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CN115477588B - A blood lipid-regulating compound based on the reverse cholesterol transport pathway, synthesis method and use - Google Patents

A blood lipid-regulating compound based on the reverse cholesterol transport pathway, synthesis method and use Download PDF

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CN115477588B
CN115477588B CN202211219995.1A CN202211219995A CN115477588B CN 115477588 B CN115477588 B CN 115477588B CN 202211219995 A CN202211219995 A CN 202211219995A CN 115477588 B CN115477588 B CN 115477588B
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向华
朱美旗
任胜楠
熊爽爽
哈斯
马露雨
齐麟
肖茂旭
骆国顺
陈德英
陈明琪
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China Pharmaceutical University
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Abstract

本发明公开了一种基于胆固醇逆转运途径的调血脂化合物、合成方法及用途，所述化合物具有通式(I)的化合物或其可药用的盐。本发明化合物可同时靶向PPARα与LXRα，发挥激动PPARα与LXRα的双重作用，通过胆固醇逆转运途径，对肝脏中的脂质蓄积有一定的缓解作用。

The invention discloses a blood lipid-regulating compound based on the reverse cholesterol transport pathway, a synthesis method and an application. The compound has a compound of general formula (I) or a pharmaceutically acceptable salt thereof. The compound of the present invention can simultaneously target PPARα and LXRα, exert dual effects of stimulating PPARα and LXRα, and have a certain alleviation effect on lipid accumulation in the liver through the reverse cholesterol transport pathway.

Description

一种基于胆固醇逆转运途径的调血脂化合物、合成方法及 用途A blood lipid regulating compound based on the reverse cholesterol transport pathway, synthesis method and use

技术领域Technical field

本发明涉及一种化合物、合成方法及用途，具体为一种基于胆固醇逆转运途径的调 血脂化合物、合成方法及用途。The present invention relates to a compound, a synthesis method and an application, specifically a blood lipid regulating compound based on the reverse cholesterol transport pathway, a synthesis method and an application.

背景技术Background technique

动脉粥样硬化(Atherosclerosis，AS)导致的心脑血管疾病严重危害人类健康和生命， 死亡率超过其他大多数疾病。而血脂异常是诱发心血管疾病(Cardiovasculardiseases， CVD)的主要因素之一。现有的调血脂药主要针对降低低密度脂蛋白胆固醇(Lowdensity lipid cholesterol，LDL-C)水平，其中他汀类药物使用最为广泛。药物治疗能减少发生心 血管事件的危险性，但仍有残留的心血管风险存在，并且调血脂药物存在的不同副作用 也不容忽视。因此，开发疗效显著、安全可靠的调血脂药物是药学工作者面临的重大挑 战。Cardiovascular and cerebrovascular diseases caused by atherosclerosis (AS) seriously endanger human health and life, and the mortality rate exceeds that of most other diseases. Dyslipidemia is one of the main factors inducing cardiovascular diseases (Cardiovascular diseases, CVD). Existing blood lipid-regulating drugs are mainly aimed at reducing low-density lipid cholesterol (LDL-C) levels, among which statins are the most widely used. Drug therapy can reduce the risk of cardiovascular events, but residual cardiovascular risks still exist, and the different side effects of lipid-lowering drugs cannot be ignored. Therefore, developing effective, safe and reliable lipid-regulating drugs is a major challenge faced by pharmaceutical workers.

胆固醇逆转运(Reverse cholesterol transport，RCT)是机体排出多余胆固醇的主要途 径，影响着AS的发生发展。研究发现肝X受体α(Liver X receptor α，LXRα)在胆固醇 从巨噬细胞流出过程中起关键作用。过氧化物酶体增殖物激活受体α(Peroxisomeproliferator activated receptor α，PPARα)在肝脏的脂质代谢中发挥重要作用，能够有效缓 解肝脏内脂质蓄积。针对RCT信号通路上的PPARα和LXRα设计双靶点激动活性的化合物，从理论上推测，不仅能促进巨噬细胞内的胆固醇逆转运过程，还能有效缓解脂质在 肝脏内的蓄积，达到理想的降脂效果。在对脂肪酸和胆固醇代谢调节过程中，PPARα和 LXRα这两个主要的核受体之间存在着密切联系，而且起着协同促进作用。因此，开发 PPARα和LXRα双靶点激动剂，能起到促进多余胆固醇的排泄，降低肝脏内脂质的沉积，发挥预防心血管疾病作用的同时，降低毒副作用。Reverse cholesterol transport (RCT) is the main way for the body to excrete excess cholesterol and affects the occurrence and development of AS. Studies have found that Liver X receptor α (LXRα) plays a key role in the efflux of cholesterol from macrophages. Peroxisome proliferator activated receptor α (PPARα) plays an important role in lipid metabolism in the liver and can effectively alleviate lipid accumulation in the liver. Designing dual-target agonistic compounds for PPARα and LXRα on the RCT signaling pathway, it is theoretically speculated that it can not only promote the reverse cholesterol transport process in macrophages, but also effectively alleviate the accumulation of lipids in the liver, achieving the ideal lipid-lowering effect. In the process of regulating fatty acid and cholesterol metabolism, there is a close connection between the two major nuclear receptors, PPARα and LXRα, and they play a synergistic role. Therefore, the development of dual-target agonists for PPARα and LXRα can promote the excretion of excess cholesterol, reduce lipid deposition in the liver, prevent cardiovascular disease, and reduce toxic and side effects.

现有的调血脂药主要针对降低低密度脂蛋白胆固醇(Low density lipidcholesterol， LDL-C)水平，其中他汀类药物使用最为广泛。药物治疗能减少发生心血管事件的危险 性，但仍有残留的心血管风险存在，并且调血脂药物存在的不同副作用也不容忽视。因 此，开发疗效显著、安全可靠的调血脂药物是药学工作者面临的重大挑战。Existing blood lipid-regulating drugs are mainly aimed at reducing low density lipoprotein cholesterol (LDL-C) levels, among which statins are the most widely used. Drug treatment can reduce the risk of cardiovascular events, but residual cardiovascular risks still exist, and the different side effects of lipid-adjusting drugs cannot be ignored. Therefore, developing effective, safe and reliable lipid-regulating drugs is a major challenge faced by pharmaceutical workers.

发明内容Contents of the invention

发明目的：本发明的目的在于提供一种基于胆固醇逆转运途径的调血脂化合物，其 可同时靶向PPARα与LXRα，发挥激动PPARα与LXRα的双重作用。Object of the invention: The object of the present invention is to provide a blood lipid-regulating compound based on the reverse cholesterol transport pathway, which can simultaneously target PPARα and LXRα and exert dual effects of stimulating PPARα and LXRα.

技术方案：本发明所述的具有通式(I)的化合物或其可药用的盐：Technical solution: the compound of general formula (I) of the present invention or a pharmaceutically acceptable salt thereof:

其中，in,

为碳数2～3的烷基链； It is an alkyl chain with 2 to 3 carbon atoms;

为碳数1～2的烷基链， It is an alkyl chain with 1 to 2 carbon atoms,

R1选自以下结构；R 1 is selected from the following structures;

R2为-CH3、-CH3或-CH3；R 2 is -CH 3 , -CH 3 or -CH 3 ;

R3为-O(CH2)zCH3、卤素、-H中的任一种，其中z为数字0～3中的任一个；R 3 is any one of -O(CH 2 )zCH 3 , halogen, and -H, where z is any one of the numbers 0 to 3;

r4为-CH2CH3或-H。r 4 is -CH 2 CH 3 or -H.

m，p是指苯环上两个取代基是间位或对位。m, p means that the two substituents on the benzene ring are in the meta or para position.

所述的化合物或其可药用的盐，所述卤素选自F、Cl、Br、I中的任一种。For the compound or a pharmaceutically acceptable salt thereof, the halogen is selected from any one of F, Cl, Br, and I.

所述的化合物或其可药用的盐，所述化合物选自如下：The compound or a pharmaceutically acceptable salt thereof, the compound is selected from the following:

2-[4-[2-[(3-(4-甲氧基苯氧基)丙基)(3-三氟甲基苯基)氨基]乙基]苯氧基]-2-甲基丙酸乙酯(A1)2-[4-[2-[(3-(4-methoxyphenoxy)propyl)(3-trifluoromethylphenyl)amino]ethyl]phenoxy]-2-methylpropyl Acid ethyl ester (A1)

2-[4-[2-[(3-(4-甲氧基苯氧基)丙基)(3-甲苯基)氨基]乙基]苯氧基]-2-甲基丙酸乙酯(A2)2-[4-[2-[(3-(4-methoxyphenoxy)propyl)(3-tolyl)amino]ethyl]phenoxy]-2-methylpropionate ( A2)

2-[4-[2-[(3-(4-甲氧基苯氧基)丙基)(苯基)氨基]乙基]苯氧基]-2-甲基丙酸乙酯(A3)2-[4-[2-[(3-(4-methoxyphenoxy)propyl)(phenyl)amino]ethyl]phenoxy]-2-methylpropionic acid ethyl ester (A3)

2-[4-[2-[(3-(4-甲氧基苯氧基)丙基)(3，5-二甲基苯基)氨基]乙基]苯氧基]-2-甲基丙酸乙酯(A4)2-[4-[2-[(3-(4-methoxyphenoxy)propyl)(3,5-dimethylphenyl)amino]ethyl]phenoxy]-2-methyl Ethyl propionate (A4)

2-[4-[2-[(3-(4-甲氧基苯氧基)丙基)(3，5-二三氟甲基苯基)氨基]乙基]苯氧基]-2-甲基丙酸乙酯(A5)2-[4-[2-[(3-(4-methoxyphenoxy)propyl)(3,5-ditrifluoromethylphenyl)amino]ethyl]phenoxy]-2- Ethyl Methylpropionate (A5)

2-[4-[2-[(3-(4-甲氧基苯氧基)丙基)(4-(8-三氟甲基喹啉))氨基]乙基]苯氧基]-2-甲基丙酸乙酯(A6)2-[4-[2-[(3-(4-methoxyphenoxy)propyl)(4-(8-trifluoromethylquinoline))amino]ethyl]phenoxy]-2 -Ethyl methylpropionate (A6)

2-[4-[2-[(3-(4-甲氧基苯氧基)丙基)(2-三氟乙基)氨基]乙基]苯氧基]-2-甲基丙酸乙酯(A7)Ethyl 2-[4-[2-[(3-(4-methoxyphenoxy)propyl)(2-trifluoroethyl)amino]ethyl]phenoxy]-2-methylpropionate Esters (A7)

2-[4-[2-[(3-(4-甲氧基苯氧基)丙基)(4-三氟甲基苯甲基)氨基]乙基]苯氧基]-2-甲基丙酸乙酯(A8)2-[4-[2-[(3-(4-methoxyphenoxy)propyl)(4-trifluoromethylbenzyl)amino]ethyl]phenoxy]-2-methyl Ethyl propionate (A8)

2-[4-[2-[(3-(4-甲氧基苯氧基)丙基)-5-喹啉氨基]乙基]苯氧基]-2-甲基丙酸(A9)2-[4-[2-[(3-(4-methoxyphenoxy)propyl)-5-quinolinino]ethyl]phenoxy]-2-methylpropionic acid (A9)

2-[4-[2-[(3-(4-甲氧基苯氧基)丙基)(3-三氟甲基苯基)氨基]乙基]苯氧基]-2-甲基丙酸(A10)2-[4-[2-[(3-(4-methoxyphenoxy)propyl)(3-trifluoromethylphenyl)amino]ethyl]phenoxy]-2-methylpropyl Acid(A10)

2-[4-[2-[(3-(4-甲氧基苯氧基)丙基)(3-甲苯基)氨基]乙基]苯氧基]-2-甲基丙酸(A11)2-[4-[2-[(3-(4-methoxyphenoxy)propyl)(3-tolyl)amino]ethyl]phenoxy]-2-methylpropionic acid (A11)

2-[4-[2-[(3-(4-甲氧基苯氧基)丙基)(苯基)氨基]乙基]苯氧基]-2-甲基丙酸(A12)2-[4-[2-[(3-(4-methoxyphenoxy)propyl)(phenyl)amino]ethyl]phenoxy]-2-methylpropionic acid (A12)

2-[4-[2-[(3-(4-甲氧基苯氧基)丙基)(3，5-二甲基苯基)氨基]乙基]苯氧基]-2-甲基丙酸(A13)2-[4-[2-[(3-(4-methoxyphenoxy)propyl)(3,5-dimethylphenyl)amino]ethyl]phenoxy]-2-methyl Propionic acid(A13)

2-[4-[2-[(3-(4-甲氧基苯氧基)丙基)(3，5-二三氟甲基苯基)氨基]乙基]苯氧基]-2-甲基丙酸(A14)2-[4-[2-[(3-(4-methoxyphenoxy)propyl)(3,5-ditrifluoromethylphenyl)amino]ethyl]phenoxy]-2- Methylpropionic acid (A14)

2-[4-[2-[(3-(4-甲氧基苯氧基)丙基)(2-三氟乙基)氨基]乙基]苯氧基]-2-甲基丙酸(A15)2-[4-[2-[(3-(4-methoxyphenoxy)propyl)(2-trifluoroethyl)amino]ethyl]phenoxy]-2-methylpropionic acid ( A15)

2-[4-[2-[(3-(4-甲氧基苯氧基)丙基)(4-三氟甲基苯甲基)氨基]乙基]苯氧基]-2-甲基丙酸(A16)2-[4-[2-[(3-(4-methoxyphenoxy)propyl)(4-trifluoromethylbenzyl)amino]ethyl]phenoxy]-2-methyl Propionic acid (A16)

2-[4-[2-[(3-(4-甲氧基苯氧基)丙基)(3，5-二三氟甲基苯基)氨基]乙基]苯氧基]-2-甲基丙酸(B1)2-[4-[2-[(3-(4-methoxyphenoxy)propyl)(3,5-ditrifluoromethylphenyl)amino]ethyl]phenoxy]-2- Methylpropionic acid (B1)

2-[4-[2-[(3-(4-甲氧基苯氧基)丙基)(3，5-二三氟甲基苯基)氨基]甲基]苯氧基]-丙酸(B2)2-[4-[2-[(3-(4-methoxyphenoxy)propyl)(3,5-ditrifluoromethylphenyl)amino]methyl]phenoxy]-propionic acid (B2)

2-[4-[2-[(3-(4-甲氧基苯氧基)丙基)(3，5-二三氟甲基苯基)氨基]乙基]苯氧基]-丁酸(B3)2-[4-[2-[(3-(4-methoxyphenoxy)propyl)(3,5-ditrifluoromethylphenyl)amino]ethyl]phenoxy]-butyric acid (B3)

2-[4-[1-[(2-(4-甲氧基苯氧基)乙基)(3，5-二三氟甲基苯基)氨基]甲基]苯氧基]-2-甲基丙酸(B4)2-[4-[1-[(2-(4-methoxyphenoxy)ethyl)(3,5-ditrifluoromethylphenyl)amino]methyl]phenoxy]-2- Methylpropionic acid (B4)

2-[4-[1-[(3-(4-氟苯氧基)丙基)(3，5-二三氟甲基苯基)氨基]甲基]苯氧基]-2-甲基丙酸(B5)2-[4-[1-[(3-(4-fluorophenoxy)propyl)(3,5-ditrifluoromethylphenyl)amino]methyl]phenoxy]-2-methyl Propionic acid (B5)

2-[4-[1-[(3-苯氧基丙基)(3，5-二三氟甲基苯基)氨基]甲基]苯氧基]-2-甲基丙酸(B6)2-[4-[1-[(3-phenoxypropyl)(3,5-ditrifluoromethylphenyl)amino]methyl]phenoxy]-2-methylpropionic acid (B6)

2-[4-[1-[(3-(4-甲氧基苯氧基)丙基)(3，5-二三氟甲基苯基)氨基]甲基]苯氧基]-2-甲基丙酸(B7)。2-[4-[1-[(3-(4-methoxyphenoxy)propyl)(3,5-ditrifluoromethylphenyl)amino]methyl]phenoxy]-2- Methylpropionic acid (B7).

所述的化合物或其可药用的盐的合成方法，包括以下步骤：The synthesis method of the compound or its pharmaceutically acceptable salt includes the following steps:

(1)化合物1与2在碱性条件下制得化合物3；(1) Compound 3 is prepared from compounds 1 and 2 under alkaline conditions;

(2)化合物3在酸性条件下脱去Boc得到化合物4；(2) Compound 3 removes Boc under acidic conditions to obtain compound 4;

(3)化合物4与5在酰胺缩合剂HATU的催化下生成酰胺缩合产物6；(3) Compounds 4 and 5 generate amide condensation product 6 under the catalysis of amide condensation agent HATU;

(4)化合物6在LiAlH4的催化下还原羰基生成仲胺化合物7；(4) Compound 6 reduces the carbonyl group under the catalysis of LiAlH 4 to generate secondary amine compound 7;

(5)化合物7与8在碱性条件下发生亲核取代反应生成化合物11；(5) Compounds 7 and 8 undergo nucleophilic substitution reaction under alkaline conditions to generate compound 11;

(6)化合物9与8在碱性条件下发生亲核取代反应生成化合物10；(6) Compounds 9 and 8 undergo nucleophilic substitution reaction under alkaline conditions to generate compound 10;

(7)化合物4与10在三乙酰氧基硼氢化钠作用下得到化合物11；(7) Compounds 4 and 10 are treated with sodium triacetoxyborohydride to obtain compound 11;

(8)化合物11与12经过亲核取代或者碳氮偶联反应(Buchwald-Hartwig反应) 生成化合物13；(8) Compounds 11 and 12 undergo nucleophilic substitution or carbon-nitrogen coupling reaction (Buchwald-Hartwig reaction) to generate compound 13;

(7)化合物13在在碱性条件下水解得到化合物14，反应式如下所示：(7) Compound 13 is hydrolyzed under alkaline conditions to obtain compound 14. The reaction formula is as follows:

所述的化合物或其可药用的盐的合成方法，反应条件分别是：(a)NaH，DMF，室温；(b)HCl，CH2Cl2，室温；(c)4-Hydroxyphenylacetic acid，DIPEA，HATU，DMF；(d)LiAlH4，CH2Cl2或THF；(e)Cs2CO3，DMF或MeCN；(f)Sodium triacetoxyborohydride，THF，室 温；(g)Chlorinated or brominated substratesubstrate，Cs2CO3，Pd2(dba)3，X-Phos， toluene，110℃；(h)KOH，THF和H2O。The synthesis method of the compound or its pharmaceutically acceptable salt, the reaction conditions are: (a) NaH, DMF, room temperature; (b) HCl, CH 2 Cl 2 , room temperature; (c) 4-Hydroxyphenylacetic acid, DIPEA , HATU, DMF; (d) LiAlH 4 , CH 2 Cl 2 or THF; (e) Cs 2 CO 3 , DMF or MeCN; (f) Sodium triacetoxyborohydride, THF, room temperature; (g) Chlorinated or brominated substrate substrate, Cs 2 CO 3 , Pd 2 (dba) 3 , X-Phos, toluene, 110°C; (h) KOH, THF and H 2 O.

药物组合物，所述的化合物或其可药用的盐、及药学上可接受的载体。Pharmaceutical composition, the compound or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

所述的化合物或其可药用的盐或所述药物组合物在制备PPARα与LXRα激动剂中的用途。The use of the compound or its pharmaceutically acceptable salt or the pharmaceutical composition in the preparation of PPARα and LXRα agonists.

所述的化合物或其可药用的盐或所述药物组合物在制备胆固醇逆转运药物中的用 途。The use of the compound or a pharmaceutically acceptable salt thereof or the pharmaceutical composition in the preparation of cholesterol reverse transport drugs.

所述的化合物或其可药用的盐或所述药物组合物在制备调血脂药物中的用途。The use of the compound or its pharmaceutically acceptable salt or the pharmaceutical composition in the preparation of blood lipid-regulating drugs.

所述的化合物或其可药用的盐或所述药物组合物在制备治疗肝脏脂质蓄积药物中 的用途。The use of the compound or a pharmaceutically acceptable salt thereof or the pharmaceutical composition in the preparation of a drug for treating liver lipid accumulation.

所述的化合物或其可药用的盐或所述药物组合物在制备治疗脂肪肝药物中的用途。The use of the compound or a pharmaceutically acceptable salt thereof or the pharmaceutical composition in preparing a drug for treating fatty liver.

本发明的技术关键点在于：合成一类药物组合物，其含有治疗有效量的一种或多种 所述的具有通式(I)的调血脂化合物或其可药用的盐，及药学上可接受的载体。The technical key point of the present invention is to synthesize a type of pharmaceutical composition, which contains a therapeutically effective amount of one or more of the blood lipid-regulating compounds of general formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition. Acceptable carrier.

有益效果：本发明与现有技术相比，具有如下优点：本发明化合物在HepG2细胞及RAW264.7细胞中有一定的脂质蓄积抑制作用。本发明化合物B4在20μM的浓度下能够 有效的降低HepG2细胞RAW264.7细胞中的总胆固醇水平，进一步说明化合物对肝脏中的脂质蓄积有一定的缓解作用。Beneficial effects: Compared with the prior art, the present invention has the following advantages: the compound of the present invention has a certain inhibitory effect on lipid accumulation in HepG2 cells and RAW264.7 cells. Compound B4 of the present invention can effectively reduce the total cholesterol level in HepG2 cells RAW264.7 cells at a concentration of 20 μM, further indicating that the compound has a certain alleviating effect on lipid accumulation in the liver.

附图说明Description of the drawings

图1为化合物在成脂肪细胞中的脂质蓄积抑制能力测试；Figure 1 shows the test of the compound’s ability to inhibit lipid accumulation in adipocytes;

图2为HepG2细胞油红染色半定量测试；Figure 2 shows the semi-quantitative test of oil red staining of HepG2 cells;

图3为HepG2细胞油红O染色成像(×20)；(A)Control组；(B)OA酯化模 型组；(C)B4给药组；(D)T0901317给药组；(E)Fenofibrate acid给药组；Figure 3 shows the Oil Red O staining imaging of HepG2 cells (×20); (A) Control group; (B) OA esterification model group; (C) B4 administration group; (D) T0901317 administration group; (E) Fenofibrate acid administration group;

图4为RAW264.7细胞油红染色半定量测试；Figure 4 shows the semi-quantitative test of oil red staining of RAW264.7 cells;

图5为RAW264.7细胞油红O染色成像(×20)；(A)Control组；(B)OA酯化 模型组；(C)T0901317给药组；(D)B4给药组；(E)Fenofibrate acid给药组；Figure 5 shows oil red O staining imaging of RAW264.7 cells (×20); (A) Control group; (B) OA esterification model group; (C) T0901317 administration group; (D) B4 administration group; (E )Fenofibrate acid administration group;

图6为细胞内总胆固醇测定，其中(A)为HepG2细胞内总胆固醇(TC)测定； (B)为RAW264.7细胞内总胆固醇(TC)测定。Figure 6 shows the measurement of total cholesterol in cells, where (A) is the measurement of total cholesterol (TC) in HepG2 cells; (B) is the measurement of total cholesterol (TC) in RAW264.7 cells.

具体实施方式Detailed ways

实施例1Example 1

2-[4-[2-[(3-(4-甲氧基苯氧基)丙基)(3-三氟甲基苯基)氨基]乙基]苯氧基]-2-甲基丙酸乙酯(A1)的合成2-[4-[2-[(3-(4-methoxyphenoxy)propyl)(3-trifluoromethylphenyl)amino]ethyl]phenoxy]-2-methylpropyl Synthesis of acid ethyl ester (A1)

合成路线：synthetic route:

步骤1：step 1:

将NaH(1.81g，75mmol)加入无水DMF(5mL)中，N2置换。在冰水浴下缓慢 滴加化合物15对羟基苯甲醚(7.81g，62.96mmol)的DMF(10mL)。0℃下搅拌20min 后，撤去冰水浴，缓慢加入N-Boc-3-氨基丙基溴(10g，41.99mmol)的DMF(10mL) 溶液。将反应在室温下搅拌直至反应结束，过程用TLC检测。反应结束后，加入100mL 水淬灭反应，加NaOH溶液调PH至碱性，用二氯甲烷(100mL×3)萃取，合并有几层， 用无水Na2SO4干燥，减压浓缩至无溶剂蒸出，得无色油状化合物粗品1611.25g，无需 纯化，直接下投。NaH (1.81g, 75mmol) was added to anhydrous DMF (5mL) and replaced with N2 . Compound 15 p-hydroxyanisole (7.81 g, 62.96 mmol) in DMF (10 mL) was slowly added dropwise under an ice-water bath. After stirring at 0°C for 20 min, the ice-water bath was removed, and a solution of N-Boc-3-aminopropyl bromide (10 g, 41.99 mmol) in DMF (10 mL) was slowly added. The reaction was stirred at room temperature until completion, and the progress was monitored by TLC. After the reaction is completed, add 100 mL of water to quench the reaction, add NaOH solution to adjust the pH to alkaline, extract with dichloromethane (100 mL The solvent was evaporated to obtain 1611.25g of crude colorless oily compound, which was directly added without purification.

步骤2：Step 2:

将上述所得化合物16(11.25g，40.17mmol)溶于20mL乙酸乙酯中，加入10mL 浓盐酸，室温下搅拌30min，并通过TLC监测反应直至完成。将反应液倒入50mL水 中，并用饱和NaHCO3溶液调节pH至中性。乙酸乙酯萃取(50mL×3)，合并有机层， 无水Na2SO4干燥，减压浓缩至干，得化合物176.62g，无需纯化直接用于下一步。1H NMR (400MHz，DMSO-d6)δ6.84(d，J＝0.8Hz，4H)，3.94(t，J＝6.4Hz，2H)，3.68(s，3H)，2.71(t， J＝6.8Hz，2H)，1.84-1.71(m，2H).Compound 16 (11.25 g, 40.17 mmol) obtained above was dissolved in 20 mL of ethyl acetate, 10 mL of concentrated hydrochloric acid was added, stirred at room temperature for 30 min, and the reaction was monitored by TLC until completion. Pour the reaction solution into 50 mL of water, and adjust the pH to neutral with saturated NaHCO solution . Extract with ethyl acetate (50 mL×3), combine the organic layers, dry over anhydrous Na 2 SO 4 , and concentrate to dryness under reduced pressure to obtain 176.62 g of compound, which can be used directly in the next step without purification. 1 H NMR (400MHz, DMSO-d 6 ) δ6.84 (d, J=0.8Hz, 4H), 3.94 (t, J=6.4Hz, 2H), 3.68 (s, 3H), 2.71 (t, J= 6.8Hz, 2H), 1.84-1.71(m, 2H).

步骤3：Step 3:

N2保护下，将对羟基苯乙酸(8.34g，54.81mmol)、N，N-二异丙基乙胺(14.25g，11.02 mmol)及HATU(20.85g，54.83mmol)加入到15mL无水DMF中，0℃反应20min 后，加入化合物17(6.62g，36.99mmol)的DMF(10mL)溶液，室温反应3h，反应 过程中用TLC监测反应的进行。反应完成后加水淬灭反应，用DCM(100mL×3)萃取， 合并有机相。有机相依次用饱和Na2CO3(100mL×3)、NaCl(100mL×3)洗涤，收集 有机相，用无水Na2SO4干燥，减压浓缩至干，经柱层析后得化合物18(8.75g，产率76％)。1H NMR(300MHz，DMSO-d6)δ9.22(s，1H)，7.03(d，J＝8.6Hz，2H)，6.82(d，J＝2.2Hz， 4H)，6.66(d，J＝8.5Hz，2H)，3.87(t，J＝6.3Hz，2H)，3.69(s，3H)，3.25(s，2H)，3.17(q，J＝ 6.4Hz，2H)，1.79(p，J＝6.6Hz，2H).MS ESI m/z316.1[M+H]+.Under N protection, add p-hydroxyphenylacetic acid (8.34g, 54.81mmol), N,N-diisopropylethylamine (14.25g, 11.02mmol) and HATU (20.85g, 54.83mmol) to 15mL anhydrous DMF After reacting at 0°C for 20 min, a solution of compound 17 (6.62 g, 36.99 mmol) in DMF (10 mL) was added, and the reaction was carried out at room temperature for 3 h. The progress of the reaction was monitored by TLC during the reaction. After the reaction is completed, add water to quench the reaction, extract with DCM (100 mL×3), and combine the organic phases. The organic phase was washed with saturated Na 2 CO 3 (100 mL × 3) and NaCl (100 mL × 3) in sequence. The organic phase was collected, dried over anhydrous Na 2 SO 4 , and concentrated to dryness under reduced pressure. Compound 18 was obtained after column chromatography. (8.75g, yield 76%). 1 H NMR (300MHz, DMSO-d 6 ) δ9.22 (s, 1H), 7.03 (d, J=8.6Hz, 2H), 6.82 (d, J=2.2Hz, 4H), 6.66 (d, J= 8.5Hz, 2H), 3.87 (t, J=6.3Hz, 2H), 3.69 (s, 3H), 3.25 (s, 2H), 3.17 (q, J= 6.4Hz, 2H), 1.79 (p, J= 6.6Hz, 2H).MS ESI m/z316.1[M+H] + .

步骤4：Step 4:

将LiAlH4(1.68g，44.27mmol)加入DCM(5mL)中，0℃下缓慢将化合物18(3.36 g，11.16mmol)的DCM(10mL)溶液。滴加完毕后反应液于38℃回流反应，TLC检 测直至反应完成。后处理：用冰水在0℃下淬灭反应，过滤除去不溶物，用DCM(5mL×2) 洗涤滤饼，滤液用DCM(30mL×3)萃取，合并有机相，无水Na2SO4干燥后减压浓缩， 经柱层析后得化合物19(1.44g，产率43％)。1H NMR(300MHz，DMSO-d6)δ9.36(s，1H)， 7.04(d，J＝8.0Hz，2H)，6.86(s，4H)，6.72(d，J＝8.0Hz，2H)，3.98(t，J＝6.1Hz，2H)，3.69 (s，3H)，3.02(p，J＝5.0，4.2Hz，4H)，2.83(dd，J＝10.3，6.0Hz，2H)，2.11-1.95(m，2H).MS ESI m/z 302.2[M+H]+.LiAlH 4 (1.68 g, 44.27 mmol) was added to DCM (5 mL), and a solution of compound 18 (3.36 g, 11.16 mmol) in DCM (10 mL) was slowly added at 0°C. After the dropwise addition, the reaction solution was refluxed at 38°C, and TLC was detected until the reaction was completed. Post-treatment: Quench the reaction with ice water at 0°C, filter to remove insoluble matter, wash the filter cake with DCM (5mL×2), extract the filtrate with DCM (30mL×3), combine the organic phases, and add anhydrous Na 2 SO 4 After drying and concentration under reduced pressure, compound 19 (1.44g, yield 43%) was obtained through column chromatography. 1 H NMR (300MHz, DMSO-d 6 ) δ9.36 (s, 1H), 7.04 (d, J=8.0Hz, 2H), 6.86 (s, 4H), 6.72 (d, J=8.0Hz, 2H) , 3.98 (t, J=6.1Hz, 2H), 3.69 (s, 3H), 3.02 (p, J=5.0, 4.2Hz, 4H), 2.83 (dd, J=10.3, 6.0Hz, 2H), 2.11- 1.95(m,2H).MS ESI m/z 302.2[M+H] + .

步骤5：Step 5:

将化合物19(0.34g，1.13mmol)、2-溴-2-甲基丙酸乙酯(0.28g，1.44mmol)、Cs2CO3(0.74g，2.27mmol)加入无水DMF(5mL)中，室温下反应搅拌过夜。后处理：用水淬灭反应，DCM(10mL×3)萃取，合并有机相，无水Na2SO4干燥后减压浓缩，经柱 层析后得化合物20(0.27g，产率57％)。1H NMR(300MHz，Methanol-d4)δ7.15(d，J＝ 8.5Hz，2H)，6.81(s，4H)，6.78(d，J＝8.5Hz，2H)，4.18(q，J＝7.1Hz，2H)，3.99(t，J＝5.8 Hz，2H)，3.71(s，3H)，3.14-3.06(m，4H)，2.89(t，J＝6Hz，2H)，2.08(p，J＝6.1Hz，2H)， 1.52(s，6H)，1.21(t，J＝7.1Hz，3H).MSESI m/z 416.2[M+H]+.Compound 19 (0.34g, 1.13mmol), ethyl 2-bromo-2-methylpropionate (0.28g, 1.44mmol), Cs 2 CO 3 (0.74g, 2.27mmol) were added to anhydrous DMF (5mL) , the reaction was stirred overnight at room temperature. Post-treatment: quench the reaction with water, extract with DCM (10 mL×3), combine the organic phases, dry over anhydrous Na 2 SO 4 and concentrate under reduced pressure, and obtain compound 20 (0.27 g, yield 57%) after column chromatography. 1 H NMR (300MHz, Methanol-d 4 ) δ7.15 (d, J= 8.5Hz, 2H), 6.81 (s, 4H), 6.78 (d, J=8.5Hz, 2H), 4.18 (q, J= 7.1Hz, 2H), 3.99 (t, J=5.8 Hz, 2H), 3.71 (s, 3H), 3.14-3.06 (m, 4H), 2.89 (t, J=6Hz, 2H), 2.08 (p, J ＝6.1Hz, 2H), 1.52(s, 6H), 1.21(t, J＝7.1Hz, 3H).MSESI m/z 416.2[M+H] + .

步骤6：Step 6:

将化合物20(1g，2.41mmol)、3-碘三氟甲苯(1.32g，4.85mmol)加入甲苯(6mL) 中，随后加入Cs2CO3(1.56g，4.79mmol)、X-phos(58mg，0.12mmol)、Pd2(dba)3(110 mg，0.12μmol)，N2置换。反应液110℃反应过夜。后处理：加水淬灭反应，EA(5mL×3) 萃取，无水Na2SO4干燥后减压浓缩，经柱层析后得橙黄色油状化合物A1(0.44g，产率33％)。1H NMR(300MHz，Chloroform-d)δ7.30(t，J＝8.0Hz，1H)，7.06(d，J＝8.5Hz，2H)， 6.96-6.87(m，3H)，6.85(s，4H)，6.81(d，J＝8.4Hz，2H)，4.25(q，J＝7.1Hz，2H)，3.92(t，J ＝5.7Hz，2H)，3.78(s，3H)，3.55(t，J＝7.5Hz，2H)，3.46(t，J＝7.0Hz，2H)，2.81(t，J＝7.5 Hz，2H)，1.98(p，J＝6.2Hz，2H)，1.60(s，6H)，1.27(t，J＝7.1Hz，3H).13C NMR(101MHz， Methanol-d4)δ175.70，155.37，155.28，154.31，149.33，134.79，132.46(q，J＝31.3Hz，1C)， 130.91，130.61，126.09(q，J＝272.7Hz，1C)，120.82，116.43，116.27，115.68，112.74(q，J＝ 4.0Hz，1C)，108.93(q，J＝4.0Hz，1C)，80.36，66.58，62.50，56.08，53.92，48.76，33.38，28.05， 25.71，14.37.HRMS(ESI)m/z calcd for C31H36F3No5[M+H]+560.2618，found 560.2615.Compound 20 (1g, 2.41mmol) and 3-iodotrifluorotoluene (1.32g, 4.85mmol) were added to toluene (6mL), followed by Cs 2 CO 3 (1.56g, 4.79mmol), X-phos (58mg, 0.12mmol), Pd 2 (dba) 3 (110 mg, 0.12 μmol), N 2 replacement. The reaction solution was reacted at 110°C overnight. Post-treatment: add water to quench the reaction, extract with EA (5mL×3), dry over anhydrous Na 2 SO 4 and concentrate under reduced pressure. After column chromatography, an orange-yellow oily compound A1 (0.44g, yield 33%) was obtained. 1 H NMR (300MHz, Chloroform-d) δ7.30 (t, J=8.0Hz, 1H), 7.06 (d, J=8.5Hz, 2H), 6.96-6.87 (m, 3H), 6.85 (s, 4H ), 6.81 (d, J = 8.4Hz, 2H), 4.25 (q, J = 7.1Hz, 2H), 3.92 (t, J = 5.7Hz, 2H), 3.78 (s, 3H), 3.55 (t, J =7.5Hz, 2H), 3.46 (t, J = 7.0Hz, 2H), 2.81 (t, J = 7.5 Hz, 2H), 1.98 (p, J = 6.2Hz, 2H), 1.60 (s, 6H), 1.27 (t, J=7.1Hz, 3H). 13 C NMR (101MHz, Methanol-d 4 ) δ 175.70, 155.37, 155.28, 154.31, 149.33, 134.79, 132.46 (q, J=31.3Hz, 1C), 130.91 , 130.61, 126.09 (q, J = 272.7Hz, 1C), 120.82, 116.43, 116.27, 115.68, 112.74 (q, J = 4.0Hz, 1C), 108.93 (q, J = 4.0Hz, 1C), 80.36, 66.58 , 62.50, 56.08, 53.92, 48.76, 33.38, 28.05, 25.71, 14.37.HRMS(ESI)m/z calcd for C 31 H 36 F 3 No 5 [M+H] + 560.2618, found 560.2615.

实施例2Example 2

2-[4-[2-[(3-(4-甲氧基苯氧基)丙基)(3-三氟甲基苯基)氨基]乙基]苯氧基]-2-甲基丙酸 (A10)的合成2-[4-[2-[(3-(4-methoxyphenoxy)propyl)(3-trifluoromethylphenyl)amino]ethyl]phenoxy]-2-methylpropyl Synthesis of acid (A10)

步骤1：step 1:

将化合物A1(0.28g，0.5mmol)溶于3mL THF以及3mL水的混合溶液中，加入 KOH(0.14g，2.5mmol)至反应液中，室温反应，TLC监测至反应结束。后处理：加入 稀盐酸调pH至中性，减压浓缩至无旋出液，剩余溶液用EA(6mL×3)萃取，合并有 机相，无水Na2SO4干燥后减压浓缩，柱层析后得化合物A10(0.13g，产率48％)。化 合物A10为淡黄色油状物。1H NMR(300MHz，Methanol-d4)δ7.15(t，J＝7.9Hz，1H)， 6.91(d，J＝8.1Hz，2H)，6.78(d，J＝11.8Hz，2H)，6.71(d，J＝9.6Hz，7H)，3.72(t，J＝5.6 Hz，2H)，3.58(s，3H)，3.38(t，J＝7.4Hz，2H)，3.28(t，J＝7.1Hz，2H)，2.63(t，J＝7.3Hz， 2H)，1.79-1.66(m，2H)，1.40(s，6H).13C NMR(101MHz，Methanol-d4)δ177.89，155.36， 155.27，154.30，149.33，134.77，132.46(q，J＝31.3Hz，1C)，130.90，130.56，126.08(q，J＝ 272.7Hz，1C)，121.09，116.44，116.27，115.68，112.76(q，J＝4.0Hz，1C)，108.94(q，J＝4.0 Hz，1C)，80.24，66.59，56.08，53.94，33.38，28.06，25.71.HRMS(ESI)m/z calcd for C29H32F3NO5[M+H]+532.2305，found532.2303.Compound A1 (0.28g, 0.5mmol) was dissolved in a mixed solution of 3mL THF and 3mL water, KOH (0.14g, 2.5mmol) was added to the reaction solution, and the reaction was carried out at room temperature. TLC monitored until the reaction was completed. Post-processing: Add dilute hydrochloric acid to adjust the pH to neutral, concentrate under reduced pressure until there is no spin-off solution, extract the remaining solution with EA (6mL×3), combine the organic phases, dry over anhydrous Na 2 SO 4 and concentrate under reduced pressure, column layer After analysis, compound A10 (0.13g, yield 48%) was obtained. Compound A10 is a light yellow oil. 1 H NMR (300MHz, Methanol-d 4 ) δ7.15 (t, J=7.9Hz, 1H), 6.91 (d, J=8.1Hz, 2H), 6.78 (d, J=11.8Hz, 2H), 6.71 (d, J=9.6Hz, 7H), 3.72 (t, J=5.6 Hz, 2H), 3.58 (s, 3H), 3.38 (t, J=7.4Hz, 2H), 3.28 (t, J=7.1Hz , 2H), 2.63 (t, J=7.3Hz, 2H), 1.79-1.66 (m, 2H), 1.40 (s, 6H). 13 C NMR (101MHz, Methanol-d 4 ) δ 177.89, 155.36, 155.27 , 154.30, 149.33, 134.77, 132.46 (q, J=31.3Hz, 1C), 130.90, 130.56, 126.08 (q, J= 272.7Hz, 1C), 121.09, 116.44, 116.27, 115.68, 112.76 (q, J= 4. 0 Hz, 1C), 108.94 (q, J=4.0 Hz, 1C), 80.24, 66.59, 56.08, 53.94, 33.38, 28.06, 25.71.HRMS (ESI) m/z calcd for C 29 H 32 F 3 NO 5 [M +H] +532.2305 , found532.2303.

实施例3Example 3

A2的合成Synthesis of A2

参照实施例1的合成方法，将3-碘三氟甲苯替换成间溴甲苯，其他条件不变。1HNMR(300MHz，Methanol-d4)δ7.05-6.97(m，3H)，6.81(s，4H)，6.76-6.70(m，2H)，6.52 (d，J＝6.5Hz，2H)，6.41(d，J＝7.4Hz，1H)，4.18(q，J＝7.1Hz，2H)，3.85(t，J＝5.7Hz，2H)， 3.71(s，3H)，3.45(t，J＝7.4Hz，2H)，3.34(t，J＝6.9Hz，2H)，2.74(t，J＝7.3Hz，2H)，2.19(s，3H)，1.86(p，J＝6.1Hz，2H)，1.50(s，6H)，1.21(t，J＝7.1Hz，3H).Referring to the synthesis method of Example 1, 3-iodotrifluorotoluene was replaced with m-bromotoluene, and other conditions remained unchanged. 1 HNMR (300MHz, Methanol-d 4 ) δ7.05-6.97 (m, 3H), 6.81 (s, 4H), 6.76-6.70 (m, 2H), 6.52 (d, J=6.5Hz, 2H), 6.41 (d, J=7.4Hz, 1H), 4.18 (q, J=7.1Hz, 2H), 3.85 (t, J=5.7Hz, 2H), 3.71 (s, 3H), 3.45 (t, J=7.4Hz , 2H), 3.34 (t, J=6.9Hz, 2H), 2.74 (t, J=7.3Hz, 2H), 2.19 (s, 3H), 1.86 (p, J=6.1Hz, 2H), 1.50 (s , 6H), 1.21 (t, J=7.1Hz, 3H).

实施例4Example 4

A3的合成Synthesis of A3

参照实施例1的合成方法，将3-碘三氟甲苯替换成溴苯，其他条件不变。1H NMR(300MHz，Methanol-d4)δ7.17-7.07(m，2H)，7.05-6.97(m，2H)，6.80(s，4H)，6.76-6.67 (m，4H)，6.58(t，J＝7.2Hz，1H)，4.17(q，J＝7.1Hz，2H)，3.85(t，J＝5.7Hz，2H)，3.70(s， 3H)，3.45(t，J＝7.4Hz，2H)，3.35(t，J＝7.0Hz，2H)，2.74(t，J＝7.4Hz，2H)，1.87(p，J＝6.1 Hz，2H)，1.50(s，6H)，1.20(t，J＝7.1Hz，3H).Referring to the synthesis method of Example 1, 3-iodotrifluorotoluene was replaced with bromobenzene, and other conditions remained unchanged. 1 H NMR (300MHz, Methanol-d 4 ) δ7.17-7.07 (m, 2H), 7.05-6.97 (m, 2H), 6.80 (s, 4H), 6.76-6.67 (m, 4H), 6.58 (t , J=7.2Hz, 1H), 4.17 (q, J=7.1Hz, 2H), 3.85 (t, J=5.7Hz, 2H), 3.70 (s, 3H), 3.45 (t, J=7.4Hz, 2H ), 3.35 (t, J=7.0Hz, 2H), 2.74 (t, J=7.4Hz, 2H), 1.87 (p, J=6.1 Hz, 2H), 1.50 (s, 6H), 1.20 (t, J =7.1Hz, 3H).

实施例5Example 5

A4的合成Synthesis of A4

参照实施例1的合成方法，将3-碘三氟甲苯替换成3，5-二甲基溴苯，其他条件不变。1H NMR(300MHz，Chloroform-d)δ7.07(d，J＝8.5Hz，2H)，6.85(s，4H)，6.81(d，J＝8.5Hz，2H)，6.39(d，J＝7.3Hz，3H)，4.26(q，J＝7.1Hz，2H)，3.94(t，J＝5.7Hz，2H)，3.79(s，3H)，3.50(dd，J＝8.9，6.5Hz，2H)，3.43(t，J＝6.9Hz，2H)，2.86-2.76(m，2H)，2.27(s，6H)，1.99(p，J＝6.2Hz，2H)，1.60(s，6H)，1.27(d，J＝7.1Hz，3H).Referring to the synthesis method of Example 1, 3-iodotrifluorotoluene was replaced with 3,5-dimethylbromobenzene, and other conditions remained unchanged. 1 H NMR (300MHz, Chloroform-d) δ7.07 (d, J=8.5Hz, 2H), 6.85 (s, 4H), 6.81 (d, J=8.5Hz, 2H), 6.39 (d, J=7.3 Hz, 3H), 4.26 (q, J=7.1Hz, 2H), 3.94 (t, J=5.7Hz, 2H), 3.79 (s, 3H), 3.50 (dd, J=8.9, 6.5Hz, 2H), 3.43 (t, J=6.9Hz, 2H), 2.86-2.76 (m, 2H), 2.27 (s, 6H), 1.99 (p, J=6.2Hz, 2H), 1.60 (s, 6H), 1.27 (d , J=7.1Hz, 3H).

实施例6Example 6

A5的合成Synthesis of A5

参照实施例1的合成方法，将3-碘三氟甲苯替换成3，5-双三氟甲基溴苯，其他条件不变。1H NMR(400MHz，DMSO-d6)δ7.11(d，J＝8.5Hz，4H)，7.06(s，1H)，6.85(s，4H)， 6.73-6.68(m，2H)，4.14(q，J＝7.1Hz，2H)，3.92(t，J＝5.7Hz，2H)，3.68(s，3H)，3.62(t，J ＝7.4Hz，2H)，3.50(t，J＝7.1Hz，2H)，2.75(t，J＝7.3Hz，2H)，1.89(t，J＝6.9Hz，2H)，1.46(s，6H)，1.15(t，J＝7.1Hz，3H).Referring to the synthesis method of Example 1, 3-iodotrifluorotoluene was replaced with 3,5-bistrifluoromethylbromobenzene, and other conditions remained unchanged. 1 H NMR (400MHz, DMSO-d 6 ) δ7.11 (d, J=8.5Hz, 4H), 7.06 (s, 1H), 6.85 (s, 4H), 6.73-6.68 (m, 2H), 4.14 ( q, J=7.1Hz, 2H), 3.92 (t, J=5.7Hz, 2H), 3.68 (s, 3H), 3.62 (t, J=7.4Hz, 2H), 3.50 (t, J=7.1Hz, 2H), 2.75 (t, J=7.3Hz, 2H), 1.89 (t, J=6.9Hz, 2H), 1.46 (s, 6H), 1.15 (t, J=7.1Hz, 3H).

实施例7Example 7

A6的合成Synthesis of A6

参照实施例1的合成方法，将3-碘三氟甲苯替换成4-溴-8-三氟甲基喹啉，其他条件不变。1H NMR(300MHz，Methanol-d4)δ8.56(d，J＝5.3Hz，1H)，7.99(dd，J＝22.1，7.9 Hz，2H)，7.37(t，J＝7.8Hz，1H)，7.04(d，J＝5.1Hz，1H)，6.90(d，J＝8.1Hz，2H)，6.70(m， 2H)，6.65-6.58(m，4H)，4.13(q，J＝7.2Hz，2H)，3.78(t，J＝5.7Hz，2H)，3.65(d，J＝1.1 Hz，3H)，3.58-3.45(m，4H)，2.77(t，J＝7.1Hz，2H)，1.91(p，J＝6.3Hz，2H)，1.46(s，6H)， 1.16(t，J＝7.1Hz，3H).Referring to the synthesis method of Example 1, 3-iodotrifluorotoluene was replaced with 4-bromo-8-trifluoromethylquinoline, and other conditions remained unchanged. 1 H NMR (300MHz, Methanol-d 4 ) δ8.56 (d, J=5.3Hz, 1H), 7.99 (dd, J=22.1, 7.9 Hz, 2H), 7.37 (t, J=7.8Hz, 1H) , 7.04 (d, J=5.1Hz, 1H), 6.90 (d, J=8.1Hz, 2H), 6.70 (m, 2H), 6.65-6.58 (m, 4H), 4.13 (q, J=7.2Hz, 2H), 3.78 (t, J=5.7Hz, 2H), 3.65 (d, J=1.1 Hz, 3H), 3.58-3.45 (m, 4H), 2.77 (t, J=7.1Hz, 2H), 1.91 ( p, J=6.3Hz, 2H), 1.46 (s, 6H), 1.16 (t, J=7.1Hz, 3H).

实施例8Example 8

A7的合成Synthesis of A7

参照实施例1的合成方法，将3-碘三氟甲苯替换成2-碘-1，1，1-三氟乙烷，其他条件 不变。1H NMR(300MHz，Methanol-d4)δ7.03(d，J＝8.6Hz，2H)，6.80(s，4H)，6.73(d，J＝8.5Hz，2H)，4.18(q，J＝7.1Hz，2H)，3.86(t，J＝6.0Hz，2H)，3.71(s，3H)，3.15(q，J＝9.7Hz，2H)，2.87-2.74(m，4H)，2.65(dd，J＝8.9，5.9Hz，2H)，1.80(p，J＝6.5Hz，2H)，1.51(s，6H)，1.21(t，J＝7.1Hz，3H).Referring to the synthesis method of Example 1, 3-iodotrifluorotoluene was replaced with 2-iodo-1,1,1-trifluoroethane, and other conditions remained unchanged. 1 H NMR (300MHz, Methanol-d 4 ) δ7.03 (d, J=8.6Hz, 2H), 6.80 (s, 4H), 6.73 (d, J=8.5Hz, 2H), 4.18 (q, J= 7.1Hz, 2H), 3.86 (t, J=6.0Hz, 2H), 3.71 (s, 3H), 3.15 (q, J=9.7Hz, 2H), 2.87-2.74 (m, 4H), 2.65 (dd, J=8.9, 5.9Hz, 2H), 1.80 (p, J=6.5Hz, 2H), 1.51 (s, 6H), 1.21 (t, J=7.1Hz, 3H).

实施例9Example 9

A8的合成Synthesis of A8

参照实施例1的合成方法，将3-碘三氟甲苯替换成4-(三氟甲基)溴苄，其他条件不变。1H NMR(300MHz，Methanol-d4)δ7.41(dd，J＝8.1，5.7Hz，2H)，7.37-7.28(m，2H)， 6.98(dd，J＝8.4，5.8Hz，2H)，6.85-6.76(m，2H)，6.74-6.64(m，4H)，f4.18(p，J＝7.1Hz， 2H)，3.79(q，J＝6.0Hz，2H)，3.72(d，J＝5.9Hz，3H)，3.65(d，J＝5.5Hz，2H)，2.72-2.63 (m，4H)，2.61(d，J＝6.3Hz，2H)，1.89-1.73(m，2H)，1.55-1.48(d，J＝6Hz，6H)，1.26- 1.16(m，3H).Referring to the synthesis method of Example 1, 3-iodotrifluorotoluene was replaced with 4-(trifluoromethyl)benzyl bromide, and other conditions remained unchanged. 1 H NMR (300MHz, Methanol-d 4 ) δ7.41 (dd, J=8.1, 5.7Hz, 2H), 7.37-7.28 (m, 2H), 6.98 (dd, J=8.4, 5.8Hz, 2H), 6.85-6.76 (m, 2H), 6.74-6.64 (m, 4H), f4.18 (p, J=7.1Hz, 2H), 3.79 (q, J=6.0Hz, 2H), 3.72 (d, J= 5.9Hz, 3H), 3.65 (d, J=5.5Hz, 2H), 2.72-2.63 (m, 4H), 2.61 (d, J=6.3Hz, 2H), 1.89-1.73 (m, 2H), 1.55- 1.48(d, J=6Hz, 6H), 1.26- 1.16(m, 3H).

实施例10Example 10

A9的合成Synthesis of A9

参照实施例1的合成方法，将3-碘三氟甲苯替换成5-溴喹啉，其他条件不变。1HNMR (300MHz，Acetone-d6)δ8.74(dd，J＝4.1，1.8Hz，1H)，8.26(d，J＝8.6Hz，1H)，7.68(d，J＝ 8.4Hz，1H)，7.63-7.54(m，1H)，7.34(d，J＝7.4Hz，1H)，7.22(dd，J＝8.5，4.1Hz，1H)，6.93 (d，J＝8.6Hz，2H)，6.75-6.66(m，4H)，6.62(d，J＝8.6Hz，2H)，4.08(q，J＝7.1Hz，2H)，3.83(t，J＝6.1Hz，2H)，3.63(s，3H)，3.35(t，J＝6.9Hz，2H)，3.30-3.22(m，2H)，2.72- 2.63(m，2H)，1.82(p，J＝6.5Hz，2H)，1.43(s，6H)，1.10(t，J＝7.1Hz，3H).Referring to the synthesis method of Example 1, 3-iodotrifluorotoluene was replaced with 5-bromoquinoline, and other conditions remained unchanged. 1 HNMR (300MHz, Acetone-d 6 ) δ8.74 (dd, J=4.1, 1.8Hz, 1H), 8.26 (d, J=8.6Hz, 1H), 7.68 (d, J=8.4Hz, 1H), 7.63-7.54 (m, 1H), 7.34 (d, J=7.4Hz, 1H), 7.22 (dd, J=8.5, 4.1Hz, 1H), 6.93 (d, J=8.6Hz, 2H), 6.75-6.66 (m, 4H), 6.62 (d, J=8.6Hz, 2H), 4.08 (q, J=7.1Hz, 2H), 3.83 (t, J=6.1Hz, 2H), 3.63 (s, 3H), 3.35 (t, J=6.9Hz, 2H), 3.30-3.22 (m, 2H), 2.72- 2.63 (m, 2H), 1.82 (p, J=6.5Hz, 2H), 1.43 (s, 6H), 1.10 ( t, J=7.1Hz, 3H).

实施例11Example 11

A11的合成Synthesis of A11

参照实施例2的合成方法，由A3水解得到。1H NMR(300MHz，Chloroform-d)δ7.16 -7.10(m，1H)，7.10-7.05(m，2H)，6.91-6.85(m，2H)，6.84(s，4H)，6.56(q，J＝7.5，6.7Hz，3H)，3.92(t，J＝5.7Hz，2H)，3.78(s，3H)，3.49(t，J＝7.7Hz，2H)，3.43(t，J＝7.0Hz，2H)，2.84-2.77(m，2H)，2.30(s，3H)，2.02-1.91(m，2H)，1.59(s，6H).Referring to the synthesis method of Example 2, it is obtained by hydrolysis of A3. 1 H NMR (300MHz, Chloroform-d) δ7.16 -7.10 (m, 1H), 7.10-7.05 (m, 2H), 6.91-6.85 (m, 2H), 6.84 (s, 4H), 6.56 (q, J=7.5, 6.7Hz, 3H), 3.92 (t, J=5.7Hz, 2H), 3.78 (s, 3H), 3.49 (t, J=7.7Hz, 2H), 3.43 (t, J=7.0Hz, 2H), 2.84-2.77(m, 2H), 2.30(s, 3H), 2.02-1.91(m, 2H), 1.59(s, 6H).

实施例12Example 12

A12的合成Synthesis of A12

参照实施例2的合成方法，由A4水解得到。1H NMR(300MHz，Methanol-d4)δ6.99(dd，J＝8.5，7.0Hz，2H)，6.83(d，J＝8.3Hz，2H)，6.67(d，J＝8.5Hz，2H)，6.62(s，4H)，6.56(d，J＝8.3Hz，2H)，6.45(t，J＝7.2Hz，1H)，3.63(t，J＝5.7Hz，2H)，3.51(s，3H)，3.25(t，J＝7.5Hz，2H)，3.17(t，J＝7.0Hz，2H)，2.54(t，J＝7.4Hz，2H)，1.67(p，J＝6.2Hz，2H)，1.35(s，6H).Referring to the synthesis method of Example 2, it was obtained by hydrolysis of A4. 1 H NMR (300MHz, Methanol-d 4 ) δ6.99 (dd, J=8.5, 7.0Hz, 2H), 6.83 (d, J=8.3Hz, 2H), 6.67 (d, J=8.5Hz, 2H) , 6.62 (s, 4H), 6.56 (d, J = 8.3Hz, 2H), 6.45 (t, J = 7.2Hz, 1H), 3.63 (t, J = 5.7Hz, 2H), 3.51 (s, 3H) , 3.25 (t, J=7.5Hz, 2H), 3.17 (t, J=7.0Hz, 2H), 2.54 (t, J=7.4Hz, 2H), 1.67 (p, J=6.2Hz, 2H), 1.35 (s,6H).

实施例13Example 13

A13的合成Synthesis of A13

参照实施例2的合成方法，由A5水解得到。1H NMR(300MHz，Methanol-d4)δ6.86 (d，J＝8.6Hz，2H)，6.69(d，J＝8.5Hz，2H)，6.65(s，4H)，6.26(s，2H)，6.18(s，1H)，3.65(t，J ＝5.7Hz，2H)，3.54(s，3H)，3.27(t，J＝7.5Hz，2H)，3.24-3.16(m，2H)，2.56(t，J＝7.4Hz，2H)，2.04(s，6H)，1.69(p，J＝6.1Hz，2H)，1.39(s，6H).Referring to the synthesis method of Example 2, it was obtained by hydrolysis of A5. 1 H NMR (300MHz, Methanol-d 4 ) δ6.86 (d, J=8.6Hz, 2H), 6.69 (d, J=8.5Hz, 2H), 6.65 (s, 4H), 6.26 (s, 2H) , 6.18 (s, 1H), 3.65 (t, J = 5.7Hz, 2H), 3.54 (s, 3H), 3.27 (t, J = 7.5Hz, 2H), 3.24-3.16 (m, 2H), 2.56 ( t, J=7.4Hz, 2H), 2.04 (s, 6H), 1.69 (p, J=6.1Hz, 2H), 1.39 (s, 6H).

实施例14Example 14

A14的合成Synthesis of A14

参照实施例2的合成方法，由A6水解得到。1H NMR(300MHz，Methanol-d4)δ7.04 (s，2H)，6.97(d，J＝8.6Hz，3H)，6.75(m，6H)，3.80(t，J＝5.5Hz，2H)，3.64(s，3H)，3.52(t，J ＝7.2Hz，2H)，3.40(t，J＝7.1Hz，2H)，2.72(t，J＝7.2Hz，2H)，1.82(p，J＝6.1Hz，2H)，1.42(s，6H).Referring to the synthesis method of Example 2, it is obtained by hydrolysis of A6. 1 H NMR (300MHz, Methanol-d 4 ) δ7.04 (s, 2H), 6.97 (d, J=8.6Hz, 3H), 6.75 (m, 6H), 3.80 (t, J=5.5Hz, 2H) , 3.64 (s, 3H), 3.52 (t, J = 7.2Hz, 2H), 3.40 (t, J = 7.1Hz, 2H), 2.72 (t, J = 7.2Hz, 2H), 1.82 (p, J = 6.1Hz, 2H), 1.42(s, 6H).

实施例15Example 15

A15的合成Synthesis of A15

参照实施例2的合成方法，由A7水解得到。1H NMR(300MHz，Methanol-d4)δ6.99 (d，J＝8.5Hz，2H)，6.77-6.71(m，6H)，3.83(t，J＝6.0Hz，2H)，3.67(s，3H)，3.12(q，J＝9.8 Hz，2H)，2.77(t，J＝7.2Hz，4H)，2.62(m，2H)，1.77(p，J＝6.4Hz，2H)，1.46(s，6H).Referring to the synthesis method of Example 2, it was obtained by hydrolysis of A7. 1 H NMR (300MHz, Methanol-d 4 ) δ6.99 (d, J=8.5Hz, 2H), 6.77-6.71 (m, 6H), 3.83 (t, J=6.0Hz, 2H), 3.67 (s, 3H), 3.12 (q, J=9.8 Hz, 2H), 2.77 (t, J=7.2Hz, 4H), 2.62 (m, 2H), 1.77 (p, J=6.4Hz, 2H), 1.46 (s, 6H).

实施例16Example 16

A16的合成Synthesis of A16

参照实施例2的合成方法，由A9水解得到。1H NMR(300MHz，Methanol-d4)δ7.56 (s，4H)，6.89(d，J＝8.5Hz，2H)，6.81-6.65(m，6H)，4.09(s，2H)，3.86(t，J＝5.7Hz，2H)， 3.68(s，3H)，3.01-2.94(m，2H)，2.93-2.86(m，2H)，2.77(dd，J＝10.2，5.6Hz，2H)，1.99 (dd，J＝9.0，5.7Hz，2H)，1.49(s，6H).Referring to the synthesis method of Example 2, it was obtained by hydrolysis of A9. 1 H NMR (300MHz, Methanol-d 4 ) δ7.56 (s, 4H), 6.89 (d, J=8.5Hz, 2H), 6.81-6.65 (m, 6H), 4.09 (s, 2H), 3.86 ( t, J=5.7Hz, 2H), 3.68 (s, 3H), 3.01-2.94 (m, 2H), 2.93-2.86 (m, 2H), 2.77 (dd, J=10.2, 5.6Hz, 2H), 1.99 (dd, J=9.0, 5.7Hz, 2H), 1.49 (s, 6H).

实施例17Example 17

B1的合成Synthesis of B1

参照实施例14的合成方法，将对羟基苯乙酸替换成间羟基苯乙酸，其他条件不变。1H NMR(400MHz，Methanol-d4)δ7.06(t，J＝7.8Hz，1H)，6.99(s，2H)，6.94(s，1H)，6.76-6.69(m，5H)，6.64(d，J＝8.0Hz，2H)，3.78(t，J＝5.4Hz，2H)，3.61(s，3H)，3.53(t，J＝7.0Hz，2H)，3.38(t，J＝7.1Hz，2H)，2.71(t，J＝6.9Hz，2H)，1.81(p，J＝5.9Hz，2H)，1.37(s，6H).Referring to the synthesis method of Example 14, p-hydroxyphenylacetic acid was replaced with m-hydroxyphenylacetic acid, and other conditions remained unchanged. 1 H NMR (400MHz, Methanol-d 4 ) δ7.06 (t, J=7.8Hz, 1H), 6.99 (s, 2H), 6.94 (s, 1H), 6.76-6.69 (m, 5H), 6.64 ( d, J=8.0Hz, 2H), 3.78 (t, J=5.4Hz, 2H), 3.61 (s, 3H), 3.53 (t, J=7.0Hz, 2H), 3.38 (t, J=7.1Hz, 2H), 2.71 (t, J=6.9Hz, 2H), 1.81 (p, J=5.9Hz, 2H), 1.37 (s, 6H).

实施例18Example 18

B2的合成Synthesis of B2

参照实施例14的合成方法，将2-溴-2-甲基丙酸乙酯替换成2-溴丙酸乙酯，其他条件不变。1H NMR(400MHz，DMSO-d6)δ7.14(d，J＝5.4Hz，3H)，7.10(d，J＝11.2Hz，2H)，6.86(s，4H)，6.79(d，J＝8.1Hz，2H)，4.71(q，J＝6.7Hz，1H)，3.92(t，J＝5.7Hz，2H)，3.69 (s，3H)，3.60(t，J＝7.5Hz，2H)，3.53(t，J＝7.3Hz，2H)，2.74(t，J＝7.5Hz，2H)，1.90(t，J＝ 6.7Hz，2H)，1.46(d，J＝6.7Hz，3H).Referring to the synthesis method of Example 14, 2-bromo-2-methylpropionic acid ethyl ester was replaced with 2-bromopropionic acid ethyl ester, and other conditions remained unchanged. 1 H NMR (400MHz, DMSO-d 6 ) δ7.14 (d, J=5.4Hz, 3H), 7.10 (d, J=11.2Hz, 2H), 6.86 (s, 4H), 6.79 (d, J= 8.1Hz, 2H), 4.71 (q, J=6.7Hz, 1H), 3.92 (t, J=5.7Hz, 2H), 3.69 (s, 3H), 3.60 (t, J=7.5Hz, 2H), 3.53 (t, J=7.3Hz, 2H), 2.74 (t, J=7.5Hz, 2H), 1.90 (t, J= 6.7Hz, 2H), 1.46 (d, J=6.7Hz, 3H).

实施例19Example 19

B3的合成Synthesis of B3

参照实施例1的合成方法，将2-溴-2-甲基丙酸乙酯替换成2-溴丁酸乙酯，其他条件不变。1H NMR(400MHz，Methanol-d4)δ7.10(d，J＝1.4Hz，2H)，7.08-7.04(m，2H)， 7.03(s，1H)，6.85-6.78(m，6H)，4.49(dd，J＝7.1，5.0Hz，1H)，3.88(t，J＝5.5Hz，2H)，3.71 (s，3H)，3.59(t，J＝7.2Hz，2H)，3.49(t，J＝7.1Hz，2H)，2.78(t，J＝7.2Hz，2H)，2.00-1.84 (m，4H)，1.05(t，J＝7.4Hz，3H).Referring to the synthesis method of Example 1, 2-bromo-2-methylpropionic acid ethyl ester was replaced with 2-bromobutyric acid ethyl ester, and other conditions remained unchanged. 1 H NMR (400MHz, Methanol-d 4 ) δ7.10 (d, J=1.4Hz, 2H), 7.08-7.04 (m, 2H), 7.03 (s, 1H), 6.85-6.78 (m, 6H), 4.49 (dd, J=7.1, 5.0Hz, 1H), 3.88 (t, J=5.5Hz, 2H), 3.71 (s, 3H), 3.59 (t, J=7.2Hz, 2H), 3.49 (t, J =7.1Hz, 2H), 2.78 (t, J = 7.2Hz, 2H), 2.00-1.84 (m, 4H), 1.05 (t, J = 7.4Hz, 3H).

实施例20Example 20

2-[4-[2-[(2-(4-甲氧基苯氧基)乙基)(3，5-二三氟甲基苯基)氨基]甲基]苯氧基]-2-甲基 丙酸(B4)的合成2-[4-[2-[(2-(4-methoxyphenoxy)ethyl)(3,5-ditrifluoromethylphenyl)amino]methyl]phenoxy]-2- Synthesis of Methylpropionic Acid (B4)

合成路线：synthetic route:

步骤1：step 1:

将NaH(1.81g，75mmol)加入无水DMF(5mL)中，N2置换。在冰水浴下缓慢滴 加化合物21对羟基苯甲醚(7.81g，62.96mmol)的DMF(10mL)。0℃下搅拌20min 后，撤去冰水浴，缓慢加入N-Boc-2-氨基乙基溴(10g，41.99mmol)的DMF(10mL) 溶液。将反应在室温下搅拌直至反应结束，过程用TLC检测。反应结束后，加入100mL 水淬灭反应，加NaOH溶液调PH至碱性，用二氯甲烷(100mL×3)萃取，合并有几层， 用无水Na2SO4干燥，减压浓缩至无溶剂蒸出，得无色油状化合物粗品22(11.25g)， 无需纯化，直接下投。NaH (1.81g, 75mmol) was added to anhydrous DMF (5mL) and replaced with N2 . Compound 21 p-hydroxyanisole (7.81 g, 62.96 mmol) in DMF (10 mL) was slowly added dropwise under an ice-water bath. After stirring at 0°C for 20 min, the ice-water bath was removed, and a solution of N-Boc-2-aminoethyl bromide (10 g, 41.99 mmol) in DMF (10 mL) was slowly added. The reaction was stirred at room temperature until completion, and the progress was monitored by TLC. After the reaction is completed, add 100 mL of water to quench the reaction, add NaOH solution to adjust the pH to alkaline, extract with dichloromethane (100 mL The solvent was evaporated to obtain a colorless oily crude compound 22 (11.25g), which was directly added without purification.

步骤2：Step 2:

将上述所得无色油状物22(11.25g，40.17mmol)溶于20mL乙酸乙酯中，加入10mL浓盐酸，室温下搅拌30min，并通过TLC监测反应直至完成。将反应液倒入50mL水 中，并用饱和NaHCO3溶液调节pH至中性。乙酸乙酯萃取(50mL×3)，合并有机层， 无水Na2SO4干燥，减压浓缩至干，得化合物236.62g，无需纯化直接用于下一步。1H NMR (400MHz，DMSO-d6)δ6.89-6.82(m，4H)，3.84(t，J＝5.8Hz，2H)，3.69(s，3H)，2.84(t，J ＝5.8Hz，2H).The colorless oil 22 (11.25 g, 40.17 mmol) obtained above was dissolved in 20 mL of ethyl acetate, 10 mL of concentrated hydrochloric acid was added, stirred at room temperature for 30 min, and the reaction was monitored by TLC until completion. Pour the reaction solution into 50 mL of water, and adjust the pH to neutral with saturated NaHCO solution . Extract with ethyl acetate (50 mL×3), combine the organic layers, dry over anhydrous Na 2 SO 4 , and concentrate to dryness under reduced pressure to obtain 236.62 g of compound, which can be used directly in the next step without purification. 1 H NMR (400MHz, DMSO-d 6 ) δ6.89-6.82 (m, 4H), 3.84 (t, J = 5.8Hz, 2H), 3.69 (s, 3H), 2.84 (t, J = 5.8Hz, 2H).

步骤3：Step 3:

将化合物24(4g，32.78mmol)和K2Co3(9.1g，65.84mmol)加入乙腈(10mL) 中，随后加入2-溴-2-甲基丙酸乙酯(9.5g，48.97mmol)，回流反应6h。纯化后得化合 物25(2g，产率26％)。1H NMR(400MHz，DMSO-d6)δ9.87(s，1H)，7.88-7.79(m，2H)， 6.96-6.88(m，2H)，4.18(q，J＝7.1Hz，2H)，1.61(s，6H)，1.14(t，J＝7.1Hz，3H).Compound 24 (4g, 32.78mmol) and K 2 Co 3 (9.1g, 65.84mmol) were added to acetonitrile (10mL), followed by ethyl 2-bromo-2-methylpropionate (9.5g, 48.97mmol). Reflux reaction for 6 hours. After purification, compound 25 (2 g, yield 26%) was obtained. 1 H NMR (400MHz, DMSO-d 6 ) δ9.87 (s, 1H), 7.88-7.79 (m, 2H), 6.96-6.88 (m, 2H), 4.18 (q, J=7.1Hz, 2H), 1.61(s, 6H), 1.14(t, J=7.1Hz, 3H).

步骤4：Step 4:

将化合物25(1g，4.24mmol)、化合物23(0.99g，5.92mmol)加入10mL无水THF 溶液中，加入三乙酰氧基硼氢化钠(2.6g，12.27mmol)，N2置换。上述反应液于室温 下反应12h。后处理：10mL饱和NaHCO3溶液淬灭反应，用EA(10mL×3)萃取，合 并有机相后用无水Na2SO4干燥，减压浓缩，纯化后得化合物26(0.64g，产率39％)。1H NMR(300MHz，Methanol-d4)δ7.30-7.23(m，2H)，6.88-6.81(m，6H)，4.23(q，J＝7.1 Hz，2H)，4.04(t，J＝5.4Hz，2H)，3.78(s，2H)，3.76(s，3H)，2.93(t，J＝5.4Hz，2H)，1.58(s， 6H)，1.26(t，J＝7.1Hz，3H).Compound 25 (1g, 4.24mmol) and compound 23 (0.99g, 5.92mmol) were added to 10 mL of anhydrous THF solution, and sodium triacetoxyborohydride (2.6g, 12.27mmol) was added, and N2 was substituted. The above reaction solution was reacted at room temperature for 12 h. Post-treatment: Quench the reaction with 10 mL saturated NaHCO solution , extract with EA (10 mL %). 1 H NMR (300MHz, Methanol-d 4 ) δ7.30-7.23 (m, 2H), 6.88-6.81 (m, 6H), 4.23 (q, J=7.1 Hz, 2H), 4.04 (t, J=5.4 Hz, 2H), 3.78 (s, 2H), 3.76 (s, 3H), 2.93 (t, J=5.4Hz, 2H), 1.58 (s, 6H), 1.26 (t, J=7.1Hz, 3H).

步骤5Step 5

将化合物26(0.33g，0.79mmol)、3，5-二三氟甲基溴苯(0.46g，1.59mmol)、Cs2CO3(0.52g，1.59mmol)、X-phos(19mg，0.04mmol)、Pd2(dba)3(36mg，0.04mmol)加 入3mL甲苯中，110℃反应过夜。后处理：加3mL水后用EA(5mL×3)萃取，合并 有机相，无水Na2SO4干燥，减压浓缩，柱层析纯化，得化合物27(0.13g，产率27％)。1H NMR(300MHz，Methanol-d4)δ7.89-7.82(m，1H)，7.18-7.08(m，3H)，6.98-6.93(m， 1H)，6.82(d，J＝2.1Hz，4H)，6.81-6.77(m，2H)，4.74(s，2H)，4.28-4.21(m，2H)，4.19- 4.14(m，2H)，3.98(t，J＝5.1Hz，2H)，3.75(s，3H)，1.55(s，6H)，1.23(t，J＝7.1Hz，3H).MS EsI m/z 622.20[M+Na]+.Compound 26 (0.33g, 0.79mmol), 3,5-ditrifluoromethylbenzene bromide (0.46g, 1.59mmol), Cs 2 CO 3 (0.52g, 1.59mmol), X-phos (19mg, 0.04mmol) ), Pd 2 (dba) 3 (36 mg, 0.04 mmol) were added to 3 mL of toluene, and the reaction was carried out at 110°C overnight. Post-treatment: Add 3 mL of water and extract with EA (5 mL × 3). Combine the organic phases, dry over anhydrous Na 2 SO 4 , concentrate under reduced pressure, and purify by column chromatography to obtain compound 27 (0.13 g, yield 27%). 1 H NMR (300MHz, Methanol-d 4 ) δ7.89-7.82 (m, 1H), 7.18-7.08 (m, 3H), 6.98-6.93 (m, 1H), 6.82 (d, J=2.1Hz, 4H ), 6.81-6.77(m, 2H), 4.74(s, 2H), 4.28-4.21(m, 2H), 4.19- 4.14(m, 2H), 3.98(t, J=5.1Hz, 2H), 3.75( s, 3H), 1.55 (s, 6H), 1.23 (t, J=7.1Hz, 3H).MS EsI m/z 622.20[M+Na] + .

步骤6：Step 6:

将化合物27(0.15g，0.24mmol)溶于3mLTHF以及3mL水的混合溶液中，加入 KOH(0.13g，2.4mmol)至反应液中，室温反应，TLC监测至反应结束。后处理：加入 稀盐酸调pH至中性，减压浓缩至无旋出液，剩余溶液用EA(6mL×3)萃取，合并有 机相，无水Na2SO4干燥后减压浓缩，化合物B4为淡黄色油状物。1H NMR(400MHz， Methanol-d4)δ7.22(d，J＝1.4Hz，2H)，7.12-7.06(m，3H)，6.85(d，J＝8.6Hz，2H)，6.82- 6.76(m，4H)，4.69(s，2H)，4.15(t，J＝5.1Hz，2H)，3.92(t，J＝5.1Hz，2H)，3.71(s，3H)，1.49 (s，6H).13C NMR(101MHz，Methanol-d4)δ179.92，156.22，155.55，154.04，150.93，133.11 (q，J＝32.3Hz，2C)，131.81，128.27，125.20(q，J＝272.7Hz，2C)，121.02，116.30，115.68， 113.24，109.47(m，1C)，80.85，67.87，56.03，55.07，52.15，25.86.HRMS(ESI)m/z calcd for C28H27F6NO5[M+Na]+594.1686，found 594.1691.Compound 27 (0.15g, 0.24mmol) was dissolved in a mixed solution of 3mLTHF and 3mL water, KOH (0.13g, 2.4mmol) was added to the reaction solution, and the reaction was carried out at room temperature. TLC monitored until the reaction was completed. Post-processing: Add dilute hydrochloric acid to adjust the pH to neutral, concentrate under reduced pressure until there is no spin-off liquid, extract the remaining solution with EA (6mL×3), combine the organic phases, dry over anhydrous Na 2 SO 4 and concentrate under reduced pressure, compound B4 It is light yellow oily substance. 1 H NMR (400MHz, Methanol-d 4 ) δ 7.22 (d, J = 1.4Hz, 2H), 7.12-7.06 (m, 3H), 6.85 (d, J = 8.6Hz, 2H), 6.82- 6.76 ( m, 4H), 4.69 (s, 2H), 4.15 (t, J=5.1Hz, 2H), 3.92 (t, J=5.1Hz, 2H), 3.71 (s, 3H), 1.49 (s, 6H). 13 C NMR (101MHz, Methanol-d 4 ) δ 179.92, 156.22, 155.55, 154.04, 150.93, 133.11 (q, J=32.3Hz, 2C), 131.81, 128.27, 125.20 (q, J=272.7Hz, 2C) , 121.02, 116.30, 115.68, 113.24, 109.47(m, 1C), 80.85, 67.87, 56.03, 55.07, 52.15, 25.86.HRMS(ESI)m/z calcd for C 28 H 27 F 6 NO 5 [M+Na] +594.1686 , found 594.1691.

实施例21Example 21

B5的合成Synthesis of B5

参照实施例20的合成方法，将对羟基苯甲醚替换成4-氟苯酚，将N-Boc-2-氨基乙基溴替换成3-(Boc-氨基)丙基溴，其他条件不变。1H NMR(400MHz，Methanol-d4)δ7.07 (d，J＝1.4Hz，2H)，7.02(d，J＝8.6Hz，2H)，6.97(s，1H)，6.91-6.85(m，2H)，6.81-6.76(m， 4H)，4.51(s，2H)，3.90(t，J＝5.6Hz，2H)，3.65(t，J＝7.1Hz，2H)，2.00(dt，J＝12.7，5.7Hz，2H)，1.44(s，6H).Refer to the synthesis method of Example 20, replace p-hydroxyanisole with 4-fluorophenol, replace N-Boc-2-aminoethyl bromide with 3-(Boc-amino)propyl bromide, and keep other conditions unchanged. 1 H NMR (400MHz, Methanol-d 4 ) δ7.07 (d, J=1.4Hz, 2H), 7.02 (d, J=8.6Hz, 2H), 6.97 (s, 1H), 6.91-6.85 (m, 2H), 6.81-6.76 (m, 4H), 4.51 (s, 2H), 3.90 (t, J=5.6Hz, 2H), 3.65 (t, J=7.1Hz, 2H), 2.00 (dt, J=12.7 ,5.7Hz,2H),1.44(s,6H).

实施例22Example 22

B6的合成Synthesis of B6

参照实施例20的合成方法，将对羟基苯甲醚替换成苯酚，将N-Boc-2-氨基乙基溴替换成3-(Boc-氨基)丙基溴，其他条件不变。1H NMR(400MHz，Methanol-d4)δ7.22- 7.16(m，2H)，7.10(d，J＝1.5Hz，2H)，7.05(d，J＝8.6Hz，2H)，6.99(s，1H)，6.87-6.83(m， 3H)，6.83-6.79(m，2H)，4.55(s，2H)，3.97(t，J＝5.6Hz，2H)，3.70(t，J＝7.2Hz，2H)，2.04 (dt，J＝12.2，5.7Hz，2H)，1.47(s，6H).Refer to the synthesis method of Example 20, replace p-hydroxyanisole with phenol, replace N-Boc-2-aminoethyl bromide with 3-(Boc-amino)propyl bromide, and keep other conditions unchanged. 1 H NMR (400MHz, Methanol-d 4 ) δ7.22- 7.16 (m, 2H), 7.10 (d, J=1.5Hz, 2H), 7.05 (d, J=8.6Hz, 2H), 6.99 (s, 1H), 6.87-6.83(m, 3H), 6.83-6.79(m, 2H), 4.55(s, 2H), 3.97(t, J=5.6Hz, 2H), 3.70(t, J=7.2Hz, 2H ), 2.04 (dt, J=12.2, 5.7Hz, 2H), 1.47 (s, 6H).

实施例23Example 23

B7的合成Synthesis of B7

参照实施例20的合成方法，将N-Boc-2-氨基乙基溴替换成3-(Boc-氨基)丙基溴，其 他条件不变。1H NMR(400MHz，Methanol-d4)δ7.15(s，2H)，7.10(d，J＝8.6Hz，2H)，7.05(s，1H)，6.87(dd，J＝8.4，1.5Hz，2H)，6.82(d，J＝2.0Hz，4H)，4.60(s，2H)，3.97(d，J＝5.1Hz，2H)，3.76-3.69(m，5H)，2.06(p，J＝6.2Hz，2H)，1.52(s，6H).Referring to the synthesis method of Example 20, N-Boc-2-aminoethyl bromide was replaced with 3-(Boc-amino)propyl bromide, and other conditions remained unchanged. 1 H NMR (400MHz, Methanol-d 4 ) δ7.15 (s, 2H), 7.10 (d, J=8.6Hz, 2H), 7.05 (s, 1H), 6.87 (dd, J=8.4, 1.5Hz, 2H), 6.82 (d, J=2.0Hz, 4H), 4.60 (s, 2H), 3.97 (d, J=5.1Hz, 2H), 3.76-3.69 (m, 5H), 2.06 (p, J=6.2 Hz, 2H), 1.52 (s, 6H).

实施例24Example 24

活性测试activity test

以下活性测试所用到的仪器来源。Source of instruments used in the following activity tests.

表1仪器来源Table 1 Instrument sources

仪器名称equipment name 生产厂家Manufacturer 仪器名称equipment name 生产厂家Manufacturer 万分之一天平One ten thousandth scale 上海上平仪器有限公司Shanghai Shangping Instrument Co., Ltd. 震荡仪Oscillator 常州国华电器有限公司Changzhou Guohua Electric Co., Ltd. CO2培养箱 CO2 incubator ESCOESCO 离心机centrifuge 湖南湘仪实验室仪器开发有Hunan Xiangyi Laboratory Instrument Development Co., Ltd. 安全柜Safety cabinet 苏州净化设备有限公司Suzhou Purification Equipment Co., Ltd. 脱色摇床Decolorizing shaker ServicebioServicebio 倒置显微镜Inverted microscope 尼康公司Nikon Corporation 电泳仪Electrophoresis 北京六一仪器厂Beijing Liuyi Instrument Factory 酶标仪microplate reader 北京普朗新技术有限公Beijing Pulang New Technology Co., Ltd. 暗匣camera obscura 广东粤华医疗器械厂有限公Guangdong Yuehua Medical Equipment Factory Co., Ltd. 高压蒸汽灭菌锅High pressure steam sterilizer 上海博讯实业有限公司Shanghai Boxun Industrial Co., Ltd. 扫描仪scanner EPSONEPSON 涡旋机scroll machine 上海沪西分析仪器厂有Shanghai Huxi Analytical Instrument Factory

(一)成脂肪细胞油红染色实验(1) Oil red staining experiment of adipocytes

1、实验方法1. Experimental methods

(1)油红染色步骤(1) Oil red dyeing step

培养好的细胞吸弃原有培养基，用PBS清洗一遍(5min)，加入4％甲醛固定细胞20-30min，PBS清洗两次，每次5min。油红染色工作液孵育样本30min，PBS清洗2 次，每次5min。每孔加入150μL异丙醇溶液，置于振荡器上低速振荡20min后，于 492nm测定吸光值。阳性药T0901317、Fenofibrate acid作为对照。脂质蓄积抑制率(％) 按下式计算：Aspirate the original culture medium of the cultured cells, wash them once with PBS (5 minutes), add 4% formaldehyde to fix the cells for 20-30 minutes, and wash them twice with PBS, 5 minutes each time. The samples were incubated with oil red staining working solution for 30 min, and washed twice with PBS for 5 min each time. Add 150 μL isopropyl alcohol solution to each well, place it on a oscillator and oscillate at low speed for 20 min, and then measure the absorbance value at 492 nm. Positive drugs T0901317 and Fenofibrate acid were used as controls. Lipid accumulation inhibition rate (%) is calculated according to the following formula:

(2)细胞成像(2) Cell imaging

油红染色工作液染色经PBS清洗后，加入甘油：PBS＝1：1溶液，封片保存，镜下 观察。After staining with oil red staining working solution and washing with PBS, add glycerol:PBS = 1:1 solution, seal the slide and store it for observation under the microscope.

(3)实验结果(3)Experimental results

成脂肪细胞株油红染色实验结果见图1。研究结果表明，本发明化合物B4在成脂肪细胞中脂质抑制活性最好，对其做进一步研究。The results of oil red staining experiments on adipogenic cell lines are shown in Figure 1. The research results show that compound B4 of the present invention has the best lipid inhibitory activity in adipocytes, and further research is conducted on it.

(二)HepG2以及RAW264.7细胞株油红染色实验(2) Oil red staining experiment of HepG2 and RAW264.7 cell lines

1、实验方法1. Experimental methods

(1)HepG2细胞酯化模型的建立(1) Establishment of HepG2 cell esterification model

选择处于对数生长期的HepG2细胞，接种于96孔细胞培养板中，细胞密度为8000个/孔。5％CO2、37℃培养24h后，弃去原细胞培养液，换成DMEM完全培养基和终 浓度为0.5mM油酸(OA)，在5％CO2、37℃培养箱中共同孵育24h，使脂质在HepG2 细胞中沉积。HepG2 cells in the logarithmic growth phase were selected and seeded in a 96-well cell culture plate at a cell density of 8000 cells/well. After culturing for 24 hours at 5% CO 2 and 37°C, discard the original cell culture medium, replace it with DMEM complete medium and a final concentration of 0.5mM oleic acid (OA), and incubate for 24 hours in an incubator with 5% CO 2 and 37°C. , causing lipid deposition in HepG2 cells.

(2)RAW264.7细胞酯化模型的建立(2) Establishment of RAW264.7 cell esterification model

选择处于对数生长期的RAW264.7细胞，接种于96孔细胞培养板中，细胞密度为5000个/孔。5％CO2、37℃培养24h后，弃去原细胞培养液，换成DMEM完全培养基 和终浓度为100μg/mL氧化型低密度脂蛋白(Ox-LDL)，在5％CO2，37℃培养箱中共同孵 育24h，使脂质在RAW264.7细胞中沉积。RAW264.7 cells in the logarithmic growth phase were selected and seeded in a 96-well cell culture plate at a cell density of 5000 cells/well. After culturing for 24 hours at 5% CO 2 and 37°C, discard the original cell culture medium and replace it with DMEM complete medium and a final concentration of 100 μg/mL oxidized low-density lipoprotein (Ox-LDL) at 5% CO 2 and 37 Incubate together for 24 hours in a ℃ incubator to allow lipids to deposit in RAW264.7 cells.

(3)实验分组(3)Experimental grouping

实验共分为3组(每组设6个复孔)，分别为①空白对照组：不做其他处理，将 HepG2细胞用DMEM完全培养基培养24h后换新DMEM完全培养基继续培养24h； ②细胞酯化模型组：在细胞液中加入终浓度为0.5mM的OA或者100μg/mL Ox-LDL， 共同孵育24h后更换不含酯化试剂的培养基继续培养24h；③给药组：细胞制成酯化 模型后，加入含不同药物的培养基继续孵育24h。The experiment was divided into 3 groups (each group had 6 duplicate wells), which were ① blank control group: without other treatment, HepG2 cells were cultured in DMEM complete medium for 24 hours and then replaced with new DMEM complete medium and continued to be cultured for 24 hours; ② Cell esterification model group: Add OA with a final concentration of 0.5mM or 100μg/mL Ox-LDL to the cell solution, incubate together for 24 hours, then replace the medium without esterification reagent and continue culturing for 24 hours; ③ Administration group: Cell preparation After forming the esterification model, medium containing different drugs was added and incubated for 24 hours.

(4)油红染色步骤(4) Oil red dyeing step

培养好的细胞吸弃原有培养基，用PBS清洗一遍(5min)，加入4％甲醛固定细胞20-30min，PBS清洗两次，每次5min。油红染色工作液孵育样本30min，PBS清洗2 次，每次5min。每孔加入150μL异丙醇溶液，置于振荡器上低速振荡20min后，于492 nm测定吸光值。脂质蓄积抑制率(％)按下式计算：Aspirate the original culture medium of the cultured cells, wash them once with PBS (5 minutes), add 4% formaldehyde to fix the cells for 20-30 minutes, and wash them twice with PBS, 5 minutes each time. The samples were incubated with oil red staining working solution for 30 min, and washed twice with PBS for 5 min each time. Add 150 μL isopropyl alcohol solution to each well, place it on a oscillator and oscillate at low speed for 20 min, and then measure the absorbance value at 492 nm. Lipid accumulation inhibition rate (%) is calculated according to the following formula:

(5)细胞成像(5) Cell imaging

油红染色工作液染色经PBS清洗后，加入甘油：PBS＝1：1溶液，封片保存，镜下 观察。After staining with oil red staining working solution and washing with PBS, add glycerol:PBS = 1:1 solution, seal the slide and store it for observation under the microscope.

2、实验结果2. Experimental results

HepG2以及RAW264.7细胞株油红染色实验结果见图2-5。研究结果表明，本发明 化合物B4在HepG2细胞及RAW264.7细胞中有一定的脂质蓄积抑制作用。The results of oil red staining experiments on HepG2 and RAW264.7 cell lines are shown in Figure 2-5. Research results show that compound B4 of the present invention has a certain inhibitory effect on lipid accumulation in HepG2 cells and RAW264.7 cells.

(三)、细胞内总胆固醇(TC)测定(3) Determination of intracellular total cholesterol (TC)

1、实验方法1. Experimental methods

将对数生长期的细胞制备为细胞悬液(细胞悬液浓度约为1×105个细胞/mL)接种于25mm直径培养皿中。在5％CO2、37℃的细胞培养箱培养12h后给药，实验分组 ①空白对照组：不做其他处理，将细胞用DMEM完全培养基培养24h后换新DMEM 完全培养基继续培养，间隔24h后更换新的DMEM完全培养基；②细胞酯化模型组： 在细胞液中加入终浓度为0.5mM的OA(HepG2细胞)或100μg/mL Ox-LDL(RAW264.7 细胞)，共同孵育24h后更换不含酯化试剂的培养基继续培养24h；③给药组：细胞 制成酯化模型后，加入含药物的培养基继续孵育24h。吸弃原培养基并用PBS清洗2 遍，而后用细胞刮将细胞刮下，收集至离心管中，离心后弃去上清液并用玻璃匀浆管将 细胞破碎3-4min，用BCA法进行蛋白定量后，使用总胆固醇检测试剂盒对细胞内的总胆固醇进行定量检测。Cells in the logarithmic growth phase were prepared into a cell suspension (the concentration of the cell suspension was approximately 1×105 cells/mL) and seeded in a 25 mm diameter culture dish. After culturing for 12 hours in a cell culture incubator with 5% CO 2 and 37°C, the drug was administered. Experimental groups: ① Blank control group: No other treatment was performed. The cells were cultured in DMEM complete medium for 24 hours and then replaced with new DMEM complete medium to continue culturing at intervals. Replace with new DMEM complete culture medium after 24 hours; ② Cell esterification model group: Add a final concentration of 0.5 mM OA (HepG2 cells) or 100 μg/mL Ox-LDL (RAW264.7 cells) to the cell solution and incubate together for 24 hours. Then replace the culture medium without esterification reagent and continue to culture for 24 hours; ③ Medication group: After the cells are made into esterification model, add the culture medium containing drug and continue to incubate for 24 hours. Aspirate away the original culture medium and wash it twice with PBS. Then use a cell scraper to scrape off the cells and collect them into a centrifuge tube. After centrifugation, discard the supernatant and use a glass homogenizer tube to disrupt the cells for 3-4 minutes. Use the BCA method to perform protein synthesis. After quantification, use the total cholesterol detection kit to quantitatively detect the total cholesterol in the cells.

2、实验结果2. Experimental results

结果如图6所示，B4在20μM的浓度下能够有效的降低HepG2细胞、RAW264.7 细胞中的总胆固醇水平，进一步说明化合物对肝脏中的脂质蓄积有一定的缓解作用，对 脂肪肝的治疗有良好的应用前景。The results are shown in Figure 6. B4 can effectively reduce the total cholesterol levels in HepG2 cells and RAW264.7 cells at a concentration of 20 μM, further indicating that the compound has a certain alleviation effect on lipid accumulation in the liver and has a certain effect on fatty liver disease. The treatment has good application prospects.

Claims (6)

1.一种化合物或其可药用的盐，其特征在于，所述化合物选自如下：1. A compound or a pharmaceutically acceptable salt thereof, characterized in that the compound is selected from the following: 2.一种药物组合物，其特征在于，包含权利要求1所述的化合物或其可药用的盐、及药学上可接受的载体。2. A pharmaceutical composition, characterized by comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. 3.根据权利要求1所述的化合物或其可药用的盐或权利要求2所述药物组合物在制备胆固醇逆转运药物中的用途。3. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 2 in the preparation of cholesterol reverse transport drugs. 4.根据权利要求1所述的化合物或其可药用的盐或权利要求2所述药物组合物在制备调血脂药物中的用途。4. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 2 in the preparation of blood lipid-regulating drugs. 5.根据权利要求l所述的化合物或其可药用的盐或权利要求2所述药物组合物在制备治疗肝脏脂质蓄积药物中的用途。5. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 2 in the preparation of a drug for treating liver lipid accumulation. 6.根据权利要求1所述的化合物或其可药用的盐或权利要求2所述药物组合物在制备治疗脂肪肝药物中的用途。6. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 2 in the preparation of a drug for treating fatty liver.
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JP2006232703A (en) * 2005-02-23 2006-09-07 Dai Ichi Seiyaku Co Ltd Phenyl ether derivative and medicine by using the same
JP2007246474A (en) * 2006-03-17 2007-09-27 Mitsubishi Pharma Corp Benzene derivatives
EP1887005A1 (en) * 2005-05-31 2008-02-13 Kowa Co., Ltd. Processes for production of optically active ppar-activating compounds and intermediates for production thereof
WO2022081350A2 (en) * 2020-09-30 2022-04-21 Duke University Methods for identification, stratification, and treatment of cns diseases
WO2022089606A1 (en) * 2020-10-30 2022-05-05 Westlake Therapeutics (Hangzhou) Co. Limited Methods for preparing mature red blood cells in vitro

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006232703A (en) * 2005-02-23 2006-09-07 Dai Ichi Seiyaku Co Ltd Phenyl ether derivative and medicine by using the same
EP1887005A1 (en) * 2005-05-31 2008-02-13 Kowa Co., Ltd. Processes for production of optically active ppar-activating compounds and intermediates for production thereof
JP2007246474A (en) * 2006-03-17 2007-09-27 Mitsubishi Pharma Corp Benzene derivatives
WO2022081350A2 (en) * 2020-09-30 2022-04-21 Duke University Methods for identification, stratification, and treatment of cns diseases
WO2022089606A1 (en) * 2020-10-30 2022-05-05 Westlake Therapeutics (Hangzhou) Co. Limited Methods for preparing mature red blood cells in vitro

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