CN114948950A - A kind of pharmaceutical composition and its antiviral use - Google Patents

Abstract

The invention provides a pharmaceutical composition consisting of a novel deuterated cyano compound or an isomer thereof or a prodrug thereof or a solvate thereof or a pharmaceutically acceptable salt thereof, and ritonavir or an isomer thereof or a prodrug thereof or a solvate thereof or a pharmaceutically acceptable salt thereof, and antiviral application thereof. Compared with other existing antiviral drug compositions, the drug composition has better pharmacokinetic characteristics, longer half-life period, higher concentration in vivo, high safety, reduced toxic and side effects of drugs, reduced drug resistance of viruses and the like.

Description

Translated fromChinese

一种药物组合物及其抗病毒用途A kind of pharmaceutical composition and its antiviral use

技术领域technical field

本发明属于药物制剂领域，具体涉及一种由新型氘代氰基类化合物或其异构体或其前药或其溶剂化物或其药学上可接受的盐、以及利托那韦或其异构体或其前药或其溶剂化物或其药学上可接受的盐组成的药物组合物及其抗病毒用途。The invention belongs to the field of pharmaceutical preparations, in particular to a novel deuterated cyano compound or its isomer or its prodrug or its solvate or its pharmaceutically acceptable salt, and ritonavir or its isomer A pharmaceutical composition composed of the body or its prodrug or its solvate or its pharmaceutically acceptable salt and its antiviral use.

背景技术Background technique

人体内的冠状病毒最早于1960年代在英国被分离出来，病毒因其表面皇冠状的突起物而得名。它可能与人、猪、猫、狗、鼠和鸡的呼吸系统感染相关。The human coronavirus was first isolated in the UK in the 1960s, and the virus is named for the crown-like protrusions on its surface. It may be associated with respiratory infections in humans, pigs, cats, dogs, mice and chickens.

SARS病毒属于套式病毒目、冠状病毒科、冠状病毒属，为β属B亚群冠状病毒。病毒粒子多呈圆形，有囊膜，外周有冠状排列的纤突,分布于细胞浆中，呈圆形，病毒直径在80～120nm之间。SARS是一种起病急、传播快、病死率高的传染病，被传染的病人多数都与患者直接或间接接触，或生活在流行区内。The SARS virus belongs to the order Nesteviridae, the family Coronaviridae, and the genus Coronavirus, and is a β-genus B subgroup coronavirus. Most of the virus particles are round, with a capsule, and there are coronal fibers around the periphery. They are distributed in the cytoplasm and are round. The diameter of the virus is between 80 and 120 nm. SARS is an infectious disease with rapid onset, rapid spread and high case fatality rate. Most of the infected patients have direct or indirect contact with patients, or live in endemic areas.

MERS病毒是一种β属C亚群冠状病毒，全名为中东呼吸综合征冠状病毒(MiddleEast Respiratory Syndrome Coronavirus，简称MERS-CoV)，感染后引发中东呼吸综合征(Middle East Respiratory Syndrome，简称MERS)。MERS-CoV最早于2012年9月在沙特被发现，早期因与SARS临床症状相似得名“类SARS病毒”，也成为第6种已知的人类冠状病毒，也是过去10年内被分离出来的第3种。MERS virus is a β genus C subgroup coronavirus, the full name is Middle East Respiratory Syndrome Coronavirus (MERS-CoV), which causes Middle East Respiratory Syndrome (MERS) after infection. . MERS-CoV was first discovered in Saudi Arabia in September 2012. It was named "SARS-like virus" because of its clinical symptoms similar to SARS. It has also become the sixth known human coronavirus and the first isolated in the past 10 years 3 types.

新型冠状病毒SARS-CoV-2是以前从未在人体中发现的冠状病毒新毒株，2019年首次被发现并报道，至今仍然在全球多个国家流行肆虐，并在很多国家区域并未得到很好的控制。The new coronavirus SARS-CoV-2 is a new strain of coronavirus that has never been found in humans before. It was first discovered and reported in 2019, and it is still prevalent in many countries around the world, and has not been well received in many countries and regions. good control.

人感染了冠状病毒后常见体征有呼吸道症状、发热、咳嗽、气促和呼吸困难等。在较严重病例中，感染可导致肺炎、严重急性呼吸综合征、肾衰竭，甚至死亡，而且目前对于新型冠状病毒所致疾病没有特异治疗方法。Common signs of people infected with coronavirus include respiratory symptoms, fever, cough, shortness of breath, and difficulty breathing. In more severe cases, infection can lead to pneumonia, severe acute respiratory syndrome, kidney failure, and even death, and there is currently no specific treatment for the disease caused by the novel coronavirus.

新型氘代氰基类化合物是由上海谷森医药有限公司独自研发的一种小分子3CL蛋白酶抑制剂，其结构为：

体外实验证实，其具有惊人的抗SARS-CoV-2活性，能有效抑制病毒的复制，更让人意外的是，该化合物在病毒抑菌活性相当的基础上，实现了比辉瑞、默克所研发的口服抗新冠药更优的药动学性质和治疗效果，目前，上海谷森医药有限公司已经准备进行临床试验用于治疗新冠病毒病患，一旦获得成功，市场前景非常巨大。而且，上海谷森医药有限公司针对新型氘代氰基类化合物申请了发明专利(申请号CN202111234708X)。The new deuterated cyano compound is a small molecule 3CL protease inhibitor independently developed by Shanghai Gusen Pharmaceutical Co., Ltd. Its structure is:

In vitro experiments have confirmed that it has amazing anti-SARS-CoV-2 activity and can effectively inhibit the replication of the virus. What is even more surprising is that the compound has a similar antibacterial activity to the virus. The oral anti-COVID-19 drug developed has better pharmacokinetic properties and therapeutic effect. At present, Shanghai Gusen Pharmaceutical Co., Ltd. is preparing to conduct clinical trials for the treatment of patients with COVID-19. Once successful, the market prospect is very huge. Moreover, Shanghai Gusen Pharmaceutical Co., Ltd. applied for an invention patent (application number CN202111234708X) for new deuterated cyano compounds.

联合用药是全球新冠特效药研发的一大热点，如CN111135166A公开了一种药物组合物，它由GC376与GS-441524组成，体外细胞试验结果表明：该组合物能抑制新型冠状病毒SARS-CoV-2在细胞中增殖，且药效优于单独用药，具有显著的协同增效作用；WO2021/203055A1涉及请涉包含蛋白酶抑制剂和RNA聚合酶抑制剂的抗病毒药物组合物，能够显著降低单药的EC5值。Combination medication is a hot spot in the research and development of new coronavirus specific drugs in the world. For example, CN111135166A discloses a pharmaceutical composition, which consists of GC376 and GS-441524. The results of in vitro cell tests show that the composition can inhibit the new coronavirus SARS-CoV- 2. Proliferate in cells, and the efficacy is better than that of single drug, and has a significant synergistic effect; WO2021/203055A1 relates to an antiviral drug composition comprising a protease inhibitor and an RNA polymerase inhibitor, which can significantly reduce single drug use. the EC5 value.

尽管新型氘代氰基类化合物在体外具有很好的抗病毒活性，但其药代动力学特征和毒副作用有待改进，而本发明公开的药物组合物恰好解决了上述问题。Although the novel deuterated cyano compounds have good antiviral activity in vitro, their pharmacokinetic characteristics and toxic and side effects need to be improved, and the pharmaceutical composition disclosed in the present invention just solves the above problems.

发明内容SUMMARY OF THE INVENTION

一方面，本发明提供了一种药物组合物，由新型氘代氰基类化合物或其异构体或其前药或其溶剂化物或其药学上可接受的盐、以及利托那韦或其异构体或其前药或其溶剂化物或其药学上可接受的盐组成。In one aspect, the present invention provides a pharmaceutical composition comprising a novel deuterated cyano compound or its isomer or its prodrug or its solvate or its pharmaceutically acceptable salt, and ritonavir or its Isomers or their prodrugs or their solvates or their pharmaceutically acceptable salts.

其中，所述新型氘代氰基类化合物的结构式为：Wherein, the structural formula of described novel deuterated cyano compound is:

所述利托那韦的结构式为：The structural formula of ritonavir is:

本发明药学上可接受的盐包括相应化合物的有机盐和无机盐，可以是本领域技术人员所熟知的所有盐，优选但非限制性实例是盐酸盐、氢溴酸盐、硫酸盐、硝酸盐、乙酸盐、己二酸盐、天冬氨酸盐、苯甲酸盐等。The pharmaceutically acceptable salts of the present invention include organic and inorganic salts of the corresponding compounds, and can be all salts well known to those skilled in the art, preferred but non-limiting examples are hydrochloride, hydrobromide, sulfate, nitric acid salts, acetates, adipates, aspartates, benzoates, etc.

进一步地，化合物可以以前药的形式施用。前药可包含共价键合的载体，其施用于哺乳动物对象时释放活性母体药物。前药可以通过以这种方式修饰化合物中存在的功能基团使得修饰在例行操作中或在体内裂解为母体化合物。Further, the compounds can be administered in the form of prodrugs. A prodrug can comprise a covalently bonded carrier that releases the active parent drug when administered to a mammalian subject. Prodrugs can be modified by modifying functional groups present in the compound in such a way that the modification is cleaved to the parent compound during routine manipulation or in vivo.

本发明的溶剂化物是指一个或多个溶剂分子与本发明形成的缔合物，形成溶剂化物的溶剂包括但不限于水、异丙醇、乙醇、甲醇、二甲亚砜等。The solvate of the present invention refers to the association formed by one or more solvent molecules and the present invention, and the solvent that forms the solvate includes but is not limited to water, isopropanol, ethanol, methanol, dimethyl sulfoxide and the like.

本发明的异构体包括立体异构、互变异构体。Isomers in the present invention include stereoisomers and tautomers.

此外，本发明的化合物包括其晶体和非晶体形式，其中晶体包括单晶、多晶和共晶。In addition, the compounds of the present invention include crystalline and amorphous forms thereof, wherein crystals include single crystals, polycrystals and co-crystals.

优选地，新型氘代氰基类化合物或其异构体或其前药或其溶剂化物或其药学上可接受的盐、与利托那韦或其异构体或其前药或其溶剂化物或其药学上可接受的盐的质量比为0.5-10：1。优选地，它们的质量比为0.5-6：1，优选地，新型氘代氰基类化合物与利托那韦的质量比为10：1、9：1、8：1、7：1、6：1、5：1、4：1、3：1、2.5：1、2：1、1：1、0.5：1。Preferably, the novel deuterated cyano compound or its isomer or its prodrug or its solvate or its pharmaceutically acceptable salt, and ritonavir or its isomer or its prodrug or its solvate The mass ratio of its pharmaceutically acceptable salt is 0.5-10:1. Preferably, their mass ratio is 0.5-6:1, preferably, the mass ratio of the novel deuterated cyano compound to ritonavir is 10:1, 9:1, 8:1, 7:1, 6 : 1, 5:1, 4:1, 3:1, 2.5:1, 2:1, 1:1, 0.5:1.

优选地，本文公开的药物组合物包含约100mg至约1000mg新型氘代氰基类化合物和25mg-100mg利托那韦。优选地，药物组合物包含约400mg新型氘代氰基类化合物和50mg-100mg利托那韦，优选地，药物组合物包含约300mg新型氘代氰基类化合物和100mg利托那韦。优选地，药物组合物包含约300mg新型氘代氰基类化合物和80mg利托那韦。优选地，药物组合物包含约300mg新型氘代氰基类化合物和60mg利托那韦。优选地，药物组合物包含约300mg新型氘代氰基类化合物和50mg利托那韦。Preferably, the pharmaceutical compositions disclosed herein comprise from about 100 mg to about 1000 mg of the novel deuterated cyano compound and 25 mg to 100 mg of ritonavir. Preferably, the pharmaceutical composition comprises about 400 mg of the novel deuterated cyano compound and 50 mg to 100 mg of ritonavir, preferably, the pharmaceutical composition comprises about 300 mg of the novel deuterated cyano compound and 100 mg of ritonavir. Preferably, the pharmaceutical composition comprises about 300 mg of the novel deuterated cyano compound and 80 mg of ritonavir. Preferably, the pharmaceutical composition comprises about 300 mg of the novel deuterated cyano compound and 60 mg of ritonavir. Preferably, the pharmaceutical composition comprises about 300 mg of the novel deuterated cyano compound and 50 mg of ritonavir.

应当理解，本发明药物组合物可以在相同或不同药物组合物中同时给药，或者可按顺序给药，也即新型氘代氰基类化合物与利托那韦可以混合在一起形成单一的给药单元，也可以分别独立形成给药单元。如果是独立给药，给第二种活性组分的间隔时间不应当使活性组分联合所能产生的协同疗效的益处失去。优选新型氘代氰基类化合物与利托那韦分别独立形成给药单元。It should be understood that the pharmaceutical compositions of the present invention may be administered simultaneously in the same or different pharmaceutical compositions, or may be administered sequentially, that is, the novel deuterated cyano compounds and ritonavir may be mixed together to form a single administration The drug unit can also be independently formed into a dosing unit. If administered independently, the interval between administration of the second active ingredient should not be such that the benefit of the synergistic effect produced by the combination of the active ingredients would be lost. Preferably, the novel deuterated cyano compound and ritonavir form a drug delivery unit independently.

所述组合物还包括药学上可接受的载体，所述载体由被认为是安全且有效的材料组成，并且可以对个体施用而不会引起不良的生物学副作用或不期望的相互作用。载体是药物制剂中存在的除活性成分以外的所有成分。如本文通常所使用的，“载体”包括但不限于调节剂、黏合剂、润滑剂、崩解剂、填充剂、着色剂、增塑剂、膜衣材料、有机溶剂、增溶剂、调味剂、表面活性剂等。The composition also includes a pharmaceutically acceptable carrier composed of materials believed to be safe and effective, and which can be administered to an individual without causing adverse biological side effects or undesired interactions. A carrier is all ingredients other than the active ingredient present in the pharmaceutical formulation. As generally used herein, "carrier" includes, but is not limited to, modifiers, binders, lubricants, disintegrants, fillers, colorants, plasticizers, film coating materials, organic solvents, solubilizers, flavoring agents, Surfactants, etc.

稀释剂，也被称为“填充剂”，其通常是增加固体剂型的体积从而提供用于压制片剂或形成珠子和颗粒的实用大小所必需的。合适的稀释剂包括但不限于磷酸二钙二水合物、硫酸钙、乳糖、蔗糖、甘露醇、山梨糖醇、纤维素、微晶纤维素、高岭土、氯化钠、干淀粉、水解淀粉、预胶化淀粉、二氧化硅、氧化钛、硅酸铝镁和糖粉。Diluents, also known as "fillers", are often necessary to increase the bulk of a solid dosage form to provide a practical size for compressing tablets or forming beads and granules. Suitable diluents include, but are not limited to, dicalcium phosphate dihydrate, calcium sulfate, lactose, sucrose, mannitol, sorbitol, cellulose, microcrystalline cellulose, kaolin, sodium chloride, dry starch, hydrolyzed starch, pre- Gelatinized starch, silicon dioxide, titanium oxide, magnesium aluminum silicate and powdered sugar.

黏合剂被用于赋予固体剂型黏结性，从而确保剂型形成后片剂或珠子或颗粒保持完整。合适的黏合剂材料包括但不限于淀粉、预胶化淀粉、明胶、糖类(包括蔗糖、葡萄糖、右旋糖、乳糖和山梨糖醇)、聚乙二醇、蜡、天然树胶和合成树胶。Binders are used to impart cohesiveness to solid dosage forms, thereby ensuring that tablets or beads or granules remain intact after the dosage form has been formed. Suitable binder materials include, but are not limited to, starch, pregelatinized starch, gelatin, sugars (including sucrose, glucose, dextrose, lactose, and sorbitol), polyethylene glycols, waxes, natural and synthetic gums.

润滑剂用于促进片剂的制造。合适的润滑剂的实例包括但不限于硬脂酸镁、硬脂酸钙、硬脂酸、山嵛酸甘油酯、聚乙二醇、滑石粉和矿物油。Lubricants are used to facilitate the manufacture of tablets. Examples of suitable lubricants include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, glyceryl behenate, polyethylene glycol, talc, and mineral oil.

崩解剂用于在施用后促进剂型崩解或“分解”，并且通常包括但不限于淀粉、淀粉乙醇酸钠、羧甲基淀粉钠、羧甲基纤维素钠、羟丙基纤维素、预胶化淀粉、黏土、纤维素、海藻酸盐、胶或交联的聚合物。Disintegrants are used to facilitate disintegration or "breakdown" of the dosage form after administration and typically include, but are not limited to, starch, sodium starch glycolate, sodium carboxymethyl starch, sodium carboxymethyl cellulose, hydroxypropyl cellulose, pre- Gelatinized starch, clay, cellulose, alginate, gum or cross-linked polymer.

稳定剂用于抑制或延缓药物分解反应，例如包括氧化反应。Stabilizers are used to inhibit or delay drug decomposition reactions, including, for example, oxidation reactions.

表面活性剂可以是阴离子表面活性剂、阳离子表面活性剂、两性表面活性剂或非离子表面活性剂。合适的阴离子表面活性剂包括但不限于含有羧酸根、磺酸根和硫酸根离子的那些。阴离子表面活性剂的实例包括长链烷基磺酸和烷基芳基磺酸的钠盐、钾盐、铵盐，例如十二烷基苯磺酸钠；二烷基磺基琥珀酸钠。The surfactant can be an anionic surfactant, a cationic surfactant, an amphoteric surfactant or a nonionic surfactant. Suitable anionic surfactants include, but are not limited to, those containing carboxylate, sulfonate, and sulfate ions. Examples of anionic surfactants include sodium, potassium, ammonium salts of long chain alkyl sulfonic acids and alkyl aryl sulfonic acids, eg, sodium dodecylbenzenesulfonate; sodium dialkylsulfosuccinate.

典型的增塑剂的实例包括聚乙二醇、丙二醇、三醋精、邻苯二甲酸二甲酯、邻苯二甲酸二乙酯、邻苯二甲酸二丁酯、癸二酸二丁酯、柠檬酸三乙酯、柠檬酸三丁酯、乙酰柠檬酸三乙酯、蓖麻油和乙酰化的单甘油酯。Examples of typical plasticizers include polyethylene glycol, propylene glycol, triacetin, dimethyl phthalate, diethyl phthalate, dibutyl phthalate, dibutyl sebacate, Triethyl citrate, tributyl citrate, acetyl triethyl citrate, castor oil, and acetylated monoglycerides.

有机溶剂可以为醇类，如异丙醇，丙二醇，聚乙二醇，聚丙烯乙二醇，甘油和聚氧乙烯醇等。The organic solvent can be alcohols, such as isopropanol, propylene glycol, polyethylene glycol, polypropylene glycol, glycerol and polyoxyethylene alcohol.

可以配制本文所述的组合物以用于调节释放或控制释放。控释剂型的实例包括延长释放剂型、延迟释放剂型、脉冲释放剂型及其组合。The compositions described herein can be formulated for modified release or controlled release. Examples of controlled release dosage forms include extended release dosage forms, delayed release dosage forms, pulsed release dosage forms, and combinations thereof.

优选地，化合物或药物组合物通过口服施用、静脉内施用，口服包括片剂或者口服液。Preferably, the compound or pharmaceutical composition is administered orally, intravenously, including tablets or oral solutions.

另一方面，本发明提供一种药物组合物在制备用于治疗或预防病毒感染的药物中的用途，所述病毒可以为新冠病毒SARS-CoV-2、SARS冠状病毒、MERS冠状病毒、甲型流感病毒和乙型流感病毒、东部马脑炎病毒、西部马脑炎病毒、委内瑞拉马脑炎病毒、奇昆古尼亚热病毒、正黏病毒科病毒或副黏病毒科病毒、RSV病毒、埃博拉病毒。优选地，所述病毒为新冠病毒SARS-CoV-2、SARS冠状病毒和MERS冠状病毒。优选地，所述病毒为新冠病毒SARS-CoV-2。On the other hand, the present invention provides the use of a pharmaceutical composition in the preparation of a medicine for the treatment or prevention of viral infection, and the virus can be the new coronavirus SARS-CoV-2, SARS coronavirus, MERS coronavirus, type A Influenza virus and influenza B virus, Eastern equine encephalitis virus, Western equine encephalitis virus, Venezuelan equine encephalitis virus, Chikungunya virus, Orthomyxoviridae or Paramyxoviridae, RSV virus, Egypt bola virus. Preferably, the virus is the new coronavirus SARS-CoV-2, SARS coronavirus and MERS coronavirus. Preferably, the virus is the new coronavirus SARS-CoV-2.

本发明的药物组合物具有以下优点和有益效果：The pharmaceutical composition of the present invention has the following advantages and beneficial effects:

(1)新型氘代氰基类化合物

是由上海谷森医药有限公司独自研发的一种小分子3CL蛋白酶抑制剂，其具有惊人的抗SARS-CoV-2活性，更让人意外的是，该化合物在病毒抑菌活性相当的基础上，实现了比其他口服抗新冠药更优的药动学特征，其抗新冠病毒活性数据在CN202111234708X公开。(1) Novel deuterated cyano compounds

It is a small molecule 3CL protease inhibitor independently developed by Shanghai Gusen Pharmaceutical Co., Ltd. It has amazing anti-SARS-CoV-2 activity. What is even more surprising is that this compound has comparable antibacterial activity to the virus. It has achieved better pharmacokinetic characteristics than other oral anti-new crown drugs, and its anti-new crown virus activity data is disclosed in CN202111234708X.

(2)本发明的药物组合物相较于其他现有的抗病毒药物组合物具有更优的药动学特征，其半衰期更长，在体内能保持较高的浓度，具有安全性高、降低药物毒副作用、降低病毒耐药性等优点。(2) Compared with other existing antiviral pharmaceutical compositions, the pharmaceutical composition of the present invention has better pharmacokinetic characteristics, has a longer half-life, can maintain a higher concentration in the body, has high safety, reduces The advantages of drug toxicity and side effects, reduction of virus resistance, etc.

具体实施方式：Detailed ways:

以下结合具体实施例对本发明做进一步详细的说明。The present invention will be described in further detail below with reference to specific embodiments.

以下详细的说明都仅是示例性和解释性的，而非限制性的。The following detailed description is exemplary and explanatory only and not restrictive.

以下实施例，除非另外指出，否则使用的所有溶剂和试剂都是商购得到并且以原样使用。In the following examples, unless otherwise indicated, all solvents and reagents used were obtained commercially and were used as received.

实施例1：新型氘代氰基类化合物(以下简称化合物1)的合成路线Embodiment 1: the synthetic route of novel deuterated cyano compound (hereinafter referred to as compound 1)

1)中间体H、K、M的制备方法可参见在先申请CN202111234708X。1) The preparation methods of intermediates H, K and M can be found in the previous application CN202111234708X.

2)新型氘代氰基类化合物(以下简称化合物1)的合成2) Synthesis of novel deuterated cyano compounds (hereinafter referred to as compound 1)

在室温下，将M(8g，21.1mmol)溶于THF(80ml)和MeOH(80ml)中。加入氢氧化锂(1.5g，62.8mmol)的水溶液(15ml)，室温搅拌2小时。调温至0-10℃，加入乙酸乙酯后使用1NHCl调反应液至酸性，分层，水相使用EA萃取两次，合并有机相后无水硫酸镁干燥，浓缩至干得到粗品，柱层析纯化得到M水解化合物。M (8 g, 21.1 mmol) was dissolved in THF (80 ml) and MeOH (80 ml) at room temperature. An aqueous solution (15 ml) of lithium hydroxide (1.5 g, 62.8 mmol) was added, and the mixture was stirred at room temperature for 2 hours. Adjust the temperature to 0-10°C, add ethyl acetate, adjust the reaction solution to acidity with 1N HCl, separate the layers, extract the aqueous phase twice with EA, combine the organic phases, dry over anhydrous magnesium sulfate, and concentrate to dryness to obtain the crude product. Analytical purification gave M hydrolyzed compound.

将上一步所得的水解化合物和DMF(100ml)投入反应瓶中，调温至0-10℃。在此温度下，依次加入EDCI(4.9g，25.6mmol)，HOBt(3.4g，25.2mmol)，NMM(4.3g，42.5mmol)，并在此温度下搅拌30分钟。在此温度下，分批加入H(4.0g，21mmol)，加完后在室温下搅拌过夜。TLC检测反应转化基本完全，加入水(80ml)后用EA萃取三次，合并有机相，依次使用0.5mol/L HCl，5％NaHCO3，水洗涤，无水硫酸镁干燥，浓缩至干得到粗品1，柱层析得到类白色固体产物8.4g，即新型氘代氰基类化合物(以下简称化合物1)，收率79.7％。The hydrolyzed compound obtained in the previous step and DMF (100 ml) were put into a reaction flask, and the temperature was adjusted to 0-10°C. At this temperature, EDCI (4.9 g, 25.6 mmol), HOBt (3.4 g, 25.2 mmol), NMM (4.3 g, 42.5 mmol) were added sequentially and stirred at this temperature for 30 minutes. At this temperature, H (4.0 g, 21 mmol) was added in portions and stirred overnight at room temperature after the addition was complete. TLC detected that the reaction conversion was basically complete. After adding water (80ml), it was extracted three times with EA. The organic phases were combined, and 0.5mol/L HCl, 5% NaHCO was used successively, washed with water, dried over anhydrous magnesium sulfate, and concentrated to dryness to obtain crude product 1, Column chromatography obtained 8.4 g of an off-white solid product, namely a novel deuterated cyano compound (hereinafter referred to as compound 1), with a yield of 79.7%.

LC-MS(ESI，m/z，C23H30D2F3N5O4，502.2，M+H)LC-MS (ESI, m/z, C23H30D2F3N5O4, 502.2, M+H)

1H NMR(400MHz,DMSO)δ:8.32(s,1H),8.18(s,1H),7.79(s,1H),4.50-4.24(m,3H),3.52-3.27(m,2H),2.2-1.9(m,5H),1.27(m,1H),0.97-0.89(m,16H)。1H NMR (400MHz, DMSO)δ: 8.32(s,1H), 8.18(s,1H), 7.79(s,1H), 4.50-4.24(m,3H), 3.52-3.27(m,2H), 2.2- 1.9 (m, 5H), 1.27 (m, 1H), 0.97-0.89 (m, 16H).

实施例2：新型氘代氰基类化合物的体外抗SARS-CoV-2病毒实验Example 2: In vitro anti-SARS-CoV-2 virus experiments of novel deuterated cyano compounds

细胞铺板:将Vero E6细胞以3×10⁵个细胞/孔接种到12孔板中，加入10％FBS的DMEM培养基，并置于37℃、5％CO₂培养箱培养过夜。药物作用：去除12孔板中Vero细胞的培养基，用PBS缓冲液清洗VeroE6细胞；化合物(终浓度100nM)加到50μL/孔细胞液中，置于37℃、5％CO₂培养箱中孵育1小时，设置50μL/孔培养基作为对照。病毒感染细胞：用SARS-CoV-2病毒感染细胞2小时，去除感染的病毒、药物混合液,加入10％FBS的DMEM培养基，置于37℃、5％CO₂培养箱中培养2～3天。PCR测定：收集培养基的上清液，置于56℃培养箱保留30min，用病毒RNA提取试剂盒抽提病毒RNA，用病毒核酸检测试剂盒按照说明书进行PCR反应，通过PCR仪器显示的CT值计算2^-ΔCT值病毒复制抑制率的计算公式为:(1-2^-ΔCT)×100％，其中，2^-ΔCT值为药物组与对照组(单宁酸)的相对病毒复制率，结果见表1。Cell plating: Vero E6 cells were seeded into 12-well plates at 3×10⁵ cells/well, added with 10% FBS in DMEM medium, and placed in a 37°C, 5% CO₂ incubator overnight. Drug action: Remove the medium of Vero cells in the 12-well plate, wash VeroE6 cells with PBS buffer; add the compound (final concentration 100nM) to 50μL/well of cell fluid and incubate in a 37°C, 5%_CO2 incubator For 1 hour, set 50 μL/well medium as a control. Virus-infected cells: Infect cells with SARS-CoV-2 virus for 2 hours, remove the infected virus and drug mixture, add 10% FBS DMEM medium, and place in a 37°C, 5% CO₂ incubator for 2-3 days sky. PCR assay: collect the supernatant of the culture medium, place it in a 56°C incubator for 30 minutes, extract viral RNA with a viral RNA extraction kit, perform PCR reaction with a viral nucleic acid detection kit according to the instructions, and use the PCR instrument to display the CT value. The formula for calculating the 2^-ΔCT value of the virus replication inhibition rate is: (1-2^-ΔCT )×100%, where the 2^-ΔCT value is the relative virus replication rate between the drug group and the control group (tannic acid). The results are shown in Table 1.

由表1可知，实施例1合成得到的新型氘代氰基类化合物(以下简称化合物1)具有很好的体外抗SARS-CoV-2病毒活性。It can be seen from Table 1 that the novel deuterated cyano compound (hereinafter referred to as compound 1) synthesized in Example 1 has good anti-SARS-CoV-2 virus activity in vitro.

表1药物组与对照组病毒复制率结果Table 1 Results of virus replication rate between drug group and control group

实施例3：大鼠药代动力学研究Example 3: Rat Pharmacokinetic Study

1)试验方法：1) Test method:

对实施例1所述新型氘代氰基类化合物(简称化合物1)+RTV(利托那韦)、新型氘代氰基类化合物(简称化合物1)、PF-07321332(辉瑞研制)+RTV(利托那韦)的药代动力学进行研究。For the new deuterated cyano compounds described in Example 1 (referred to as compound 1) + RTV (ritonavir), new deuterated cyano compounds (referred to as compound 1), PF-07321332 (developed by Pfizer) + RTV ( The pharmacokinetics of ritonavir) were studied.

在7-10周龄，雄性Wistar-Hannover大鼠进行药代动力学研究。在药代动力学研究期间，所有动物都单独饲养。自由摄取食物和水(动物在进食状态下给药)。动物禁食过夜并在给药后4小时进食。在灌胃给药(分别为化合物1+RTV(利托那韦)、化合物1、PF-07321332(辉瑞研制)+RTV(利托那韦))后的预定时间点(0、6、12、18、24小时)通过颈静脉插管收集血样。在研究完成时，通过过量吸入麻醉随后放血对动物实施安乐死。将血样收集到含有K₂EDTA的试管中并储存在冰上，直至离心获得血浆，将其储存在-20℃冰箱。Pharmacokinetic studies were performed in male Wistar-Hannover rats at 7-10 weeks of age. All animals were housed individually during the pharmacokinetic study. Food and water were given ad libitum (animals were dosed in a fed state). Animals were fasted overnight and fed 4 hours after dosing. Predetermined time points (0, 6, 12, 18, 24 hours) blood samples were collected via jugular vein cannula. At study completion, animals were euthanized by overdose inhalation anesthesia followed by exsanguination. Blood samples were collected into tubes containing_K2EDTA and stored on ice until centrifugation to obtain plasma, which was stored in a -20°C freezer.

血浆样品的LC-MS/MS分析：血浆样品使用蛋白质沉淀法进行处理，使用含有500:50的乙腈:甲醇，含有propranolol(50ng/ml)作为内标，然后根据空白血浆中制备的标准曲线(0.1-2500ng/ml)进行定量。使用LC-MS/MS对血浆样品中的分析物进行定量。简而言之，使用与配备Sciex 6500三重四极杆质谱仪联用的Waters ACQUITY超高效液相色谱系统。使用Waters Acquity UPLC BEH C18柱(1.7m，2.1 50mm)完成色谱分离。优化流动相以实现分析物之间的良好分离。通常，溶剂A由0.025％甲酸和1mM乙酸铵的水/乙腈溶液(95:5v/v)组成，溶剂B包括0.025％甲酸和1mM乙酸铵的水/乙腈溶液(5:95v/v)。梯度一般从3-30％B开始，直到大约1.2分钟，然后增加到50-65％B到1.6分钟，然后降低到10-30％B，直到约1.7-1.9分钟。Analyst 1.7软件用于峰积分和标准曲线回归。LC-MS/MS analysis of plasma samples: Plasma samples were processed using protein precipitation using 500:50 acetonitrile:methanol containing propranolol (50 ng/ml) as internal standard and then based on a standard curve prepared in blank plasma ( 0.1-2500ng/ml) for quantification. Analytes in plasma samples were quantified using LC-MS/MS. Briefly, a Waters ACQUITY ultra-performance liquid chromatography system coupled to a Sciex 6500 triple quadrupole mass spectrometer was used. Chromatography was accomplished using a Waters Acquity UPLC BEH C18 column (1.7m, 2.1 50mm). Optimize mobile phases for good separation between analytes. Typically, solvent A consists of 0.025% formic acid and 1 mM ammonium acetate in water/acetonitrile (95:5 v/v) and solvent B consists of 0.025% formic acid and 1 mM ammonium acetate in water/acetonitrile (5:95 v/v). The gradient typically starts at 3-30% B until about 1.2 minutes, then increases to 50-65% B for about 1.6 minutes, then decreases to 10-30% B until about 1.7-1.9 minutes. Analyst 1.7 software was used for peak integration and standard curve regression.

药代动力学分析：使用非房室分析(Watson v.7.5，Thermo Scientific)计算药代动力学参数。使用线性梯形法则估计从t＝0到无穷大(AUC_0-∞)的血浆浓度-时间曲线下面积。在某些情况下，药代动力学计算是使用线性对数线性梯形规则生成的，C₀是使用公式计算：

Pharmacokinetic analysis: Pharmacokinetic parameters were calculated using non-compartmental analysis (Watson v. 7.5, Thermo Scientific). The area under the plasma concentration-time curve from t=0 to infinity (AUC_0-∞ ) was estimated using the linear trapezoidal rule. In some cases, pharmacokinetic calculations were generated using the linear-log-linear trapezoidal rule, and_C0 was calculated using the formula:

其中Vb是血容量(大鼠，69.0ml/min/kg)，BPR是血液与血浆的比率。终末速率常数(kel)通过对数-线性浓度-时间曲线终末期的线性回归计算，终末消除t_1/2计算如下：

where Vb is blood volume (rat, 69.0 ml/min/kg) and BPR is the blood to plasma ratio. The terminal rate constant (kel) was calculated by linear regression of the terminal phase of the log-linear concentration-time curve, and the terminal elimination t_1/2 was calculated as follows:

1)试验结果：1) Test results:

化合物1+RTV(利托那韦)、化合物1、PF-07321332(辉瑞研制)+RTV(利托那韦)的药代动力学结果见表2。The pharmacokinetic results of compound 1+RTV (ritonavir), compound 1, PF-07321332 (developed by Pfizer)+RTV (ritonavir) are shown in Table 2.

由表2可知，一方面利托那韦可以减缓新型氘代氰基类化合物的代谢或分解，改善药物的药动学特征。It can be seen from Table 2 that, on the one hand, ritonavir can slow down the metabolism or decomposition of new deuterated cyano compounds and improve the pharmacokinetic characteristics of the drug.

另一方面，本发明的药物组合物相较于其他现有的抗病毒药物组合物(如辉瑞)具有更优的药动学特征，其半衰期更长，在体内能保持较高的浓度，具有安全性高、降低药物毒副作用、降低病毒耐药性等优点。On the other hand, compared with other existing antiviral pharmaceutical compositions (such as Pfizer), the pharmaceutical composition of the present invention has better pharmacokinetic characteristics, has a longer half-life, can maintain a higher concentration in the body, and has It has the advantages of high safety, reduced drug toxicity and side effects, and reduced viral resistance.

表2：化合物1+RTV(利托那韦)、化合物1、PF-07321332(辉瑞研制)+RTV的药代动力学研究Table 2: Pharmacokinetic study of compound 1+RTV (ritonavir), compound 1, PF-07321332 (developed by Pfizer)+RTV

以上所述实施例仅表达了本发明的几种实施方式，其描述较为具体和详细，但并不能因此而理解为对发明专利范围的限制。应当指出的是，对于本领域的普通技术人员来说，在不脱离本发明构思的前提下，还可以做出若干变形和改进，这些都属于本发明的保护范围。因此，本发明专利的保护范围应以所附权利要求为准。The above-mentioned embodiments only represent several embodiments of the present invention, and the descriptions thereof are specific and detailed, but should not be construed as a limitation on the scope of the invention patent. It should be pointed out that for those of ordinary skill in the art, without departing from the concept of the present invention, several modifications and improvements can also be made, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention should be subject to the appended claims.

Claims (10)

Translated fromChinese

1.一种药物组合物，其特征在于：由新型氘代氰基类化合物或其异构体或其前药或其溶剂化物或其药学上可接受的盐、以及利托那韦或其异构体或其前药或其溶剂化物或其药学上可接受的盐组成；1. a pharmaceutical composition is characterized in that: by novel deuterated cyano compound or its isomer or its prodrug or its solvate or its pharmaceutically acceptable salt and ritonavir or its isoform Construct or its prodrug or its solvate or its pharmaceutically acceptable salt composition;其中，所述新型氘代氰基类化合物的结构式为：Wherein, the structural formula of described novel deuterated cyano compound is:

所述利托那韦的结构式为：The structural formula of ritonavir is:

2.根据权利要求1所述的药物组合物，其特征在于：新型氘代氰基类化合物或其异构体或其前药或其溶剂化物或其药学上可接受的盐、与利托那韦或其异构体或其前药或其溶剂化物或其药学上可接受的盐的质量比为0.5-10:1。2. pharmaceutical composition according to claim 1 is characterized in that: novel deuterated cyano compound or its isomer or its prodrug or its solvate or its pharmaceutically acceptable salt, and ritona The mass ratio of Wei or its isomer or its prodrug or its solvate or its pharmaceutically acceptable salt is 0.5-10:1.3.根据权利要求2所述的药物组合物，其特征在于：新型氘代氰基类化合物或其异构体或其前药或其溶剂化物或其药学上可接受的盐、与利托那韦或其异构体或其前药或其溶剂化物或其药学上可接受的盐的质量比为0.5-6:1。3. pharmaceutical composition according to claim 2 is characterized in that: novel deuterated cyano compound or its isomer or its prodrug or its solvate or its pharmaceutically acceptable salt, and ritona The mass ratio of Wei or its isomer or its prodrug or its solvate or its pharmaceutically acceptable salt is 0.5-6:1.4.根据权利要求1-3任一项所述的药物组合物，其特征在于：还包括一种或多种药学上可接受的载体。4. The pharmaceutical composition according to any one of claims 1-3, further comprising one or more pharmaceutically acceptable carriers.5.根据权利要求4所述的药物组合物，其特征在于：药学上可接受的载体包括调节剂、黏合剂、润滑剂、崩解剂、填充剂、着色剂、增塑剂、膜衣材料、有机溶剂、增溶剂、调味剂。5. The pharmaceutical composition according to claim 4, wherein the pharmaceutically acceptable carrier comprises a regulator, a binding agent, a lubricant, a disintegrating agent, a filler, a coloring agent, a plasticizer, and a film coating material , organic solvent, solubilizer, flavoring agent.6.根据权利要求1-5任一项所述的药物组合物，其特征在于：新型氘代氰基类化合物或其异构体或其前药或其溶剂化物或其药学上可接受的盐、与利托那韦或其异构体或其前药或其溶剂化物或其药学上可接受的盐可以混合在一起形成单一的给药单元，也可以分别独立形成给药单元。6. The pharmaceutical composition according to any one of claims 1-5, characterized in that: a novel deuterated cyano compound or its isomer or its prodrug or its solvate or its pharmaceutically acceptable salt , and ritonavir or its isomers or its prodrugs or its solvates or its pharmaceutically acceptable salts can be mixed together to form a single administration unit, or can be independently formed into a single administration unit.7.根据权利要求6所述的药物组合物，其特征在于：新型氘代氰基类化合物或其异构体或其前药或其溶剂化物或其药学上可接受的盐、与利托那韦或其异构体或其前药或其溶剂化物或其药学上可接受的盐分别独立形成给药单元。7. pharmaceutical composition according to claim 6 is characterized in that: novel deuterated cyano compound or its isomer or its prodrug or its solvate or its pharmaceutically acceptable salt, and ritona Wei or its isomer or its prodrug or its solvate or its pharmaceutically acceptable salt, respectively, independently form an administration unit.8.权利要求1-7任一项所述药物组合物在制备抗新冠病毒SARS-CoV-2、SARS冠状病毒、MERS冠状病毒、甲型流感病毒和乙型流感病毒、东部马脑炎病毒、西部马脑炎病毒、委内瑞拉马脑炎病毒、奇昆古尼亚热病毒、正黏病毒科病毒或副黏病毒科病毒、RSV病毒、埃博拉病毒的药物中的用途。8. The pharmaceutical composition described in any one of claims 1-7 is in the preparation of anti-new coronavirus SARS-CoV-2, SARS coronavirus, MERS coronavirus, influenza A virus and influenza B virus, eastern equine encephalitis virus, Use in medicine for western equine encephalitis virus, Venezuelan equine encephalitis virus, Chikungunya virus, Orthomyxoviridae or Paramyxoviridae, RSV virus, Ebola virus.9.根据权利要求8所述的用途，其特征在于：病毒优选为新冠病毒SARS-CoV-2、SARS冠状病毒和MERS冠状病毒。9. purposes according to claim 8 is characterized in that: virus is preferably new coronavirus SARS-CoV-2, SARS coronavirus and MERS coronavirus.10.根据权利要求9所述的用途，其特征在于：病毒优选为新冠病毒SARS-CoV-2。10. The use according to claim 9, wherein the virus is preferably the new coronavirus SARS-CoV-2.