TW202140009A - 3-carboxamido-4-hydroxynaltrexone deuterated derivative, preparation method therefor and application thereof - Google Patents

Abstract

The present invention relates to a 3-carboxamido-4-hydroxynaltrexone deuterated derivative, a preparation method therefor and the application thereof in medicine. In particular, the present invention relates to a 3-carboxamido-4-hydroxynaltrexone deuterated derivative represented by the general formula (I), a preparation method thereof, and a pharmaceutical composition containing the derivative, and as an opioid receptor modulator in the treatment of cough, diarrhea, drug dependence, gastrointestinal diseases, irritable bowel syndrome, obesity, respiratory depression, convulsions, pain, allergies, pruritus and other diseases, as well as central nervous diseases or/ and mental illness and other related diseases. Wherein, each substituent in the general formula (I) has the same definition as in the specification.

Description

3-甲醯胺基-4-羥基納曲酮氘代衍生物、其製備方法及其在醫藥上的應用 3-methamido-4-hydroxynaltrexone deuterated derivative, its preparation method and its application in medicine

本發明屬於醫藥領域，具體涉及3-甲醯胺基-4-羥基納曲酮氘代衍生物、其製備方法及含有該衍生物的醫藥組成物在醫藥研究上的應用。 The invention belongs to the field of medicine, and specifically relates to a deuterated 3-methamido-4-hydroxynaltrexone derivative, a preparation method thereof, and the application of a pharmaceutical composition containing the derivative in medical research.

精神***症是患病率最高的一種精神病，病程緩慢，容易反復發作、加重或惡化，對患者及患者家屬所造成了嚴重的疾病負擔和不良後果。精神病患者會出現妄想，幻覺，思想語言和行為紊亂等陽性症狀，無情緒和表情，言語貧乏，缺乏愉悅感等陰性症狀以及認知障礙等症狀。雖然在過去的幾十年中，抗精神***症藥物的研發和臨床應用有了很大發展，但傳統抗精神病藥(一代)(氟哌啶醇、氟哌利多、甲硫噠嗪等)及非典型抗精神病藥(二代)(氯氮平、利培酮、奧氮平、阿立哌唑等)都存在各種各樣的安全性問題。奧氮平是第二代精神病藥物的典型代表，它具有突出的抗精神***症陽性症狀藥效，臨床應用廣泛，但同時也具有較嚴重的體重增加的臨床副作用。開發一個藥物保持奧氮平藥效同時降低其體重增加副作用仍是臨床急需解決的問題。 Schizophrenia is a kind of mental illness with the highest prevalence. It has a slow course and is prone to recurring, aggravating or worsening. It causes a serious burden of disease and adverse consequences for patients and their families. Psychiatric patients will have positive symptoms such as delusions, hallucinations, thoughts, language and behavior disorders, lack of emotion and expression, poor speech, lack of pleasure and other negative symptoms, as well as symptoms such as cognitive impairment. Although in the past few decades, the development and clinical application of anti-schizophrenia drugs have been greatly developed, traditional antipsychotics (first generation) (haloperidol, droperidol, thioridazine, etc.) and Atypical antipsychotics (second generation) (clozapine, risperidone, olanzapine, aripiprazole, etc.) have various safety issues. Olanzapine is a typical representative of the second generation of psychiatric drugs. It has outstanding anti-schizophrenia-positive symptoms and has a wide range of clinical applications, but it also has serious clinical side effects of weight gain. The development of a drug to maintain the efficacy of olanzapine while reducing the side effects of weight gain is still an urgent clinical problem.

Samidorphan也叫ALKS-33，是阿片受體調節劑，主要作用於μ阿片受體，目前Alkermers公司正在開發其用於重度抑鬱和其它精神領域疾病的治療。Samidorphan主要是μ阿片受體拮抗劑，對μ阿片受體抑制IC₅₀為0.88nM，同時Samidorphan還具有κ和δ阿片受體的部分激動和拮抗活性。 Samidorphan, also called ALKS-33, is an opioid receptor modulator, mainly acting on mu opioid receptors. Alkermers is currently developing its treatment for major depression and other mental illnesses. Samidorphan mainly μ opioid receptor antagonist, for the μ opioid receptor inhibitory IC ₅₀ of 0.88 nm, while also having a portion Samidorphan δ and κ opioid receptor agonist and antagonist activity.

本發明的目的在於提供一種通式(I)所示的化合物、其立體異構體或其藥學上可接受鹽結構如下： The purpose of the present invention is to provide a compound represented by the general formula (I), its stereoisomer or its pharmaceutically acceptable salt structure is as follows:

其中： in:

R₁、R₂、R₃和R₄各自獨立地選自氫、氘或烷基； R ₁ , R ₂ , R ₃ and R ₄ are each independently selected from hydrogen, deuterium or alkyl;

且，R₁、R₂、R₃或R₄中至少有一個為氘。 In addition, at least one of _{R 1} , R ₂ , R ₃ or R _{4 is deuterium.}

在本發明還提供了一種較佳方案，該化合物、其立體異構體或其藥學上可接受鹽，其具體結構如通式(II)所示： The present invention also provides a preferred solution. The specific structure of the compound, its stereoisomer or its pharmaceutically acceptable salt is as shown in the general formula (II):

本發明還提供了一種較佳方案，任一項所述的化合物、其立體異構體或其藥學上可接受鹽，其中，選自如下化合物： The present invention also provides a preferred solution. The compound, stereoisomer or pharmaceutically acceptable salt thereof according to any one of them is selected from the following compounds:

本發明的另一方面涉及一種醫藥組成物，其含有治療有效劑量的各通式所示的化合物、其立體異構體或其藥學上可接受鹽，以及一種或多種藥學上可接受的載體、稀釋劑或賦形劑。 Another aspect of the present invention relates to a pharmaceutical composition, which contains a therapeutically effective dose of a compound represented by each general formula, its stereoisomer or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, Diluent or excipient.

本發明還涉及一種製備上述組成物的方法，其包括將各通式所示的化合物、其立體異構體或其藥學上可接受鹽與藥學上可接受的載體、稀釋劑或賦形劑相混合。 The present invention also relates to a method for preparing the above-mentioned composition, which comprises combining the compound represented by each general formula, its stereoisomer or its pharmaceutically acceptable salt with a pharmaceutically acceptable carrier, diluent or excipient mix.

本發明進一步涉及通式(I)所示的化合物、其立體異構體或其藥學上可接受鹽，或包含其的醫藥組成物在製備用於預防和/或治療預防作為阿片受體調節劑在治療咳嗽、腹瀉、藥物依賴性、胃腸道疾病、過敏性腸綜合症、肥胖、呼吸抑制、驚厥、疼痛、過敏、瘙癢等疾病和中樞神經類疾病或/和精神類疾病等相關疾病藥物中的用途。阿片受體調節劑可以用於咳嗽、腹瀉、藥物依賴性、胃腸道疾病、過敏性腸綜合症、肥胖、呼吸抑制、驚厥、疼痛、過敏、瘙癢等疾病和中樞神經類疾病或/和精神類疾病等相關疾病，該中樞神經疾病或/和精神類疾病選自：精神***症、睡眠障礙、心境障礙、精神***症譜系障礙、痙攣性障礙、記憶障礙和/或認知障礙、運動障礙、人格障礙、自閉症譜系障礙、外傷性腦損傷、血管疾病、物質濫用障礙和/或戒斷綜合症、耳鳴、抑鬱症、潮熱、經前綜合症、創傷後應激障礙、神經性貪食症、強迫症、廣泛性 焦慮症、藥癮、自閉症、老年癡呆症、阿爾茲海默症、癲癇發作、神經痛、重度抑鬱症和狂躁症等疾病；該藥癮包括酒精、尼古丁、阿片調節劑和***成癮。 The present invention further relates to a compound represented by the general formula (I), its stereoisomers or pharmaceutically acceptable salts thereof, or a pharmaceutical composition containing the same in preparation for prevention and/or treatment prevention as an opioid receptor modulator In the treatment of cough, diarrhea, drug dependence, gastrointestinal diseases, irritable bowel syndrome, obesity, respiratory depression, convulsions, pain, allergies, pruritus and other diseases and central nervous diseases or/and mental diseases and other related diseases the use of. Opioid receptor modulators can be used for cough, diarrhea, drug dependence, gastrointestinal diseases, irritable bowel syndrome, obesity, respiratory depression, convulsions, pain, allergies, pruritus and other diseases and central nervous diseases or/and mental health Diseases and other related diseases, the central nervous disease or/and mental diseases are selected from: schizophrenia, sleep disorder, mood disorder, schizophrenia spectrum disorder, spastic disorder, memory disorder and/or cognitive disorder, movement disorder, personality Disorders, autism spectrum disorder, traumatic brain injury, vascular disease, substance abuse disorder and/or withdrawal syndrome, tinnitus, depression, hot flashes, premenstrual syndrome, post-traumatic stress disorder, bulimia nervosa , Obsessive-compulsive disorder, pervasive Anxiety, drug addiction, autism, Alzheimer's disease, Alzheimer's disease, epileptic seizures, neuralgia, major depression and mania; drug addiction includes alcohol, nicotine, opioid modifiers and ***e addiction .

本發明進一步涉及通式(I)所示的化合物、其立體異構體或其藥學上可接受鹽，醫藥組成物在製備阿片受體調節劑藥物中的應用。 The present invention further relates to the application of the compound represented by the general formula (I), its stereoisomer or pharmaceutically acceptable salt, and the pharmaceutical composition in the preparation of opioid receptor modulator drugs.

本發明進一步涉及通式(I)所示的化合物、其立體異構體或其藥學上可接受鹽，醫藥組成物在製備治療咳嗽、腹瀉、藥物依賴性、胃腸道疾病、過敏性腸綜合症、肥胖、呼吸抑制、驚厥、疼痛、過敏、瘙癢等疾病和中樞神經類疾病或/和精神類疾病等相關疾病藥物中的應用；該中樞神經疾病或/和精神類疾病選自：精神***症、睡眠障礙、心境障礙、精神***症譜系障礙、痙攣性障礙、記憶障礙和/或認知障礙、運動障礙、人格障礙、自閉症譜系障礙、外傷性腦損傷、血管疾病、物質濫用障礙和/或戒斷綜合症、耳鳴、抑鬱症、潮熱、經前綜合症、創傷後應激障礙、神經性貪食症、強迫症、廣泛性焦慮症、藥癮、自閉症、老年癡呆症、阿爾茲海默症、癲癇發作、神經痛、重度抑鬱症和狂躁症等疾病；該藥癮包括酒精、尼古丁、阿片調節劑和***成癮。 The present invention further relates to the compound represented by the general formula (I), its stereoisomer or its pharmaceutically acceptable salt, and the pharmaceutical composition is used in the preparation of the treatment of cough, diarrhea, drug dependence, gastrointestinal disease, irritable bowel syndrome , Obesity, respiratory depression, convulsions, pain, allergies, itching and other diseases and central nervous diseases or/and mental diseases and other related diseases; the central nervous disease or/and mental diseases are selected from: schizophrenia , Sleep disorder, mood disorder, schizophrenia spectrum disorder, spastic disorder, memory disorder and/or cognitive disorder, movement disorder, personality disorder, autism spectrum disorder, traumatic brain injury, vascular disease, substance abuse disorder and/or Or withdrawal syndrome, tinnitus, depression, hot flashes, premenstrual syndrome, post-traumatic stress disorder, bulimia nervosa, obsessive-compulsive disorder, generalized anxiety disorder, drug addiction, autism, Alzheimer's disease, Alzheimer's disease Diseases such as Zheimer’s disease, seizures, neuralgia, major depression, and mania; drug addiction includes addiction to alcohol, nicotine, opioid modifiers, and ***e.

本發明還涉及一種治療預防和/或治療預防阿片受體介導的病理學特徵的疾病的方法，其包括向患者施用治療有效劑量的通式(I)所示的化合物、其立體異構體或其藥學上可接受鹽，或包含其的醫藥組成物。阿片受體調節劑可以用於治療咳嗽、腹瀉、藥物依賴性、胃腸道疾病、過敏性腸綜合症、肥胖、呼吸抑制、驚厥、疼痛、過敏、瘙癢等疾病和中樞神經類疾病或/和精神類疾病等相關疾病。 The present invention also relates to a method for treating and/or treating and preventing diseases with opioid receptor-mediated pathological characteristics, which comprises administering to a patient a therapeutically effective dose of a compound represented by general formula (I) and its stereoisomers Or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same. Opioid receptor modulators can be used to treat cough, diarrhea, drug dependence, gastrointestinal diseases, irritable bowel syndrome, obesity, respiratory depression, convulsions, pain, allergies, pruritus and other diseases and central nervous system diseases or/and mental illness Related diseases.

本發明另一方面涉及一種治療咳嗽、腹瀉、藥物依賴性、胃腸道疾病、過敏性腸綜合症、肥胖、呼吸抑制、驚厥、疼痛、過敏、瘙癢等疾病 的方法，該方法包括向患者施用治療有效劑量的本發明的通式(I)所示的化合物、其立體異構體或其藥學上可接受鹽。 Another aspect of the present invention relates to a treatment for diseases such as cough, diarrhea, drug dependence, gastrointestinal diseases, irritable bowel syndrome, obesity, respiratory depression, convulsions, pain, allergies, pruritus, etc. The method comprises administering to the patient a therapeutically effective dose of the compound represented by the general formula (I) of the present invention, its stereoisomers, or a pharmaceutically acceptable salt thereof.

本發明另一方面涉及一種治療中樞神經疾病或/和精神類疾病的方法，該方法包括向患者施用治療有效劑量的本發明的通式(I)所示的化合物、其立體異構體或其藥學上可接受鹽。該方法顯示出突出的療效和較少的副作用，該中樞神經疾病或/和精神類疾病選自：精神***症、睡眠障礙、心境障礙、精神***症譜系障礙、痙攣性障礙、記憶障礙和/或認知障礙、運動障礙、人格障礙、自閉症譜系障礙、外傷性腦損傷、血管疾病、物質濫用障礙和/或戒斷綜合症、耳鳴、抑鬱症、潮熱、經前綜合症、創傷後應激障礙、神經性貪食症、強迫症、廣泛性焦慮症、藥癮、自閉症、老年癡呆症、阿爾茲海默症、癲癇發作、神經痛、重度抑鬱症和狂躁症等疾病；該藥癮包括酒精、尼古丁、阿片調節劑和***成癮。 Another aspect of the present invention relates to a method for the treatment of central nervous system diseases or/and mental diseases, the method comprising administering to a patient a therapeutically effective dose of the compound represented by the general formula (I) of the present invention, its stereoisomers or Pharmaceutically acceptable salt. The method shows outstanding efficacy and fewer side effects. The central nervous disease or/and mental disease is selected from: schizophrenia, sleep disorder, mood disorder, schizophrenia spectrum disorder, spastic disorder, memory disorder and/ Or cognitive impairment, movement disorder, personality disorder, autism spectrum disorder, traumatic brain injury, vascular disease, substance abuse disorder and/or withdrawal syndrome, tinnitus, depression, hot flashes, premenstrual syndrome, post-traumatic Diseases such as stress disorder, bulimia nervosa, obsessive-compulsive disorder, generalized anxiety disorder, drug addiction, autism, Alzheimer's disease, Alzheimer's disease, epileptic seizures, neuralgia, major depression and mania; the Drug addiction includes addiction to alcohol, nicotine, opioid modifiers, and ***e.

本發明發現一類Samidorphan氘代化合物，具有強效的μ阿片受體拮抗活性，與Samidorphan相比具有顯著優勢，能顯著提高口服暴露量和生物利用度。該類Samidorphan氘代化合物除應用於精神***症和抑鬱症治療外，也可應用於鎮痛止癢、抗驚厥、鎮咳、降低食欲、減肥、藥物成癮、呼吸抑制、疼痛等精神類和胃腸道疾病治療，具有巨大的臨床應用價值。 The present invention finds that a class of Samidorphan deuterated compounds has potent mu opioid receptor antagonistic activity, has significant advantages compared with Samidorphan, and can significantly improve oral exposure and bioavailability. This type of Samidorphan deuterated compound is not only used in the treatment of schizophrenia and depression, but also used in analgesia and itching, anticonvulsant, antitussive, appetite reduction, weight loss, drug addiction, respiratory depression, pain and other mental and gastrointestinal tract Disease treatment has great clinical application value.

發明的詳細說明Detailed description of the invention

除非有相反陳述，在說明書和申請專利範圍中使用的術語具有下述含義。 Unless stated to the contrary, the terms used in the specification and the scope of the patent application have the following meanings.

術語“烷基”指飽和脂肪族烴基團，其為包含1至20個碳原子的直鏈或支鏈基團，較佳含有1至8個碳原子的烷基，更佳1至6個碳原子的烷基，最更佳1至3個碳原子的烷基。非限制性實例包括甲基、乙基、正丙 基、異丙基、正丁基、異丁基、第三丁基、第二丁基、正戊基、1,1-二甲基丙基、1,2-二甲基丙基、2,2-二甲基丙基、1-乙基丙基、2-甲基丁基、3-甲基丁基、正己基、1-乙基-2-甲基丙基、1,1,2-三甲基丙基、1,1-二甲基丁基、1,2-二甲基丁基、2,2-二甲基丁基、1,3-二甲基丁基、2-乙基丁基、2-甲基戊基、3-甲基戊基、4-甲基戊基、2,3-二甲基丁基、正庚基、2-甲基己基、3-甲基己基、4-甲基己基、5-甲基己基、2,3-二甲基戊基、2,4-二甲基戊基、2,2-二甲基戊基、3,3-二甲基戊基、2-乙基戊基、3-乙基戊基、正辛基、2,3-二甲基己基、2,4-二甲基己基、2,5-二甲基己基、2,2-二甲基己基、3,3-二甲基己基、4,4-二甲基己基、2-乙基己基、3-乙基己基、4-乙基己基、2-甲基-2-乙基戊基、2-甲基-3-乙基戊基、正壬基、2-甲基-2-乙基己基、2-甲基-3-乙基己基、2,2-二乙基戊基、正癸基、3,3-二乙基己基、2,2-二乙基己基，及其各種支鏈異構體等。更佳的是含有1至6個碳原子的低級烷基，非限制性實施例包括甲基、乙基、正丙基、異丙基、正丁基、異丁基、第三丁基、第二丁基、正戊基、1,1-二甲基丙基、1,2-二甲基丙基、2,2-二甲基丙基、1-乙基丙基、2-甲基丁基、3-甲基丁基、正己基、1-乙基-2-甲基丙基、1,1,2-三甲基丙基、1,1-二甲基丁基、1,2-二甲基丁基、2,2-二甲基丁基、1,3-二甲基丁基、2-乙基丁基、2-甲基戊基、3-甲基戊基、4-甲基戊基、2,3-二甲基丁基等。烷基可以是取代的或非取代的，當被取代時，取代基可以在任何可使用的連接點上被取代，該取代基較佳為一個或多個以下基團，其獨立地選自烷基、烯基、炔基、烷氧基、烷硫基、烷基胺基、鹵素、巰基、羥基、硝基、氰基、環烷基、雜環烷基、芳基、雜芳基、環烷氧基、雜環烷氧基、環烷硫基、雜環烷硫基、側氧基、羧基或羧酸酯基。 The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 8 carbon atoms, more preferably 1 to 6 carbons An alkyl group having 1 to 3 carbon atoms is most preferred. Non-limiting examples include methyl, ethyl, n-propyl Base, isopropyl, n-butyl, isobutyl, tertiary butyl, second butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2, 2-Dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2- Trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethyl Butyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl Base, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2 -Dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethyl 2-methylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl Base, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched isomers. More preferably, it is a lower alkyl group containing 1 to 6 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tertiary butyl, and Dibutyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl Group, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2- Dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methyl Benzylpentyl, 2,3-dimethylbutyl, etc. Alkyl groups may be substituted or unsubstituted. When substituted, the substituents may be substituted at any available attachment point. The substituents are preferably one or more of the following groups, which are independently selected from alkanes Group, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, ring Alkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, pendant oxy, carboxy, or carboxylate.

“DMSO”指二甲基亞碸。 "DMSO" refers to dimethyl sulfoxide.

“THF”指四氫呋喃。 "THF" means tetrahydrofuran.

“EtOAc”指乙酸乙酯。 "EtOAc" refers to ethyl acetate.

“AcOH”指乙醇。 "AcOH" refers to ethanol.

“DMF”指N,N-二甲基甲醯胺。 "DMF" refers to N,N-dimethylformamide.

“DIPEA”指二異丙基乙胺。 "DIPEA" refers to diisopropylethylamine.

“TFA”指三氟乙酸。 "TFA" refers to trifluoroacetic acid.

“MeCN”指乙晴。 "MeCN" means Otoharu.

“DMA”指N,N-二甲基乙醯胺。 "DMA" refers to N,N-dimethylacetamide.

“Et₂O”指***。 "Et ₂ O" refers to diethyl ether.

“DCE”指1,2二氯乙烷。 "DCE" refers to 1,2 dichloroethane.

“DIPEA”指N,N-二異丙基乙胺。 "DIPEA" refers to N,N-diisopropylethylamine.

“NBS”指N-溴琥珀醯亞胺。 "NBS" refers to N-bromosuccinimide.

“NIS”指N-碘丁二醯亞胺。 "NIS" refers to N-iodosuccinimide.

“Cbz-Cl”指氯甲酸苄酯。 "Cbz-Cl" refers to benzyl chloroformate.

“Pd₂(dba)₃”指三(二亞苄基丙酮)二鈀。 "Pd ₂ (dba) ₃ "refers to tris(dibenzylideneacetone) dipalladium.

“Dppf”指1,1’-雙二苯基膦二茂鐵。 "Dppf" refers to 1,1'-bisdiphenylphosphinoferrocene.

“HATU”指2-(7-氧化苯并***)-N,N,N’,N’-四甲基脲六氟磷酸鹽。 "HATU" refers to 2-(7-oxybenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate.

“KHMDS”指六甲基二矽基胺基鉀。 "KHMDS" refers to potassium hexamethyldisilazide.

“LiHMDS”指雙三甲基矽基胺基鋰。 "LiHMDS" refers to lithium bistrimethylsilylamide.

“MeLi”指甲基鋰。 "MeLi" refers to methyl lithium.

“n-BuLi”指正丁基鋰。 "N-BuLi" refers to n-butyl lithium.

“NaBH(OAc)₃”指三乙醯氧基硼氫化鈉。 "NaBH(OAc) ₃ "means sodium triacetoxyborohydride.

“X選自A、B、或C”、“X選自A、B和C”、“X為A、B或C”、“X為A、B和C”等不同用語均表達了相同的意義，即表示X可以是A、B、C中的任意一種或幾種。 "X is selected from A, B, or C", "X is selected from A, B and C", "X is A, B or C", "X is A, B and C" and other terms all express the same Meaning, which means that X can be any one or more of A, B, and C.

“立體異構”包含幾何異構(順反異構)、旋光異構、構象異構三類。 "Stereoisomerism" includes geometric isomerism (cis-trans isomerism), optical isomerism, and conformational isomerism.

本發明所述的氫原子均可被其同位素氘所取代，本發明涉及的實施例化合物中的任一氫也均可被氘取代。 The hydrogen atoms described in the present invention can be replaced by its isotope deuterium, and any hydrogen in the example compounds of the present invention can also be replaced by deuterium.

“任選”或“任選地”意味著隨後所描述的事件或環境可以但不必發生，該說明包括該事件或環境發生或不發生的場合。例如，“任選被烷基取代的雜環基團”意味著烷基可以但不必須存在，該說明包括雜環基團被烷基取代的情形和雜環基團不被烷基取代的情形。 "Optional" or "optionally" means that the event or environment described later can but does not have to occur, and the description includes occasions where the event or environment occurs or does not occur. For example, "heterocyclic group optionally substituted by an alkyl group" means that an alkyl group may but does not have to be present. The description includes the case where the heterocyclic group is substituted by an alkyl group and the case where the heterocyclic group is not substituted by an alkyl group. .

“取代的”指基團中的一個或多個氫，較佳為最多5個，更佳為1~3個氫彼此獨立地被相應數目的取代基取代。不言而喻，取代基僅處在它們的可能的化學位置，本領域技術人員能夠在不付出過多努力的情況下確定(藉由實驗或理論)可能或不可能的取代。例如，具有游離氫的胺基或羥基與具有不飽和(如烯屬)鍵的碳原子結合時可能是不穩定的。 "Substituted" refers to one or more hydrogens in the group, preferably at most 5, and more preferably 1 to 3 hydrogens independently of each other replaced by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without too much effort. For example, an amine group or a hydroxyl group having free hydrogen may be unstable when combined with a carbon atom having an unsaturated (e.g., olefinic) bond.

“醫藥組成物”表示含有一種或多種本文所述化合物或其生理學上/可藥用的鹽或前體藥物與其他化學組分的混合物，以及其他組分例如生理學/可藥用的載體和賦形劑。醫藥組成物的目的是促進對生物體的給藥，利於活性成分的吸收進而發揮生物活性。 "Pharmaceutical composition" means a mixture containing one or more of the compounds described herein or their physiologically/pharmaceutically acceptable salts or prodrugs and other chemical components, as well as other components such as physiological/pharmaceutically acceptable carriers And excipients. The purpose of the medicinal composition is to promote the administration of the organism, which is conducive to the absorption of the active ingredient and thus the biological activity.

“可藥用鹽”是指本發明化合物的鹽，這類鹽用於哺乳動物體內時具有安全性和有效性，且具有應有的生物活性。 "Pharmaceutically acceptable salt" refers to the salt of the compound of the present invention, which is safe and effective when used in mammals, and has due biological activity.

以下結合實施例進一步描述本發明，但這些實施例並非限制著本發明的範圍。 The present invention is further described below in conjunction with examples, but these examples do not limit the scope of the present invention.

實施例 Example

化合物的結構是藉由核磁共振(NMR)或質譜(MS)來確定的。NMR的測定是用Bruker AVANCE-400核磁儀，測定溶劑為氘代二甲基亞碸(DMSO-d ₆)、氘代氯仿(CDCl₃)、氘代甲醇(CD₃OD)，內標為四甲基矽烷(TMS)，化學位移是以10^-6(ppm)作為單位給出。 The structure of the compound is determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS). NMR was measured with Bruker AVANCE-400 nuclear magnetic instrument, and the solvent was deuterated dimethyl sulfoxide (DMSO- d ₆ ), deuterated chloroform (CDCl ₃ ), deuterated methanol (CD ₃ OD), and the internal standard was four For methyl silane (TMS), the chemical shift is ^{given in units of 10 -6} (ppm).

MS的測定用FINNIGAN LCQAd(ESI)質譜儀(生產商：Thermo，型號：Finnigan LCQ advantage MAX)。 The MS was measured with a FINNIGAN LCQAd (ESI) mass spectrometer (manufacturer: Thermo, model: Finnigan LCQ advantage MAX).

HPLC的測定使用安捷倫1200DAD高壓液相色譜儀(Sunfire C18 150×4.6mm色譜管柱)和Waters 2695-2996高壓液相色譜儀(Gimini C18 150×4.6mm色譜管柱)。 HPLC determination uses Agilent 1200DAD high pressure liquid chromatograph (Sunfire C18 150×4.6mm chromatographic column) and Waters 2695-2996 high pressure liquid chromatograph (Gimini C18 150×4.6mm chromatographic column).

激酶平均抑制率及IC₅₀值的測定用NovoStar酶標儀(德國BMG公司)。 The average value of ₅₀ measured kinase inhibition rate and IC NovoStar using a microplate reader (BMG, Germany).

薄層層析矽膠板使用煙臺黃海HSGF254或青島GF254矽膠板，薄層色譜法(TLC)使用的矽膠板採用的規格是0.15mm~0.2mm，薄層層析分離純化產物採用的規格是0.4mm~0.5mm矽膠板。 The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate, the size of the silica gel plate used in thin layer chromatography (TLC) is 0.15mm~0.2mm, and the size of the thin layer chromatography separation and purification product is 0.4mm. ~0.5mm silicon rubber sheet.

管柱層析一般使用煙臺黃海200~300目矽膠為載體。 Column chromatography generally uses Yantai Huanghai 200~300 mesh silica gel as the carrier.

本發明的已知的起始原料可以採用或按照本領域已知的方法來合成，或可購買自ABCR GmbH & Co.KG，Acros Organnics，Aldrich Chemical Company，韶遠化學科技(Accela ChemBio Inc)、達瑞化學品等公司。 The known starting materials of the present invention can be synthesized by or according to methods known in the art, or can be purchased from ABCR GmbH & Co.KG, Acros Organnics, Aldrich Chemical Company, Accela ChemBio Inc, Companies such as Darui Chemicals.

實施例中如無特殊說明，反應均在氬氣氛或氮氣氛下進行。 Unless otherwise specified in the examples, the reactions are all carried out under an argon atmosphere or a nitrogen atmosphere.

氬氣氛或氮氣氛是指反應瓶連接一個約1L容積的氬氣或氮氣氣球。 The argon atmosphere or nitrogen atmosphere means that the reaction flask is connected to an argon or nitrogen balloon with a volume of about 1L.

氫氣氛是指反應瓶連接一個約1L容積的氫氣氣球。 The hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon with a volume of about 1L.

加壓氫化反應使用Parr 3916EKX型氫化儀和清藍QL-500型氫氣發生器或HC2-SS型氫化儀。 The pressure hydrogenation reaction uses Parr 3916EKX hydrogenator and Qinglan QL-500 hydrogen generator or HC2-SS hydrogenator.

氫化反應通常抽真空，充入氫氣，重複操作3次。 The hydrogenation reaction is usually evacuated and filled with hydrogen, and the operation is repeated 3 times.

微波反應使用CEM Discover-S 908860型微波反應器。 The microwave reaction uses a CEM Discover-S 908860 microwave reactor.

實施例中如無特殊說明，反應中的溶液是指水溶液。 Unless otherwise specified in the examples, the solution in the reaction refers to an aqueous solution.

實施例中如無特殊說明，反應的溫度為室溫。 Unless otherwise specified in the examples, the reaction temperature is room temperature.

室溫為最適宜的反應溫度，溫度範圍是20℃~30℃。 Room temperature is the most suitable reaction temperature, and the temperature range is 20℃~30℃.

實施例中的反應進程的監測採用薄層色譜法(TLC)，反應所使用的展開劑的體系有：A：二氯甲烷和甲醇體系，B：正己烷和乙酸乙酯體系，C：石油醚和乙酸乙酯體系，D：丙酮，溶劑的體積比如化合物的極性不同而進行調節。 The monitoring of the reaction progress in the examples adopts thin-layer chromatography (TLC). The developing reagent system used in the reaction includes: A: dichloromethane and methanol system, B: n-hexane and ethyl acetate system, C: petroleum ether And ethyl acetate system, D: acetone, the volume of the solvent, such as the polarity of the compound, is adjusted.

純化化合物採用的管柱層析的沖提劑的體系和薄層色譜法的展開劑的體系包括：A：二氯甲烷和甲醇體系，B：正己烷和乙酸乙酯體系，C：正己烷、乙酸乙酯和二氯甲烷體系，D：石油醚和乙酸乙酯體系，E：乙酸乙酯，溶劑的體積比如化合物的極性不同而進行調節，也可以加入少量的三乙胺和酸性或鹼性試劑等進行調節。 The eluent system of column chromatography and the developing agent system of thin layer chromatography used to purify compounds include: A: dichloromethane and methanol system, B: n-hexane and ethyl acetate system, C: n-hexane, Ethyl acetate and dichloromethane system, D: petroleum ether and ethyl acetate system, E: ethyl acetate, the volume of the solvent is adjusted such as the polarity of the compound, and a small amount of triethylamine and acidic or alkaline can also be added Reagents, etc. are adjusted.

實施例1Example 1

(4bR,8aS,9R)-11-(環丙基甲基-d2)-4,8a-二羥基-6-側氧-6,7,8,8a,9,10-六氫-5H-9,4b-(表胺基乙烷)菲-3-甲醯胺的製備(4bR,8aS,9R)-11-(cyclopropylmethyl-d2)-4,8a-dihydroxy-6-oxo-6,7,8,8a,9,10-hexahydro-5H-9 Preparation of 4b-(epiaminoethane)phenanthrene-3-formamide

第一步：(4R,4aS,7aR,12bS)-3-(環丙基甲基-d2)-4a,9-二羥基-2,3,4,4a,5,6-六氫-1H-4,12-甲基苯并呋喃并[3,2-e]異喹啉-7(7aH)-酮(中間體1a)的製備 The first step: (4R,4aS,7aR,12bS)-3-(cyclopropylmethyl-d2)-4a,9-dihydroxy-2,3,4,4a,5,6-hexahydro-1H- Preparation of 4,12-methylbenzofuro[3,2-e]isoquinoline-7(7aH)-one (Intermediate 1a)

在0-5℃溫度下，將甲磺醯氯(5.31mL)添加到環丙基甲基-d2-醇(5.71mL)和三乙胺(20.13mL)的DMF(200mL)溶液中。該溶液在0-5℃溫度下保持3.0小時。將溴化鋰(6.22g)添加到反應混合物中並繼續在0-5℃下攪拌3.0小時。將(-)-去甲氧嗎啡酮(6.18g)在0-5℃下加入上述反應混合物中，並在1.0h內將溫度升高至65-70℃。反應混合物在65-70℃下保持14-15小時。反應完成後，在20℃以下的純化水中(200mL)對反應進行淬滅。將水溶液鹼化至pH>9.0，並用甲苯(200mL X 3)萃取。用去氧水(300mL)然後用10%鹽水溶液(300mL)洗滌合併的甲苯層，無水硫酸鈉乾燥後濃縮，得到固體產物，用甲醇再結晶，得到中間體1a(5.11g)。 At a temperature of 0-5°C, methanesulfonyl chloride (5.31 mL) was added to a solution of cyclopropylmethyl-d2-ol (5.71 mL) and triethylamine (20.13 mL) in DMF (200 mL). The solution was kept at a temperature of 0-5°C for 3.0 hours. Lithium bromide (6.22g) was added to the reaction mixture and stirring was continued at 0-5°C for 3.0 hours. (-)-Demethoxymorphone (6.18g) was added to the above reaction mixture at 0-5°C, and the temperature was increased to 65-70°C within 1.0h. The reaction mixture is kept at 65-70°C for 14-15 hours. After the reaction was completed, the reaction was quenched in purified water (200 mL) below 20°C. The aqueous solution was basified to pH>9.0, and extracted with toluene (200 mL×3). The combined toluene layer was washed with deoxygenated water (300 mL) and then with 10% saline solution (300 mL), dried over anhydrous sodium sulfate and concentrated to obtain a solid product, which was recrystallized from methanol to obtain Intermediate 1a (5.11 g).

MS m/z(ESI)：344.2[M+H]⁺. MS m/z(ESI): 344.2[M+H] ⁺ .

第二步：(4'R,4a'S,7a'R,12b'S)-3'-(環丙基甲基-d2)-1',2',3',4',5',6'-六氫-4a'H,7a'H-螺環[[1,3]二氧雜環戊烷-2,7'-[4,12]甲基苯并呋喃并[3,2-e]異喹啉]-4a',9'-二醇(中間體1b)的製備 The second step: (4'R,4a'S,7a'R,12b'S)-3'-(cyclopropylmethyl-d2)-1',2',3',4',5',6'-hexa Hydrogen-4a'H,7a'H-spiro[[1,3]dioxolane-2,7'-[4,12]methylbenzofuro[3,2-e]isoquine Preparation of 4a',9'-diol (Intermediate 1b)

在氮氣保護下，將甲苯(70mL)、中間體1a(7.0g)、乙二醇(31.6g)和對甲苯磺酸(4.1g)組成的混合溶液加熱至110℃，並攪拌16小時。根據高效液相色譜法鑒定反應完成後，將反應混合物冷卻至18-20℃。分離各層並將底層轉移至乾淨的容器中。加入純化水(125mL)並冷卻至5至8℃。藉由添加固體碳酸鈉(Na₂CO₃)將pH調整至9-10。將所得懸浮液在室溫攪拌45分鐘，分離出固體沉澱物。用純化水洗滌沉澱，並在30-35℃下真空乾燥，得到中間體1b(7.5g)。 Under the protection of nitrogen, a mixed solution consisting of toluene (70 mL), intermediate 1a (7.0 g), ethylene glycol (31.6 g) and p-toluenesulfonic acid (4.1 g) was heated to 110° C. and stirred for 16 hours. After the completion of the reaction was identified by high performance liquid chromatography, the reaction mixture was cooled to 18-20°C. Separate the layers and transfer the bottom layer to a clean container. Add purified water (125 mL) and cool to 5 to 8°C. Adjust the pH to 9-10 by adding solid sodium carbonate (Na ₂ CO _{3 ).} The resulting suspension was stirred at room temperature for 45 minutes, and a solid precipitate was separated. The precipitate was washed with purified water and dried under vacuum at 30-35°C to obtain Intermediate 1b (7.5 g).

MS m/z(ESI)：388.2[M+H]⁺. MS m/z(ESI): 388.2[M+H] ⁺ .

第三步：(4'R,4a'S,7a'R,12b'S)-3'-(環丙基甲基-d2)-4a'-羥基-2',3',4',4a',5',6'-六氫-1'H,7a'-H-螺環[[1,3]二氧雜環戊烷-2,7'-[4,12]甲基苯并呋喃并[3,2-e]異喹啉]-9'-基三氟甲磺酸酯(中間體1c)的製備 The third step: (4'R,4a'S,7a'R,12b'S)-3'-(cyclopropylmethyl-d2)-4a'-hydroxy-2',3',4',4a',5' ,6'-hexahydro-1'H,7a'-H-spiro[[1,3]dioxolane-2,7'-[4,12]methylbenzofuro[3, Preparation of 2-e]isoquinoline]-9'-yl trifluoromethanesulfonate (Intermediate 1c)

在惰性氣氛下將二氯甲烷(200mL)和中間體1b(7.2g)的混合物攪拌並冷卻至-15℃以下，將二異丙基乙胺(3.62g)緩慢添加至反應器中，然後添加三氟甲磺酐(5.7g)。將反應混合物保持在-15℃和0℃之間持續攪拌至少15分鐘。根據HPLC鑒定反應完成後，向反應器中加入20%氯化銨溶液(50mL)和甲基第三丁基醚(100mL)，攪拌至少15分鐘，然後靜置分離。依次用20%氯化銨溶液(50mL)，水(50mL)和鹽水(50mL)洗滌有機層，然後用硫酸鎂乾燥。將溶液過濾並在真空下濃縮，並用矽膠管柱層析純化，得到中間體1c(7.8g)。 Under an inert atmosphere, the mixture of dichloromethane (200mL) and intermediate 1b (7.2g) was stirred and cooled to below -15°C, diisopropylethylamine (3.62g) was slowly added to the reactor, and then added Trifluoromethanesulfonic anhydride (5.7g). Keep the reaction mixture between -15°C and 0°C with continuous stirring for at least 15 minutes. After the completion of the reaction was identified by HPLC, 20% ammonium chloride solution (50 mL) and methyl tert-butyl ether (100 mL) were added to the reactor, stirred for at least 15 minutes, and then left to stand for separation. The organic layer was washed sequentially with 20% ammonium chloride solution (50 mL), water (50 mL) and brine (50 mL), and then dried over magnesium sulfate. The solution was filtered and concentrated under vacuum, and purified by silica gel column chromatography to obtain Intermediate 1c (7.8 g).

MS m/z(ESI)：520.2[M+H]⁺. MS m/z(ESI): 520.2[M+H] ⁺ .

第四步：(4'R,4a'S,7a'R,12b'S)-3'-(環丙基甲基-d2)-4a'-羥基-2',3',4',4a',5',6'-六氫-1'H,7a'H-螺環[[1,3]二氧雜環戊烷-2,7'-[4,12]甲基苯并呋喃并[3,2-e]異喹啉]-9'-碳腈(中間體1d)的製備 The fourth step: (4'R,4a'S,7a'R,12b'S)-3'-(cyclopropylmethyl-d2)-4a'-hydroxy-2',3',4',4a',5' ,6'-hexahydro-1'H,7a'H-spiro[[1,3]dioxolane-2,7'-[4,12]methylbenzofuro[3,2 Preparation of -e]isoquinoline]-9'-carbonitrile (Intermediate 1d)

在惰性氣氛下，將N,N-二甲基甲醯胺(DMF；50mL)，中間體1c(8.8g)，醋酸鈀(200mg)，和1,1'-二(二苯基膦基)二茂鐵(DPPF；943mg)混合攪拌並緩慢加熱至約65℃，達到65℃後，加入氰化鋅(1.2g)。將反應繼續加熱至80℃並攪拌至少1小時。根據HPLC判斷反應完成後，反應器的內容物被冷卻到室溫。向反應器中注入純化水(230mL)，然後控制添加氨水以將pH值調整至10.5至11。將懸浮液加熱至>15℃，過濾分離得到固體。固體用矽膠管柱層析純化得到中間體1d(4.1g)。 Under an inert atmosphere, N,N-dimethylformamide (DMF; 50mL), intermediate 1c (8.8g), palladium acetate (200mg), and 1,1'-bis(diphenylphosphino) Ferrocene (DPPF; 943 mg) was mixed and stirred and slowly heated to about 65°C. After reaching 65°C, zinc cyanide (1.2 g) was added. The reaction was continued to be heated to 80°C and stirred for at least 1 hour. After judging the completion of the reaction according to HPLC, the contents of the reactor were cooled to room temperature. Inject purified water (230 mL) into the reactor, and then control the addition of ammonia water to adjust the pH to 10.5 to 11. The suspension was heated to >15°C, and the solid was separated by filtration. The solid was purified by silica gel column chromatography to obtain Intermediate 1d (4.1g).

MS m/z(ESI)：397.2[M+H]⁺. MS m/z(ESI): 397.2[M+H] ⁺ .

第五步：(4'R,4a'S,7a'R,12b'S)-3'-(環丙基甲基-d2)-4a'-羥基-2',3',4',4a',5',6'-六氫-1'H,7a'-H-螺環[[1,3]二氧雜環戊烷-2,7'-[4,12]甲基苯并呋喃并[3,2-e]異喹啉]-9'-甲醯胺(中間體1e)的製備 The fifth step: (4'R,4a'S,7a'R,12b'S)-3'-(cyclopropylmethyl-d2)-4a'-hydroxy-2',3',4',4a',5' ,6'-hexahydro-1'H,7a'-H-spiro[[1,3]dioxolane-2,7'-[4,12]methylbenzofuro[3, Preparation of 2-e]isoquinoline]-9'-formamide (Intermediate 1e)

在惰性氣氛下，將第三丁醇(60mL)、氯化鈉(12g)和中間體1d(6.0g)混合攪拌，然後將氫氧化鉀(3.0g)緩慢加入。將反應混合物加熱至80℃並攪拌。用HPLC監測反應完成後，將反應混合物冷卻至20℃至30℃，加入2-甲基四氫呋喃(80mL)。向反應器中注入15%鹽水溶液(60mL)，並攪拌至少20分鐘。分離各層並用2-甲基四氫呋喃(30mL)進一步萃取取水層。將有機層合併，用15%的鹽水洗三次，再用硫酸鎂乾燥處理。將溶液過濾並在真空下濃縮，然後用矽膠管柱層析進行純化，得到中間體1e(4.2g)。 Under an inert atmosphere, tertiary butanol (60 mL), sodium chloride (12 g) and intermediate 1d (6.0 g) were mixed and stirred, and then potassium hydroxide (3.0 g) was slowly added. The reaction mixture was heated to 80°C and stirred. After monitoring the completion of the reaction by HPLC, the reaction mixture was cooled to 20°C to 30°C, and 2-methyltetrahydrofuran (80 mL) was added. Inject 15% brine solution (60 mL) into the reactor and stir for at least 20 minutes. The layers were separated and the aqueous layer was further extracted with 2-methyltetrahydrofuran (30 mL). The organic layers were combined, washed three times with 15% brine, and dried over magnesium sulfate. The solution was filtered and concentrated under vacuum, and then purified by silica gel column chromatography to obtain Intermediate 1e (4.2g).

MS m/z(ESI)：415.2[M+H]⁺. MS m/z(ESI): 415.2[M+H] ⁺ .

第六步：(4R,4aS,7aR,12bS)-3-(環丙基甲基-d2)-4a-羥基-7-側氧-2,3,4,4a,5,6,7,7a-八氫-1H-4,12-甲基苯并呋喃[3,2-e]異喹啉-9-甲醯胺(中間體1f)的製備 The sixth step: (4R,4aS,7aR,12bS)-3-(cyclopropylmethyl-d2)-4a-hydroxy-7-side oxygen-2,3,4,4a,5,6,7,7a Preparation of -octahydro-1H-4,12-methylbenzofuran[3,2-e]isoquinoline-9-carboxamide (Intermediate 1f)

在惰性氣氛下，將純化水(75mL)和37%鹽酸(8mL)緩慢混合。冷卻後添加中間體1e(5.0g)和甲苯(50mL)。將雙相混合物加熱至80℃。根據HPLC判斷反應完成後，將反應混合物冷卻至15℃至20℃。分離各層並將二氯甲烷(DCM；25mL)添加至水層，並將混合物攪拌至少15分鐘。將各層分離，將水相過濾到乾淨的容器中，並將濾液冷卻至0℃。藉由向反應容器中加入氨水，將反應混合物的pH值調整為9至10。將所得懸浮液在20℃以下攪拌45分鐘。藉由過濾分離固體並用水(4 X 15mL)沖洗。固體在30-35℃的真空烘箱中乾燥，得到中間體1f(3.1g)。 Under an inert atmosphere, purified water (75 mL) and 37% hydrochloric acid (8 mL) were slowly mixed. After cooling, intermediate 1e (5.0 g) and toluene (50 mL) were added. The biphasic mixture was heated to 80°C. After judging the completion of the reaction according to HPLC, the reaction mixture was cooled to 15°C to 20°C. The layers were separated and dichloromethane (DCM; 25 mL) was added to the aqueous layer, and the mixture was stirred for at least 15 minutes. Separate the layers, filter the aqueous phase into a clean container, and cool the filtrate to 0°C. The pH value of the reaction mixture is adjusted to 9-10 by adding ammonia water to the reaction vessel. The resulting suspension was stirred below 20°C for 45 minutes. The solid was separated by filtration and rinsed with water (4 X 15 mL). The solid was dried in a vacuum oven at 30-35°C to obtain Intermediate 1f (3.1 g).

MS m/z(ESI)：371.2[M+H]⁺. MS m/z(ESI): 371.2[M+H] ⁺ .

第七步：(4bR,8aS,9R)-11-(環丙基甲基-d2)-4,8a-二羥基-6-側氧-6,7,8,8a,9,10-六氫-5H-9,4b-(表胺基乙烷)菲-3-甲醯胺(實施例1)的製備 The seventh step: (4bR,8aS,9R)-11-(cyclopropylmethyl-d2)-4,8a-dihydroxy-6-oxo-6,7,8,8a,9,10-hexahydro Preparation of -5H-9,4b-(epiaminoethane)phenanthrene-3-methanamide (Example 1)

回流狀態下，將鋅粉(114mg)緩慢分批添加到中間體1f(122mg)在HCl(37%，0.2mL)和AcOH(2mL)中的溶液中。回流加熱15分鐘後，添加冰/水(20mL)來冷卻反應，然後用氨水將pH調為9。用DCM(3 x 20mL)萃取所得混合物，有機相經鹽水洗滌、乾燥和濃縮，然後用HPLC純化，得到實施例1(55mg)。 Under reflux, zinc powder (114 mg) was slowly added in batches to the solution of intermediate 1f (122 mg) in HCl (37%, 0.2 mL) and AcOH (2 mL). After heating under reflux for 15 minutes, ice/water (20 mL) was added to cool the reaction, and then the pH was adjusted to 9 with ammonia water. The resulting mixture was extracted with DCM (3 x 20 mL), the organic phase was washed with brine, dried and concentrated, and then purified by HPLC to obtain Example 1 (55 mg).

MS m/z(ESI)：373.2[M+H]⁺. MS m/z(ESI): 373.2[M+H] ⁺ .

實施例1也可根據如下方法獲得： Embodiment 1 can also be obtained according to the following method:

第一步：環丙基甲烷-d2-醇(中間體1a)的製備 Step 1: Preparation of cyclopropylmethane-d2-ol (Intermediate 1a)

冰鹽浴下，四氘代鋁鋰(5.04g,120mmol)緩慢加入到四氫呋喃(50mL)中，在零下10℃，氬氣保護下，緩慢加入環丙基甲酸甲酯(12.00g,120mmol)的四氫呋喃溶液(20mL)。上述反應液在攝氏零度下攪拌反應一個小時，隨後在此溫度下，用十水硫酸鈉焠滅反應。過濾除掉固體，用四氫呋喃(50mL)洗固體兩次，合併的有機層在室溫下水泵減壓旋蒸去大部分溶劑得到中間體1a(6.76g，無色液體)，產率：76.1%。 Under ice-salt bath, tetradeuterated aluminum lithium (5.04g, 120mmol) was slowly added to tetrahydrofuran (50mL), at minus 10℃, under the protection of argon, slowly added methyl cyclopropyl formate (12.00g, 120mmol) Tetrahydrofuran solution (20 mL). The above reaction solution was stirred and reacted at zero degrees Celsius for one hour, and then at this temperature, the reaction was quenched with sodium sulfate decahydrate. The solid was removed by filtration, and the solid was washed twice with tetrahydrofuran (50 mL). The combined organic layer was rotated at room temperature with a water pump under reduced pressure to remove most of the solvent to obtain Intermediate 1a (6.76 g, colorless liquid). Yield: 76.1%.

¹H NMR(400MHz,CDCl₃)δ 1.93-1.89(m,1H),1.12-0.99(m,1H),0.57-0.39(m,2H),0.24-0.09(m,2H). ¹ H NMR (400MHz, CDCl ₃ ) δ 1.93-1.89 (m, 1H), 1.12-0.99 (m, 1H), 0.57-0.39 (m, 2H), 0.24-0.09 (m, 2H).

第二步：対甲基苯磺酸-1,1-二氘代環丙基甲酯(中間體1b)的製備 Step 2: Preparation of 1,1-dideuterated cyclopropyl methyl benzenesulfonic acid (Intermediate 1b)

將中間體1a(2.16g，29mmol)，對二甲胺吡啶(366mg，3mmol)和三乙胺(15mL)的二氯甲烷(10mL)溶液緩慢加入到-10℃(冰鹽浴)冷卻對甲苯磺醯氯(5.51g,29mmol)的二氯甲烷(30mL)懸濁液中。在-10℃到4℃ 之間溫度攪拌反應12小時。反應液緩慢倒入到攪拌30%的硫酸碎冰(100mL)中，用二氯甲烷(20mLX3)萃取，合併有機層，飽和食鹽水(20mL)洗一次(上述操作保持溫度不高於15℃)，用無水硫酸鈉乾燥，減壓濃縮(室溫)対甲基苯磺酸-1,1-二氘代環丙基甲酯(3.60g，冰箱冷凍後是淡黃色固體)，產率：54.5% Intermediate 1a (2.16g, 29mmol), p-dimethylaminopyridine (366mg, 3mmol) and triethylamine (15mL) in dichloromethane (10mL) were slowly added to -10°C (ice-salt bath) to cool p-toluene Sulfonyl chloride (5.51g, 29mmol) in dichloromethane (30mL) suspension. At -10°C to 4°C The reaction was stirred at the temperature for 12 hours. The reaction solution was slowly poured into 30% sulfuric acid crushed ice (100mL) with stirring, extracted with dichloromethane (20mLX3), combined the organic layers, washed with saturated brine (20mL) once (the above operation keeps the temperature not higher than 15℃) , Dried with anhydrous sodium sulfate, concentrated under reduced pressure (room temperature) 1,1-dideuterated cyclopropyl methyl benzenesulfonic acid (3.60g, light yellow solid after freezing in the refrigerator), yield: 54.5 %

¹H NMR(400MHz,CDCl₃)δ 7.83-7.75(m,2H),7.35(d,2H),2.45(s,3H),1.16-1.03(m,1H),0.65-0.50(m,2H),0.29-0.18(m,2H). ¹ H NMR (400MHz, CDCl ₃ ) δ 7.83-7.75 (m, 2H), 7.35 (d, 2H), 2.45 (s, 3H), 1.16-1.03 (m, 1H), 0.65-0.50 (m, 2H) ,0.29-0.18(m,2H).

第三步：(4R,4aS,7aR,12bS)-4a,9-二羥基-2,3,4,4a,5,6-六氫-1H-4,12-甲氧基苯并呋喃[3,2-e]異喹啉-7(7aH)-酮鹽酸鹽(中間體1c)的製備 The third step: (4 R ,4a S ,7a R ,12b S )-4a,9-dihydroxy-2,3,4,4a,5,6-hexahydro-1H-4,12-methoxybenzene Preparation of nitrofuran[3,2-e]isoquinoline-7(7aH)-one hydrochloride (Intermediate 1c)

將(4R,4aS,7aR,12bS)-4a,9-二羥基-2,3,4,4a,5,6-六氫-1H-4,12-甲氧基苯并呋喃并[3,2-e]異喹啉-7(7aH)-酮甲酸鹽(1.20g，3.60mmol)溶於鹽酸(50mL,5%/wt)中，室溫下攪拌，溶解，冷凍乾燥得到中間體1c，產率：97.0%。 Add (4 R ,4a S ,7a R ,12b S )-4a,9-dihydroxy-2,3,4,4a,5,6-hexahydro-1H-4,12-methoxybenzofuro [3,2-e]Isoquinoline-7(7aH)-ketocarboxylate (1.20g, 3.60mmol) was dissolved in hydrochloric acid (50mL, 5%/wt), stirred at room temperature, dissolved, and freeze-dried to obtain Intermediate 1c, yield: 97.0%.

¹H NMR(400MHz,DMSO-d₆)δ 9.45(s,1H),9.04(s,1H),6.65(q,2H),6.42(s,1H),4.93(s,1H),4.10(dd,1H),3.67(t,1H),3.26(s,1H),3.09(d,1H),3.07-3.02(m,1H),3.00(d,1H),2.92(d,1H),2.56(d,1H),2.11(d,1H),1.93(d,1H),1.52-1.37(m,2H). ¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.45 (s, 1H), 9.04 (s, 1H), 6.65 (q, 2H), 6.42 (s, 1H), 4.93 (s, 1H), 4.10 (dd ,1H), 3.67(t, 1H), 3.26(s, 1H), 3.09(d, 1H), 3.07-3.02(m, 1H), 3.00(d, 1H), 2.92(d, 1H), 2.56( d, 1H), 2.11 (d, 1H), 1.93 (d, 1H), 1.52-1.37 (m, 2H).

第四步：((4R,4aS,7aR,12bS)-3-(環丙基二氘代甲基)-4a,9-二羥基-2,3,4,4a,5,6-六氫-1H-4,12-甲氧基苯并呋喃并[3,2-e]異喹啉-7(7aH)-酮(中間體1d)的製備 The fourth step: ((4R,4aS,7aR,12bS)-3-(cyclopropyl di-deuterated methyl)-4a,9-dihydroxy-2,3,4,4a,5,6-hexahydro- Preparation of 1H-4,12-methoxybenzofuro[3,2-e]isoquinoline-7(7aH)-one (Intermediate 1d)

將中間體1c(1.20g，3.72mmol)溶於甲基吡咯烷酮(6mL)和水(1mL)的混合溶劑中，加入三乙胺(1.6mL，11.14mmol)和碘化鉀(1.27g，5.58mmol)，室溫攪拌5分鐘，向上述懸濁液中加入環丙基二氘代甲基-4-甲基苯磺酸酯(1.27g,5.58mmol)，氬氣保護下70℃加熱反應12小時。減壓濃縮去除揮發性物質，剩餘物加入到二氯甲烷(100mL)中，用飽和氯化鈉水溶液(30mL)洗一次，水相用二氯甲烷(20mL)萃取一次，合併有機層，用飽和食鹽水(20mL)再洗一次，無水硫酸鈉乾燥，過濾，減壓濃縮得到中間體1d(1.20g，粗品棕色液體含有NMP)，收率：93.8%。直接進入下步反應。 Intermediate 1c (1.20g, 3.72mmol) was dissolved in a mixed solvent of methylpyrrolidone (6mL) and water (1mL), triethylamine (1.6mL, 11.14mmol) and potassium iodide (1.27g, 5.58mmol) were added, Stir at room temperature for 5 minutes, add cyclopropyl dideuteromethyl-4-methylbenzenesulfonate (1.27 g, 5.58 mmol) to the above suspension, and heat the reaction at 70° C. for 12 hours under the protection of argon. Concentrate under reduced pressure to remove volatile substances, add the residue to dichloromethane (100mL), wash once with saturated sodium chloride aqueous solution (30mL), extract the aqueous phase with dichloromethane (20mL) once, combine the organic layers, and use saturated Wash with brine (20 mL) again, dry with anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain intermediate 1d (1.20 g, crude brown liquid containing NMP), yield: 93.8%. Go directly to the next step reaction.

MS m/z(ESI)：344.1[M+1]⁺。 MS m/z (ESI): 344.1 [M+1] ⁺ .

第五步：(4R,4aS,7aR,12bS)-3-(環丙基二氘代甲基)-4a-羥基-7-側氧-2,3,4,4a,5,6,7,7a-八氫-1H-4,12-甲氧基苯并呋喃并[3,2-e]異喹啉-9-基三氟甲基磺酸酯(中間體1e)的製備 The fifth step: (4R,4aS,7aR,12bS)-3-(cyclopropyl di-deuterated methyl)-4a-hydroxy-7-side oxygen-2,3,4,4a,5,6,7, Preparation of 7a-octahydro-1H-4,12-methoxybenzofuro[3,2-e]isoquinolin-9-yl trifluoromethanesulfonate (Intermediate 1e)

將中間體1d(1.20g,3.49mmol)溶解在二氯甲烷(30mL)中，加入三乙胺(528mg,5.23mmol)和對二甲胺吡啶(42mg,0.35mmol。0℃加入1,1,1- 三氟-N-苯基-N-((三氟甲基)磺醯基)甲磺醯胺(1.25g,3.49mmol)，室溫攪拌反應兩個小時。加入二氯甲烷(50mL)，用飽和氯化鈉水溶液(20mL)洗一次，無水硫酸鈉乾燥，過濾，減壓濃縮得到剩餘物，過矽膠管柱層析(石油醚：乙酸乙酯=2：3)得到中間體1e(720mg，白色固體)，產率：43.4%。 Intermediate 1d (1.20g, 3.49mmol) was dissolved in dichloromethane (30mL), and triethylamine (528mg, 5.23mmol) and p-dimethylaminopyridine (42mg, 0.35mmol) were added to add 1,1, 1- Trifluoro-N-phenyl-N-((trifluoromethyl)sulfonyl)methanesulfonamide (1.25g, 3.49mmol) was stirred at room temperature and reacted for two hours. Add dichloromethane (50mL), wash once with saturated sodium chloride aqueous solution (20mL), dry with anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain the residue, which is subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 2: 3) Intermediate 1e (720 mg, white solid) was obtained, with a yield of 43.4%.

MS m/z(ESI)：476.0[M+1]⁺。 MS m/z (ESI): 476.0 [M+1] ⁺ .

第六步：(4R,4aS,7aR,12bS)-3-(環丙基甲基-d2)-4a-羥基-7-側氧-2,3,4,4a,5,6,7,7a-八氫-1H-4,12-甲基苯并呋喃并[3,2-e]異喹啉-9-腈(中間體1f)的製備 The sixth step: (4R,4aS,7aR,12bS)-3-(cyclopropylmethyl-d2)-4a-hydroxy-7-side oxygen-2,3,4,4a,5,6,7,7a -Preparation of octahydro-1H-4,12-methylbenzofuro[3,2-e]isoquinoline-9-carbonitrile (Intermediate 1f)

將中間體1e(480mg，1.01mmol)溶於DMF(2mL)，加入四三苯基膦鈀(115mg，0.10mmol)和氰化鋅(236mg，2.02mmol)。氬氣保護下，微波120℃反應兩個小時。冷卻至室溫，加入到乙酸乙酯(30mL)和水(20mL)中，攪拌分液，水層用乙酸乙酯(20mL)再萃取一次。合併有機層，用飽和氯化鈉水溶液(20mL)洗一次，乾燥，過濾，減壓濃縮得到固體，在石油醚和乙酸乙酯混合溶液中(3：1)打漿得到1f(520mg，白色固體)，產率：100%。 Intermediate le (480 mg, 1.01 mmol) was dissolved in DMF (2 mL), and palladium tetrakistriphenylphosphine (115 mg, 0.10 mmol) and zinc cyanide (236 mg, 2.02 mmol) were added. Under the protection of argon, microwave at 120°C for two hours. It was cooled to room temperature, added to ethyl acetate (30 mL) and water (20 mL), stirred for liquid separation, and the aqueous layer was extracted once more with ethyl acetate (20 mL). The organic layers were combined, washed once with saturated sodium chloride aqueous solution (20 mL), dried, filtered, and concentrated under reduced pressure to obtain a solid, which was slurried in a mixture solution of petroleum ether and ethyl acetate (3:1) to obtain 1f (520 mg, white solid) , Yield: 100%.

MS m/z(ESI)：353.0[M+1]⁺。 MS m/z (ESI): 353.0 [M+1] ⁺ .

第七步：(4R,4aS,7aR,12bS)-3-(環丙基二氘代甲基)-4a-羥基-7-側氧-2,3,4,4a,5,6,7,7a-八氫-1H-4,12-甲基苯并呋喃并[3,2-e]異喹啉-9-甲醯胺(中間體1g)的製備 The seventh step: (4R,4aS,7aR,12bS)-3-(cyclopropyl di-deuterated methyl)-4a-hydroxy-7-side oxygen-2,3,4,4a,5,6,7, Preparation of 7a-octahydro-1H-4,12-methylbenzofuro[3,2-e]isoquinoline-9-carboxamide (Intermediate 1g)

將中間體1f(100mg，0.28mmol)溶於DMSO(1mL)和乙醇(1mL)，加入碳酸鉀(195mg，1.42mmol)，冰浴下攪拌加入雙氧水(0.2mL，30%wt)，50攝氏度氬氣保護下反應12個小時。冷卻至室溫，加入乙酸乙酯(20mL)，用飽和氯化鈉溶液(5mLX 3)洗，有機層用無水硫酸鈉乾燥，過濾，減壓濃縮得到固體，用石油醚和乙酸乙酯的混合溶劑(3：1)打漿過濾出固體，乾燥得到中間體1g(30mg，白色固體)，產率：28.5%。 Intermediate 1f (100mg, 0.28mmol) was dissolved in DMSO (1mL) and ethanol (1mL), potassium carbonate (195mg, 1.42mmol) was added, and hydrogen peroxide (0.2mL, 30%wt) was added with stirring under ice bath, argon at 50 degrees Celsius React for 12 hours under air protection. Cool to room temperature, add ethyl acetate (20mL), wash with saturated sodium chloride solution (5mL×3), dry the organic layer with anhydrous sodium sulfate, filter, concentrate under reduced pressure to obtain a solid, mix with petroleum ether and ethyl acetate The solvent (3:1) was slurried and the solid was filtered out, and dried to obtain 1 g (30 mg, white solid) of the intermediate. Yield: 28.5%.

MS m/z(ESI)：371.1[M+1]⁺。 MS m/z (ESI): 371.1 [M+1] ⁺ .

第八步：(4bR,8aS,9R)-11-(環丙基甲基-d2)-4,8a-二羥基-6-側氧-6,7,8,8a,9,10-六氫-5H-9,4b-(表胺基乙醇)菲-3-甲醯胺(實施例1)的製備 The eighth step: (4bR,8aS,9R)-11-(cyclopropylmethyl-d2)-4,8a-dihydroxy-6-oxo-6,7,8,8a,9,10-hexahydro Preparation of -5H-9,4b-(epiaminoethanol)phenanthrene-3-methanamide (Example 1)

將中間體1g(50mg,0.14mmol)，經過稀鹽酸活化的鋅粉(31mg,0.47mmol)，氯化銨(25mg,0.47mmol)溶於1mL乙醇，將混合物在氮氣保護下攪拌3個小時，反應結束後，冷卻至室溫，將反應液濃縮後再溶於DMF中，藉由Prep-HPLC純化，得到實施例1(32mg，白色固體)，產率：59.8%。 The intermediate 1g (50mg, 0.14mmol), zinc powder (31mg, 0.47mmol) activated by dilute hydrochloric acid, and ammonium chloride (25mg, 0.47mmol) were dissolved in 1mL of ethanol, and the mixture was stirred for 3 hours under the protection of nitrogen. After the reaction, it was cooled to room temperature, the reaction solution was concentrated and then dissolved in DMF, and purified by Prep-HPLC to obtain Example 1 (32 mg, white solid), yield: 59.8%.

MS m/z(ESI)：373.2[M+1]⁺。 MS m/z (ESI): 373.2 [M+1] ⁺ .

¹H NMR(400MHz,MeOD)δ 7.52(d,J=8.2Hz,1H),6.67(d,J=8.2Hz,1H),3.60(q,J=7.1Hz,1H),3.10(s,2H),2.92(s,1H),2.76(td,J=13.7,7.5Hz,2H),2.23-2.02(m,3H),1.93(ddd,J=27.9Hz,14.1,6.6Hz,2H),1.71(d,J=11.6Hz,1H),1.17(t,J=7.0Hz,1H),0.94(s,1H),0.59(s,2H),0.23(s,2H). ¹ H NMR(400MHz, MeOD)δ 7.52(d,J=8.2Hz,1H), 6.67(d,J=8.2Hz,1H), 3.60(q,J=7.1Hz,1H), 3.10(s,2H) ), 2.92(s,1H), 2.76(td,J=13.7,7.5Hz,2H),2.23-2.02(m,3H),1.93(ddd,J=27.9Hz,14.1,6.6Hz,2H),1.71 (d,J=11.6Hz,1H),1.17(t,J=7.0Hz,1H),0.94(s,1H),0.59(s,2H),0.23(s,2H).

實施例2Example 2

(4bR,8aS,9R)-11-(環丙基甲基)-4,8a-二羥基-6-側氧-6,7,8,8a,9,10-六氫-5H-9,4b-(表胺基乙烷)菲-1,2-d2-3-甲醯胺的製備(4bR,8aS,9R)-11-(cyclopropylmethyl)-4,8a-dihydroxy-6-oxo-6,7,8,8a,9,10-hexahydro-5H-9,4b -(Epiaminoethane) phenanthrene-1,2-d2-3-formamide preparation

第一步：(4'R,4a'S,7a'R,12b'S)-3'-(環丙基甲基)-1',2',3',4',5',6'-六氫-4a'H,7a'H-螺環[[1,3]二氧雜環戊烷-2,7'-[4,12]甲基苯并呋喃并[3,2-e]異喹啉]-4a',9'-二醇(中間體2a)的製備 The first step: (4'R,4a'S,7a'R,12b'S)-3'-(cyclopropylmethyl)-1',2',3',4',5',6'-hexahydro- 4a'H,7a'H-spiro[[1,3]dioxolane-2,7'-[4,12]methylbenzofuro[3,2-e]isoquinoline] Preparation of -4a',9'-diol (Intermediate 2a)

參考實施例1第二步的方法，用納曲酮代替中間體1a與乙二醇進行反應，製備中間體2a。 Referring to the method of the second step in Example 1, naltrexone was used instead of intermediate 1a to react with ethylene glycol to prepare intermediate 2a.

MS m/z(ESI)：386.2[M+H]⁺. MS m/z(ESI): 386.2[M+H] ⁺ .

第二步：(4'R,4a'S,7a'R,12b'S)-10',11'-二溴-3'-(環丙基甲基)-1',2',3',4',5',6'-六氫-4a'H,7a'H-螺環[[1,3]二氧雜環戊烷-2,7'-[4,12]甲基苯并呋喃并[3,2-e]異喹啉]-4a',9'-二醇(中間體2b)的製備 The second step: (4'R,4a'S,7a'R,12b'S)-10',11'-dibromo-3'-(cyclopropylmethyl)-1',2',3',4', 5',6'-hexahydro-4a'H,7a'H-spiro[[1,3]dioxolane-2,7'-[4,12]methylbenzofuro[3 ,2-e]isoquinoline]-4a',9'-diol (Intermediate 2b) preparation

室溫下將中間體2a(612mg)溶於0.1N TFA(100mL)。然後在攪拌下一次加入N-溴琥珀醯亞胺(850mg)。將反應混合物在室溫下攪拌24小時，加入二氯甲烷(100mL)，並用胺水將pH調到9。萃取分離後，水相用DCM再次萃取(2 X 100mL)，合併的有機相用硫酸鈉乾燥，濃縮並用矽膠管柱層析純化，得到中間體2b(310mg)。 Intermediate 2a (612 mg) was dissolved in 0.1N TFA (100 mL) at room temperature. Then N-bromosuccinimide (850 mg) was added under stirring. The reaction mixture was stirred at room temperature for 24 hours, dichloromethane (100 mL) was added, and the pH was adjusted to 9 with amine water. After extraction and separation, the aqueous phase was re-extracted with DCM (2×100 mL), and the combined organic phase was dried over sodium sulfate, concentrated and purified by silica gel column chromatography to obtain Intermediate 2b (310 mg).

MS m/z(ESI)：544.0[M+H]⁺. MS m/z(ESI): 544.0[M+H] ⁺ .

第三步：(4'R,4a'S,7a'R,12b'S)-3'-(環丙基甲基)-1',2',3',4',5',6'-六氫-4a'H,7a'H-螺環[[1,3]二氧雜環戊烷-2,7'-[4,12]甲基苯并呋喃并[3,2-e]異喹啉]-10',11'-d2-4a',9'-二醇(中間體2c)的製備 The third step: (4'R,4a'S,7a'R,12b'S)-3'-(cyclopropylmethyl)-1',2',3',4',5',6'-hexahydro- 4a'H,7a'H-spiro[[1,3]dioxolane-2,7'-[4,12]methylbenzofuro[3,2-e]isoquinoline] Preparation of -10',11'-d2-4a',9'-diol (Intermediate 2c)

將中間體2b(310mg)用5mL無水四氫呋喃溶解，加入200μL無水三乙胺，50mg 10% Pd-C，將反應瓶連接於高真空系統，通氘氣，常溫攪拌反應24小時。濾去固體，減壓蒸乾，用矽膠管柱層析分離，得到中間體2c(170mg)。 Intermediate 2b (310mg) dissolved in 5mL of anhydrous tetrahydrofuran, was added 200 μ L of anhydrous triethylamine, 50mg 10% Pd-C, and the reaction flask is connected to a high vacuum system through deuterium gas, the reaction was stirred at room temperature for 24 hours. The solid was filtered off, evaporated to dryness under reduced pressure, and separated by silica gel column chromatography to obtain Intermediate 2c (170 mg).

MS m/z(ESI)：388.2[M+H]⁺. MS m/z(ESI): 388.2[M+H] ⁺ .

第四步：(4'R,4a'S,7a'R,12b'S)-3'-(環丙基甲基)-4a'-羥基-2',3',4',4a',5',6'-六氫-1'H,7a'H-螺環[[1,3]二氧雜環戊烷-2,7'-[4,12]甲基苯并呋喃并[3,2-e]異喹啉]-9'-基-10',11'-d2三氟甲烷磺酸酯(中間體2d)的製備 The fourth step: (4'R,4a'S,7a'R,12b'S)-3'-(cyclopropylmethyl)-4a'-hydroxy-2',3',4',4a',5',6 '-Hexahydro-1'H,7a'H-spiro[[1,3]dioxolane-2,7'-[4,12]methylbenzofuro[3,2-e ]Isoquinoline]-9'-yl-10',11'-d2 Trifluoromethanesulfonate (Intermediate 2d) Preparation

參考實施例1第三步的方法，用中間體2c代替中間體1b與三氟甲磺酸酐進行反應，得到中間體2d。 Referring to the method of the third step in Example 1, Intermediate 2c was used instead of Intermediate 1b to react with trifluoromethanesulfonic anhydride to obtain Intermediate 2d.

MS m/z(ESI)：520.2[M+H]⁺. MS m/z(ESI): 520.2[M+H] ⁺ .

第四步：(4'R,4a'S,7a'R,12b'S)-3'-(環丙基甲基)-4a'-羥基-2',3',4',4a',5',6'-六氫-1'H,7a'H-螺環[[1,3]二氧雜環戊烷-2,7'-[4,12]甲基苯并呋喃并[3,2-e]異喹啉]-9'-碳腈-10',11'-d2(中間體2e)的製備 The fourth step: (4'R,4a'S,7a'R,12b'S)-3'-(cyclopropylmethyl)-4a'-hydroxy-2',3',4',4a',5',6 '-Hexahydro-1'H,7a'H-spiro[[1,3]dioxolane-2,7'-[4,12]methylbenzofuro[3,2-e ]Isoquinoline]-9'-carbonitrile-10', 11'-d2 (Intermediate 2e) preparation

參考實施例1第四步的方法，用中間體2d代替中間體1c與氰化鋅進行反應，得到中間體2e。 Referring to the method of the fourth step in Example 1, intermediate 2d was used instead of intermediate 1c to react with zinc cyanide to obtain intermediate 2e.

MS m/z(ESI)：397.2[M+H]⁺. MS m/z(ESI): 397.2[M+H] ⁺ .

第五步：(4'R,4a'S,7a'R,12b'S)-3'-(環丙基甲基)-4a'-羥基-2',3',4',4a',5',6'-六氫-1'H,7a'H-螺環[[1,3]二氧雜環戊烷-2,7'-[4,12]甲基苯并呋喃并[3,2-e]異喹啉]-10',11'-d2-9'-甲醯胺(中間體2f)的製備 The fifth step: (4'R,4a'S,7a'R,12b'S)-3'-(cyclopropylmethyl)-4a'-hydroxy-2',3',4',4a',5',6 '-Hexahydro-1'H,7a'H-spiro[[1,3]dioxolane-2,7'-[4,12]methylbenzofuro[3,2-e ]Isoquinoline]-10',11'-d2-9'-formamide (Intermediate 2f) Preparation

參考實施例1第五步的方法，用中間體2e代替中間體1d與氫氧化鉀進行反應，得到中間體2f。 With reference to the method in the fifth step of Example 1, the intermediate 2e was used instead of the intermediate 1d to react with potassium hydroxide to obtain the intermediate 2f.

MS m/z(ESI)：415.2[M+H]⁺. MS m/z(ESI): 415.2[M+H] ⁺ .

第六步：(4R,4aS,7aR,12bS)-3-(環丙基甲基)-4a-羥基-7-側氧-2,3,4,4a,5,6,7,7a-八氫-1H-4,12-甲基苯并呋喃并[3,2-e]異喹啉-10,11-d2-9-甲醯胺(中間體2g)的製備 The sixth step: (4R,4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-side oxygen-2,3,4,4a,5,6,7,7a-eight Preparation of hydrogen-1H-4,12-methylbenzofuro[3,2-e]isoquinoline-10,11-d2-9-methanamide (Intermediate 2g)

參考實施例1第六步的方法，用中間體2f代替中間體1e與鹽酸進行反應，得到中間體2g。 With reference to the method in the sixth step of Example 1, the intermediate 2f was used instead of the intermediate 1e to react with hydrochloric acid to obtain the intermediate 2g.

MS m/z(ESI)：371.2[M+H]⁺. MS m/z(ESI): 371.2[M+H] ⁺ .

第七步：(4bR,8aS,9R)-11-(環丙基甲基)-4,8a-二羥基-6-側氧-6,7,8,8a,9,10-六氫-5H-9,4b-(表胺基乙烷)菲-1,2-d2-3-甲醯胺(實施例2)的製備 The seventh step: (4bR,8aS,9R)-11-(cyclopropylmethyl)-4,8a-dihydroxy-6-oxo-6,7,8,8a,9,10-hexahydro-5H Preparation of -9,4b-(epiaminoethane)phenanthrene-1,2-d2-3-formamide (Example 2)

參考實施例1第七步的方法，用中間體2g代替中間體1f與鋅粉、鹽酸進行反應，得到實施例2。 Referring to the method in the seventh step of Example 1, using intermediate 2g instead of intermediate 1f to react with zinc powder and hydrochloric acid to obtain Example 2.

MS m/z(ESI)：373.2[M+H]⁺. MS m/z(ESI): 373.2[M+H] ⁺ .

生物學評價Biological evaluation

以下結合測試例進一步描述解釋本發明，但這些實施例並非意味著限制本發明的範圍。 The following further describes and explains the present invention in conjunction with test examples, but these examples are not meant to limit the scope of the present invention.

1.放射性配體受體結合實驗1. Radioligand receptor binding experiment

測試例1、本發明化合物對μ阿片受體結合能力的測定Test Example 1. Determination of the binding ability of the compounds of the present invention to μ opioid receptors

1.實驗目的： 1. The purpose of the experiment:

該測試例的目的是測量化合物和μ阿片受體的親和力。 The purpose of this test case is to measure the affinity of the compound to the mu opioid receptor.

2.實驗儀器和試劑： 2. Experimental instruments and reagents:

2.1 實驗儀器： 2.1 Experimental equipment:

Unifilter-96 GF/C filter plates,Perkin Elmer(Cat#6005174)； Unifilter-96 GF/C filter plates, Perkin Elmer(Cat#6005174);

384-well PP ECHO plate,Labcyte(Cat#P-05525)； 384-well PP ECHO plate, Labcyte (Cat#P-05525);

96 well conical polypropylene plates,Agilent(Cat#5042-1385)； 96 well conical polypropylene plates,Agilent(Cat#5042-1385);

TopSeal-A sealing film,Perkin Elmer(Cat#6050185)； TopSeal-A sealing film, Perkin Elmer(Cat#6050185);

Scint-tube,6mL(PerkinElmer,Cat：6000192)； Scint-tube, 6mL (PerkinElmer, Cat: 6000192);

ECHO 555(Labcyte)； ECHO 555 (Labcyte);

Bravo(Agilent)； Bravo(Agilent);

MicroBeta2(PerkinElmer)； MicroBeta2(PerkinElmer);

Tri-Carb 2910 TR(PerkinElmer)； Tri-Carb 2910 TR (PerkinElmer);

Cell harvest C961961,(Perkin Elmer)。 Cell harvest C961961, (Perkin Elmer).

2.2 實驗試劑： 2.2 Experimental reagents:

[3H]DAMG(PE-NET902250UC)； [3H]DAMG(PE-NET902250UC);

1M HEPES(Invitrogen,Cat：15630130)； 1M HEPES (Invitrogen, Cat: 15630130);

Tris base(Sigma,Cat：T1503-1KG),prepare 1M stock and adjust pH to 7.4. Tris base(Sigma,Cat:T1503-1KG),prepare 1M stock and adjust pH to 7.4.

0.5M EDTA(Invitrogen,Cat：15575-038)； 0.5M EDTA (Invitrogen, Cat: 15575-038);

1M MgCl₂(Sigma,Cat：63020)； 1M MgCl ₂ (Sigma, Cat: 63020);

BSA(Sigma,Cat：A3059-100G)； BSA (Sigma, Cat: A3059-100G);

Ascorbic acid(TCI-A0537)； Ascorbic acid (TCI-A0537);

DTT(Sigma,Cat：43815)； DTT (Sigma, Cat: 43815);

Dextran(Sigma,Cat：D1662)。 Dextran (Sigma, Cat: D1662).

2.3 受試化合物： 2.3 Test compound:

本發明實施例化合物，自製。 The example compounds of the present invention are self-made.

3.實驗方法： 3. Experimental method:

1)細胞膜蛋白配製：用實驗緩衝液稀釋細胞膜蛋白至指定濃度。 1) Preparation of cell membrane protein: Dilute cell membrane protein to the specified concentration with experimental buffer.

2)放射性同位素配體配製：用實驗緩衝液稀釋放射性同位素配體至指定濃度。 2) Preparation of radioisotope ligand: Dilute the radioisotope ligand with experimental buffer to the specified concentration.

3)轉移1μL配製好的非特異性結合化合物和DMSO至反應板中，分別作為非特異性結合孔(低信號對照：LC)和總結合孔(高信號對照：HC)。 3) Transfer 1 μ L formulated and non-specific binding compound in DMSO to the reaction plate, respectively, as non-specific binding hole (low control signals: LC) and total binding hole (high control signal: HC).

4)根據反應板的排布，加入100μL指定濃度的細胞膜蛋白。 4) The reaction plate arrangement, the indicated concentrations was added 100 μ L of cell membrane proteins.

5)再加入100μL指定濃度的放射性同位素。 5) was added 100 μ L of the indicated concentrations of radioisotopes.

6)封板後在搖床上和指定溫度條件下孵育指定的時間。 6) After sealing the plate, incubate the plate for a specified time on a shaker and at a specified temperature.

7)用50μL的0.3%PEI溶液在室溫下浸泡GF/C板半小時以上。 7) with 50 μ L of a solution of 0.3% PEI soaked GF / C plate more than half an hour at room temperature.

8)待反應結束之後，將細胞膜用cell harvest收集到GF/C過濾板上，用冷的洗板緩衝液洗4次。 8) After the reaction is over, the cell membrane is collected on the GF/C filter plate with cell harvest, and washed 4 times with cold plate washing buffer.

9)GF/C板置於50℃烘箱中烘乾1小時。 9) Place the GF/C board in an oven at 50°C for 1 hour.

10)把乾燥好的GF/C過濾板底部封膜，每孔加入50μL閃爍液，並密封。 10) The dried well GF / C filter bottom plate sealing film, was added 50 μ L per well of scintillation fluid and sealed.

11)用MicroBeta2讀數。 11) Use MicroBeta2 to read.

12)計算公式：%抑制率=100 x[1-(樣品孔讀值-非特異性結合孔讀值)/(總結合孔讀值-非特異性結合孔讀值)]。 12) Calculation formula:% inhibition rate=100 x [1-(sample well reading value-non-specific binding well reading value)/(total binding well reading value-non-specific binding well reading value)].

13)數據用GraphPad Prism 5.0中模型“log(inhibitor)vs.response--Variable slope”進行分析。 13) The data is analyzed using the model "log(inhibitor) vs.response--Variable slope" in GraphPad Prism 5.0.

4.實驗數據處理方法： 4. Experimental data processing method:

藉由MicroBeta2讀取閃爍值。根據High control(DMSO對照物)和Low control(100nM陽性化合物)實驗組的讀值計算的百分比抑制[3H]DAMG結合的數據{%抑制率=100 x[1-(樣品孔讀值-非特異性結合孔讀值)/(總結合 孔讀值-非特異性結合孔讀值)]}。使用GraphPad prism擬合百分比抑制率和多點濃度數據至參數非線性邏輯公式計算出化合物的IC₅₀值。 Read the flicker value by MicroBeta2. Percentage inhibition of [3H]DAMG binding data calculated based on the readings of High control (DMSO control) and Low control (100nM positive compound) experimental group {% inhibition rate=100 x[1-(sample well reading-non-specific Reading of sex binding wells)/(Readings of total binding wells-Readings of non-specific binding wells)]}. Use GraphPad prism to fit percent inhibition rate and multi-point concentration data to a parameter nonlinear logic formula to calculate the IC ₅₀ value of the compound.

5.實驗結果： 5. Experimental results:

藉由以上方案得出本發明所示的化合物在放射性配體受體結合試驗中顯示出約0.01nM至20nM(IC₅₀)的生物活性。 Based on the above scheme, it is concluded that the compound of the present invention shows a biological activity _{of about 0.01 nM to 20 nM (IC 50) in the radioligand receptor binding test.}

在一些實施方案中，本發明的化合物對於μ阿片受體結合的IC₅₀小於約10nM、較佳化合物小於約5nM、進一步較佳小於約1nM、更佳小於約0.5nM、本發明所列示的化合物中最佳小於0.1nM。 In some embodiments, the compounds of this invention for μ opioid receptor binding IC ₅₀ of less than about 10 nM, preferably less than about compounds of 5 nM, more preferably less than about 1 nM, more preferably less than about of 0.5 nM, as listed in the present invention The best compound is less than 0.1 nM.

6.實驗結論： 6. Experimental conclusion:

本發明實施例化合物與μ阿片受體有良好的親和力。 The compounds of the examples of the present invention have good affinity with mu opioid receptors.

測試例2、本發明化合物對MOPR受體結合能力的測定Test Example 2. Determination of the binding ability of the compound of the present invention to MOPR receptor

1.實驗目的： 1. The purpose of the experiment:

測量化合物對MOPR受體的親和力。 The affinity of the compound to the MOPR receptor is measured.

2.實驗儀器和試劑： 2. Experimental instruments and reagents:

2.1 儀器： 2.1 Instrument:

渦旋混合器(IKA；MS3 basic)； Vortex mixer (IKA; MS3 basic);

電熱恆溫培養箱(上海一恒；DHP-9032)； Electric heating constant temperature incubator (Shanghai Yiheng; DHP-9032);

微板振動篩(VWR；12620-928)； Microplate vibrating screen (VWR; 12620-928);

TopCount(PerkinElmer；NTX)； TopCount(PerkinElmer; NTX);

Universal Harvester(PerkinElmer；UNIFILTER-96)。 Universal Harvester (PerkinElmer; UNIFILTER-96).

2.2 試劑： 2.2 Reagents:

DAMGO,[Tyrosyl-3,5-3H(N)](Perkin Elmer；NET902250UC)； DAMGO,[Tyrosyl-3,5-3H(N)](Perkin Elmer;NET902250UC);

Opioid Mu(OP3)(human)membrane(Perkin Elmer；ES-542-M400UA)； Opioid Mu(OP3)(human)membrane(Perkin Elmer; ES-542-M400UA);

DAMGO(MCE；78123-71-4 2)； DAMGO(MCE; 78123-71-4 2);

ULTIMA GOLD(Perkin Elmer；77-16061)； ULTIMA GOLD (Perkin Elmer; 77-16061);

96 round deep well plate 1.1mL(Perkin Elmer；P-DW-11-C)； 96 round deep well plate 1.1mL (Perkin Elmer; P-DW-11-C);

UNIFILTER-96 GF/B filter plate(PerkinElmer；6005177)； UNIFILTER-96 GF/B filter plate (PerkinElmer; 6005177);

聚乙烯亞胺，branched(Sigma；408727)； Polyethyleneimine, branched (Sigma; 408727);

離心管(BD，352096；352070)； Centrifuge tube (BD, 352096; 352070);

Loading slot(JET BIOFIL；LTT001050)； Loading slot(JET BIOFIL; LTT001050);

移液槍頭(Axygen；T-300-R-S，T-200-Y-R-S，T-1000-B-R-S)； Pipette tips (Axygen; T-300-R-S, T-200-Y-R-S, T-1000-B-R-S);

Tris-base(Sigma；77-86-1)； Tris-base (Sigma; 77-86-1);

Bovine Serum AIbumin(Sigma；9048-46-8)； Bovine Serum AIbumin (Sigma; 9048-46-8);

HEPES(Sigma；7365-45-9)。 HEPES (Sigma; 7365-45-9).

3.實驗方法： 3. Experimental method:

1.緩衝液製備：50mM HEPES+0.025% BSA。 1. Buffer preparation: 50mM HEPES+0.025% BSA.

清洗液：50mM Tris-HCL。 Washing solution: 50mM Tris-HCL.

0.5% PEI溶液：0.5ml PEI+100ml ddH20 0.5% PEI solution: 0.5ml PEI+100ml ddH20

2.在96孔-試驗板中加入0.5-5μL待測化合物(0.005nM-100nM，共10個濃度)及100μL緩衝液。每孔中加入0.5μL細胞膜及300μL緩衝液。緩衝液中加入[³H]-DAMGO，27℃孵育1h。將加入0.5%PEI預先孵育1h的UNIFILTER-96 GF/B filter plate用1mL/well清洗緩衝液洗滌2次，並將細胞膜混懸液加入UNIFILTER-96 GF/B filter plate，並洗滌4次，55℃烘10min。每孔加入40μL ULTIMA GOLD，進行液閃計數。 2. 96 - test compound was added 0.5-5 μ L assay plates (0.005nM-100nM, a total of 10 concentrations) and 100 μ L buffer. Each well was added 0.5 μ L membrane and 300 μ L buffer. Add [ ³ H]-DAMGO to the buffer and incubate at 27°C for 1 hour. Wash the UNIFILTER-96 GF/B filter plate pre-incubated with 0.5% PEI for 1 hour with 1mL/well washing buffer twice, and add the cell membrane suspension to the UNIFILTER-96 GF/B filter plate, and wash 4 times, 55 Bake at ℃ for 10min. Each well was added 40 μ L ULTIMA GOLD, for liquid scintillation counting.

4.實驗數據處理方法： 4. Experimental data processing method:

藉由TopCount讀取CPM(Counts per minute)值。根據High control(DMSO對照物)和Low control(100nM陽性化合物)實驗組的讀值計算的百分比抑制[³H]-methylspiperone結合的數據{%抑制率=(CPM_sample-CPM_{low control})/(CPM_{high control}-CPM_{low control})×100}。化合物的濃度經過反應體系稀釋3倍後的10個濃度為100nM至0.005nM。使用GraphPad prism擬合百分比抑制率和十點濃度數據至參數非線性邏輯公式計算出化合物的IC₅₀值。 Read CPM (Counts per minute) value by TopCount. ^{Percentage inhibition of [3} H]-methylspiperone binding data calculated based on the readings of the High control (DMSO control) and Low control (100nM positive compounds) experimental group {% inhibition rate=(CPM _sample -CPM _{low control} )/(CPM _{high control} -CPM _{low control} )×100}. The concentration of the compound was diluted by 3 times in the reaction system, and the 10 concentrations were 100 nM to 0.005 nM. Use GraphPad prism to fit percent inhibition rate and ten-point concentration data to a parameter nonlinear logic formula to calculate the IC ₅₀ value of the compound.

5.實驗結果： 5. Experimental results:

6.實驗結論： 6. Experimental conclusion:

從表中數據可以看出，本發明所示的實施例化合物在MOPR受體結合能力實驗中顯示出良好的受體結合能力作用。 It can be seen from the data in the table that the compound of the example shown in the present invention shows a good receptor binding ability effect in the MOPR receptor binding ability experiment.

二、細胞功能實驗2. Cell function experiment

測試例1、本發明化合物在穩定表達MOPR受體細胞對cAMP含量影響的Test Example 1. The effect of the compound of the present invention on cAMP content in cells stably expressing MOPR receptors 測定Determination

1.實驗目的： 1. The purpose of the experiment:

測量化合物對MOPR受體的拮抗作用。 The antagonistic effect of the compound on the MOPR receptor is measured.

2.實驗儀器和試劑： 2. Experimental instruments and reagents:

2.1 儀器： 2.1 Instrument:

384孔-試驗板(Perkin Elmer；6007680)； 384-well test plate (Perkin Elmer; 6007680);

96-well conical btm PP Plt nature RNASE/Dnase-free plate(ThermoFisher；249944)； 96-well conical btm PP Plt nature RNASE/Dnase-free plate (ThermoFisher; 249944);

EnVision(Perkin Elmer)。 EnVision (Perkin Elmer).

2.2 試劑： 2.2 Reagents:

Fetal Bovine Serum(Gibco，10999141C)； Fetal Bovine Serum (Gibco, 10999141C);

Ham's F-12K(Kaighn's)Medium(Hyclone；SH30526.01)； Ham's F-12K (Kaighn's) Medium (Hyclone; SH30526.01);

Penicillin-Streptomycin，Liquid(Gibco；15140122)； Penicillin-Streptomycin, Liquid (Gibco; 15140122);

Hygromycin B(ThermoFisher；10687010)； Hygromycin B (ThermoFisher; 10687010);

Forskolin(Selleck，S2449)； Forskolin (Selleck, S2449);

BSA stabilizer(Perkin Elmer；CR84-100)； BSA stabilizer (Perkin Elmer; CR84-100);

cAMP kit(Perkin Elmer；TRF0263)； cAMP kit (Perkin Elmer; TRF0263);

IBMX(Sigma；I5879)； IBMX (Sigma; I5879);

HEPES(Gibco；15630080)； HEPES (Gibco; 15630080);

HBSS(Gibco；14025076)； HBSS (Gibco; 14025076);

Endomorphin 1(MCE，HY-P0185)。 Endomorphin 1 (MCE, HY-P0185).

3.實驗方法： 3. Experimental method:

1.緩衝液製備：1* HBSS+20mM HEPES+0.1% BSA+500μM IBMX。 1. Preparation of Buffer: 1 * HBSS + 20mM HEPES + 0.1% BSA + 500 μ M IBMX.

完全培養基：Ham's F12K+10% Fetal Bovine Serum+1* Penicillin-Streptomycin+800μg/mL Hygromycin B。 Complete medium: Ham's F12K + 10% Fetal Bovine Serum + 1 * Penicillin-Streptomycin + 800 μ g / mL Hygromycin B.

2.將CHO-MOPR細胞株(來自康龍化成)培養於37℃，5% CO₂環境下的完全培養基中；消化處理後將細胞重新懸浮於實驗buffer中，種到384細胞培養板中，接種密度8000每孔，每孔接種體積為15μL。 2. Cultivate the CHO-MOPR cell line (from Kanglong Chemical) in _{a complete medium at 37°C and 5% CO 2} ; after digestion, resuspend the cells in the experimental buffer and plant them in a 384 cell culture plate. The seeding density is 8000 per well, and the seeding volume per well is 15 μL .

3.將化合物用實驗緩衝液稀釋；每孔加入2.5μL的化合物，37℃培養10分鐘；用實驗緩衝液稀釋8*Endomorphin 1(80nM)和8*forskolin至(16μM)混合液；加入2.5μL稀釋好的混合液，於37℃孵育30分鐘；凍融Eu-cAMP tracer和Ulight-anti-cAMP，用lysis buffer將其稀釋；加入10μL Eu-cAMP tracer至實驗孔，然後 加入10μL Ulight-anti-cAMP至實驗孔中；將反應板於室溫200g離心30s，25℃靜置1h後，利用Envision收集數據 3. Compound diluted with assay buffer; L 2.5 μ compound was added to each well, cultivate 37 ℃ 10 min; diluted 8 * Endomorphin 1 (80nM) and with assay buffer to 8 * forskolin (16 μ M) mixture; was added 2.5 μ L of diluted mixture was incubated at 37 ℃ 30 min; Eu-cAMP tracer and freeze-thaw Ulight-anti-cAMP, and diluted with lysis buffer; L Eu-cAMP tracer were added to the experimental wells 10 μ, and then Add 10 μ L Ulight-anti-cAMP to experimental wells; the reaction plate was centrifuged at 200g at room temperature 30s, 25 deg.] C was allowed to stand after 1h, data collection using Envision

實驗數據處理方法： Experimental data processing method:

1)% inhibition計算 1)% inhibition calculation

% inhibition=100-(Signalcmpd-SignalAve_PC)/(SignalAve_VC-SignalAve_PC)×100. % inhibition=100-(Signalcmpd-SignalAve_PC)/(SignalAve_VC-SignalAve_PC)×100.

2)利用GraphPad非線性擬合公式計算化合物IC₅₀： 2) Calculate compound IC ₅₀ using GraphPad nonlinear fitting formula:

Y=Bottom+(Top-Bottom)/(1+10^((LogIC₅₀-X)*HillSlope)) Y=Bottom+(Top-Bottom)/(1+10^((LogIC ₅₀ -X)*HillSlope))

X：化合物濃度log值；Y：Inhibition % X: compound concentration log value; Y: Inhibition%

4.實驗結果： 4. Experimental results:

5.實驗結論： 5. Experimental conclusion:

從表中數據可以看出，本發明所示的實施例化合物在穩定表達MOPR受體細胞對cAMP影響實驗中顯示出良好的拮抗作用。 It can be seen from the data in the table that the compounds of the examples shown in the present invention show a good antagonistic effect in the experiment of stably expressing the MOPR receptor cells on cAMP.

測試例2、本發明化合物在穩定表達OPRD受體細胞對cAMP含量影響的測定Test Example 2. Determination of the effect of the compound of the present invention on the content of cAMP in cells stably expressing OPRD receptors

1.實驗目的： 1. The purpose of the experiment:

測量化合物對OPRD和OPRK受體的激動作用。 The agonistic effect of the compound on OPRD and OPRK receptors is measured.

2.實驗儀器和試劑： 2. Experimental instruments and reagents:

2.1 儀器： 2.1 Instrument:

384孔-試驗板(Perkin Elmer；6007680)； 384-well test plate (Perkin Elmer; 6007680);

96-well conical btm PP Plt nature RNASE/Dnase-free plate(ThermoFisher；249944)； 96-well conical btm PP Plt nature RNASE/Dnase-free plate (ThermoFisher; 249944);

EnVision(Perkin Elmer)。 EnVision (Perkin Elmer).

2.2 試劑： 2.2 Reagents:

Fetal Bovine Serum(Gibco，10999141C)； Fetal Bovine Serum (Gibco, 10999141C);

DMEM(500ML)(Hyclone；11965-092)； DMEM (500ML) (Hyclone; 11965-092);

Penicillin-Streptomycin，Liquid(Gibco；15140122)； Penicillin-Streptomycin, Liquid (Gibco; 15140122);

Hygromycin B(ThermoFisher；10687010)； Hygromycin B (ThermoFisher; 10687010);

Forskolin(Selleck，S2449)； Forskolin (Selleck, S2449);

BSA stabilizer(Perkin Elmer；CR84-100)； BSA stabilizer (Perkin Elmer; CR84-100);

cAMP kit(Perkin Elmer；TRF0263)； cAMP kit (Perkin Elmer; TRF0263);

IBMX(Sigma；I5879)； IBMX (Sigma; I5879);

HEPES(Gibco；15630080)； HEPES (Gibco; 15630080);

HBSS(Gibco；14025076)。 HBSS (Gibco; 14025076).

3.實驗方法： 3. Experimental method:

1.緩衝液製備：1* HBSS+20mM HEPES+0.1% BSA+500μM IBMX。 1. Preparation of Buffer: 1 * HBSS + 20mM HEPES + 0.1% BSA + 500 μ M IBMX.

完全培養基：DMEM+10% Fetal Bovine Serum+1* Penicillin-Streptomycin+200μg/mL Hygromycin B。 Complete medium: DMEM + 10% Fetal Bovine Serum + 1 * Penicillin-Streptomycin + 200 μ g / mL Hygromycin B.

2.將293-OPRD或293-OPRK細胞株(來自康龍化成)培養於37℃，5% CO₂環境下的完全培養基中；消化處理後將細胞重新懸浮於實驗buffer中，種到384細胞培養板中，接種密度20000每孔，每孔接種體積為15μL。 2. Cultivate the 293-OPRD or 293-OPRK cell line (from Kanglong Chemicals) in _{a complete medium at 37°C and 5% CO 2} ; after digestion, resuspend the cells in the experimental buffer to grow 384 cells In the culture plate, the seeding density is 20,000 per well, and the seeding volume per well is 15 μL .

3.將化合物用實驗緩衝液稀釋；每孔加入2.5μL的化合物，37℃培養10分鐘；用實驗緩衝液將forskolin稀釋至16μM(8*)；加入2.5μL稀釋好的 forskolin，於37℃孵育30分鐘；凍融Eu-cAMP tracer和Ulight-anti-cAMP，用lysis buffer將其稀釋；加入10μL Eu-cAMP tracer至實驗孔，然後加入10μL Ulight-anti-cAMP至實驗孔中；將反應板於室溫200g離心30s，25℃靜置1h後，利用Envision收集數據 3. Compound diluted with assay buffer; μ L of compound 2.5 was added to each well, cultivate 37 ℃ 10 min; the forskolin was diluted to 16 μ M (8 *) with assay buffer; 2.5 μ L forskolin dilution was added good, incubation at 37 ℃ 30 min; Eu-cAMP tracer and freeze-thaw Ulight-anti-cAMP, which was diluted with lysis buffer; L Eu-cAMP tracer were added to the experimental wells 10 μ, followed by addition of 10 μ L Ulight-anti-cAMP to In the experimental wells; centrifuge the reaction plate at 200g at room temperature for 30s and let it stand for 1 hour at 25°C, then collect data with Envision

實驗數據處理方法： Experimental data processing method:

1)%Activity計算 1)%Activity calculation

% Activity=(Signalcmpd-SignalAve_VC)/(SignalAve_PC-SignalAve_VC)×100. % Activity=(Signalcmpd-SignalAve_VC)/(SignalAve_PC-SignalAve_VC)×100.

2)利用GraphPad非線性擬合公式計算化合物EC₅₀： 2) non-linear fitting using GraphPad compound of formula EC _50:

Y=Bottom+(Top-Bottom)/(1+10^((LogEC₅₀-X)*HillSlope)) Y=Bottom+(Top-Bottom)/(1+10^((LogEC ₅₀ -X)*HillSlope))

X：化合物濃度log值；Y：Activity % X: compound concentration log value; Y: Activity%

4.實驗結果： 4. Experimental results:

5.實驗結論： 5. Experimental conclusion:

從表中數據可以看出，本發明所示的實施例化合物在穩定表達OPRD和OPRK受體細胞對cAMP影響實驗中顯示出良好的激動作用。 It can be seen from the data in the table that the compounds of the examples shown in the present invention show good agonistic effects in the experiment of stably expressing OPRD and OPRK receptor cells on cAMP.

三、大鼠藥物代謝動力學評價試驗3. Rat pharmacokinetic evaluation test

1.研究目的： 1. Research purpose:

以SD大鼠為受試動物，研究本發明化合物，在5mg/kg劑量下口服給藥在大鼠體內(血漿)的藥物代謝動力學行為。 SD rats were used as test animals to study the pharmacokinetic behavior of the compound of the present invention, administered orally at a dose of 5 mg/kg in rats (plasma).

2.實驗方案： 2. Experimental program:

2.1 實驗藥品： 2.1 Experimental drugs:

本發明實施例化合物，自製。 The example compounds of the present invention are self-made.

2.2 實驗動物： 2.2 Experimental animals:

SD大鼠每組3隻，雄性，上海傑思捷實驗動物有限公司，動物生產許可證號(SCXK(滬)2013-0006 N0.311620400001794)。 Each group of 3 SD rats, male, Shanghai Jiesjie Experimental Animal Co., Ltd., animal production license number (SCXK (Shanghai) 2013-0006 N0.311620400001794).

2.3 製劑處方： 2.3 Formulation prescription:

0.5%CMC-Na(1%Tween80)，超聲溶解，配製為澄清溶液或均一混懸液。 0.5% CMC-Na (1% Tween80), dissolve by ultrasound, prepare as a clear solution or homogeneous suspension.

2.4 給藥： 2.4 Administration:

大鼠禁食一夜後分別p.o.； The rats were fasted for one night and p.o. respectively;

p.o.劑量為5mg/kg，給藥體積10mL/kg。 The p.o. dose is 5 mg/kg, and the administration volume is 10 mL/kg.

2.5 樣品採集： 2.5 Sample collection:

大鼠口服給藥後，在0.25、0.5、1、2、4、6、8和24小時； After oral administration to rats, at 0.25, 0.5, 1, 2, 4, 6, 8 and 24 hours;

頸靜脈採血0.2mL，置於EDTA-K₂試管中，4℃ 6000rpm離心6min分離血漿，於-80℃保存。 0.2 mL blood was collected from the jugular vein, placed in an EDTA-K ₂ test tube, centrifuged at 6000 rpm at 4°C for 6 min to separate the plasma, and stored at -80°C.

2.6 樣品處理： 2.6 Sample processing:

1)血漿樣品40μL加入160μL乙腈沉澱，混合後3500×g離心5~20分鐘。 1) plasma sample was added 40 μ L 160 μ L acetonitrile precipitation, 3500 × g centrifugation after mixing 5 to 20 minutes.

2)取處理後上清溶液進行LC/MS/MS分析待測化合物的濃度，LC/MS/MS分析儀器：AB Sciex API 4000 Qtrap。 2) Take the processed supernatant solution and analyze the concentration of the test compound by LC/MS/MS. LC/MS/MS analysis instrument: AB Sciex API 4000 Qtrap.

2.7 液相分析： 2.7 Liquid phase analysis:

●液相條件：Shimadzu LC-20AD泵 ●Liquid phase conditions: Shimadzu LC-20AD pump

●色譜管柱：Agilent ZORBAX XDB-C18(50×2.1mm,3.5μm)移動相：A液為0.1%甲酸水溶液，B液為乙腈 ●Chromatographic column: Agilent ZORBAX XDB-C18 (50×2.1mm, 3.5 μ m) Mobile phase: A solution is 0.1% formic acid aqueous solution, B solution is acetonitrile

●流速：0.4mL/min ●Flow rate: 0.4mL/min

●沖提時間：0-4.0分鐘，沖提液如下： ●The extraction time: 0-4.0 minutes, the extraction solution is as follows:

3.試驗結果與分析 3. Test results and analysis

藥物代謝動力學主要參數用WinNonlin 6.1計算得到，大鼠藥物代謝實驗結果見下表4所示： The main parameters of pharmacokinetics were calculated with WinNonlin 6.1. The results of the rat drug metabolism experiment are shown in Table 4 below:

註：AUC；暴露量；C_max：口服給藥後化合物濃度最大值；t_max：達到C_max的時間；F：生物利用度。 Note: AUC; exposure; _Cmax : maximum compound concentration after oral administration; _tmax : time to reach _Cmax ; F: bioavailability.

3.4 實驗結論： 3.4 Experimental conclusion:

表中數據顯示，在大鼠藥物代謝動力學評價實驗中，本發明實施例1化合物口服給藥後顯示出較高的暴露量和生物利用度。 The data in the table shows that in the rat pharmacokinetic evaluation experiment, the compound of Example 1 of the present invention showed higher exposure and bioavailability after oral administration.

四、肝微粒體穩定性試驗方案Fourth, liver microsome stability test protocol

1.實驗目的 1. The purpose of the experiment

本實驗的目的是檢測實施例化合物在大鼠肝微粒體中的穩定性情況。 The purpose of this experiment is to test the stability of the compounds of the examples in rat liver microsomes.

2.實驗儀器與材料 2. Experimental equipment and materials

2.1 儀器： 2.1 Instrument:

2.2 試劑： 2.2 Reagents:

3.實驗步驟 3. Experimental steps

3.1 配製化合物工作液 3.1 Preparation of compound working solution

化合物的工作液配製：將化合物儲備溶液加入磷酸緩衝液，終濃度為20μM。 Compound working solution preparation: add the compound stock solution to phosphate buffer, the final concentration is 20μM.

3.2 配製肝微粒體工作液 3.2 Preparation of working fluid for liver microsomes

用100mM磷酸緩衝液稀釋至終濃度為0.625mg/mL。 Dilute with 100mM phosphate buffer to a final concentration of 0.625mg/mL.

3.3 準備NADPH和UDPGA 3.3 Prepare NADPH and UDPGA

稱取NADPH和UDPGA，加入100mM磷酸緩衝液，終濃度均為20mM。 Weigh NADPH and UDPGA, add 100mM phosphate buffer, the final concentration is 20mM.

3.4 準備打孔劑(Alamethicin) 3.4 Prepare punch (Alamethicin)

稱取1mg Alamethicin加入200μL DMSO，配製成5mg/mL的溶液。再用磷酸緩衝液稀釋至終濃度為50μg/mL。 Weigh 1 mg Alamethicin and add 200 μL DMSO to prepare a 5 mg/mL solution. Dilute with phosphate buffer to a final concentration of 50μg/mL.

3.5 配製反應終止液 3.5 Preparation of reaction stop solution

終止液：含有100ng/mL的鹽酸拉貝洛爾和400ng/mL甲苯磺丁脲為內標的冷乙腈。 Stop solution: cold acetonitrile containing 100ng/mL labetalol hydrochloride and 400ng/mL tolbutamide as internal standards.

3.6 孵育流程 3.6 Incubation process

在96孔板中依次加入400μL配製好的肝微粒體、25μL化合物工作液和25μL Alamethicin，於37℃預孵育10min。隨後加入50μL配製好的NADPH/UDPGA啟動反應，37℃孵育，反應體系的總體積為500μL，各成分最終含量如下： Add 400 μL of prepared liver microsomes, 25 μL of compound working solution, and 25 μL of Alamethicin to a 96-well plate in sequence, and pre-incubate at 37°C for 10 min. Then add 50μL of the prepared NADPH/UDPGA to start the reaction, and incubate at 37°C. The total volume of the reaction system is 500μL. The final content of each component is as follows:

3.7 樣品分析 3.7 Sample analysis

3.7.1 色譜條件 3.7.1 Chromatographic conditions

儀器：島津LC-30 AD Instrument: Shimadzu LC-30 AD

色譜管柱：XBridge® C18(50*4.6mm，5μm粒徑)； Chromatographic column: XBridge® C18 (50*4.6mm, 5μm particle size);

流動相：A：0.1%甲酸溶液，B：甲醇； Mobile phase: A: 0.1% formic acid solution, B: methanol;

沖洗梯度：0.2~1.6min 5%A到95%A，3.0~3.1min 95%A到5%A Washing gradient: 0.2~1.6min 5%A to 95%A, 3.0~3.1min 95%A to 5%A

運行時間：4.0min Running time: 4.0min

3.7.2 質譜條件 3.7.2 Mass spectrometry conditions

儀器：API5500型液相色譜質譜聯用儀，AB Sciex公司； Instrument: API5500 Liquid Chromatography Mass Spectrometer, AB Sciex Company;

離子源：電噴霧離子化源(ESI) Ion source: Electrospray ionization source (ESI)

乾燥氣體：N₂，溫度500℃ Dry gas: N ₂ , temperature 500℃

電噴霧電壓：5000V Electrospray voltage: 5000V

檢測方式：正離子檢測 Detection method: positive ion detection

掃描方式：反應監測(MRM)方式 Scanning mode: reaction monitoring (MRM) mode

4.實驗結果： 4. Experimental results:

註： Note:

5.實驗結論： 5. Experimental conclusion:

以上數據顯示，本發明實施例1在大鼠肝微粒體中為中等代謝。 The above data shows that Example 1 of the present invention is moderately metabolized in rat liver microsomes.

Claims (7)

一種通式(I)所示的化合物、其立體異構體或其藥學上可接受的鹽， A compound represented by general formula (I), its stereoisomers or a pharmaceutically acceptable salt thereof,

其中： in: R₁、R₂、R₃和R₄各自獨立地選自氫、氘或烷基； R ₁ , R ₂ , R ₃ and R ₄ are each independently selected from hydrogen, deuterium or alkyl; 且，R₁、R₂、R₃或R₄中至少有一個為氘。 In addition, at least one of _{R 1} , R ₂ , R ₃ or R _{4 is deuterium.} 如請求項1所述所示的化合物、其立體異構體或其藥學上可接受的鹽，其中，該通式(I)進一步如通式(II)所示： The compound, its stereoisomer or its pharmaceutically acceptable salt as described in claim 1, wherein the general formula (I) is further represented by the general formula (II):

如請求項1或2所述的化合物、其立體異構體或其藥學上可接受的鹽，其中，選自如下化合物： The compound according to claim 1 or 2, its stereoisomer or pharmaceutically acceptable salt thereof, wherein the compound is selected from the following compounds:

一種醫藥組成物，其包括治療有效劑量的請求項1至3中任一項所述的化合物、其立體異構體或其藥學上可接受的鹽以及一種或多種藥學上可接受的載體、稀釋劑或賦形劑。 A pharmaceutical composition, which comprises a therapeutically effective dose of the compound described in any one of claims 1 to 3, its stereoisomers or pharmaceutically acceptable salts thereof, and one or more pharmaceutically acceptable carriers and dilutions Agents or excipients. 一種如請求項1至3中任一項所述的化合物、其立體異構體或其藥學上可接受的鹽或請求項4所述的醫藥組成物在製備阿片受體受體調節劑藥物中的應用，該阿片受體調節劑包括阿片受體κ部分激動劑、阿片受體μ拮抗劑、阿片受體μ激動劑、阿片受體μ部分激動劑和阿片受體δ部分激動劑。 A compound according to any one of claims 1 to 3, its stereoisomers or pharmaceutically acceptable salts thereof, or the pharmaceutical composition according to claim 4 is used in the preparation of opioid receptor receptor modulator drugs The opioid receptor modulator includes an opioid receptor κ partial agonist, an opioid receptor μ antagonist, an opioid receptor μ agonist, an opioid receptor μ partial agonist, and an opioid receptor δ partial agonist. 一種如請求項1至3中任一項所述的化合物、其立體異構體或其藥學上可接受的鹽或如請求項4所述的醫藥組成物在製備治療咳嗽、腹瀉、藥物依賴性、胃腸道疾病、過敏性腸綜合症、肥胖、呼吸抑制、驚厥、疼痛、過敏、瘙癢疾病和中樞神經類疾病或/和精神類疾病等相關疾病藥物中的應用。 A compound as described in any one of claims 1 to 3, its stereoisomers or pharmaceutically acceptable salts thereof, or the pharmaceutical composition as described in claim 4 is used in the preparation of the treatment of cough, diarrhea, drug dependence , Gastrointestinal diseases, irritable bowel syndrome, obesity, respiratory depression, convulsions, pain, allergies, pruritus diseases and central nervous diseases or/and mental diseases and other related diseases. 如請求項6所述的應用，其中，該中樞神經類疾病或/和精神類疾病選自精神***症、睡眠障礙、心境障礙、精神***症譜系障礙、痙攣性障礙、記憶障礙和/或認知障礙、運動障礙、人格障礙、自閉症譜系障礙、外傷性腦損傷、血管疾病、物質濫用障礙和/或戒斷綜合症、耳鳴、抑鬱症、潮熱、經前綜合症、創傷後應激障礙、神經性貪食症、強迫症、廣泛性焦慮症、藥癮、自閉症、老年癡呆症、阿爾茲海默症、癲癇發作、神經痛、重度抑鬱症 和/或狂躁症等疾病；該藥癮包括酒精、尼古丁、阿片調節劑和/或***成癮。 The application according to claim 6, wherein the central nervous system disease or/and mental disease is selected from schizophrenia, sleep disorder, mood disorder, schizophrenia spectrum disorder, spastic disorder, memory disorder and/or cognition Disorders, movement disorders, personality disorders, autism spectrum disorders, traumatic brain injury, vascular diseases, substance abuse disorders and/or withdrawal syndromes, tinnitus, depression, hot flashes, premenstrual syndrome, post-traumatic stress Disorders, bulimia nervosa, obsessive-compulsive disorder, generalized anxiety disorder, drug addiction, autism, Alzheimer's disease, Alzheimer's disease, seizures, neuralgia, major depression And/or diseases such as mania; the drug addiction includes alcohol, nicotine, opioid modifier and/or ***e addiction.