a disease or condition responsive to _A2A antagonism.
如权利要求4所述的用途，其特征在于，所述疾病或病症包含神经退行性疾病、锥体外综合征、抑郁、多动综合征、睡眠障碍、焦虑症、糖尿病或肿瘤中的任意一种或多种。The use according to claim 4, wherein the disease or condition comprises any one of a neurodegenerative disease, an extrapyramidal syndrome, depression, hyperactivity syndrome, sleep disorder, anxiety disorder, diabetes or tumor. Or a variety.
如权利要求5所述的用途，其特征在于，所述肿瘤为肺癌、卵巢癌、胰腺癌、胃癌、乳腺癌、胶质母细胞瘤、黑色素瘤、肾细胞癌、三阴性乳腺癌、结直肠癌、头颈部癌、膀胱癌、***癌、肝细胞肝癌或胆管癌。The use according to claim 5, wherein the tumor is lung cancer, ovarian cancer, pancreatic cancer, gastric cancer, breast cancer, glioblastoma, melanoma, renal cell carcinoma, triple negative breast cancer, colorectal Cancer, head and neck cancer, bladder cancer, prostate cancer, hepatocellular carcinoma or cholangiocarcinoma.
如权利要求6所述的用途，其特征在于，所述肺癌为非小细胞肺癌。The use according to claim 6, wherein the lung cancer is non-small cell lung cancer.
一种药物组合物，其特征在于，其包含如权利要求1-3中任意一项所述的作为腺苷A _2A受体拮抗剂的4-氨基嘧啶衍生物，及其药学上可接受的赋形剂。 A pharmaceutical composition comprising the 4-aminopyrimidine derivative as an adenosine A _2A receptor antagonist according to any one of claims 1 to 3, and a pharmaceutically acceptable compound thereof Shape agent.
一种复方药物，其特征在于，包含如权利要求1-3中任意一项所述的4-氨基嘧啶衍生物以及能够与其联用的药物。A compound drug comprising the 4-aminopyrimidine derivative according to any one of claims 1 to 3 and a drug capable of being used in combination therewith.
如权利要求9所述的复方药物，其特征在于，所述联用的药物为用于治疗以下疾病或病症的化合物：神经退行性疾病、锥体外综合征、抑郁、多动综合征、睡眠障碍、焦虑症、糖尿病或肿瘤。The combination drug according to claim 9, wherein the combination drug is a compound for treating a disease or a condition: a neurodegenerative disease, an extrapyramidal syndrome, depression, hyperactivity syndrome, sleep disorder , anxiety, diabetes or cancer.