Cycloalkane-substituted amine derivative, preparation method therefor and use thereof

Abstract

Disclosed in the present invention are a cycloalkane-substituted amine derivative, and a preparation method therefor and the use thereof. Specifically disclosed is a compound as shown in formula (I) or a pharmaceutically acceptable salt thereof. The cycloalkane-substituted amine derivative provided in the present invention has good 5-HT2A receptor agonistic activity and wide application prospects.

Description

Cycloalkane substituted amine derivatives, preparation method and application thereof

[0001] This application claims priority to Chinese patent application No. 202311217884.1, filed September 20, 2023. This application incorporates the entire text of the aforementioned Chinese patent application. Technical Field

[0002] The present invention relates to cycloalkane substituted amine derivatives, preparation methods and applications thereof. Background Art

[0003] Serotonin 5-HT 2A The receptor is a 5-HT receptor, a member of the 5-HT2 receptor subfamily. 2A The receptor belongs to G protein coupled receptor (GPCR), which is widely distributed in the central nervous system and peripheral nervous system. Its endogenous ligand 5-hydroxytryptamine is a monoamine neurotransmitter. 2A 5-HT is an endogenous agonist of 5-HT receptors and participates in a wide range of biological functions in the body, such as regulating emotions and consciousness in the central nervous system, as well as cardiovascular function and peripheral intestinal motility. Disorders in 5-HT secretion in the body can cause central and peripheral nervous system diseases, such as depression, schizophrenia, and cardiovascular diseases (Alison G Paquette et al.; J Cell Biochem, 2014, 11512, 2065-2072).

[0004] Exogenous 5-HT has been reported 2A Receptor agonists are mainly derived from natural extracts or semi-synthetic analogues, such as psilocybin, lysergic acid diethylamide (LSD), mescaline, etc., and are mainly used in medical applications (David E. Nichols; Pharmacol Rev, 2016, 68, 264-355). In early clinical trials, it was found that this type of compound has antidepressant and sedative effects. However, it was soon reported that most of the natural extracts and related synthetic derivatives of this type of 5-HT 2A When it is a receptor agonist, it is very easy to produce hallucinogenic effects in the human body, which has a huge negative impact on people's life and production, thus greatly limiting its application in medicinal value (Antoni R. Blaazer et al.; ChemMedChem, 2008, 3, 1299-1309).

[0005] In recent years, due to the exogenous 5-HT 2AReceptor agonists have been shown to have significant clinical antidepressant effects (Robin L. Carhart-Harris et al.; N Engl J Med 2021, 384(15), 1402-1411), so 5-HT 2A The research and development of 5-HT receptor agonists has become a hot topic in drug development. They have been widely studied and applied in both the central and peripheral nervous systems, such as antidepressant, sedative, anti-addiction, treatment of obsessive-compulsive disorder and neuralgic headache in the central nervous system, and in regulating intraocular pressure and cardiovascular disease in the peripheral nervous system. 2A The chemical structure of 5-HT receptor agonists is still mainly concentrated in the three types of mother nuclei of natural extracts: tryptamine, ergoline and phenylethylamine. Based on these three types of mother nuclei, there are a large number of 5-HT receptor agonists. 2A However, due to toxic side effects and poor drug properties, the small molecules approved for marketing and clinical research are still concentrated on natural extracts discovered decades ago, and there is no new chemical structure of 5-HT 2A Receptor agonists are being further developed clinically.

[0006] Pimavanserin is a known 5-HT 2A A highly selective antagonist of 5-HT 2A The receptor affinity is pKi=9.70, and the antagonist activity is pIC 50 =8.73 (Kimberly E. Vanover et al., J Pharmacol Exp Ther, 2006, 317(2), 910-918).

[0007] The structure of Pimavanserin is Modifying it to improve its pharmaceutical properties, such as making it exhibit agonist activity, is of great significance for the treatment of neurological diseases.

[0008] Summary of the Invention

[0009] In order to solve the above technical problems, the present invention provides cycloalkane substituted amine derivatives, their preparation methods and applications. The cycloalkane substituted amine derivatives provided by the present invention have 5-HT 2A Receptor agonist activity, and has broad application prospects.

[0010] The present invention provides a compound represented by formula I or a pharmaceutically acceptable salt thereof;

[0011] in,

[0012] m and n are each independently 1, 2 or 3;

[0013] R 1 and R 2 Each independently represents hydrogen, C1-C6 alkyl or substituted by 1, 2 or 3 R 1-1 Substituted C1-C6 alkyl;

[0014] R 1-1 Each independently is C6-C 10 Aryl or 1, 2 or 3 R 1-1a Substituted C6-C 10 aryl;

[0015] R 1-1a is halogen, hydroxy or C1-C6 alkyl;

[0016] R 3 is hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl or 1, 2 or 3 R 3-1 Substituted C1-C6 alkyl;

[0017] R 3-1 are independently halogen, hydroxyl or 1, 2 or 3 R 3-1a Substituted C6-C 10 aryl;

[0018] R 3-1a Each is independently a C1-C4 alkoxy group;

[0019] X is NH or a single bond;

[0020] Y is a single bond, -CH2-, -(CH2)2-, -(CH2)3-, -CH=CH-、-CH=CH-CH2-、 or carbonyl;

[0021] Ar is C6-C 10 Aryl, C5-C 12 Heteroaryl, 1, 2 or 3 R 5 Substituted C6-C 10 Aryl or 1, 2 or 3 R 6 Substituted C5-C 12 Heteroaryl, the C5-C 12 In the heteroaryl group, the heteroatom is selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3;

[0022] R 5 and R 6 Each independently represents halogen, hydroxyl, C1-C 10 Alkyl, C1-C 10 Alkoxy, C1-C 10 Alkylthio or

[0023] R N-1 and R N-2 Each is independently H or C1-C6 alkyl.

[0024] In one embodiment, certain groups in the compound represented by Formula I and its pharmaceutically acceptable salts are defined as follows. Groups not mentioned are the same as those described in any embodiment of the present invention (referred to as "one embodiment").

[0025] In one scheme, R 1 and R 2 wherein the C1-C6 alkyl group is replaced by 1, 2 or 3 R 1-1 The C1-C6 alkyl groups in the substituted C1-C6 alkyl groups are each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example, methyl.

[0026] In one scheme, R 1-1 In the C6-C 10 Aryl and 1, 2 or 3 R 1-1a Substituted C6-C 10 C6-C 10 Each aryl group is independently phenyl or naphthyl, for example phenyl.

[0027] In one scheme, R 1-1a wherein the C1-C6 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl.

[0028] In one scheme, R 1-1a wherein the halogen is fluorine, chlorine or bromine.

[0029] In one scheme, R 3 wherein the C1-C6 alkyl group is replaced by 1, 2 or 3 R 3-1 The C1-C6 alkyl groups in the substituted C1-C6 alkyl groups are each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example, methyl, ethyl or isopropyl.

[0030] In one scheme, R 3 wherein the C3-C6 cycloalkyl group is a cyclopropyl group, a cyclobutyl group, a cyclopentyl group or a cyclohexyl group, for example a cyclopropyl group, and for example a cyclopropyl group or a cyclobutyl group.

[0031] In one scheme, R 3-1 wherein the halogen is fluorine, chlorine or bromine, for example fluorine.

[0032] In one scheme, R 3-1 wherein said 1, 2 or 3 R 3-1aSubstituted C6-C 10 C6-C 10 Aryl is phenyl or naphthyl, for example phenyl.

[0033] In one scheme, R 3-1a wherein the C1-C4 alkoxy group is methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy or tert-butoxy, for example, methoxy.

[0034] In one embodiment, in Ar, the C6-C 10 Aryl and 1, 2 or 3 R 5 Substituted C6-C 10 C6-C 10 Each aryl group is independently phenyl or naphthyl, for example phenyl.

[0035] In one embodiment, in Ar, the C5-C 12 Heteroaryl and 1, 2 or 3 R 6 Substituted C5-C 12 C5-C 12 The heteroaryl groups are each independently C5-C9 heteroaryl groups, wherein the heteroatom is preferably N and the number of heteroatoms is preferably 1. For example, the C5-C 12 Heteroaryl is

[0036] In one scheme, R 5 and R 6 wherein the halogen is fluorine, chlorine or bromine, for example fluorine.

[0037] In one scheme, R 5 and R 6 In the C1-C 10 The alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl or n-decyl.

[0038] In one scheme, R 5 and R 6 In the C1-C 10 Alkoxy is methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, n-pentoxy, n-hexoxy, n-heptoxy, n-octoxy, n-nonoxy or n-decoxy, for example n-hexoxy, isobutoxy, n-decoxy, n-heptoxy, n-octoxy or methoxy.

[0039] In one scheme, R 5 and R 6 In the C1-C 10Alkylthio is methylthio, ethylthio, n-propylthio, isopropylthio, n-butylthio, isobutylthio, tert-butylthio, n-pentylthio, n-hexylthio, n-heptylthio, n-octylthio, n-nonylthio or n-decylthio.

[0040] In one scheme, R N-1 and R N-2 wherein the C1-C6 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl.

[0041] In one embodiment, when m and n are not equal, the compound represented by Formula I is a compound represented by Formula I-1a, I-2a, I-3a or I-4a:

[0042] Among them, R 1 、R 2 、R 3 , X, Ar, Y, m and n are each independently as described in any one of the present invention.

[0043] In one embodiment, when m and n are equal, the compound represented by Formula I is a compound represented by Formula I-5a or I-6a:

[0044] Among them, R 1 、R 2 、R 3 , X, Ar, Y, m and n are each independently as described in any one of the present invention.

[0045] In one embodiment, m and n are each independently 1 or 2.

[0046] In one scheme, R 1 and R 2 Each independently represents hydrogen, C1-C6 alkyl or substituted by 1, 2 or 3 R 1-1 Substituted C1-C6 alkyl; R 1-1 For 1, 2 or 3 R 1-1a Substituted C6-C 10 Aryl; R 1-1a is a hydroxyl group; preferably, R 1 and R 2 are each independently hydrogen.

[0047] In one scheme, R 3 is hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl or 1, 2 or 3 R 3-1 Substituted C1-C6 alkyl; R 3-1 are each independently halogen or 1 R 3-1a Substituted C6-C 10 Aryl; R 3-1aEach independently is a C1-C4 alkoxy group; preferably, R 3 is hydrogen, C1-C6 alkyl or 1, 2 or 3 (e.g. 1) R 3-1 Substituted C1-C6 alkyl (e.g., substituted by 1, 2 or 3 R 3-1 The substituted C1-C6 alkyl group is );R 3-1 are each independently substituted by 1, 2 or 3 (eg 1) R 3-1a Substituted C6-C 10 Aryl (e.g., phenyl); R 3-1a are each independently a C1-C6 alkoxy group (such as a methoxy group); further preferably, R 3 is hydrogen or C1-C6 alkyl, for example, hydrogen or linear C1-C6 alkyl, and further, R 3 is a straight-chain C1-C6 alkyl group (eg, methyl).

[0048] In one embodiment, X and Y are not both single bonds.

[0049] In one embodiment, Y is a single bond, -CH2-, -(CH2)2-, -CH=CH- or -C(=O)-; preferably, Y is -CH2-, -(CH2)2-, -CH=CH- or -C(=O)-; for example, Y is -CH2- or -C(=O)-.

[0050] In one embodiment, when X is a single bond and Y is -CH=CH-, for

[0051] In one embodiment, when X is a single bond and Y is -(CH2)2-, for

[0052] In one embodiment, Ar is substituted by 1, 2 or 3 R 5 Substituted C6-C 10 Aryl or 1, 2 or 3 R 6 Substituted C5-C 12 Heteroaryl, the C5-C 12 In the heteroaryl group, the heteroatom is selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3; preferably, Ar is C6-C 10 Aryl or 1, 2 or 3 R 5 Substituted C6-C 10 Aryl, for example, with 1, 2 or 3 R 5 Substituted C6-C 10 Aryl.

[0053] In one scheme, R 5 and R 6 Each independently is C1-C 10 Alkoxy; preferably, when X is NH, Y is -CH2-, R 5 and R 6 Each independently a linear C1-C 10 Alkoxy.

[0054] In one scheme, R 1 and R 2 are each independently hydrogen, or methyl.

[0055] In one plan, -NH2,

[0056] In one scheme, R 3 Methyl, hydrogen, Isopropyl, ethyl, or cyclopropane, for example methyl or H.

[0057] In one scheme, R 3 Methyl, hydrogen, Isopropyl, ethyl, Cyclopropyl or cyclobutane.

[0058] In one embodiment, X is NH or a single bond.

[0059] In one embodiment, Y is -CH2-, -CH=CH-, Single bond or carbonyl

[0060] In a certain scheme, -XY- is -CH=CH-, Single bond or carbonyl

[0061] In one embodiment, Ar is

[0062] In one scheme, R 5 for

[0063] In one scheme, R 6 for

[0064] In one embodiment, the compound represented by formula I is any of the following compounds:

[0065] In one embodiment, the pharmaceutically acceptable salt of the compound represented by formula I is the hydrochloride salt of the compound represented by formula I.

[0066] The present invention provides a method for preparing the compound represented by the above formula I, which is any of the following schemes:

[0067] Scheme 1: The compound represented by Formula Ia is hydrolyzed (for example, under the action of hydrochloric acid) in an organic solvent (for example, a 1,4-dioxane solution) to obtain the compound represented by Formula I;

[0068] W is an amino protecting group, such as tert-butyloxycarbonyl (BOC) or benzyloxycarbonyl (CBz), and another example is tert-butyloxycarbonyl;

[0069] Scheme 2: In an organic solvent (such as methanol), under the action of a catalyst (such as sodium cyanoborohydride), ammonia reagent Ic and the compound represented by formula Ib are coupled to obtain the compound represented by formula I

[0070] Among them, R 1 、R 2 、R 3 , X, Ar, Y, m and n are each independently as described in any one of the present invention.

[0071] The present invention also provides compounds represented by the following formula Ia or Ib:

[0072] Among them, R 2 、R 3 , X, Ar, Y, m and n are as described in any one of the present invention.

[0073] In one embodiment, the compound represented by Formula Ia is any of the following compounds:

[0074] In one embodiment, the compound represented by Formula Ib is any of the following compounds:

[0075] The present invention provides a pharmaceutical composition comprising:

[0076] (1) any one of the compounds of formula I of the present invention or a pharmaceutically acceptable salt thereof, and

[0077] (2) Pharmaceutically acceptable excipients.

[0078] The present invention provides a use of a compound as shown in formula I, a pharmaceutically acceptable salt thereof or the above-mentioned pharmaceutical composition in the preparation of a drug for treating and / or preventing 5-HT 2A Receptor-related diseases.

[0079] In one embodiment, the 5-HT 2A Diseases related to 5-HT receptors 2A β-arrestin2 downstream of the receptor recruits signaling pathways and / or 5-HT 2A Diseases related to the activation of Gq protein signaling pathways downstream of the receptor, such as those related to 5-HT 2A A receptor-related disease is depression.

[0080] The present invention provides a compound as shown in formula I, a pharmaceutically acceptable salt thereof or the above pharmaceutical composition for preparing 5-HT 2A The use of 5-HT receptor agonists 2A 5-HT receptor agonist 2A β-arrestin2 downstream of the receptor recruits signaling pathway agonists and / or 5-HT 2A Gq proteins downstream of the receptor activate signaling pathway agonists, such as 5-HT 2A β-arrestin2 downstream of the receptor recruits signaling pathway agonists.

[0081] Unless otherwise specified, the terms used in this invention have the following meanings:

[0082] It will be understood by those skilled in the art that the structural formulas used in the present invention to describe groups are based on the conventions used in the art. It means that the corresponding group is connected to other fragments and groups in the compound through this site.

[0083] Unless otherwise specified, use a solid wedge key. and dotted wedge key Indicates the absolute configuration of a stereocenter.

[0084] Unless otherwise specified, when a compound contains a double bond structure, such as a carbon-carbon double bond, and each atom on the double bond is connected to two different substituents, if a wavy line is used between the atom on the double bond and the substituent in the compound, If connected, it represents the (Z) isomer, (E) isomer or a mixture of the two isomers of the compound. For example, the following formula (A) represents that the compound exists as a single isomer of formula (A-1) or formula (A-2) or as a mixture of the two isomers of formula (A-1) and formula (A-2);

[0085] The term "pharmaceutically acceptable" means relatively non-toxic, safe, and suitable for use by patients.

[0086] The term "pharmaceutically acceptable salt" refers to a salt obtained by reacting a compound with a pharmaceutically acceptable acid or base. When the compound contains a relatively acidic functional group, a base addition salt can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable base in a suitable inert solvent. When the compound contains a relatively basic functional group, an acid addition salt can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable acid in a suitable inert solvent. For details, see Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl, Camille G. Wermuth, 2011, 2nd Revised Edition).

[0087] The expression "a group substituted with one, two or three substituents" means that one, two or three hydrogen atoms in a group are independently replaced by the substituents. When multiple substituents appear at the same time, unless otherwise specified, their definitions are independent of each other and do not affect each other. Therefore, if a group is replaced with one, two or three R 1-1 group substituted, that is, the group may be replaced by up to 3 R 1-1 Replace, the position R 1-1 Definition and other positions R 1-1 In addition, combinations of substituents and / or variables are permissible only if such combinations result in stable compounds.

[0088] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0089] The term "alkyl" refers to a group having a specified number of carbon atoms (e.g., C1-C 10 , C1-C6), linear or branched (e.g., linear), saturated monovalent hydrocarbon group. Alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, or n-decyl.

[0090] The term "alkoxy" refers to the group -OR X , R X The same definition as the term "alkyl". Alkoxy includes, but is not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, and the like.

[0091] The term "alkylthio" refers to the group -SR X , R XThe same definition as the term "alkyl". Alkoxy includes, but is not limited to, methylthio, ethylthio, n-propylthio, isopropylthio, n-butylthio, isobutylthio, tert-butylthio, and the like.

[0092] The term "aryl" refers to a group having the specified number of carbon atoms (e.g., C6-C 10 ) is a ring consisting only of carbon atoms, which is monocyclic or polycyclic, and each ring is aromatic (in accordance with Huckel's rule). Aryl includes but is not limited to phenyl and naphthyl.

[0093] The term "heteroaryl" refers to a cyclic, unsaturated, monovalent group having a specified number of ring atoms (e.g., 5-12 members, or 6-10 members), a specified number of heteroatoms (e.g., 1, 2, or 3), and a specified type of heteroatom (1, 2, or 3 of N, O, and S). It is monocyclic or bicyclic, with the two rings sharing two atoms and one bond, and each ring being aromatic (in accordance with Huckel's rule). A heteroaryl group is attached to the rest of the molecule through a carbon atom or a heteroatom; a heteroaryl group is attached to the rest of the molecule through a ring containing heteroatoms or a ring without heteroatoms.

[0094] The term "cycloalkyl" refers to a group having the specified number of ring carbon atoms (e.g., C 3-6 ), saturated monocyclic, spirocyclic, bridged, or paracyclic groups whose ring atoms consist solely of carbon atoms. Cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0095] Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain preferred embodiments of the present invention.

[0096] The reagents and raw materials used in the present invention are commercially available.

[0097] The positive progress of the present invention is that the compound represented by formula I provided by the present invention has improved pharmaceutical properties, can exhibit agonist activity, and has broad application prospects. DETAILED DESCRIPTION

[0098] The present invention is further illustrated by way of examples below, but the present invention is not limited to the scope of the examples. Experimental methods in the following examples where specific conditions are not specified were performed according to conventional methods and conditions, or selected according to the product specifications.

[0099] Example 1 Preparation of (1-(4-aminocyclohexyl)-3-(4-n-hexyloxy)benzyl)-1-methylurea) (Compound I-1)

[0100] Benzaldehyde (0.2 mL, 2 mmol) and tert-butyl (4-aminocyclohexyl)carbamate (428 mg, 2 mmol) were placed in a reaction flask, and methanol (10 mL) was added and stirred at room temperature to dissolve. Ten minutes later, sodium cyanoborohydride (138 mg, 2.2 mmol) was added to the system in batches and continued to stir at room temperature. The reaction progress was monitored by TLC. After 4 hours, the reaction was completed and quenched by adding water. Most of the solvent was removed by distillation under reduced pressure, and the mixture was extracted three times with ethyl acetate. The organic phases were combined and dried over anhydrous sodium sulfate. The crude product was spin-dried and purified on a silica gel column (dichloromethane: methanol = 20:1) to obtain a light yellow oil DW03112 (427 mg, yield: 78%). HRMS (ESI) C 18 H 29 N2O2 + ([M+H] + )Calculated value: 305.2224; measured value: 305.2248. 1 H NMR (800MHz, CDCl3) δ7.35-7.31(m,5H),2.71(s,1H),2.01(t,J=12.4Hz,1H),1.71(d,J=10.1Hz,4H),1.64-1.55(m,2H),1.51(s,2H),1.43(s,9H).

[0101] Step 2: Preparation of tert-butyl (4-(benzyl(methyl)amino)cyclohexyl)carbamate (compound DW03113)

[0102] 37% formaldehyde (0.3 mL, 3.3 mmol) and DW03112 (900 mg, 3 mmol) were placed in a reaction flask, and methanol (12 mL) was added and stirred at room temperature to dissolve. Ten minutes later, sodium cyanoborohydride (226 mg, 3.6 mmol) was added to the system in batches and continued to stir at room temperature. TLC was used to monitor the reaction progress. After 4 hours, the reaction was completed and water was added to quench the reaction. Most of the solvent was removed by distillation under reduced pressure, and the mixture was extracted three times with ethyl acetate. The organic phases were combined and dried over anhydrous sodium sulfate. The crude product was spin-dried to obtain a light yellow oily substance DW03113 (746 mg, yield: 78%). HRMS (ESI) C 19 H 31 N2O2 + ([M+H] + )Calculated value: 3319.2380; measured value: 319.2390.

[0103] Step 3: Preparation of (tert-butyl(4-methylamino)cyclohexyl)carbamate (Compound DW03115)

[0104] Compound DW03113 (700 mg, 2.2 mmol) was placed in a reaction flask and dissolved in methanol (10 mL). Palladium hydroxide (100 mg) was added and the gas was replaced with hydrogen. The system was then heated to 60°C and stirred for reaction. The reaction progress was monitored by TLC. After 5 hours, the reaction was complete. The palladium hydroxide was filtered off and the filtrate was distilled under reduced pressure to remove methanol, yielding DW03115 (620 mg, yield: 88%) as a colorless oil. HRMS (ESI) C 12 H 25 N2O2 + ([M+H] + )Calculated value: 229.1911; measured value: 229.1923.

[0105] Step 4: Preparation of 4-n-hexyloxybenzylcyanide (compound DW03067)

[0106] The compound p-fluorobenzyl cyanide (1.2 g, 10 mmol) and n-hexanol (1.1 g, 10 mmol) were placed in a dry reaction flask, and anhydrous tetrahydrofuran (20 mL) was added and dissolved under stirring at room temperature. 60% sodium hydride (360 mg, 15 mmol) was added to the reaction flask in batches, and the reaction was continued by stirring at room temperature. The reaction progress was monitored by TLC. After 4 hours, the reaction was completed. Water was added to quench the reaction, and most of the solvent was removed by distillation under reduced pressure. The product was extracted three times with ethyl acetate, and the organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was concentrated to obtain a crude product, which was purified by silica gel column (petroleum ether: ethyl acetate = 90:10) to obtain a colorless oil DW03067 (1.3 g, yield: 65%). HRMS (ESI) C 13 H 18 NO + ([M+H] + )Calculated value: 204.1383; measured value: 204.1401. 1 H NMR (800MHz, CDCl3) δ7.57(d,J=8.8Hz,2H),6.93(d,J=8.8Hz,2H),4.00(t,J=6.5Hz, 2H),1.83-1.77(m,2H),1.50-1.41(m,2H),1.38-1.31(m,4H),0.91(t,J=6.9Hz,3H).

[0107] Step 5: Preparation of 4-n-hexyloxyphenethylamine (compound DW03073)

[0108] Compound DW03067 (1.3 g, 6.4 mmol) and palladium on carbon (10%, 500 mg) were placed in a reaction flask, methanol (15 mL) was added, and the mixture was stirred at room temperature. 2M hydrochloric acid (0.5 mL) was added to the reaction system to replace the atmosphere with hydrogen. The reaction was heated to 50°C with stirring, and the reaction progress was monitored by TLC. After 4 hours, the reaction was complete. The palladium on carbon was filtered out, and the filtrate was concentrated under reduced pressure to obtain DW03073 (1.3 g, 83%) as a colorless oil.

[0109] Step 6: Preparation of N-(4-(n-hexyloxy)benzyl)-1H-imidazole-1-carboxamide (Compound DW03075)

[0110] Compound DW03073 (1.3 g, 6.3 mmol) and 1,1'-carbonyldiimidazole (2.1 g, 12.6 mmol) were placed in a reaction flask, and anhydrous DMF (20 mL) was added and stirred at room temperature. The reaction progress was monitored by TLC. The reaction was stirred overnight and the reaction was complete. Most of the solvent was removed by distillation under reduced pressure, and an appropriate amount of water was added and extracted three times with ethyl acetate. The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated to obtain a crude product, which was purified on a silica gel column (petroleum ether:ethyl acetate = 1:1) to obtain a light yellow oil DW03075 (1.3 g, yield: 71%). HRMS (ESI) C 17 H 24 N3O2 + ([M+H] + )Calculated value: 302.1863; measured value: 302.1883.

[0111] Step 7: Preparation of tert-butyl (4-(3-(4-(n-hexyloxy)-1-methylureido)cyclohexyl)carbamate) (Compound DW03116)

[0112] Compound DW03115 (600 mg, 2.6 mmol), DW03075 (800 mg, 2.6 mmol), and potassium carbonate (718.6 mg, 5.2 mmol) were placed in a reaction flask, and anhydrous DMF (10 mL) was added and stirred at room temperature. The reaction progress was monitored by TLC, and the reaction was complete after 4 hours. Most of the solvent was removed by vacuum distillation, and an appropriate amount of water was added and extracted three times with ethyl acetate. The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated to obtain a crude product, which was purified on a silica gel column (dichloromethane:methanol = 15:1) to obtain a light yellow oil DW03116 (744 mg, yield: 62%). HRMS (ESI) C 21 H 36 N3O2 + ([M+H] + )Calculated value: 462.3326; measured value: 462.3351.

[0113] Step 8: Preparation of (1-(4-aminocyclohexyl)-3-(4-n-hexyloxy)benzyl)-1-methylurea (Compound I-1)

[0114] Compound DW03116 (744 mg, 1.6 mmol) was placed in a reaction flask and 5 mL of 2N hydrochloric acid in dioxane was added to dissolve the mixture at room temperature. The reaction was continued with stirring at room temperature. TLC was used to monitor the reaction progress. After 2 hours, the reaction was complete. The solvent was removed by distillation under reduced pressure to obtain the crude product, which was purified on a silica gel column (dichloromethane:methanol = 15:1) to obtain I-1 (521 mg) as a white solid in a 92% yield. HRMS (ESI) revealed C 21 H 36 N3O2 + ([M+H] + )Calculated value: 362.2802; measured value: 362.2806. 1 H NMR (800MHz, CDCl3) δ8.35 (s, 1H), 7.21 (d, J = 8.4Hz, 2H), 6.85 (d, J = 8.4Hz, 2H), 4. 59-4.57(m,1H),4.34(d,J=4.9Hz,2H),4.25-4.23(m,1H),3.94-3.92(m,2H),3.46( s,1H),2.79(s,3H),2.05(d,J=13.6Hz,1H),1.87-1.85(m,2H),1.83-1.73(m,4H),1 .59(d,J=11.5Hz,2H),1.48-1.42(m,2H),1.37-1.31(m,4H),0.90(t,J=6.7Hz,3H).

[0115] Example 2: Preparation of 1-(4-amino-cis-cyclohexyl)-3-(4-hexyloxy)benzyl-1-methylurea (Compound I-2)

[0116] The preparation method is similar to that of I-1, except that tert-butyl (4-aminocyclohexyl)carbamate is replaced with 1-N-Boc-cis-1,4-cyclohexanediamine to obtain colorless oil I-2 (yield 92%). HRMS (ESI) C 21 H 36 N3O2 + ([M+H] + )Calculated value: 362.2802; measured value: 362.2802. 1H NMR (800MHz, CDCl3) δ7.22(d,J=8.3Hz,2H),6.85(d,J=8.3Hz,2H),4.62(s,1H),4.34(d,J=5.3Hz,2H),4.12(s,1H),3.93(t,J=6.6Hz,2H),2.8 4(s,1H),2.71(d,J=22.7Hz,3H),2.12-2.01(m,2H),1.78-1.74(m,2H) ,1.73(s,2H),1.52-1.42(m,6H),1.36-1.31(m,4H),0.92-0.88(m,3H).

[0117] Example 3: Preparation of 1-(4-amino-trans-cyclohexyl)-3-(4-hexyloxy)benzyl-1-methylurea (Compound I-3)

[0118] The preparation method is similar to that of I-1, except that tert-butyl (4-aminocyclohexyl)carbamate is replaced with 1-N-Boc-trans-1,4-cyclohexanediamine to obtain white solid I-3 (yield 90%).

[0119] HRMS(ESI)C 21 H 36 N3O2 + ([M+H] + )Calculated value: 362.2802; measured value: 362.2805. 1 H NMR (800MHz, CDCl3) δ7.23(d,J=7.8Hz,2H),6.85(d,J=7.8Hz,2H),4.53(s,1H),4.36(d,J=5.2Hz,2H),4.11(s,1H),3.94(t,J =6.5Hz,2H),3.25(s,1H),2.76(s,3H),1.83-1.68(m,8H),1.49-1.42(m,4H),1.33(s,4H),1.25(s,2H),0.90(t,J=6.1Hz,3H).

[0120] Example 4: Preparation of 1-(4-amino-trans-cyclohexyl)-3-(4-isobutoxybenzyl)-1-methylurea (Compound I-4)

[0121] The preparation method is similar to that of I-3, except that n-hexanol is replaced with 2-tert-butanol to obtain white solid I-4 (yield 90%).

[0122] HRMS(ESI)C 19 H 32N3O2 + ([M+H] + )Calculated value: 334.2489; measured value: 334.2493. 1 H NMR (800MHz, CD3OD) δ7.19(d,J=8.1Hz,2H),6.83(d,J=8.1Hz,2H),4.29(s,2H),4.09(s,1H),3.93(t,J=6.3Hz,2H),3.08(s,1H),2.7 8(s,3H),2.10(d,J=10.8Hz,2H),1.79-1.71(m,4H),1.58-1.50(m,2H),1.50-1.42(m,2H),1.40-1.24(m,14H),0.90(t,J=7.1Hz,3H).

[0123] Example 5: Preparation of 1-trans-4-amino-cyclohexyl-3-(4-decyloxy)benzyl-1-methylurea (Compound I-5)

[0124] The preparation method is similar to that of I-3, except that n-hexanol is replaced with n-decanol to obtain white solid I-5 (yield 92%). HRMS (ESI) C 25 H 44 N3O2 + ([M+H] + Calculated value: 418.3428; measured value: 418.3426. 1 H NMR(800MHz,CD3OD)δ7.19(d,J=8.1Hz,2H),6.83(d,J=8.1Hz,2H),4.29(s,2 H),4.09(s,1H),3.93(t,J=6.3Hz,2H),3.33(d,J=31.0Hz,2H),3.08(s,1H),2 .78(s,3H),2.10(d,J=10.8Hz,2H),1.79-1.71(m,4H),1.70-1.63(m,2H),1.5 8-1.50(m,2H),1.50-1.42(m,2H),1.40-1.24(m,14H),0.90(t,J=7.1Hz,3H).

[0125] Example 6: Preparation of 1-(4-amino-trans-cyclohexyl)-3-(4-heptyloxy)benzyl-1-methylurea (Compound I-6)

[0126] The preparation method is similar to that of I-3, except that n-hexanol is replaced with n-heptanol to obtain white solid I-6 (yield 88%). HRMS (ESI) C22 H 37 N3O2 + ([M+H] + Calculated value: 376.2959; measured value: 376.2965. 1 H NMR(800MHz,d6-DMSO)δ8.08(s,3H),7.14(d,J=8.0Hz,2H),6.83(d,J=8.1Hz,2H),4.14(d,J=5.1Hz,2H),3.91(t,J=6.1Hz,2H),2.92(s ,1H),2.65(s,3H),1.98(d,J=10.5Hz,2H),1.71-1.64(m,2H),1.52(s,4H),1.45-1.35(m,4H),1.33-1.24(m,6H),0.86(t,J=6.6Hz,3H).

[0127] Example 7: Preparation of 1-(4-amino-trans-cyclohexyl)-1-methyl-3-(4-octyloxy)benzylurea (Compound I-7)

[0128] The preparation method is similar to that of I-3, except that n-hexanol is replaced with n-octanol to obtain a white solid (yield 90%). HRMS (ESI) C 23 H 40 N3O2 + ([M+H] + )Calculated value: 390.3115; measured value: 390.3118. 1 H NMR(800MHz,d6-DMSO)δ7.14(d,J=8.6Hz,2H),6.84-6.82(m,2H),4.14(d,J=5.8Hz,2H),3.91(t,J=6.5Hz,3H),2.96-2.89(m,1H),2. 65(s,3H),1.99(d,J=12.3Hz,2H),1.71-1.64(m,2H),1.56-1.50(m,4H),1.47-1.36(m,4H),1.33-1.22(m,8H),0.86(t,J=7.1Hz,3H).

[0129] Example 8: Preparation of 1-(4-amino-cis-cyclohexyl)-3-(4-heptyloxy)benzyl-1-methylurea (Compound I-8)

[0130] The preparation method is similar to that of I-2, except that n-hexanol is replaced with n-heptanol to obtain white solid I-8 (yield 68%). HRMS (ESI) C 22H 37 N3O2 + ([M+H] + Calculated value: 376.2959; Measured value: 376.2958. 1 H NMR (800MHz, d6-DMSO) δ7.15(d,J=8.5Hz,2H),6.83(d,J=8.6Hz,2H),4.15(d,J=5.8Hz,2H),3.93(d,J=12.1Hz,1H),3.91(t,J=6.5Hz,2H),3.32 (s,1H),2.77(s,3H),1.90(d,J=14.2Hz,2H),1.87-1.79(m,2H),1.73-1 .64(m,4H),1.43-1.35(m,2H),1.35-1.25(m,8H),0.86(t,J=7.0Hz,3H).

[0131] Example 9: Preparation of 1-(3-amino-trans-cyclobutyl)-3-(4-hexyloxy)benzyl-1-methylurea (Compound I-9)

[0132] The preparation method is similar to that of I-1, except that (4-aminocyclohexyl)carbamic acid tert-butyl ester is replaced with (trans-3-aminocyclobutyl)carbamic acid tert-butyl ester to obtain white solid I-9 (yield 92%). HRMS (ESI) C 19 H 32 N3O2 + ([M+H] + )Calculated value: 334.2489; measured value: 334.2486. 1 H NMR(800MHz,d6-DMSO)δ7.16(d,J=8.4Hz,2H),6.88(t,J=5.7Hz,1H),6.83(d,J=8.5Hz,2H),4.14(d,J=5.7Hz,2H),3.91(t,J=6.5Hz,2H),3.5 7(s,1H),2.79(s,3H),2.55-2.45(m,3H),2.26-2.16(m,2H),1.70-1.6 4(m,2H),1.43-1.35(m,2H),1.33-1.27(m,4H),0.87(t,J=6.6Hz,3H).

[0133] Example 10: Preparation of 1-(4-amino-trans-cyclohexyl)-3-(4-hexyloxy)benzylurea (Compound I-10)

[0134] The preparation method is similar to that of I-3 to obtain a light yellow solid I-10 (yield 92%). HRMS (ESI) C 20 H 34 N3O2 + ([M+H] + )Calculated value: 348.2646; measured value: 348.2652. 1 H NMR (800MHz, CDCl3) δ7.18 (d, J=8.5Hz, 2H), 6.87-6.80 (m, 2H), 4.64 (s, 1H), 4.25 (d, J= 5.3Hz,2H),4.23(d,J=7.9Hz,1H),3.92(t,J=6.6Hz,2H),3.52-3.44(m,1H),2.64-2.57 (m,1H),1.97(d,J=11.3Hz,2H),1.84(d,J=12.2Hz,2H),1.78-1.75(m,2H),1.46-1.43( m,2H),1.26-1.24(m,2H),1.22-1.15(m,2H),1.14-1.06(m,2H),0.90(t,J=6.7Hz,3H).

[0135] Example 11: Preparation of 1-(4-amino-trans-cyclohexyl)-1-benzyl-3-(4-hexyloxy)benzylurea (Compound I-11)

[0136] Step 1: Preparation of trans-tert-butyl 4-(1-benzyl-3-(4-(hexyloxy)benzyl)ureidocyclohexyl)carbamate) (Compound DW04018-1)

[0137] The preparation method is similar to that of DW03116, and white solid DW04018-1 is obtained (yield 68%). HRMS (ESI) C 32 H 47 N3O4 + ([M+H] + )Calculated value: 538.3689; measured value: 538.3699. 1H NMR (800MHz, CDCl3) δ7.31(t,J=7.5Hz,2H),7.25(d,J=7.4Hz,1H),7.22(d,J=7.5Hz,2H),6 .95(d,J=8.5Hz,2H),6.75(t,J=5.7Hz,2H),4.51(s,1H),4.41-4.29(m,4H),4.25(d,J=2.5H z,2H),3.95-3.85(m,2H),3.33(s,1H),2.03(t,J=12.1Hz,2H),1.83(d,J=11.6Hz,2H),1.78 -1.72(m,2H),1.48-1.40(m,11H),1.37-1.30(m,4H),1.30-1.23(m,2H),0.93-0.88(m,3H).

[0138] Step 2: Preparation of 1-(4-amino-trans-cyclohexyl)-1-benzyl-3-(4-hexyloxy)benzylurea (Compound I-11):

[0139] The preparation method is similar to that of I-1, except that DW03116 is replaced by DW04018-1 to obtain white solid I-11 (yield 85%). HRMS (ESI) C 27 H 40 N3O2 + Calculated value: 438.3115; measured value: 438.3117. 1 H NMR (800MHz, CDCl3) δ7.29 (t, J=7.5Hz, 2H), 7.22 (dd, J=13.0, 5.6Hz, 1H), 7.20 (d, J= 7.4Hz,2H),6.94(d,J=8.1Hz,2H),6.73(d,J=8.4Hz,2H),4.32(s,2H),4.25(s,1H),4 .22(s,2H),3.92-3.86(m,2H),3.04(s,1H),2.22(d,J=8.7Hz,2H),1.86(d,J=9.5Hz, 2H),1.78-1.69(m,4H),1.52-1.39(m,4H),1.34-1.31(m,4H),0.90(t,J=6.9Hz,3H).

[0140] Example 12: Preparation of 1-(4-amino-trans-cyclohexyl)-1-(4-fluorobenzyl)-3-(4-hexyloxy)benzyl)urea (Compound I-12)

[0141] The preparation method is similar to that of I-11, except that benzyl bromide is replaced with 4-fluorobenzyl bromide to obtain white solid I-12 (yield 86%). HRMS (ESI) C 27 H 39 N3O2 + ([M+H] + )Calculated value: 456.3021; ​​measured value: 456.3038. 1 H NMR (800MHz, CDCl3) δ7.17(s,2H),6.98(dd,J=20.2,8.2Hz,4H),6.75(d,J=8.2Hz,2H),4.29(s,2H),4.23(s,3H),3.89(t,J=6.5Hz,2H),3.11( td,J=12.1,7.3Hz,1H),3.06(s,1H),2.23(s,2H),1.83(s,2H),1.77-1. 71(m,4H),1.45-1.39(m,4H),1.34-1.32(m,4H),0.90(t,J=6.7Hz,3H).

[0142] Example 13: Preparation of 1-(4-amino-trans-cyclohexyl)-3-(4-hexyloxy)benzyl-1-(2-methoxybenzyl)urea (Compound I-13)

[0143] The preparation method is similar to that of I-11, except that benzyl bromide is replaced with 2-methoxybenzyl bromide to obtain white solid I-13 (yield 92%). HRMS (ESI) C 28 H 42 N3O3 + ([M+H] + )Calculated value: 468.3221; measured value: 468.3223. 1 HNMR(800MHz, CDCl3)δ7.21(t,J=7.8Hz,1H),7.16(d,J=7.3Hz,1H),7.03(d,J=8.5Hz,2H), 6.91(t,J=7.4Hz,1H),6.80(d,J=8.2Hz,1H),6.77(d,J=8.6Hz,2H),4.25(d,J=5.8Hz,4H), 3.90(t,J=6.6Hz,2H),3.73(s,3H),2.22(d,J=10.5Hz,2H),1.85(d,J=11.2Hz,2H),1.78-1 .70(m,4H),1.55-1.47(m,2H),1.46-1.41(m,4H),1.34-1.31(m,4H),0.90(t,J=6.9Hz,3H).

[0144] Example 14: Preparation of 1-(4-amino-trans-cyclohexyl)-3-(4-hexyloxy)benzyl)-1-(3-methoxybenzyl)urea (Compound I-14)

[0145] The preparation method is similar to that of I-11, except that benzyl bromide is replaced with 3-methoxybenzyl bromide to obtain white solid I-14 (yield 90%). HRMS (ESI) C 28 H 42 N3O3 + ([M+H] + )Calculated value: 468.3221; measured value: 468.3221. 1 HNMR (800MHz, CDCl3) δ7.20(t,J=7.8Hz,1H),6.96(d,J=8.0Hz,2H),6.78(d,J=6.9Hz,1H),6.74(dd,J=18.1,9.8Hz,4H),4.34-4.25(m,3H),4.25-4 .19(m,2H),3.93-3.86(m,2H),3.72(s,3H),2.23(s,2H),1.85(s,2H),1. 77-1.69(m,4H),1.49-1.39(m,4H),1.36-1.31(m,4H),0.91-0.88(m,3H).

[0146] Example 15: Preparation of (3-(4-(hexyloxy)benzyl)-1-trans-4-(2-hydroxybenzyl)amino)cyclohexyl)-1-methylurea) (Compound I-15)

[0147] The preparation method is the same as that of DW03112, except that tert-butyl (4-aminocyclohexyl)carbamate is replaced by I-3 and benzaldehyde is replaced by 2-hydroxybenzaldehyde to obtain a light yellow solid I-15 (yield: 45%). HRMS (ESI) C 28 H 42 N3O3 + ([M+H] + )Calculated value: 468.3221; measured value: 468.3232. 1H NMR (800MHz, CDCl3) δ7.05(t,J=9.0Hz,3H),7.01(d,J=6.9Hz,1H),6.83(d,J=7.2Hz,1H),6.74( d,J=8.2Hz,2H),6.66(t,J=7.1Hz,1H),4.85(s,1H),4.06(s,2H),3.99(s,1H),3.94(s,2H),3.8 3(t,J=6.5Hz,2H),2.71(s,1H),2.57(s,3H),2.05(s,2H),1.71-1.64(m,2H),1.57(s,2H),1.48 (d,J=9.2Hz,2H),1.39-1.33(m,2H),1.29(d,J=11.4Hz,2H),1.25(s,4H),0.82(t,J=6.3Hz,3H).

[0148] Example 16: Preparation of 3-(4-hexyloxy)benzyl)-1-methyl-1-(4-(methylamino)cyclohexyl)urea (Compound I-16)

[0149] Step 1: Preparation of 3-(4-hexyloxy)benzyl)-1-methyl-1-(4-oxocyclohexyl)urea (Compound DW04108):

[0150] The preparation method is the same as that of DW03116, except that DW03115 is replaced by 4-(methylamino)cyclohexyl-1-one, resulting in a pale yellow oil with a yield of 56%. HRMS (ESI) C 21 H 33 N2O3 + ([M+H] + )Calculated value: 361.2486; measured value: 361.2497.

[0151] Step 2: Preparation of compound I-16:

[0152] Compound DW04108 (100 mg, 0.28 mmol) and 33% methylamine solution (0.06 mL, 0.28 mmol) were placed in a reaction flask. 5 mL of methanol was stirred at room temperature to dissolve the mixture. Sodium cyanoborohydride (23 mg, 0.34 mmol) was then added to the reaction system and the mixture was stirred at room temperature. TLC confirmed the reaction was complete after 4 hours. A small amount of water was added to the reaction system to quench the reaction. The reaction was extracted three times with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified on a silica gel column (dichloromethane:methanol = 20:1) to afford I-16 as a pale yellow oil in a 56% yield. HRMS (ESI) revealed C 22 H 38N3O2 + ([M+H] + )Calculated value: 376.2959; measured value: 376.2966. 1 H NMR (800MHz, DMSO) δ7.17-7.12(m,2H),6.86-6.81(m,2H),4.18-4.13(m,2H),3.94-3.89(m,2H),3.17(d,J=4.4Hz,1H),2.69-2.65(m,2H) ),2.56-2.52(m,2H),2.16-2.03(m,2H),1.71-1.66(m,2H),1.62-1.50(m,2H),1.41-1.37(m,2H),1.31-1.28(m,4H),0.88-0.86(m,3H).

[0153] Example 17: Preparation of N-(4-amino-trans-cyclohexyl)-3-(4-hexyloxy)phenyl-N-methylacrylamide (Compound I-17):

[0154] Step 1: Preparation of methyl 3-(4-(hexyloxy)phenyl)acrylate (compound DW04093):

[0155] 4-n-Hexyloxybenzaldehyde (2.1 g, 10 mmol) and methoxymethylenetriphenylphosphine (3.7 g, 11 mmol) were placed in a reaction flask, 30 mL of dichloromethane was added, and the mixture was stirred at room temperature to dissolve. The mixture was heated to 50°C and refluxed with stirring. The reaction progress was monitored by TLC. The reaction was allowed to proceed overnight, and the solvent was removed by distillation under reduced pressure. The mixture was purified on a silica gel column (petroleum ether:ethyl acetate = 3:1) to obtain DW04093 (2.1 g, yield: 82%) as a white solid. HRMS (ESI) C 16 H 23 O3 + ([M+H] + )Calculated value: 263.1642; measured value: 263.1663.

[0156] Step 2: Preparation of 3-(4-(hexyloxy)phenyl)acrylic acid (compound DW04094):

[0157] Compound DW04093 (2.6 g, 10 mmol) was placed in a reaction flask and dissolved in 20 mL of methanol. 10 mL of 2N sodium hydroxide solution was added while stirring at room temperature, and then heated to 90°C and refluxed with stirring for reaction. The reaction progress was monitored by TLC. After 1 hour, the reaction was complete. The pH was adjusted to 3.0 with 6N hydrochloric acid, and most of the solvent was removed by distillation under reduced pressure. A white solid precipitated and was filtered to obtain white solid DW04094 (2.2 g, yield: 88%). HRMS (ESI) C15 H 19 O3 - ([MH] - )Calculated value: 247.1340; measured value: 247.1335.

[0158] Step 3: Preparation of tert-butyl 4-(3-(4-(hexyloxy)phenyl)-N-methylacrylamido)cyclohexyl)carbamate (Compound DW04095):

[0159] Compound DW04094 (500 mg, 2 mmol), trans-4-(methylamino)cyclohexylcarbamic acid tert-butyl ester (460 mg, 2 mmol), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (HATU) (1.1 g, 3 mmol), and diisopropylethylamine (1 mL, 6 mmol) were placed in a reaction flask and dissolved in 10 mL of dry N,N-dimethylformamide. The reaction was stirred at room temperature. TLC was used to monitor the reaction progress. The reaction was stirred overnight and the reaction was complete. Most of the solvent was removed by distillation under reduced pressure. An appropriate amount of water was added and the mixture was extracted three times with ethyl acetate. The organic phases were combined and dried over anhydrous sodium sulfate. The crude product was concentrated and purified on a silica gel column (petroleum ether:ethyl acetate = 1:1) to obtain DW04095 (322 mg, 70% yield) as a white solid. HRMS (ESI) C 27 H 43 N2O4 + ([M+H] + )Calculated value: 459.3217; measured value: 459.3238. 1 H NMR (600MHz, CDCl3) δ7.64(d,J=15.3Hz,1H),7.48-7.44(m,2H),6.91-6.85(m,2H),6.70(d,J=15.3Hz,1H),4.40(d,J=6.9Hz,1H),3.99-3.96(m,2 H),3.41(s,1H),2.96(s,3H),2.14-2.05(m,2H),1.82-1.72(m,4H),1.70 -1.52(m,2H),1.50-1.39(m,10H),1.37-1.23(m,7H),0.94-0.86(m,3H).

[0160] Step 4: Preparation of N-(4-amino-trans-cyclohexyl)-3-(4-hexyloxy)phenyl-N-methylacrylamide (Compound I-17):

[0161] The preparation method is the same as that of I-1, except that DW03116 is replaced by DW04095, resulting in a white solid with a yield of 75%. HRMS (ESI) C22 H 35 N2O2 + ([M+H] + )Calculated value: 359.2693; measured value: 359.2692. 1 H NMR(600MHz,CD3OD)δ7.63-7.49(m,3H),7.02-6.88(m,3H),4.50-4.41(m,1H), 4.03-3.96(m,2H),3.32-3.30(m,3H),3.14(t,J=11.8Hz,1H),3.08(d,J=13.7H z,2H),2.14(d,J=11.7Hz,2H),1.87(s,1H),1.81-1.73(m,4H),1.62-1.53(m,2 H),1.51-1.45(m,2H),1.40-1.35(m,4H),1.33-1.27(m,9H),0.96-0.88(m,3H).

[0162] Example 18: Preparation of N-(4-amino-trans-cyclohexyl)-3-(4-hexyloxyphenyl)-N-methylpropionamide (Compound I-18):

[0163] Step 1: Preparation of 3-(4-(hexyloxy)phenyl)propanoic acid (compound DW04096):

[0164] Compound DW04094 (800 mg, 3.2 mmol) was placed in a reaction flask and dissolved in 10 mL of methanol. 10% palladium on carbon (100 mg) was then added, and the gas was replaced with hydrogen. The reaction was heated to 50°C and stirred. The reaction progress was monitored by TLC. Stirring overnight indicated that the reaction was complete. The palladium on carbon was filtered out, and the filtrate was evaporated to dryness under reduced pressure to obtain the crude product. Yield: 83%. HRMS (ESI): C 15 H 23 O3 + ([M+H] + )Calculated value: 251.1642; measured value: 251.1661.

[0165] Step 2: Preparation of tert-butyl 4-(3-(4-(hexyloxy)phenyl)-N-methylpropionamido)cyclohexyl)-trans-carbamate (Compound DW04097):

[0166] The preparation method is the same as that of compound DW04095. Pale yellow solid, yield: 67%. HRMS (ESI) C 27 H 45 N2O4 + ([M+H] +)Calculated value: 461.3374; measured value: 461.3381. 1 H NMR(600MHz, CDCl3) δ7.10(t,J=10.8Hz,2H),6.81(d,J=8.2Hz,2H),4.46(d,J=11.7Hz,1H) ,4.43-4.29(m,1H),3.92(t,J=6.6Hz,2H),3.36(s,1H),2.94-2.88(m,2H),2.81-2.74(m,1H ),2.71(s,2H),2.63-2.54(m,2H),2.08-2.01(m,2H),1.91(s,2H),1.80-1.73(m,2H),1.66 -1.60(m,2H),1.59-1.51(m,1H),1.50-1.41(m,11H),1.38-1.31(m,4H),0.93-0.85(m,3H).

[0167] Step 3: Preparation of N-(4-amino-trans-cyclohexyl)-3-(4-hexyloxyphenyl)-N-methylpropionamide (Compound I-18):

[0168] The preparation method is the same as that of compound I-17, except that DW04095 is replaced by DW04097, resulting in a pale yellow solid with a yield of 89%. HRMS (ESI) C 22 H 37 N2O2 + ([M+H] + )Calculated value: 361.2850; measured value: 361.2851. 1 H NMR(600MHz,CD3OD)δ7.14-7.08(m,2H),6.81(dd,J=13.0,8.6Hz,2H),4.36-4.33(m,1H),3.92(t, J=6.5Hz,2H),3.60-3.53&3.21(dd,J=14.6,7.3Hz,1H),3.10-3.01(m,1H),2.88-2.83(m,2H),2.79 -2.74(m,3H),2.68(t,J=7.4Hz,1H),2.63(t,J=7.5Hz,1H),2.11-2.07(m,1H),2.06-2.00(m,1H), 1.76-1.71(m,2H),1.70-1.62(m,3H),1.53-1.44(m,4H),1.38-1.33(m,4H),0.92(t,J=6.8Hz,3H).

[0169] Example 19: Preparation of N-(4-amino-cis-cyclohexyl)-3-(4-hexyloxy)phenyl-N-methylacrylamide (Compound I-19):

[0170] The preparation method is the same as that of compound I-17, except that 1-N-Boc-trans-1,4-cyclohexanediamine is replaced by 1-N-Boc-cis-1,4-cyclohexanediamine, resulting in a pale yellow solid with a yield of 67%. HRMS (ESI) C 22 H 35 N2O2 + ([M+H] + )Calculated value: 359.2693; measured value: 359.2702. 1 H NMR(800MHz,d6-DMSO)δ7.67-7.58(m,2H),7.45-7.38(m,1H),7.02(dd,J=61.1,15.3H z,1H),6.94(d,J=8.8Hz,2H),4.43-4.35&4.10-4.01(m,1H),3.99(t,J=6.5Hz,2H),3. 42-3.34(m,2H),3.07(s,3H),1.93-1.84(m,4H),1.75(d,J=13.2Hz,1H),1.73-1.69(m ,2H),1.49(d,J=10.0Hz,1H),1.43-1.39(m,2H),1.32-1.29(m,4H),0.90-0.85(m,3H).

[0171] Example 20: Preparation of N-(4-amino-cis-cyclohexyl)-3-(4-hexyloxyphenyl)-N-methylpropionamide (Compound I-20):

[0172] The preparation method is the same as that of compound I-18, except that 1-N-Boc-trans-1,4-cyclohexanediamine is replaced with 1-N-Boc-cis-1,4-cyclohexanediamine. The product is a pale yellow solid with a yield of 67%. HRMS (ESI) C 22 H 37 N2O2 + ([M+H] + )Calculated value: 361.2850; measured value: 361.2857. 1H NMR(800MHz,d6-DMSO)δ7.12(d,J=8.4Hz,2H),6.81(d,J=8.5Hz,2H),4.33-4. 26&3.61-3.55(m,1H),3.90(t,J=6.5Hz,2H),3.36-3.28(m,1H),2.81(s,3H), 2.74-2.69(m,2H),2.60-2.51(m,2H),1.95-1.80(m,4H),1.75-1.64(m,4H),1 .43-1.36(m,2H),1.34(d,J=8.8Hz,1H),1.32-1.29(m,4H),0.91-0.84(m,3H).

[0173] Example 21: Preparation of N-(4-amino-trans-cyclohexyl)-2-(4-hexyloxy)phenyl-N-methylacetamide (Compound I-21):

[0174] Step 1: Preparation of methyl 2-(4-(hexyloxy)phenyl)acetate (compound DW04100):

[0175] Place methyl 4-hydroxyphenylacetate (1.66 g, 10 mmol), cesium carbonate (4.9 g, 15 mmol), and n-bromohexane (1.6 g, 10 mmol) in a reaction flask, add 15 mL of dichloromethane, stir and dissolve at room temperature, and continue stirring to react. TLC monitors the reaction progress, and the reaction is complete after 6 hours. Add appropriate amount of water and extract with dichloromethane three times, combine the organic phases, and dry over anhydrous sodium sulfate. Concentrate the organic phase to obtain a crude product, which is purified on a silica gel column (petroleum ether: ethyl acetate = 1:1) to obtain a colorless oil (1.75 g, yield: 70%). HRMS (ESI) C 15 H 23 O3 + ([M+H] + )Calculated value: 251.1642; measured value: 251.1653. 1 H NMR(800MHz, CDCl3)δ7.18(d,J=8.6Hz,2H),6.88-6.81(m,2H),3.93(t,J=6.6Hz,2H),3.68(s, 3H),3.56(s,2H),1.80-1.73(m,2H),1.49-1.42(m,2H),1.39-1.28(m,4H),0.93-0.85(m,3H).

[0176] Step 2: Preparation of 2-(4-(hexyloxy)phenyl)acetic acid (compound DW04101):

[0177] The preparation method is the same as that of DW04094, except that DW04093 is replaced by DW04100, resulting in a white solid with a yield of 89%. HRMS (ESI) C 14 H 21 O3 + ([M+H] + )Calculated value: 237.1485; measured value: 237.1493.

[0178] Step 3: Preparation of tert-butyl 4-(2-(4-(hexyloxy)phenyl)-N-methylacetylamino)cyclohexyl)-trans-carbamate (compound DW04102):

[0179] The preparation method is the same as that of DW04095, except that DW04094 is replaced by DW04101, resulting in a white solid with a yield of 72%. HRMS (ESI) C 26 H 43 N2O4 + ([M+H] + )Calculated value: 447.3217; measured value: 447.3233. 1 H NMR (800MHz, CDCl3) δ7.13(t,J=7.9Hz,2H),6.83(d,J=8.4Hz,2H),4.48(s,1H),4.37(d,J=39.9Hz,1H),3.92(t,J=6.6Hz,2H),3.71-3.61(m,2H),2.7 8(s,3H),2.06-1.98(m,2H),1.78-1.74(m,2H),1.71-1.64(m,6H),1.61-1 .52(m,1H),1.44(d,J=18.8Hz,11H),1.35-1.31(m,4H),0.95-0.87(m,3H).

[0180] Step 4: Preparation of N-(4-amino-trans-cyclohexyl)-2-(4-hexyloxy)phenyl-N-methylacetamide (Compound I-21):

[0181] The preparation method is the same as that of I-17, except that DW04095 is replaced by DW04102, resulting in a white solid with a yield of 90%. HRMS (ESI) C 21 H 35 N2O2 + ([M+H] + )Calculated value: 347.2693; measured value: 347.2693. 1H NMR (800MHz, d6-DMSO) δ7.10 (dd, J=21.2, 8.5Hz, 2H), 6.85 (t, J=8.0Hz, 2H), 4.27-4. 18&3.67-3.64(m,1H),3.94-3.88(m,2H),3.69-3.67(m,1H),3.58(s,1H),2.97-2.88 (m,1H),2.79&2.65(m,3H),1.99(d,J=11.8Hz,1H),1.95-1.88(m,1H),1.71-1.66(m, 2H),1.59-1.47(m,3H),1.44-1.34(m,5H),1.31-1.28(m,4H),0.87(t,J=7.0Hz,3H).

[0182] Example 22: Preparation of N-(4-amino-cis-cyclohexyl)-2-(4-hexyloxy)phenyl-N-methylacetamide (Compound I-22):

[0183] The preparation method is the same as that of compound I-21, except that 1-N-Boc-trans-1,4-cyclohexanediamine is replaced by 1-N-Boc-cis-1,4-cyclohexanediamine, resulting in a white solid with a yield of 88%. HRMS (ESI) C 21 H 35 N2O2 + ([M+H] + )Calculated value: 347.2693; measured value: 347.2693. 1 H NMR(800MHz,d6-DMSO)δ7.15-7.08(m,2H),6.84(d,J=8.4Hz,2H),4.31-4.26&3.75-3. 69(m,1H),4.15-4.11(m,1H),3.92(t,J=6.5Hz,2H),3.65(s,1H),3.58(s,1H),3.32(d ,J=15.1Hz,1H),3.17(d,J=5.2Hz,2H),2.90&2.75(s,3H),1.92-1.87(m,1H),1.85-1. 81(m,2H),1.70-1.66(m,4H),1.42-1.37(m,2H),1.32-1.29(m,5H),0.89-0.85(m,3H).

[0184] Example 23: Preparation of N-(3-amino-trans-cyclobutyl)-2-(4-hexyloxy)phenyl-N-methylacetamide (Compound I-23):

[0185] The preparation method is the same as that of compound I-21, except that 1-N-Boc-trans-1,4-cyclohexanediamine is replaced by 1-N-Boc-trans-1,3-cyclobutanediamine, resulting in a white solid with a yield of 90%. HRMS (ESI) C 19 H 31 N2O2 + ([M+H] + )Calculated value: 319.2380; measured value: 319.2381. 1 H NMR (800MHz, d6-DMSO) δ7.15(d,J=8.4Hz,1H),7.09(d,J=8.4Hz,1H),6.84(t,J=8.0Hz,1H),3.91(t,J=6.3Hz,2H),3. 63(s,1H),3.61-3.60&3.55-3.51(m,1H),3.59(s,1H),3.02&2.73(s,3H),2.55-2.46(m,2H),2.16-2.05(m,2H),1.71 -1.65(m,2H),1.43-1.36(m,2H),1.33-1.26(m,4H),0.90-0.84(m,3H).

[0186] Example 24: Preparation of N-(1S, 3S)-3-aminocyclopentyl-2-(4-hexyloxy)phenyl-N-methylacetamide (Compound I-24):

[0187] The preparation method is the same as that of compound I-21, except that 1-N-Boc-trans-1,4-cyclohexanediamine is replaced with (1S,3S)-3-aminocyclopentyl)carbamic acid tert-butyl ester, resulting in a white solid with a yield of 82%. HRMS (ESI) C 20 H 33 N2O2 + ([M+H] + )Calculated value: 333.2537; measured value: 333.2554. 1H NMR(800MHz,d6-DMSO)δ7.23-7.16(m,1H),7.09(d,J=8.2Hz,1H),6.88-6.80(m,2H),5.05-4.9 6&4.79-4.74(m,1H),3.92(t,J=6.4Hz,2H),3.74-3.63(m,1H),3.60-3.56(m,2H),2.99(d,J=7. 0Hz,1H),2.84-2.62(m,3H),2.10-2.01(m,1H),1.93-1.83(m,1H),1.82-1.74(m,2H),1.71-1. 67(m,2H),1.62-1.53(m,2H),1.43-1.36(m,2H),1.29(t,J=16.2Hz,4H),0.88(t,J=6.3Hz,3H).

[0188] Example 25: Preparation of N-(4-amino-trans-cyclohexyl)-2-(4-isobutoxyphenyl)-N-methylacetamide (Compound I-25):

[0189] The preparation method is the same as that of compound I-21, except that n-bromohexane is replaced by 1-bromo-2-methylpropyl. The product is a white solid with a yield of 90%. HRMS (ESI) C 19 H 31 N2O2 + ([M+H] + )Calculated value: 319.2380; measured value: 319.2385. 1 HNMR(800MHz,d6-DMSO)δ7.11(dd,J=20.5,8.5Hz,2H),6.85(t,J=8.3Hz,2H), 4.26-4.18&3.67-3.64(m,1H),3.72-3.69(m,2H),3.67(s,1H),3.58(s,1H),2 .97-2.87(m,1H),2.83-2.62(m,3H),2.02-1.96(m,2H),1.94-1.88(m,1H),1. 62-1.47(m,3H),1.45-1.40(m,1H),1.39-1.33(m,2H),0.97(d,J=6.7Hz,6H).

[0190] Example 26: Preparation of N-(4-amino-trans-cyclohexyl)-N-methyl-2-(4-octyloxy)phenyl)acetamide (Compound I-26):

[0191] The preparation method is the same as that of compound I-21, except that n-bromohexane is replaced by n-bromooctane, resulting in a white solid with a yield of 85%. HRMS (ESI) C 23 H 39 N2O2 + ([M+H] + )Calculated value: 375.3006; measured value: 375.3015. 1 H NMR (800MHz, d6-DMSO) δ7.10 (dd, J=25.9, 8.5Hz, 2H), 6.84 (t, J=8.4Hz, 2H), 4.25-4.1 8&3,67-3.63(m,1H),3.94-3.89(m,2H),3.68-3.62(m,1H),3.57(s,1H),2.80-2.60(m ,3H),1.87(d,J=11.7Hz,1H),1.78(d,J=11.6Hz,1H),1.71-1.66(m,2H),1.56-1.46(m ,2H),1.46-1.42(m,1H),1.42-1.36(m,2H),1.32-1.23(m,10H),0.86(t,J=7.0Hz,3H).

[0192] Example 27: Preparation of N-(4-amino-trans-cyclohexyl)-2-(4-decyloxy)phenyl-N-methylacetamide compound (I-27)

[0193] The preparation method is the same as that of compound I-21. Replace n-bromohexane with n-bromodecane, white solid, yield: 85%. HRMS (ESI) C 25 H 43 N2O2 + ([M+H] + )Calculated value: 403.3319; measured value: 403.3327. 1 H NMR (800MHz, d6-DMSO) δ7.10 (dd, J=21.0, 8.5Hz, 2H), 6.84 (t, J=7.8Hz, 2H), 4.25 -4.19&3.68-3.64(m,1H),3.91(t,J=6.5Hz,2H),3.71-3.62(m,1H),3.58(s,1H),2 .80-2.64(m,3H),1.99(d,J=11.7Hz,1H),1.92(d,J=9.3Hz,1H),1.71-1.66(m,2H) ,1.61-1.47(m,3H),1.45-1.34(m,5H),1.33-1.21(m,13H),0.86(t,J=7.1Hz,3H).

[0194] Example 28: Preparation of N-(4-amino-trans-cyclohexyl)-2-(4-methoxyphenyl)-N-methylacetamide (Compound I-28)

[0195] The preparation method is the same as that of compound I-21, except that 4-hydroxyphenylacetic acid methyl ester is replaced by 4-methoxyphenylacetic acid methyl ester, resulting in a white solid with a yield of 89%. HRMS (ESI) C 16 H 25 N2O2 + ([M+H] + )Calculated value: 277.1911; measured value: 277.1905. 1 H NMR(800MHz,d6-DMSO)δ7.12(dd,J=20.7,8.6Hz,2H),6.88-6.84(m,2H),4.25-4.19&3.68-3.63(m,1H),3.72(d,J=1.5Hz,3H ),3.69(s,1H),3.59(s,1H),2.81-2.64(m,3H),2.02-1.87(m,2H),1.62-1.48(m,3H),1.44-1.34(m,3H),1.31-1.26(m,1H).

[0196] Example 29: Preparation of N-(3-amino-trans-cyclobutyl)-2-(4-isobutoxyphenyl)-N-methylacetamide (Compound I-29)

[0197] The preparation method is the same as that of compound I-21, except that n-bromohexane is replaced by 1-bromo-2-methylpropyl, and 1-N-Boc-trans-1,4-cyclohexanediamine is replaced by 1-N-Boc-trans-1,3-cyclobutanediamine. The yield is 85%. HRMS (ESI) C 17 H 27 N2O2 + ([M+H] + )Calculated value: 291.2067; measured value: 291.2067. 1H NMR(800MHz,d6-DMSO)δ7.10(dd,J=13.8,8.5Hz,2H),6.85(dd,J=8.3,3.7Hz,2H),5.32-5.25(m,1H),5.09-5.01(m,1H),3.70(d,J =6.5Hz,2H),3.65-3.57(m,3H),2.98-2.78(m,3H),2.60-2.51(m,2H),2.20-2.10(m,2H),2.00-1.98(m,2H),0.97(d,J=6.7Hz,6H).

[0198] Example 30: Preparation of N-(1S, 3S)-3-aminocyclopentyl-2-(4-isobutoxyphenyl)-N-methylacetamide (Compound I-30)

[0199] The preparation method is the same as that of compound I-29, except that 1-N-Boc-trans-1,3-cyclobutanediamine is replaced with (1S,3S)-3-aminocyclopentyl)carbamic acid tert-butyl ester, resulting in a white solid with a yield of 82%. HRMS (ESI) C 18 H 29 N2O2 + ([M+H] + )Calculated value: 305.2224; measured value: 305.2227. 1 H NMR(800MHz,d6-DMSO)δ7.11(dd,J=27.8,8.4Hz,2H),6.85(d,J=8.5Hz,2H),5.03- 4.96&4.67-4.58(m,1H),3.70(d,J=6.5Hz,2H),3.68-3.57(m,3H),2.74(d,J=125. 5Hz,3H),2.10-2.03(m,1H),2.02-1.96(m,2H),1.93-1.75(m,2H),1.74-1.67(m,2 H),1.63-1.55(m,1H),1.54-1.45(m,1H),1.19-1.13(m,1H),0.97(d,J=6.7Hz,6H).

[0200] Example 31: Preparation of N-(4-amino-trans-cyclohexyl)-2-(5-methoxy-1H-indol-3-yl)-N-methylacetamide (Compound I-31)

[0201] The preparation method is the same as that of compound I-21, except that DW04101 is replaced with 5-methoxyindole-3-acetic acid, resulting in a white solid with a yield of 91%. HRMS (ESI) C18 H 26 N3O2 + ([M+H] + )Calculated value: 316.2020; measured value: 316.2040. 1 H NMR(800MHz,d6-DMSO)δ7.25-7.21(m,1H),7.12-7.09(m,1H),7.05(dd,J=10.3,2.3Hz,1H ),6.72(dd,J=8.7,2.4Hz,1H),4.28-4.20(m,1H),3.83-3.78(m,1H),3.78(s,1H),3.73(d, J=2.9Hz,3H),3.68(s,1H),3.09-3.04(m,1H),2.96-2.86(m,1H),2.85-2.61(m,3H),2.01 -1.84(m,2H),1.59-1.52(m,2H),1.51-1.48(m,1H),1.44-1.39(m,1H),1.37-1.27(m,2H).

[0202] Example 32: Preparation of N-(3-amino-trans-cyclobutyl)-2-(3-hexyloxy)phenyl)-N-methylacetamide (Compound I-32)

[0203] The preparation method is the same as that of compound I-23, except that 4-hydroxyphenylacetic acid methyl ester is replaced by 3-hydroxyphenylacetic acid methyl ester, resulting in a white solid with a yield of 89%. 1 H NMR (800MHz, CDCl3) δ7.13-7.07(m,1H),6.68(dd,J=20.6,7.6Hz,2H),6.63(s,1H),3.96-3.79(m,3H),3.75-3.53(m,2H),2. 89-2.80(m,3H),2.69(s,1H),2.55-2.45(m,2H),1.75-1.67(m,2H),1.45-1.38(m,2H),1.35-1.28(m,4H),0.91-0.86(m,3H).

[0204] Example 33: Preparation of N-(3-amino-trans-cyclobutyl)-N-methyl-2-(4-octyloxy)phenyl)acetamide (Compound I-33)

[0205] The preparation method is the same as that of compound I-23, except that n-bromohexane is replaced by n-bromooctane. The product is a white solid with a yield of 81%. 1H NMR(800MHz, CDCl3) δ7.12(dd,J=39.4,8.4Hz,2H),6.82(d,J=6.7Hz,2H),3.91(t,J =6.6Hz,2H),3.64(d,J=61.1Hz,3H),2.92(s,2H),2.49-2.35(m,2H),2.27-2.22(m, 1H),1.98-1.92(m,1H),1.80-1.73(m,2H),1.45-1.42(m,2H),1.34-1.27(m,10H),0.88(t,J=7.2Hz,3H).

[0206] Example 34 Preparation of N-(3-amino-trans-cyclobutyl)-2-(4-hexyloxy)phenyl-N-isopropylacetamide (Compound I-34)

[0207] The preparation method is the same as that of compound I-23, except that formaldehyde is replaced by acetone. The product is a white solid with a yield of 90%. 1 H NMR(800MHz, CDCl3) δ7.09(d,J=8.3Hz,2H),6.84(d,J=8.4Hz,2H),4.40-4.3 1(m,1H),4.13-4.07(m,1H),4.00-3.94(m,1H),3.92(t,J=6.6Hz,2H),3.62( s,2H),3.43-3.32(m,2H),2.40(t,J=10.4Hz,2H),1.79-1.73(m,2H),1.48-1 .41(m,2H),1.36-1.30(m,4H),0.98(d,J=6.5Hz,6H),0.90(t,J=6.8Hz,3H).

[0208] Example 35: Preparation of N-(3-amino-trans-cyclobutyl)-N-(4-fluorobenzyl)-2-(4-hexyloxy)phenyl)acetamide (Compound I-35)

[0209] The preparation method is the same as that of compound I-23, except that benzaldehyde is replaced by 4-fluorobenzaldehyde, resulting in a white solid with a yield of 77%. 1H NMR (800MHz, CDCl3) δ7.29-7.17(m,2H),7.03-6.86(m,4H),6.79-6.57(m,2H),4.59-4.30(m,2H),3.90-3.68(m,3H),3.46-3.30 (m,1H),2.74-2.57(m,1H),2.51-2.28(m,2H),1.75-1.64(m,2H),1.44-1.35(m,2H),1.34-1.26(m,4H),0.90(t,J=13.6Hz,3H).

[0210] Example 36: Preparation of N-(3-amino-trans-cyclobutyl)-4-hexyloxy-N-methylbenzamide (Compound I-36)

[0211] The preparation method is the same as that of compound I-23, except that 4-hydroxyphenylacetic acid methyl ester is replaced by 4-hydroxybenzoic acid methyl ester, resulting in a white solid with a yield of 92%. 1 H NMR (800MHz, CDCl3) δ7.32(d,J=8.1Hz,2H),6.83(d,J=8.3Hz,2H),3.94(t,J=6.5Hz,2H),2.98(s,3H),2.74-2.66(m,3H),1.83-1.73(m,2H),1.48- 1.41(m,2H),1.38-1.26(m,8H),0.90(t,J=6.9Hz,3H).

[0212] Example 37: Preparation of N-(4-amino-trans-cyclohexyl)-2-(4-hexyloxy)phenyl-N-methyl-2-oxoacetamide (Compound I-37)

[0213] The preparation method is the same as that of compound I-21, except that 4-hydroxyphenylacetic acid methyl ester is replaced by 2-(4-hydroxyphenyl)-2-oxoacetic acid methyl ester, resulting in a white solid with a yield of 79%. 1H NMR(800MHz,d6-DMSO)δ7.81(d,J=8.8Hz,2H),7.78(d,J=8.8Hz,2H),7.12(dd,J=8.9,3.4Hz,4H), 4.27-4.20(m,1H),4.09(td,J=6.5,3.6Hz,4H),3.15-3.09(m,1H),3.07-2.98(m,3H),2.97-2.92( m,1H),2.89(s,3H),2.72(s,3H),2.09-2.01(m,2H),1.97-1.88(m,2H),1.83-1.76(m,2H),1.76-1 .69(m,8H),1.64-1.59(m,2H),1.54-1.47(m,2H),1.45-1.38(m,4H),0.88(dt,J=7.0,3.5Hz,6H).

[0214] Example 38: Preparation of N-(3-amino-trans-cyclobutyl)-N-ethyl-2-(4-(hexyloxy)phenyl)acetamide

[0215] Step 1. In a round-bottom flask, tert-butyl (3-amino-trans-cyclobutyl)carbamate (500 mg, 2.69 mmol) and a tetrahydrofuran solution of acetaldehyde (0.65 mL, 5 M) were dissolved in methanol (10 mL), and glacial acetic acid (30 μL) was added and stirred at room temperature for 30 minutes. NaBH3CN (339 mg, 5.38 mmol) was then added and the reaction system was stirred at room temperature for 3 hours. After the reaction was completed, the solvent was removed and WHH114 (0.5 g, 88%) was purified and isolated by flash silica gel column chromatography. HRMS (ESI): C 11 H 23 N2O2 + [M+H]+ calculated: 215.1754; found: 215.1753.

[0216] Step 2. WHH114 (0.18 g, 0.84 mmol), hexyloxyphenylacetic acid (236 mg, 1 mmol), HATU (638 mg, 1.68 mmol), and DIPEA (0.5 mL) were added to a round-bottom flask and dissolved in tetrahydrofuran (8 mL). The reaction system was stirred at room temperature overnight. After the reaction, the solvent was removed and the product was purified by flash silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 2) to obtain WHH116 (0.21 g, 58%). HRMS (ESI): C 25 H 41 N2O4 + [M+H] +Calculated value: 433.3066; measured value: 433.2930.

[0217] Step 3. Add WHH116 (0.19 g, 0.44 mmol) to a round-bottom flask, followed by a 1,4-dioxane solution of hydrogen chloride (6 mL, 1 M), and stir at room temperature overnight. After the reaction is complete, filter to obtain compound I-41 (70 mg, 50%) as a white solid. 1 H NMR (800MHz, CD3OD) δ7.23-7.03(m,2H),6.87(dd,J=8.4,3.9Hz,2H),4.56(p,J=8.5Hz,1H),3. 95-3.93(m,2H),3.90-3.87(m,1H),3.71-3.68(m,2H),3.50-3.43(m,2H),2.85-2.79(m,1H),2. 67-2.62(m,1H),2.44-2.41(m,1H),2.23-2.20(m,1H),1.78-1.72(m,2H),1.49-1.45(m,2H),1 .38-1.34(m,4H),1.16(t,J=7.0Hz,1H),1.06(t,J=7.1Hz,2H),0.96-0.89(m,3H).HRMS(ESI):C 20 H 33 N2O2 + [M+H] + Calculated value: 333.2537; measured value: 333.2521.

[0218] Example 39: N-(3-amino-trans-cyclobutyl)-N-ethyl-2-(3-(hexyloxy)phenyl)acetamide

[0219] Following the preparation method of compound I-41, 4-hexyloxyphenylacetic acid was replaced with 3-hexyloxyphenylacetic acid to synthesize compound I-42 (8 mg). 1H NMR (800MHz, CD3OD) δ7.16-7.11(m,2H),6.88-6.84(m,2H),4.57(p,J=8.5Hz,1H),3.95-3.93(m,2H),3.90-3.87(m,1H),3.70-3. 68(m,2H),3.49-3.44(m,2H),2.86-2.78(m,1H),2.67-2.61(m,1H),2.45-2.40(m,1H),2.23-2.18(m,1H),1.78-1.72(m,2H),1.50 -1.45(m,2H),1.38-1.33(m,4H),1.16(t,J=7.0Hz,1H),1.06(t,J=7.1Hz,2H),0.94-0.91(m,3H).HRMS(ESI):C 20 H 33 N2O2 + [M+H] + Calculated value: 333.2537; measured value: 333.2535.

[0220] Example 40. Preparation of N-(3-aminocyclobutyl)-N-cyclopropyl-2-(3-(hexyloxy)phenyl)acetamide

[0221] Step 1. Add 3-N-Boc-cyclobutanone (0.5 g, 2.7 mmol) and cyclopropylamine (0.23 g, 4 mmol) to a round-bottom flask. Dissolve in methanol (10 mL) and add glacial acetic acid (30 μL) and stir at room temperature for 30 minutes. Then add NaBH3CN (2.28 g, 10.7 mmol). The reaction system continues to stir at room temperature overnight. After the reaction is completed, water is added to quench the reaction, and the product is extracted with ethyl acetate. The organic phases are combined, the solvent is removed, and the product is separated and purified by rapid silica gel column chromatography (eluent: methanol / dichloromethane = 10 / 90) to obtain a colorless oil WHH98 (0.56 g, 92%). HRMS (ESI) C 12 H 23 N2O2 + [M+H] + Calculated value: 227.1754; measured value: 227.1757.

[0222] Step 2. WHH98 (0.55 g, 2.43 mmol), 3-hexyloxyphenylacetic acid (0.63 g, 2.67 mmol), HATU (1.85 g, 4.86 mmol), and DIPEA (1 mL) were added to a round-bottom flask and the reaction system was stirred at room temperature overnight. After the reaction, the solvent was removed and the mixture was purified by flash silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 2) to obtain an oily compound WHH99 (0.2 g, 20%). HRMS (ESI): C 26 H 41 N2O4 + [M+H] + Calculated value: 445.3066; measured value: 445.3058.

[0223] Step 3. Add compound WHH99 (210 mg, 0.45 mmol) to a round-bottom flask and add a 2 mL, 2 M solution of hydrogen chloride in 1,4-dioxane. Stir at room temperature for 4 hours. After the reaction, remove the solvent, dissolve in MeOH, and purify by HPLC to obtain compound I-43 as an oil (50 mg, 59% yield). 1 H NMR(800MHz,CD3OD)δ7.23-7.19(m,1H),6.84-6.82(m,2H),6.81-6.79(m,1H ),3.97-3.90(m,3H),3.88(s,2H),3.49-3.43(m,1H),2.82-2.75(m,2H),2.7 1(br,1H),2.41-2.35(m,2H),1.78-1.74(m,2H),1.50-1.44(m,2H),1.39-1. 34(m,4H),1.01-0.97(m,2H),0.95-0.90(m,3H),0.78(br,2H).HRMS(ESI):C 21 H 33 N2O2 + [M+H] + Calculated value: 345.2537; measured value: 345.2539.

[0224] Example 41: Preparation of N-(3-aminocyclobutyl)-N-cyclopropyl-2-(4-(hexyloxy)phenyl)acetamide

[0225] Following the preparation of compound I-43, 3-hexyloxyphenylacetic acid was replaced with 4-hexyloxyphenylacetic acid to synthesize compound I-44 (13 mg, 10%) 1H NMR (800MHz, CD3OD) δ7.17(dd,J=8.3,6.0Hz,2H),6.88-6.84(m,2H),3.94(t,J=6.5Hz,3H),3.84(s,2H),3.46(p,J=7.9Hz,1H),2.78(br,2H),2.69(br, 1H),2.36(q,J=10.0Hz,2H),1.78-1.73(m,2H),1.51-1.44(m,2H),1.38-1. 33(m,4H),1.01-0.96(m,2H),0.96-0.89(m,3H),0.78(br,2H).HRMS(ESI):C 21 H 33 N2O2 + [M+H] + Calculated value: 345.2537; measured value: 345.2539.

[0226] Example 42: Preparation of N-(3-amino-trans-cyclobutyl)-N-cyclobutyl-2-(4-(hexyloxy)phenyl)acetamide

[0227] Step 1. Add tert-butyl (3-amino-trans-cyclobutyl)carbamate (250 mg, 1.34 mmol) to a round-bottom flask and dissolve it in methanol (8 mL). Then add cyclobutanone (113 mg, 1.61 mmol) and glacial acetic acid (30 μL) and stir at room temperature for 1 hour. Then add NaBH3CN (1.42 g, 6.7 mmol). The reaction system is stirred at room temperature overnight. After removing the solvent, the product is purified by flash silica gel column (methanol / dichloromethane = 1 / 9) to obtain a white solid compound WHH72 (0.16 g, yield 50%). HRMS (ESI) C 13 H 25 N2O2 + [M+H] + Calculated value: 241.1911; measured value: 241.19112.

[0228] Step 2. To a round-bottom flask, add WHH72 (70 mg, 0.29 mmol), 4-hexyloxyphenylacetic acid (103 mg, 0.44 mmol), and HATU (228 mg, 0.6 mmol). Dissolve the mixture in tetrahydrofuran (10 mL), followed by DIPEA (0.5 mL). Stir the reaction mixture overnight at room temperature. After completion of the reaction, remove the solvent, and purify the residue on a flash silica gel column (eluent: methanol / dichloromethane = 1 / 9) to afford WHH78 (60 mg, 45%) as a yellow oil. 1H NMR (600MHz, CDCl3) δ7.12-7.07(m,2H),6.86-6.81(m,2H),3.92(t,J=6.6Hz,2H),3.62(s,2H),2.18-2.06(m,1H),2.02-1.96(m,2H),1.84 -1.70(m,9H),1.47-1.42(m,1H),1.44(s,9H),1.38(d,J=6.7Hz,2H),1.35-1.30(m,4H),1.28-1.19(m,2H),0.92-0.87(m,3H).HRMS(ESI):C 27 H 43 N2O4 + [M+H] + Calculated value: 459.3217; measured value: 459.3217.

[0229] Step 3. Compound WHH78 (57 mg, 0.124 mmol) was added to a round-bottom flask, followed by a 1,4-dioxane solution of hydrogen chloride (5 mL, 2 M), and stirred at room temperature for 4 hours. Filtering afforded compound I-45 (24 mg, 54%) as a white solid. 1 H NMR (600MHz, CDCl3) δ7.07 (d, J = 8.1Hz, 2H), 6.86-6.80 (m, 2H), 4.59 (br, 1H), 4.3 1-4.10(m,1H),4.02(br,1H),3.92(t,J=6.6Hz,2H),3.62(s,2H),3.24(br,2H),2. 38(br,2H),2.05(d,J=12.9Hz,2H),1.95-1.89(m,3H),1.79-1.70(m,2H),1.70-1 .60(m,1H),1.50-1.39(m,2H),1.37-1.28(m,4H),0.94-0.85(m,3H).HRMS(ESI):C 22 H 35 N2O2 + [M+H] + Calculated value: 359.2693; measured value: 359.2738.

[0230] Example 43: Preparation of 1-(4-amino-trans-cyclohexyl)-1-ethyl-3-(4-(hexyloxy)phenyl)urea (Compound I-46):

[0231] Step 1. Dissolve tert-butyl (4-amino-trans-cyclohexyl)carbamate (0.5 g, 2.34 mmol) and a tetrahydrofuran solution of acetaldehyde (0.56 mL, 5 M) in methanol (10 mL). Add glacial acetic acid (30 μL) and stir at room temperature for 1 hour. Then add NaBH3CN (295 mg, 4.68 mmol) and continue stirring at room temperature overnight. After the reaction, remove the solvent and separate and purify by flash silica gel column chromatography to obtain oily compound WHI14 (0.47 g, 84%). HRMS (ESI), [M+H + ]C 13 H 27 N2O2 + Calculated value: 243.2067; measured value: 243.2068.

[0232] Step 2. To a round-bottom flask, add WHI14 (0.47 g, 1.94 mmol), N-(4-(hexyloxy)benzyl)-1H-imidazole-1-carboxamide (876 mg, 2.91 mmol), KCO (745 mg, 5.82 mmol), and DMF (8 mL). Stir the reaction at room temperature overnight. After completion of the reaction, remove the solvent, and purify by flash silica gel column chromatography to obtain WHI16-1 (0.17 g, 18%). 1 H NMR(600MHz,CD3OD)δ7.21-7.14(m,2H),6.87-6.80(m,2H),4.29(s,2H),4.03-3.96 (m,1H),3.94(t,J=6.5Hz,2H),3.28-3.24(m,1H),3.22(q,J=7.1Hz,2H),1.98-1.92 (m,2H),1.78-1.66(m,4H),1.60(qd,J=12.8,3.5Hz,2H),1.49-1.44(m,2H),1.43(s ,9H),1.39-1.25(m,6H),1.13(t,J=7.0Hz,3H),0.94-0.91(m,3H).HRMS(ESI),[M+H] + C 27 H 46 N3O4 + Calculated value: 476.3483; measured value: 476.3493.

[0233] Step 3. Add 2 mL of 3 M hydrogen chloride in 1,4-dioxane to intermediate WHI16-1 (0.15 g, 0.32 mmol) and stir overnight at room temperature. After the reaction, filter to obtain compound I-46 hydrochloride (0.1 g, 83%) as a white solid. 1H NMR(600MHz,CD3OD)δ7.19(d,J=8.3Hz,2H),6.87-6.81(m,2H),4.30(s,2H),4 .08-4.00(m,1H),3.94(t,J=6.4Hz,2H),3.23(q,J=7.1Hz,2H),3.08(tt,J=12 .3,4.1Hz,1H),2.09(d,J=12.2Hz,2H),1.86-1.60(m,6H),1.59-1.42(m,4H), 1.38-1.33(m,4H),1.14(t,J=7.0Hz,3H),0.96-0.89(m,3H).HRMS(ESI),[M+H + ]C 22 H 38 N3O2 + :Calculated value: 376.2959; measured value: 376.2969.

[0234] Comparative Example 1: Preparation of 1-(4-isobutoxybenzyl)-3-(1-methylpiperidin-4-yl)urea (Compound I-38)

[0235] Step 1: Preparation of N-(4-isobutoxybenzyl)-1H-imidazole-1-carboxamide (Compound 1)

[0236] Compound 4-isobutoxybenzylamine (1.1 g, 6.3 mmol) and 1,1'-carbonyldiimidazole (2.1 g, 12.6 mmol) were placed in a reaction flask, anhydrous DMF (20 mL) was added and stirred at room temperature. The reaction progress was monitored by TLC. After stirring overnight, the reaction was complete. Most of the solvent was removed by distillation under reduced pressure, an appropriate amount of water was added and extracted three times with ethyl acetate. The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated to obtain a crude product, which was purified on a silica gel column (petroleum ether: ethyl acetate = 1:1) to obtain a light yellow oil 1 (0.9 g, yield: 73%). HRMS (ESI) C 15 H 20 N3O2 + ([M+H] + )Calculated value: 274.1550; measured value: 274.1578.

[0237] Step 2: Preparation of 1-(4-isobutoxybenzyl)-3-(1-methylpiperidin-4-yl)urea (Compound I-38)

[0238] Compound 1 (500 mg, 2.6 mmol), 4-amino-1-methylpiperidine (800 mg, 2.6 mmol), and potassium carbonate (718.6 mg, 5.2 mmol) were placed in a reaction flask, and anhydrous DMF (10 mL) was added and stirred at room temperature. The reaction progress was monitored by TLC, and the reaction was complete after 4 hours. Most of the solvent was removed by vacuum distillation, and an appropriate amount of water was added and extracted three times with ethyl acetate. The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated to obtain a crude product, which was purified on a silica gel column (dichloromethane:methanol = 15:1) to obtain a light yellow solid I-38 (443 mg, yield: 71%). HRMS (ESI) C 18 H 30 N3O2 + ([M+H] + )Calculated value: 320.2333; measured value: 320.2337. 1 H NMR (600MHz, CDCl3) δ7.19 (d, J = 8.4Hz, 2H), 6.84 (d, J = 8.5Hz, 2H), 4.62 (t, J = 5.6Hz, 1H), 4.29 (d, J = 7.7Hz, 1H), 4.26 (d, J = 5. 5Hz,2H),3.69(d,J=6.5Hz,2H),3.64-3.54(m,1H),2.27(s,3H),2.14-1.92(m,7H),1.47-1.41(m,2H),1.01(d,J=6.7Hz,6H).

[0239] Comparative Example 2: Preparation of 3-(4-isobutoxybenzyl)-1-methyl-1-(1-methylpiperidin-4-yl)urea (Compound I-39)

[0240] The preparation method is the same as that of compound I-38, except that 4-amino-1-methylpiperidine is replaced by 1-methyl-4-(methylamino)piperidine, resulting in a pale yellow solid with a yield of 77%. HRMS (ESI) C 19 H 32 N3O2 + ([M+H] + )Calculated value: 334.2489; measured value: 334.2492. 1H NMR (800MHz, CDCl3) δ7.19(d,J=8.5Hz,2H),6.83(d,J=8.6Hz,2H),4.70(t,J=5.0Hz,1H),4.44-4.41(m,1H),4.32(d,J=5.4Hz,2H),3.68(d,J=6.6H z,2H),3.25(d,J=11.6Hz,2H),2.73(s,3H),2.58-2.50(m,5H),2.18-2.14 (m,2H),2.07-2.02(m,1H),1.70(d,J=12.7Hz,2H),0.99(d,J=6.7Hz,6H).

[0241] Comparative Example 3: Preparation of 3-(4-isobutoxybenzyl)-1-isopropyl-1-(1-methylpiperidin-4-yl)urea (Compound I-40)

[0242] Step 1: Preparation of N-isopropyl-1-methylpiperidin-4-amine (Compound 40-2)

[0243] 4-Amino-1-methylpiperidine (571 mg, 5 mmol) was dissolved in methanol (10 mL). Acetone (0.4 mL, 5 mmol) was added while stirring at room temperature and the mixture was stirred at room temperature for 1 hour. Sodium cyanoborohydride (471 mg, 7.5 mmol) was added to the reaction system and stirred at room temperature overnight. The reaction progress was monitored by TLC. After the reaction was completed, water (5 mL) was added to the reaction system to quench the reaction. The mixture was extracted three times with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure and purified on a silica gel column (dichloromethane: methanol = 95:5) to obtain compound 40-2 as a light yellow oil with a yield of 76%. HRMS (ESI) C9H 21 N2 + ([M+H] + )Calculated value: 157.1699; measured value: 157.1703.

[0244] Step 2: Preparation of 3-(4-isobutoxybenzyl)-1-isopropyl-1-(1-methylpiperidin-4-yl)urea (Compound I-40)

[0245] The preparation method is the same as that of compound I-38, except that 4-amino-1-methylpiperidine is replaced by compound 40-2, resulting in a pale yellow solid with a yield of 72%. HRMS (ESI) C 21 H 36 N3O2 + ([M+H] + )Calculated value: 362.2802; measured value: 362.2890. 1H NMR(800MHz, CDCl3) δ7.21(d,J=8.5Hz,2H),6.85(d,J=8.5Hz,2H),4.51-4.49 (m,1H),4.37(d,J=5.3Hz,2H),3.87-3.83(m,1H),3.79-3.74(m,1H),3.70(d,J =6.5Hz,2H),2.93(d,J=10.2Hz,2H),2.30(s,3H),2.10-2.04(m,3H),2.01-1.9 5(m,2H),1.64(d,J=11.9Hz,2H),1.25(d,J=7.0Hz,6H),1.01(d,J=6.7Hz,6H).

[0246] Biological test example 1: Effect of the compounds of the present invention on 5-HT 2A Receptor affinity testing

[0247] Method: The compounds of the present invention are effective for 5-HT 2A The receptor affinity was determined by radioligand competition binding assay.

[0248] The first step is to prepare 5-HT 2A 293T cells (ATCC CRL-11268) cultured in 10 cm culture dishes were treated with 5-HT 2A The receptor (10 μg) and PEI (Cat. No. 24765-1, 1 g / L, 40 μL) were transfected. After 48 hours, 5-HT was expressed on the cell membrane. 2A Receptor. Aspirate the cell culture medium, add 2mL lysis buffer (1mM HEPES, 2mM EDTA, pH 7.4) to the culture dish, transfer the cells to a 2mL centrifuge tube, centrifuge at 15000rpm at 4℃ for 5 minutes, and discard the supernatant. Add 1mL lysis buffer to the cell pellet and grind it thoroughly with a tissue homogenizer until the cells are broken. Centrifuge at 15000rpm at 4℃ for 5 minutes and discard the supernatant. Repeat twice, and the final pellet contains 5-HT 2A Cell membrane component of the receptor.

[0249] In the second step, the membrane fractions were used for ligand competition binding experiments. First, the membrane fractions were broken and resuspended in standard binding buffer (50mM HEPES, 5mM MgCl2, 0.5mM EDTA, pH 7.4), and 30μL of membrane protein suspension was added to each well of a 96-well plate. Then, 30μL of different drugs were added from left to right in the 96-well plate, using drug buffer (1×HBSS, 20mM HEPES, pH 7.4) containing 1 / 1000 BSA (ABcone), ensuring that the final drug concentration was 10 from bottom to top.-5 M, 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M, 0M, two replicates for each treatment. Then, 30 μL of approximately 3 nM [ 3 H]-LSD (Perkin Elmer). Incubate at room temperature in the dark for 2 hours. The membrane bound to the isotope was collected using a rapid vacuum filter onto GF / A filter paper (Perkin Elmer) soaked in 0.3% polyethyleneimine, and the unbound isotope was washed away with a wash buffer (50 mM Tris, pH 7.4). Solid scintillant (Perkin Elmer) was dissolved on the dry filter paper, and radioactivity was detected using a Wallac Trilux MicroBeta counter. The machine read the value of the reaction membrane bound to [ 3 The amount of [H]-LSD was determined using Graphpad Prism's "One-site-Fit K i "The formula is further processed to obtain the effect of different compounds on 5-HT 2A Receptor affinity K i value.

[0250] Results: The compounds of the present invention have an effect on 5-HT 2A Receptor affinity K i The values ​​are shown in Table 1 ("NA", no affinity).

[0251] Table 1

[0252] Biological test example 2: Effect of the compounds of the present invention on 5-HT 2A Receptor functional activity testing

[0253] Methods: Detection of 5-HT via calcium signaling 2A Receptor-mediated downstream Gq protein signaling pathway.

[0254] On the first day, cells cultured in a 6 cm culture dish were used to induce stable expression of 5-HT. 2A The recipient 293T cells were spread on a 384-well black bottom corning plate treated with 50 μg / μL poly-(l-lysine) (Sigma) at 37°C for 2 h, and 30 μL of culture medium (DMEM (Gibco) + 2% dFBS (Ausbian WS500TZ)) was added to each well. 1 / 10,000 tetracycline was added to the culture medium to induce 5-HT. 2AThe next day, the culture medium was removed from the 384-well plate and 15 μL of the dye Fluo-4 (4 μM, Invitrogen) was added. The plates were incubated for 1 hour at 37°C in a 5% CO2 incubator. Drugs were prepared using a drug buffer containing 2.5 mM probenecid (Sigma) to ensure that the final concentration of the drug added to the 384-well plate from bottom to top was 10 - 4.5 M, 10 -5 M, 10 -5.5 M, 10 -6 M, 10 -6.5 M, 10 -7 M, 10 -7.5 M, 10 -8 M, 10 -8.5 M, 10 -9 M, 10 -9.5 M, 10 -10 M, 10 -10.5 M, 10 -11 M, 10 -11.5 Each treatment was repeated three times. Drugs (7.5 μL per well) were added and the plate was read using a FLIPR Penta high-throughput real-time fluorescence detector. The fluorescence reading at 520 nm reflects the amount of intracellular calcium signal released after activation of the cellular Gq protein; an increase in the reading indicates receptor activation. The data were processed using the Graphpad Prism "log (agonist) vs. response -- Variable slope (four parameters)" formula to obtain the effects of different compounds on 5-HT. 2A EC activated by Gq receptor protein 50 value.

[0255] Detection of 5-HT by BRET 2A On the first day, 293T cells cultured in 6 cm culture dishes (DMEM + 10% FBS (ExCell Bio)) were inoculated with 500 μg of 5-HT containing algae luciferase at the C-terminus. 2A Receptors (5-HT 2A-Rluc), 500μg Gq protein-coupled receptor kinase 2 (GRK2), 3000μg N-terminal green fluorescent protein-containing β-arrestin2 (GFP2-ARRB2) and 16μL PEI were transfected. On the second day, the cells were digested and a 96-well all-white plate (Biyuntian) was laid on each 6cm culture dish, with 100μL culture medium (DMEM+2% dFBS) per well. On the third day, drug addition was performed. The culture medium was removed from the 96-well plate, and 40μL of substrate coelenterazine 400a (final concentration 5μM, Goldbio) was added to each well, followed by the addition of 20μL of drugs of different concentrations in sequence, prepared with drug buffer to ensure that the final concentration of the drug was 10 from bottom to top. -5 M, 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 Each treatment was repeated twice, and the results were detected using an LB940 Mithras plate reader. Rluc fluorescence signals were read at 410nm-480nm, and GFP fluorescence signals were read at 515nm-540nm. The ratio of the GFP / Rluc readings reflects the intracellular β-arrestin 2 surface, which is a marker of 5-HT. 2A The activation degree of the β-arrestin2 signaling pathway downstream of the receptor is measured. An increase in the ratio indicates receptor activation. The data were processed using the Graphpad Prism "log (agonist) vs. response--Variable slope (four parameters)" formula to obtain the effects of different compounds on 5-HT 2A ECs activated by receptor β-arrestin2 signaling pathway 50 value.

[0256] Results: The compounds of the present invention have an effect on 5-HT 2A The agonist activity of the receptors is shown in Table 2 ("NA", no agonist activity; "\", not tested).

[0257] Table 2

[0258] As shown in Table 2, the compounds of the present invention exhibited good 5-HT 2A Receptor agonist activity.

[0259] Biological Test Example 3:

[0260] Methods: Detection of compounds inhibiting 5-HT by calcium signaling 2A The receptor mediates the activity of the downstream Gq protein signaling pathway.

[0261] The Gq protein downstream signaling pathway was tested as described in Table 2 to determine whether the compound had agonist activity. After the machine test was completed, for compounds that did not show agonist activity (see Table 3 below), 7.5 μL of serotonin (Sigma) was added to the machine immediately to ensure that all wells treated with the compound contained a final concentration of 10 -9 M of serotonin, and read the plate, and use Graphpad Prism "log (inhibitor) vs. response--Variable slope (four parameters)" formula to process the data to obtain the different compounds for 5-HT 2A IC of Gq receptor protein inhibition 50 value.

[0262] Detection of 5-HT inhibition by compounds using BRET 2A The activity of the downstream β-arrestin2 signaling pathway mediated by the receptor was tested as shown in Table 2 to determine whether the compound had agonist activity. After the machine test was completed, for compounds that did not show agonist activity (see Table 3 below), 20 μL of serotonin was added immediately to ensure that all wells treated with the compound contained a final concentration of 10 -7 The plate was read again using a machine, and the data were processed using the Graphpad Prism "log (inhibitor) vs. response--Variable slope (four parameters)" formula to obtain the effects of different compounds on 5-HT 2A IC for inhibition of receptor β-arrestin2 signaling pathway 50 value.

[0263] Results: The compounds had an effect on 5-HT 2A The inhibitory activities of the receptors are shown in Table 3.

[0264] Table 3

[0265] As shown in Table 3, the comparative compounds showed 2A Inhibitory activity on receptor-related targets, unable to exhibit agonistic activity.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof; in, m and n are each independently 1, 2 or 3; R 1 and R 2 Each independently is hydrogen, C1-C6 alkyl or substituted by 1, 2 or 3 R 1-1 Substituted C1-C6 alkyl; R 1-1 Each independently is C6-C 10 Aryl or 1, 2 or 3 R 1-1a Substituted C6-C 10 Aryl; R 1-1a is halogen, hydroxyl or C1-C6 alkyl; R 3 is hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl or 1, 2 or 3 R 3-1 Substituted C1-C6 alkyl; R 3-1 Each independently is halogen, hydroxyl or 1, 2 or 3 R 3-1a Substituted C6-C 10 Aryl; R 3-1a Each is independently a C1-C4 alkoxy group; X is NH or a single bond; Y is a single bond, -CH2-, -(CH2)2-, -(CH2)3-, -CH=CH-、-CH=CH-CH2-、 or carbonyl; Ar is C6-C 10 Aryl, C5-C 12 Heteroaryl, 1, 2 or 3 R 5 Substituted C6-C 10 Aryl or 1, 2 or 3 R 6 Substituted C5-C 12 Heteroaryl, the C5-C 12 In the heteroaryl group, the heteroatom is selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3; R 5 and R 6 are independently halogen, hydroxyl, C1-C 10 Alkyl, C1-C 10 Alkoxy, C1-C 10 Alkylthio or R N-1 and R N-2 Each is independently H or C1-C6 alkyl.

2. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The compound represented by formula I satisfies one or more of the following conditions: (1)R 1 and R 2 wherein the C1-C6 alkyl group and 1, 2 or 3 R 1-1 The C1-C6 alkyl groups in the substituted C1-C6 alkyl groups are each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl; (2)R 1-1 In the C6-C 10 Aryl and 1, 2 or 3 R 1-1a Substituted C6-C 10 C6-C 10 Aryl is each independently phenyl or naphthyl, for example phenyl; (3)R 1-1a wherein the C1-C6 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl; (4)R 1-1a wherein the halogen is fluorine, chlorine or bromine; (5)R 3 wherein the C1-C6 alkyl group and 1, 2 or 3 R 3-1 The C1-C6 alkyl groups in the substituted C1-C6 alkyl groups are each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl, ethyl or isopropyl; (6)R 3 wherein the C3-C6 cycloalkyl group is a cyclopropyl group, a cyclobutyl group, a cyclopentyl group or a cyclohexyl group, for example, a cyclopropyl group, for example, a cyclopropyl group or a cyclobutyl group; (7)R 3-1 wherein the halogen is fluorine, chlorine or bromine, for example fluorine; (8)R 3-1 In the embodiment, the 3-1a Substituted C6-C 10 C6-C 10 Aryl is phenyl or naphthyl, for example phenyl; (9)R 3-1a wherein the C1-C4 alkoxy group is methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy or tert-butoxy, such as methoxy; (10) In Ar, the C6-C 10 Aryl and 1, 2 or 3 R 5 Substituted C6-C 10 C6-C 10 Aryl is each independently phenyl or naphthyl, for example phenyl; (11) In Ar, the C5-C 12 Heteroaryl and 1, 2 or 3 R 6 Substituted C5-C 12 C5-C 12 The heteroaryl groups are each independently C5-C9 heteroaryl groups, wherein the heteroatom is preferably N, and the number of heteroatoms is preferably 1. For example, the C5-C 12 Heteroaryl is (12)R 5 and R 6 wherein the halogen is fluorine, chlorine or bromine, for example fluorine; (13)R 5 and R 6 In the above, C1-C 10 Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl or n-decyl; (14)R 5 and R 6 In the above, C1-C 10 Alkoxy is methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, n-pentoxy, n-hexoxy, n-heptyloxy, n-octyloxy, n-nonyloxy or n-decyloxy, for example n-hexyloxy, isobutoxy, n-decyloxy, n-heptyloxy, n-octyloxy or methoxy; (15)R 5 and R 6 In the above, C1-C 10 Alkylthio is methylthio, ethylthio, n-propylthio, isopropylthio, n- Butylthio, isobutylthio, tert-butylthio, n-pentylthio, n-hexylthio, n-heptylthio, n-octylthio, n-nonylthio or n-decylthio; and (16)R N-1 and R N-2 In the above, the C1-C6 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl.

3. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The compound represented by formula I satisfies one or more of the following conditions: (1) m and n are each independently 1 or 2; (2)R 1 and R 2 Each independently is hydrogen, C1-C6 alkyl or substituted by 1, 2 or 3 R 1-1 Substituted C1-C6 alkyl; R 1-1 For 1, 2 or 3 R 1-1a Substituted C6-C 10 Aryl; R 1-1a is a hydroxyl group; preferably, R 1 and R 2 each independently is hydrogen; (3)R 3 is hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl or 1, 2 or 3 R 3-1 Substituted C1-C6 alkyl; R 3-1 are independently halogen or 1 R 3-1a Substituted C6-C 10 Aryl; R 3-1a Each is independently C1-C4 alkoxy; for example, R 3 is hydrogen, C1-C6 alkyl or 1, 2 or 3 R 3-1 Substituted C1-C6 alkyl; R 3-1 Each independently is 1, 2 or 3 R 3-1a Substituted C6-C 10 Aryl; R 3-1a are each independently C1-C6 alkoxy; preferably, R 3 is hydrogen, C1-C6 alkyl, More preferably, R 3 is hydrogen or C1-C6 alkyl, for example, hydrogen or straight-chain C1-C6 alkyl, further, R 3 is a straight chain C1-C6 alkyl group, for example, R 3 is methyl; (4) X and Y are not single bonds at the same time; (5) Y is a single bond, -CH2-, -(CH2)2-, -CH=CH- or -C(=O)-; preferably, Y is -CH2-, -(CH2)2-, -CH=CH- or -C(=O)-; for example, Y is -CH2- or -C(=O)-; Preferably, when X is a single bond and Y is -CH=CH-, for Alternatively, when X is a single bond and Y is -(CH2)2-, for (6) Ar is substituted by 1, 2 or 3 R 5 Substituted C6-C 10 Aryl or 1, 2 or 3 R 6 Substituted C5-C 12 Heteroaryl, the C5-C 12 In the heteroaryl group, the heteroatom is selected from one or more of N, O and S. The number of sub-groups is 1, 2 or 3; preferably, Ar is C6-C 10 Aryl or 1, 2 or 3 R 5 Substituted C6-C 10 Aryl, for example, with 1, 2 or 3 R 5 Substituted C6-C 10 Aryl; and (7)R 5 and R 6 Each independently is C1-C 10 Alkoxy; preferably, when X is NH, Y is -CH2-, R 5 and R 6 Each independently is a straight-chain C1-C 10 Alkoxy.

4. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The compound represented by formula I satisfies one or more of the following conditions: (1)R 1 and R 2 are independently hydrogen, or methyl; Preferably, -NH2, (2)R 3 Methyl, hydrogen, Isopropyl, Ethyl, Cyclopropane or cyclobutane; preferably, R 3 Methyl, hydrogen, Isopropyl, Ethyl, or cyclopropane, for example methyl or H; (3) X is NH or a single bond; (4)Y-CH2-、-CH=CH-、 single bond or carbonyl; Preferably, -XY- is -HC=CH-、 single bond or carbonyl; (5)Ar is (6)R 5 for and (7)R 6 for 5. The compound of formula I or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, characterized in that: When m and n are not equal, the compound represented by formula I is a compound represented by formula I-1a, I-2a, I-3a or I-4a: Alternatively, when m and n are equal, the compound represented by formula I is a compound represented by formula I-5a or I-6a; Among them, R 1 , R 2 , R 3 , X, Ar, Y, m and n are each independently as described in any one of claims 1-4.

6. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The compound represented by formula I is any of the following compounds: Preferably, the pharmaceutically acceptable salt of the compound represented by Formula I is the hydrochloride salt of the compound represented by Formula I.

7. A pharmaceutical composition comprising a compound of formula I as claimed in any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

8. A use of the compound of formula I according to any one of claims 1 to 6, a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 7; The application is (1) preparing a drug for treating and / or preventing 5-HT 2A and / or (2) preparation of drugs for 5-HT receptor-related diseases; 2A Receptor agonists; Preferably, the 5-HT 2A Diseases associated with 5-HT receptors 2A β-arrestin2 recruitment signaling pathway downstream of receptor and / or 5-HT 2A Diseases related to the activation of Gq protein signaling pathways downstream of the receptor, such as those related to 5-HT 2A Receptor-related diseases are depression; Preferably, the 5-HT 2A The receptor agonist is 5-HT 2A β-arrestin2 downstream of the receptor recruits signaling pathway agonists and / or 5-HT 2A The Gq protein downstream of the receptor activates the signaling pathway agonist, such as 5-HT 2A β-arrestin2 recruits signaling agonists downstream of the receptor.

9. A method for preparing a compound of formula I as claimed in any one of claims 1 to 6, characterized in that: It is any of the following options: Scheme 1: hydrolyzing the compound represented by formula Ia in an organic solvent to obtain the compound represented by formula I; W is an amino protecting group, such as tert-butyloxycarbonyl or benzyloxycarbonyl, for example tert-butyloxycarbonyl; Scheme 2: In an organic solvent, under the action of a catalyst, ammonia reagent Ic and the compound represented by formula Ib are coupled to obtain a compound represented by formula I Among them, R 1 , R 2 , R 3 , X, Ar, Y, m and n are each independently as described in any one of claims 1-6.

10. The compound represented by formula Ia or Ib: in, R 1 , R 2 , R 3 , X, Ar, Y, m and n are each independently as described in claim 9; Preferably, the compound represented by formula Ia is any of the following compounds: Preferably, the compound represented by formula Ib is any of the following compounds:

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