CN112716927B - Alpha-amino amide compound and preparation method and application thereof - Google Patents

Abstract

The invention discloses application of alpha-amino amide compounds as sodium ion channel (particularly Nav1.7) inhibitors in pain treatment. Specifically, the invention discloses application of an alpha-amino amide compound shown in a structural formula I, a stereoisomer, a pharmaceutically acceptable salt, a solvent compound and a prodrug thereof or a pharmaceutical composition containing the compound as an active ingredient as an analgesic drug of a sodium ion channel (particularly Nav1.7) inhibitor, and a preparation method of the alpha-amino amide compound.

Description

Alpha-amino amide compound and preparation method and application thereof

Technical Field

The invention belongs to the field of pharmaceutical chemistry, and relates to alpha-amino amide compounds, stereoisomers, pharmaceutically acceptable salts, solvate compounds, prodrugs and pharmaceutical compositions containing the compounds as active ingredients, a preparation method of the compounds and application of the compounds as sodium ion channel (particularly Nav1.7) inhibitor analgesic drugs.

Background

The International Association for The Study of Pain (IASP) defines Pain as an unpleasant sensory and emotional experience associated with actual or potential tissue damage. Pain is an important function of self-protection of the body and a major health problem, which greatly reduces the quality of life and brings high health cost and economic loss to the society. Although a considerable amount of analgesic drugs have been clinically used for pain treatment, the existing pain treatment drugs have a complex pain mechanism, strong side effects or low effectiveness, and generally can not meet the requirements of clinical pain treatment, so that the development of a new-mechanism analgesic drug with strong analgesic activity and good safety has great research significance and application value.

Ralfinamide is the only Nav1.7 inhibitor entering clinical stage III at present, has certain therapeutic effect in the II phase clinic of moderate and severe neuropathic pain, but the clinical stage III for chronic neuropathic lumbago can not reach the main end point, so the patent inventor designs and synthesizes a series of alpha-amino amide compounds to carry out activity test screening on the alpha-amino amide compounds, is favorable for developing analgesic active compound application and clinical medicine resources, and has no relevant report at present.

Disclosure of Invention

On the basis of the common general knowledge in the field, the above-mentioned preferred conditions can be combined arbitrarily without departing from the concept and the protection scope of the invention.

In order to develop and utilize the existing clinical medicine resources, the invention aims to provide a class of alpha-amino amide compounds, stereoisomers, pharmaceutically acceptable salts, solvate compounds, prodrugs and pharmaceutical compositions containing the compounds as active ingredients; the invention aims to provide the preparation method, thereby opening up a new way for searching new analgesic active compounds; another object of the present invention is to provide the use thereof as an analgesic drug of a sodium ion channel (particularly Nav1.7) inhibitor.

The invention provides an alpha-amino amide compound shown as a formula (I), a stereoisomer, a pharmaceutically acceptable salt, a solvent compound, a prodrug thereof or a pharmaceutical composition containing the compound as an active ingredient, wherein the formula (I) is as follows:

wherein X is selected from CH ₂ NH, O and S; ring A is selected from C _6- C ₁₀ Aryl of (C) _6- C ₁₀ Cycloalkyl of (C) _6- C ₁₀ Heteroaryl and C _6- C ₁₀ Said C is _6- C ₁₀ Aryl of (C) _6- C ₁₀ Cycloalkyl of (C) _6- C ₁₀ Heteroaryl and C _6- C ₁₀ Is optionally substituted by 1, 2,3,4 or 5R ₁ Substitution; each R ₁ Each independently selected from H, F, cl, br, I, CN, OH, NH ₂ 、C _1- C ₄ Alkyl of (C) _1- C ₄ Alkoxy group of (1), C _1- C ₄ Epoxy group of (b), said C _1- C ₄ Alkyl and C _1- C ₄ Is optionally substituted with 1, 2 or 3R; r is ₂ Selected from H, C ₁ ～C ₄ Alkyl of (C) ₃ ～C ₆ Cycloalkyl and C ₃ ～C ₆ Said C is ₁ ～C ₄ Alkyl of (C) ₃ ～C ₆ Cycloalkyl and C ₃ ～C ₆ Heterocycloalkyl is optionally substituted with 1, 2 or 3R; r ₃ 、R ₄ 、R ₅ And R ₆ Each independently selected from H, F, cl, br, I, CN, OH, NH ₂ 、C _1- C ₄ Alkyl and C _1- C ₄ Alkoxy of (b), said C _1- C ₄ Alkyl and C _1- C ₄ Is optionally substituted with 1, 2 or 3R; n is 1 or 2; r ₇ Selected from H, C ₁ ～C ₄ Alkyl of (C) ₃ ～C ₆ Cycloalkyl and C ₃ ～C ₆ Heterocycloalkyl radical of said C ₁ ～C ₄ Alkyl of (C) ₃ ～C ₆ Cycloalkyl and C ₃ ～C ₆ Heterocycloalkyl is optionally substituted with 1, 2 or 3R; r ₈ Selected from H, C ₁ ～C ₄ Alkyl of (C) ₃ ～C ₆ Cycloalkyl of, C ₃ ～C ₆ Heterocycloalkyl, aromatic, heteroaryl and benzyl, said C ₁ ～C ₄ Alkyl of (C) ₃ ～C ₆ The cycloalkyl or heterocycloalkyl, aromatic, heteroaryl, and benzyl groups of (a) are optionally substituted with 1, 2, or 3R; r is ₉ And R ₁₀ Each independently selected from H and C ₁ ～C ₄ Alkyl of (C) ₃ ～C ₆ Cycloalkyl and C ₃ ～C ₆ Heterocycloalkyl radical of said C ₁ ～C ₄ Alkyl of (C) ₃ ～C ₆ Cycloalkyl and C ₃ ～C ₆ Heterocycloalkyl is optionally substituted with 1, 2 or 3R; or R ₉ And R ₁₀ To the carbon atoms to which they are attached to form C ₃ ～C ₆ Cycloalkyl of (a), said C ₃ ～C ₆ Is optionally substituted with 1, 2 or 3R; r is ₁₁ And R ₁₂ Each independently selected from H and C ₁ ～C ₄ Alkyl of (C) ₃ ～C ₆ Cycloalkyl and C ₃ ～C ₆ Heterocycloalkyl radical of said C ₁ ～C ₄ Alkyl of (C) ₃ ～C ₆ Cycloalkyl and C ₃ ～C ₆ Heterocycloalkyl is optionally substituted with 1, 2 or 3R; or R ₁₁ And R ₁₂ To the carbon atoms to which they are attached to form C ₃ ～C ₆ Cycloalkyl of (b), said C ₃ ～C ₆ Is optionally substituted with 1, 2 or 3R; each R is independently selected from H, F, cl, br, I, CN, OH and NH ₂ 。

As a preferred embodiment, in the present invention, the ring A is selected from C _6- C ₁₀ Aryl of, C _6- C ₁₀ The heteroaryl group of (a); said C is _6- C ₁₀ Aryl of (C) _6- C ₁₀ Is optionally substituted by 1, 2,3,4 or 5R ₁ Substitution; each R ₁ Are respectively and independently selected from H, F, cl, br, I and C _1- C ₄ Alkoxy group of (C) _1- C ₄ Epoxy group of (b), said C _1- C ₄ Alkyl and C _1- C ₄ Is optionally substituted with 1, 2 or 3R; r ₂ Selected from H, C ₁ ～C ₄ Alkyl groups of (a); said C is ₁ ～C ₄ Is optionally substituted with 1, 2 or 3R; r ₃ 、R ₄ 、R ₅ And R ₆ Are respectively and independently selected from H, F, cl, br, I and C _1- C ₄ Alkyl and C _1- C ₄ Alkoxy of (a), said C _1- C ₄ Alkyl and C _1- C ₄ Is optionally substituted with 1, 2 or 3R; n is 1; r ₇ Selected from H, C ₁ ～C ₄ Alkyl groups of (a); said C is ₁ ～C ₄ Is optionally substituted with 1, 2 or 3R; r is ₈ Selected from H, C ₁ ～C ₄ Alkyl groups of (a); said C is ₁ ～C ₄ Is optionally substituted with 1, 2 or 3R; r ₉ And R ₁₀ Each independently selected from H and C ₁ ～C ₄ Alkyl groups of (a); said C is ₁ ～C ₄ Is optionally substituted with 1, 2 or 3R; or R ₉ And R ₁₀ To the carbon atom to which they are attached to form C ₃ ～C ₆ Cycloalkyl of (a), said C ₃ ～C ₆ Is optionally substituted with 1, 2 or 3R; r is ₁₁ And R ₁₂ Each independently selected from H, C ₁ ～C ₄ Alkyl of (C) ₁ ～C ₄ Is optionally substituted with 1, 2 or 3R; each R is independently selected from H, F, cl, br, I, CN, OH and NH ₂ 。

As a preferred embodiment, the α -aminoamide compound, stereoisomer, pharmaceutically acceptable salt, solvate, prodrug or pharmaceutical composition containing these compounds as active ingredient has a structural formula selected from:

wherein each R is ₁ Each independently selected from H, F, cl, br, I, C _1- C ₄ Alkoxy group of (C) _1- C ₄ The epoxy group of (A), the C _1- C ₄ Alkyl and C _1- C ₄ Is optionally substituted with 1, 2 or 3R; r ₂ Selected from H, CH ₃ 、CF ₃ ；R ₈ Selected from H, C ₁ ～C ₄ Alkyl groups of (a); said C is ₁ ～C ₄ Is optionally substituted with 1, 2 or 3R; r ₉ And R ₁₀ Each independently selected from H and C ₁ ～C ₄ Alkyl groups of (a); said C is ₁ ～C ₄ Is optionally substituted with 1, 2 or 3R; each R is independently selected from H, F, cl, br, I, CN, OH and NH ₂ 。

As a preferred technical solution, the invention is described inAlpha-aminoamide compounds, stereoisomers, pharmaceutically acceptable salts, solvate compounds, prodrugs or pharmaceutical compositions containing these compounds as active ingredients, wherein R ₁ Selected from H, F, cl, br, I, CN, OH, NH ₂ 、CH ₃ 、CF ₃ 、OCH ₃ And OCF ₃ ；R ₂ Is selected from H; r ₈ Selected from H, CH ₃ 、CH ₂ CH ₃ And

R ₉ and R ₁₀ Each independently selected from H and CH ₃ 、CH ₂ OH、CH ₂ CH ₃ 、CH(OH)CH ₃ 、CH(CH ₃ ) ₂ And CH ₂ CH(CH ₃ ) ₂ (ii) a Or R ₉ And R ₁₀ To the carbon atom to which they are attached to form a cyclopentyl group.

As a preferred technical scheme, the alpha-amino amide compound, stereoisomer, pharmaceutically acceptable salt, solvent compound, prodrug or pharmaceutical composition containing the compound as an active ingredient in the invention has a structural unit in a formula (I)

Selected from the group consisting of:

in formula (I)

Selected from:

as a preferred embodiment, the α -aminoamide compound, stereoisomer, pharmaceutically acceptable salt, solvate, prodrug or pharmaceutical composition containing these compounds as an active ingredient in the present invention has a structure selected from the group consisting of:

the second aspect of the invention provides an application of the alpha-amino amide compound, the stereoisomer, the pharmaceutically acceptable salt, the solvent compound, the prodrug or the pharmaceutical composition containing the compounds as the active ingredients in analgesia.

As a preferred embodiment, for the treatment of pain; the pain includes treating or relieving pain during surgery, chronic pain, neuropathic pain, cancer pain.

The third aspect of the invention provides a preparation method of the alpha-amino amide compound, the stereoisomer, the pharmaceutically acceptable salt, the solvent compound, the prodrug or the pharmaceutical composition containing the compound as the active ingredient,

the following synthetic route was used:

wherein the substituent group X is selected from the group consisting of NH, O and S and the other substituent groups are as defined in any one of claims 1 to 6.

As a preferred technical solution, the preparation method comprises the following steps:

(1) The synthesis of the target compound shown in the formula I directly takes 2-fluorobenzyl bromide as a raw material and potassium carbonate as alkali to carry out nucleophilic substitution reaction with a compound b to obtain a compound c, and then takes sodium hydroxide as alkali to carry out ester hydrolysis to obtain a compound d. And reducing the compound d by lithium aluminum hydride to obtain a compound e, and oxidizing the compound e into corresponding aldehyde by a desmartin oxidant to obtain a compound f. And (3) carrying out reductive amination reaction on the compound f and corresponding alpha-aminoamide through sodium cyanoborohydride to obtain a series of target compounds g.

(2) And (3) removing benzyl protection from the compound g through palladium carbon hydrogenation reaction to obtain a compound h, and then reacting to obtain a series of target compounds i.

(3) And (3) carrying out reductive amination reaction or substitution reaction on the compound i to obtain a series of target compounds j.

(4) Similar to the synthesis, a series of target products l are obtained by the reductive amination reaction of ketone compounds k.

(5) And (3) removing benzyl protection from the compound l through palladium-carbon hydrogenation reaction to obtain a compound m, and then reacting to obtain a series of target compounds n.

(6) And carrying out reductive amination reaction or substitution reaction on the compound n to obtain a series of target compounds I.

The fourth aspect of the present invention provides an administration dosage form of the α -aminoamide compound, the stereoisomer, the pharmaceutically acceptable salt, the solvate, the prodrug thereof, or the pharmaceutical composition containing these compounds as an active ingredient, wherein the dosage form is: solvent tablet, capsule, injection, and solution.

The fifth aspect of the present invention provides a method for administering the α -aminoamide compound, the stereoisomer, the pharmaceutically acceptable salt thereof, the solvate thereof, the prodrug thereof, or the pharmaceutical composition containing the same as an active ingredient, by an injection route or orally.

Has the advantages that:

the alpha-amino amide compound has a good inhibition effect on a sodium ion channel Nav1.7, and can be applied to treatment or alleviation of pain. The inhibitory activity is superior to Ralfinamide. In the preliminary patent medicine research, especially, the compounds I-1 and I-30 have good metabolic stability, in the preliminary in vivo analgesic experiment, the compounds I-1 and I-30 show good analgesic effect in the mouse sciatic nerve branch selective injury experiment (SNI model), and simultaneously, the compound I-1 also shows good analgesic effect in the mouse formalin model.

In the present invention, the definitions and terms in the present invention are explained in turn as follows:

unless otherwise stated or indicated, the term "C" is used ₆ -C ₁₀ The aryl group of (a) means an all-carbon monocyclic or fused polycyclic group having a conjugated pi-electron system, means an aryl group having 6 to 10 carbon atoms, preferably phenyl and naphthyl.

Unless otherwise stated or indicated, the term "C" is used _6- C ₁₀ The term "cycloalkyl" refers to a saturated or partially unsaturated cyclic alkyl group, a cyclic alkyl group containing from 6 to 10 carbon atoms, partially unsaturated refers to a group containing one or more unsaturated bonds, but not having a completely conjugated pi-electron system.

Unless otherwise stated or indicated, the term "C" is used _6-10 The heteroaryl group of (a) means an aryl group containing 6 to 10 carbon atoms, and wherein one or more atoms are selected from nitrogen, oxygen and sulfur.

Unless otherwise stated or indicated, the term "C" is used _6-10 The "heterocyclic group" of (a) means a saturated or partially unsaturated monocyclic or polycyclic cyclic alkyl group in which one or more atoms are selected from nitrogen, oxygen and sulfur, and partial unsaturation is as defined above.

Unless otherwise stated or indicated, the term "C" is used ₁ ～C ₄ The "alkyl group" of (a) means a straight or branched hydrocarbon chain having 1 to 4 carbon atoms, and includes methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl and tert-butyl.

Unless otherwise stated or indicated, the term C ₃ ～C ₆ Cycloalkyl in (b) means a saturated or partially unsaturated cyclic alkyl group, preferably cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

Unless otherwise stated or indicated, the term C _1-4 Alkoxy of (D) means O-C _1-4 Alkyl groups, including methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy and tert-butoxy.

Unless otherwise specified or indicated, the alpha-amino amide compounds, stereoisomers, pharmaceutically acceptable salts, solvate compounds, prodrugs or pharmaceutical compositions containing these compounds as active ingredients of formula I, wherein the stereoisomers may be the levo, dextro, racemic forms of chiral compounds, and mixtures thereof in any proportion;

unless otherwise stated or indicated, the alpha-aminoamide compounds, stereoisomers, pharmaceutically acceptable salts, solvate compounds, prodrugs or pharmaceutical compositions containing these compounds as active ingredients of formula I are characterized in that the salts form pharmaceutically acceptable acid addition salts from the alpha-aminoamide compounds with inorganic or organic acids.

Unless otherwise stated or indicated, the alpha-aminoamide compounds, stereoisomers, pharmaceutically acceptable salts, solvent compounds, prodrugs or pharmaceutical compositions containing these compounds as active ingredients, which are shown in the general formula I, are characterized in that the inorganic acid is one or a combination of any two or more of hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, sulfuric acid and phosphoric acid; the organic acid is one or a composition of any two or more of tartaric acid, acetic acid, mandelic acid, maleic acid, fumaric acid, benzoic acid, succinic acid, lactic acid, citric acid, gluconic acid, methanesulfonic acid, benzenesulfonic acid and p-toluenesulfonic acid.

Drawings

FIG. 1. Effect of Compound I-1 on nerve injury (SNI) -induced neuropathic pain.

Figure 2. Effect of compound I-30 on nerve injury (SNI) -induced neuropathic pain.

FIG. 3. Effect of Compound I-1 on formalin-induced inflammatory pain.

Detailed Description

The technical solutions of the present invention are described in detail below with reference to the drawings and embodiments, but the present invention is not limited to the scope of the embodiments.

EXAMPLE 1 preparation of Compound I-1

1.1 Synthesis of methyl 3- (4- ((2-fluorobenzyl) oxy) phenyl) propionate

5.41g (30.02 mmol) of methyl p-hydroxyphenylpropionate is put into a 250mL single-neck flask, 150mL of acetonitrile is added, 4.98g (36.03 mmol) of potassium carbonate and 6.24g (33.01 mmol) of 2-fluorobenzyl bromide are sequentially added, and the temperature is slowly raised to 90 ℃ for reaction for 4 hours. The reaction solution was cooled to room temperature, filtered, washed twice with acetonitrile, the organic phase was concentrated, 250mL of water was added, extracted three times with 250mL of dichloromethane, the organic phases were combined, washed twice with 50mL of saturated brine, the organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure, and purified by silica gel column chromatography to give 6.09g of a white solid with a yield of 70%.

1.2 Synthesis of 3- (4- ((2-fluorobenzyl) oxy) phenyl) propionic acid

The above 6.09g (21.12 mmol) of methyl 3- (4- ((2-fluorobenzyl) oxy) phenyl) propionate was taken in a 250mL single-neck flask, 50mL of methanol and 50mL of tetrahydrofuran were added and stirred, 51mL of a 0.62M aqueous solution of sodium hydroxide was added, the temperature was slowly raised to 60 ℃ and after reaction for 3 hours, the reaction solution was cooled to room temperature and concentrated under reduced pressure, the concentrate was dissolved in 300mL of dichloromethane and 200mL of water, pH =6 was adjusted, the aqueous phase was extracted twice with 250mL of dichloromethane, the organic phases were combined, the organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure and purified by silica gel column chromatography to obtain 5.39g of a white solid with a yield of 93%.

1.3 Synthesis of 3- (4- ((2-fluorobenzyl) oxy) phenyl) propanol

Taking 5.39g (19.65 mmol) of the 3- (4- ((2-fluorobenzyl) oxy) phenyl) propionic acid in a 250mL three-necked bottle, adding 100mL of anhydrous tetrahydrofuran, stirring, under the protection of argon, slowly and dropwise adding 39mL (39 mmol) of 1M lithium aluminum hydride tetrahydrofuran solution under the ice bath condition, slowly heating to 80 ℃, reacting for 2 hours, and cooling the reaction solution to room temperature. Under ice bath conditions, saturated ammonium chloride solution was slowly added dropwise until no bubbles were generated, filtered, and concentrated under reduced pressure, the concentrate was dissolved in 300mL of dichloromethane, washed with 200mL of water, the aqueous phase was extracted twice with 100mL of dichloromethane, the organic phases were combined, the organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure, and purified by silica gel column chromatography to give 3.05g of colorless oil with a yield of 60%.

1.4 Synthesis of 3- (4- ((2-fluorobenzyl) oxy) phenyl) propanal

The above 1.54g (5.92 mmol) of 3- (4- ((2-fluorobenzyl) oxy) phenyl) propanol was put in a 100mL single-necked flask, 60mL of methylene chloride was added thereto, and the mixture was stirred, 3.02g (7.12 mmol) of dess-martin reagent was added thereto, and the mixture was reacted at room temperature for 4 hours. 60mL of water was added, the aqueous phase was extracted twice with 60mL of dichloromethane, the organic phases were combined, the organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure, and purified by silica gel column chromatography to give 1.23g of yellow oil in 80% yield.

1.5 Synthesis of Compound I-1

2.99g (11.58 mmol) of 3- (4- ((2-fluorobenzyl) oxy) phenyl) propanal are put into a 100mL single-neck flask, 60mL of methanol are added, stirring is carried out, 1.73g (13.89 mmol) of L-alaninamide hydrochloride, 0.86g (13.91 mmol) of sodium cyanoborohydride and 6mL of glacial acetic acid are sequentially added, the temperature is raised to 60 ℃, and reaction is carried out for 4 hours. Cooled to room temperature, concentrated under reduced pressure, the concentrate was dissolved in 300mL of ethyl acetate and 200mL of water, adjusted pH =9 with ammonia, the aqueous phase was extracted three times with 50mL of ethyl acetate, the organic phases were combined, the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography to give 2.63g of a white solid with a yield of 69%. Melting point: 113.6-115.9 ℃. ¹ H NMR(400MHz,CD ₃ OD)δ7.50(s,1H),7.35(s,1H),7.14(d,J＝24.5Hz,4H),6.90(s,2H),5.09(s,2H),3.16(s,1H),2.55(d,J＝22.1Hz,4H),1.77(s,2H),1.24(s,3H).LCMS(M+H) ⁺ :331.0,HRMS(ESI,m/z):calculated for C ₁₉ H ₂₃ FN ₂ O ₂ (M+H) ⁺ 331.1816,found 331.1826.

EXAMPLE 2 preparation of Compound I-2

Taking 103mg (0.40 mmol) of 3- (4- ((2-fluorobenzyl) oxy) phenyl) propanal in a 50mL single-neck flask, adding 10mL of methanol, stirring, sequentially adding 44mg (0.40 mmol) of glycinamide hydrochloride, 50mg (0.80 mmol) of sodium cyanoborohydride and 0.20mL of glacial acetic acid, stirring at room temperature overnight, and concentrating under reduced pressureAfter condensation, the concentrate was dissolved in 10mL of ethyl acetate and 10mL of water, pH =9 was adjusted with aqueous ammonia, the aqueous phase was extracted three times with 10mL of ethyl acetate, the organic phases were combined, the organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure, and purified by silica gel column chromatography to obtain 56mg of a white solid with a yield of 44%. Melting point: 61.5-62.3 ℃. ¹ H NMR(400MHz,CD ₃ OD)δ7.49(d,J＝7.7Hz,1H),7.34(s,1H),7.24–7.03(m,4H),6.91(d,J＝8.3Hz,2H),5.08(d,J＝8.9Hz,2H),3.31(s,2H),2.63(d,J＝7.6Hz,4H),1.90–1.68(m,2H).LCMS(M+H) ⁺ :317.0,HRMS(ESI,m/z):calculated for C ₁₈ H ₂₁ FN ₂ O ₂ (M+H) ⁺ 317.1660,found 317.1668.

EXAMPLE 3 preparation of Compound I-3

The procedure of example 2 was followed, using 2-methylalaninamide instead of glycinamide hydrochloride, to give 98mg of a white solid in 71% yield. Melting point: 87.4-88.3 ℃. ¹ H NMR(400MHz,CD ₃ OD)δ7.48(t,J＝7.3Hz,1H),7.33(d,J＝5.9Hz,1H),7.14(dd,J＝22.5,7.8Hz,4H),6.90(d,J＝7.6Hz,2H),5.07(s,2H),2.59(dd,J＝15.9,8.1Hz,4H),1.89–1.67(m,2H),1.31(d,J＝25.6Hz,6H).LCMS(M+H) ⁺ :344.9,HRMS(ESI,m/z):calculated for C ₂₀ H ₂₅ FN ₂ O ₂ (M+H) ⁺ 345.1973,found 345.1982.

EXAMPLE 4 preparation of Compound I-4

The procedure of example 2 was followed using L-2-aminobutanamide hydrochloride in place of glycinamide hydrochloride to give 76mg of a white solid in 55% yield. Melting point: 102.1-101.2 ℃. ¹ H NMR(400MHz,CD ₃ OD)δ7.48(s,1H),7.33(d,J＝5.4Hz,1H),7.14(dd,J＝24.6,7.1Hz,4H),6.89(d,J＝7.1Hz,2H),5.07(s,2H),3.21(s,1H),2.74–2.47(m,4H),1.83(d,J＝5.0Hz,2H),1.69(s,2H),0.96(d,J＝6.1Hz,3H).LCMS(M+H) ⁺ :345.0,HRMS(ESI,m/z):calculated for C ₂₀ H ₂₅ FN ₂ O ₂ (M+H) ⁺ 345.1973,found345.1982.

EXAMPLE 5 preparation of Compound I-5

The procedure of example 2 was followed using L-valinamide hydrochloride instead of glycinamide hydrochloride to give 79mg of a white solid in 55% yield. Melting point: 98.1-100.2 ℃. ¹ H NMR(400MHz,CD ₃ OD)δ7.48(s,1H),7.34(s,1H),7.22–7.03(m,4H),6.89(s,2H),5.08(s,2H),2.93(d,J＝5.4Hz,1H),2.57(d,J＝21.2Hz,4H),1.91(s,1H),1.80(s,2H),0.98(d,J＝3.6Hz,6H).LCMS(M+H) ⁺ :359.0,HRMS(ESI,m/z):calculated for C ₂₁ H ₂₇ FN ₂ O ₂ (M+H) ⁺ 359.2129,found 359.2140.

EXAMPLE 6 preparation of Compound I-6

The preparation of example 2, using L-leucinamide hydrochloride instead of glycinamide hydrochloride, gave 118mg of a white solid in 79% yield. ¹ H NMR(400MHz,CD ₃ OD)δ7.49(s,1H),7.33(s,1H),7.13(d,J＝26.9Hz,4H),6.89(d,J＝7.6Hz,2H),5.08(s,2H),3.24(s,1H),2.59(d,J＝5.2Hz,4H),1.80(d,J＝7.0Hz,2H),1.68(d,J＝5.9Hz,1H),1.48(dd,J＝32.2,6.6Hz,2H),0.94(dd,J＝9.2,7.3Hz,6H).LCMS(M+H) ⁺ :373.0.

EXAMPLE 7 preparation of Compound I-7

The preparation method of the compound was the same as example 2 except that L-sersamine hydrochloride was used instead of glycinamide hydrochloride, and 110mg of a white solid was obtainedThe yield was 79%. ¹ H NMR(400MHz,CD ₃ OD)δ7.47(s,1H),7.32(s,1H),7.19–6.97(m,4H),6.88(d,J＝6.1Hz,2H),5.05(s,2H),3.72(d,J＝28.8Hz,2H),3.30(s,1H),2.61(d,J＝20.8Hz,4H),1.81(s,2H).LCMS(M+H) ⁺ :346.9.

EXAMPLE 8 preparation of Compound I-8

The preparation of example 2, using threonine amide hydrochloride instead of glycinamide hydrochloride, gave 106mg of white solid in 74% yield. ¹ H NMR(400MHz,CD ₃ OD)δ7.46(t,J＝6.7Hz,1H),7.31(s,1H),7.17–7.01(m,4H),6.88(d,J＝7.6Hz,2H),5.04(s,2H),3.91–3.74(m,1H),3.04(d,J＝5.9Hz,1H),2.59(d,J＝7.6Hz,4H),1.80(s,2H),1.22(d,J＝5.5Hz,3H).LCMS(M+H) ⁺ :361.0.

EXAMPLE 9 preparation of Compound I-9

The preparation of example 2, using 1-amino-1-cyclopentanecarboxamide instead of glycinamide hydrochloride, gave 133mg of a white solid with a yield of 90%. ¹ H NMR(400MHz,CD ₃ OD)δ7.46(s,1H),7.30(s,1H),7.09(s,4H),6.87(s,2H),5.04(s,2H),2.59(s,4H),2.08(s,2H),1.77(s,8H).LCMS(M+H) ⁺ :370.8.

EXAMPLE 10 Synthesis of Compound I-10

Taking 99mg (0.30 mmol) of (S) -2- ((3- (4- ((2-fluorobenzyl) oxy) phenyl) propyl) amino) propionamide into a 50mL single-neck flask, adding 10mL of methanol, stirring, sequentially adding 180mg (6 mmol) of formaldehyde, 378mg (6 mmol) of sodium cyanoborohydride and 0.30mL of glacial acetic acid, stirring at room temperature overnight, concentrating under reduced pressure, dissolving the concentrate in 10mL of ethyl acetateAnd 10mL of water, pH =9 with ammonia water, the aqueous phase was extracted three times with 10mL of ethyl acetate, the organic phases were combined, the organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure, and purified by silica gel column chromatography to obtain 91mg of a white solid with a yield of 88%. Melting point: 107.0-108.0 ℃. ¹ H NMR(400MHz,CD ₃ OD)δ7.29(s,1H),7.14(s,1H),6.94(d,J＝20.3Hz,4H),6.70(s,2H),4.87(s,2H),2.99(s,1H),2.32(d,J＝31.1Hz,4H),2.07(s,3H),1.58(s,2H),1.00(s,3H).LCMS(M+H) ⁺ :345.3,HRMS(ESI,m/z):calculated for C ₂₀ H ₂₅ FN ₂ O ₂ (M+H) ⁺ 345.1973,found 345.1979.

EXAMPLE 11 Synthesis of Compound I-11

The procedure is as in example 10, except that acetaldehyde is used instead of formaldehyde, to give 78mg of a pale yellow oil in 73% yield. ¹ H NMR(400MHz,CD ₃ OD)δ7.23(dd,J＝13.2,7.1Hz,1H),7.08(d,J＝5.8Hz,1H),6.85(d,J＝16.6Hz,4H),6.64(d,J＝14.1Hz,2H),4.80(d,J＝21.5Hz,2H),3.22–3.02(m,1H),2.27(d,J＝5.9Hz,6H),1.49(d,J＝6.4Hz,2H),0.91(dd,J＝14.3,7.0Hz,3H),0.77(dd,J＝14.1,6.7Hz,3H).LCMS(M+H) ⁺ :359.3.

EXAMPLE 12 Synthesis of Compound I-12

The procedure is as in example 10 except that benzaldehyde is used instead of formaldehyde, and 31mg of a white solid is obtained in a yield of 25%. ¹ H NMR(400MHz,CD ₃ OD)δ7.48(s,1H),7.40–7.20(m,6H),7.19–7.08(m,2H),7.01(d,J＝7.6Hz,2H),6.85(d,J＝8.2Hz,2H),5.06(s,2H),3.75(d,J＝13.1Hz,1H),3.46(dd,J＝53.9,10.0Hz,2H),2.51(s,4H),1.78(d,J＝6.2Hz,2H),1.20(d,J＝6.5Hz,3H).LCMS(M+H) ⁺ :421.3.

EXAMPLE 13 preparation of Compound I-13

13.1 Preparation of (S) -2- ((3- (4-hydroxyphenyl) propyl) amino) propanamide

The reaction solution was cooled to room temperature after the reaction was allowed to react overnight by adding 330mg (1 mmol) of (S) -2- ((3- (4- ((2-fluorobenzyl) oxy) phenyl) propyl) amino) propionamide to a 50mL single-neck flask, adding 25mL of ethanol, stirring, adding 320mg (0.15 mmol) of 5% palladium-carbon, replacing with hydrogen for 5 times, slowly raising the temperature to 45 ℃. Celite was filtered, washed twice with 10mL of ethanol, and concentrated under reduced pressure, followed by purification by silica gel column chromatography to give 200mg of colorless oil in 90% yield.

13.2 preparation of Compound I-13

142mg (0.54 mmol) of triphenylphosphine and 124mg (0.54 mmol) of di-tert-butyl azodicarboxylate were placed in a 10mL sealed tube, 2mL of tetrahydrofuran was added and stirred until reaching a clear solution, 77mg (0.54 mmol) of 2-chlorobenzyl alcohol was added and stirred until reaching a clear solution, 222mg (0.45 mmol) of (S) -2- ((3- (4-hydroxyphenyl) propyl) amino) propionamide was added, the temperature was slowly increased to 90 ℃ and after overnight reaction, the mixture was concentrated under reduced pressure and purified by silica gel column chromatography to obtain 51mg of a white solid with a yield of 33%. Melting point: 96.0-97.4 ℃. ¹ H NMR(400MHz,CD ₃ OD)δ7.52(s,1H),7.40(d,J＝3.1Hz,1H),7.29(d,J＝3.0Hz,2H),7.11(d,J＝7.2Hz,2H),6.89(d,J＝7.0Hz,2H),5.10(s,2H),3.36(dd,J＝13.8,4.9Hz,1H),2.61(dd,J＝11.2,5.4Hz,4H),1.89–1.74(m,2H),1.32(d,J＝6.3Hz,3H).LCMS(M+H) ⁺ :346.9,HRMS(ESI,m/z):calculated for C ₁₉ H ₂₃ ClN ₂ O ₂ (M+H) ⁺ 347.1521,found 347.1526.

EXAMPLE 14 preparation of Compound I-14

The preparation method used was the same as in example 13 except that 2-bromobenzyl alcohol was used instead of 2-chlorobenzyl alcohol, whereby 41mg of a white solid was obtained in a yield of 23%. ¹ H NMR(400MHz,CD ₃ OD)δ7.71–7.47(m,2H),7.31(dd,J＝51.9,5.9Hz,2H),7.14(d,J＝6.1Hz,2H),6.92(s,2H),5.10(s,2H),3.38(d,J＝7.5Hz,1H),2.62(s,4H),1.83(s,2H),1.32(d,J＝5.6Hz,3H).LCMS(M+H) ⁺ :390.8.

EXAMPLE 15 preparation of Compound I-15

The procedure for preparation of 2-methoxybenzyl alcohol instead of 2-chlorobenzyl alcohol is as in example 13, giving 51mg of a white solid in 33% yield. Melting point: 81.8-83.6 ℃. ¹ H NMR(400MHz,CD ₃ OD)δ7.37(d,J＝7.1Hz,1H),7.26(d,J＝7.6Hz,1H),7.08(d,J＝7.8Hz,2H),6.99–6.89(m,2H),6.86(d,J＝8.2Hz,2H),5.02(d,J＝7.0Hz,2H),3.82(d,J＝7.3Hz,3H),3.22(d,J＝6.7Hz,1H),2.55(d,J＝6.0Hz,4H),1.77(s,2H),1.26(d,J＝6.8Hz,3H).LCMS(M+H) ⁺ :343.0,HRMS(ESI,m/z):calculated for C ₂₀ H ₂₆ N ₂ O ₃ (M+H) ⁺ 343.2016,found 343.2023.

EXAMPLE 16 preparation of Compound I-16

The procedure of example 13 was repeated except for using 2-methylbenzyl alcohol instead of 2-chlorobenzyl alcohol to give 37mg of a white solid in a yield of 25%. Melting point: 108.2-109.1 ℃. ¹ H NMR(400MHz,CD ₃ OD)δ7.35(d,J＝4.9Hz,1H),7.19(s,3H),7.10(d,J＝4.8Hz,2H),6.89(d,J＝6.6Hz,2H),4.99(s,2H),3.23(d,J＝4.4Hz,1H),2.57(s,4H),2.34(d,J＝4.6Hz,3H),1.79(s,2H),1.27(d,J＝4.5Hz,3H).LCMS(M+H) ⁺ :327.0,HRMS(ESI,m/z):calculated for C ₂₀ H ₂₆ N ₂ O ₂ (M+H) ⁺ 327.2067,found 327.2078.

EXAMPLE 17 preparation of Compounds I-17

The procedure for preparation of example 13 was carried out using 2-trifluoromethoxy-benzyl alcohol instead of 2-chlorobenzyl alcohol, giving 70mg of pale white solid in 39% yield. ¹ H NMR(400MHz,CD ₃ OD)δ7.58(d,J＝5.9Hz,1H),7.40(d,J＝7.3Hz,1H),7.33(s,2H),7.10(d,J＝7.9Hz,2H),6.87(d,J＝8.1Hz,2H),5.08(s,2H),3.24(d,J＝6.6Hz,1H),2.56(d,J＝7.0Hz,4H),1.79(s,2H),1.36–1.23(m,3H).LCMS(M+H) ⁺ :397.2.

EXAMPLE 18 preparation of Compound I-18

The preparation method used was the same as in example 13 except that 2-chlorobenzyl alcohol was replaced with 2-trifluoromethylbenzyl alcohol to obtain 60mg of a white solid with a yield of 35%. ¹ H NMR(400MHz,CD ₃ OD)δ7.80–7.65(m,2H),7.61(t,J＝7.5Hz,1H),7.48(t,J＝7.5Hz,1H),7.12(d,J＝8.3Hz,2H),6.87(d,J＝8.5Hz,2H),5.21(s,2H),3.32–3.28(m,1H),2.59(d,J＝3.0Hz,4H),1.81(dd,J＝14.9,7.5Hz,2H),1.28(d,J＝6.9Hz,3H).LCMS(M+H) ⁺ :381.2.

EXAMPLE 19 preparation of Compounds I-19

The procedure of example 13 was repeated except for using 2, 3-difluorobenzyl alcohol instead of 2-chlorobenzyl alcohol to give 72mg of a white solid in a yield of 46%. ¹ H NMR(400MHz,CD ₃ OD)δ7.27(s,1H),7.21(d,J＝7.9Hz,1H),7.13(t,J＝11.6Hz,3H),6.89(d,J＝8.5Hz,2H),5.10(s,2H),3.34–3.25(m,1H),2.59(s,4H),1.81(d,J＝6.6Hz,2H),1.29(d,J＝6.9Hz,3H).LCMS(M+H) ⁺ :349.2.

EXAMPLE 20 preparation of Compound I-20

The preparation method using 2, 3-methoxybenzyl alcohol instead of 2-chlorobenzyl alcohol is the same as example 13, 80mg of pale yellow solid is obtainedBulk, yield 46%. ¹ H NMR(400MHz,CD ₃ OD)δ7.02(s,2H),7.00–6.86(m,3H),6.83(d,J＝6.6Hz,2H),4.94(d,J＝11.7Hz,2H),3.90–3.67(m,6H),3.30(d,J＝21.6Hz,1H),2.54(d,J＝23.2Hz,4H),1.76(s,2H),1.25(d,J＝12.3Hz,3H).LCMS(M+H) ⁺ :373.2.

EXAMPLE 21 preparation of Compound I-21

The preparation method used was the same as in example 13 except that 2, 4-difluorobenzyl alcohol was used instead of 2-chlorobenzyl alcohol, whereby 71mg of a white solid was obtained in a yield of 45%. Melting point: 105.9-106.2 ℃. ¹ H NMR(400MHz,CD ₃ OD)δ7.50(dd,J＝15.2,8.1Hz,1H),7.10(d,J＝8.1Hz,2H),7.02–6.82(m,4H),5.02(s,2H),3.34–3.24(m,1H),2.58(d,J＝3.8Hz,4H),1.86–1.67(m,2H),1.28(t,J＝9.2Hz,3H).LCMS(M+H) ⁺ :349.2.HRMS(ESI,m/z):calculated for C ₁₉ H ₂₂ F ₂ N ₂ O ₂ (M+H) ⁺ 349.1722,found 349.1733.

EXAMPLE 22 preparation of Compound I-22

The preparation was carried out in the same manner as in example 13 except for using 2-fluoro-4-methoxybenzyl alcohol in place of 2-chlorobenzyl alcohol to give 48mg of a white solid with a yield of 30%. ¹ H NMR(400MHz,CD ₃ OD)δ7.51(s,1H),7.25(s,2H),7.04(s,2H),6.88(s,2H),5.12(s,2H),3.93(s,3H),3.46(s,1H),2.74(s,4H),1.96(s,2H),1.44(s,3H).LCMS(M+H) ⁺ :361.2.

EXAMPLE 23 preparation of Compound I-23

The preparation method used was the same as in example 13 except that 2, 5-difluorobenzyl alcohol was used instead of 2-chlorobenzyl alcohol, whereby 36mg of a white solid was obtained in a yield of 23%. ¹ H NMR(400MHz,CD ₃ OD)δ7.24(s,1H),7.12(d,J＝6.0Hz,4H),6.90(d,J＝5.5Hz,2H),5.07(s,2H),3.27(s,1H),2.58(s,4H),1.80(s,2H),1.28(s,3H).LCMS(M+H) ⁺ :349.2.

EXAMPLE 24 preparation of Compound I-24

The preparation method used was the same as in example 13 except that 2, 6-difluorobenzyl alcohol was used instead of 2-chlorobenzyl alcohol, whereby 28mg of a white solid was obtained in 18% yield. Melting point: 98.8-99.4 ℃. ¹ H NMR(400MHz,CD ₃ OD)δ7.46–7.32(m,1H),7.09(dd,J＝17.7,8.3Hz,2H),6.99(dd,J＝16.6,9.0Hz,2H),6.93–6.83(m,2H),5.05(d,J＝17.9Hz,2H),3.29–3.13(m,1H),2.56(d,J＝6.8Hz,4H),1.79(s,2H),1.27(d,J＝6.7Hz,3H).LCMS(M+H) ⁺ :348.9.HRMS(ESI,m/z):calculated for C ₁₉ H ₂₂ F ₂ N ₂ O ₂ (M+H) ⁺ 349.1722,found349.1727.

EXAMPLE 25 preparation of Compound I-25

The preparation method used 2,4, 5-trifluorobenzyl alcohol instead of 2-chlorobenzyl alcohol was the same as example 13, 36mg of a white solid was obtained, with a yield of 22%. ¹ H NMR(400MHz,CD ₃ OD)δ7.45(d,J＝8.2Hz,1H),7.18(dd,J＝22.4,8.0Hz,3H),6.88(dd,J＝36.7,7.4Hz,2H),5.06(s,2H),3.32(d,J＝4.8Hz,1H),2.61(s,4H),1.82(d,J＝5.6Hz,2H),1.32(d,J＝7.0Hz,3H).LCMS(M+H) ⁺ :366.8.

EXAMPLE 26 preparation of Compound I-26

The preparation method used 2,4, 6-trifluorobenzyl alcohol instead of 2-chlorobenzyl alcohol was the same as example 13, and 44mg of a white solid was obtained in 27% yield. ¹ H NMR(400MHz,CD ₃ OD)δ7.11(d,J＝7.7Hz,2H),6.89(d,J＝4.6Hz,4H),5.02(s,2H),3.24(q,J＝6.5Hz,1H),2.64–2.51(m,4H),1.79(s,2H),1.27(d,J＝6.7Hz,3H).LCMS(M+H) ⁺ :367.2.

EXAMPLE 27 preparation of Compounds I-27

The preparation method was the same as in example 13 except for using 2,3,4,5, 6-pentafluorobenzyl alcohol instead of 2-chlorobenzyl alcohol, and 42mg of a white solid was obtained in a yield of 23%. ¹ H NMR(400MHz,CD ₃ OD)δ7.15(s,2H),6.92(s,2H),5.15(s,2H),3.30(s,1H),2.60(s,4H),1.81(s,2H),1.30(s,3H).LCMS(M+H) ⁺ :402.9.

EXAMPLE 28 preparation of Compound I-28

The procedure of example 13 was repeated except that benzyl alcohol was used instead of 2-chlorobenzyl alcohol to give 45mg of a white solid in a yield of 32%. ¹ H NMR(400MHz,CD ₃ OD)δ7.36(dd,J＝29.1,16.1Hz,5H),7.09(s,2H),6.88(s,2H),5.01(d,J＝7.2Hz,2H),3.31(s,1H),2.59(s,4H),1.81(s,2H),1.29(s,3H).LCMS(M+H) ⁺ :313.2.

EXAMPLE 29 preparation of Compounds I-29

The preparation was carried out in the same manner as in example 13 except that 2-chlorobenzyl alcohol was replaced by naphthalen-1-ylmethanol, to give 49mg of a white solid in a yield of 30%. ¹ H NMR(400MHz,CD ₃ OD)δ8.02(s,1H),7.90–7.77(m,2H),7.54(s,1H),7.48(s,2H),7.43(d,J＝8.1Hz,1H),7.10(d,J＝7.3Hz,2H),6.94(d,J＝7.1Hz,2H),5.41(s,2H),3.30(d,J＝4.8Hz,1H),2.58(s,4H),1.80(s,2H),1.29(d,J＝5.5Hz,3H).LCMS(M+H) ⁺ :363.0.

EXAMPLE 30 preparation of Compound I-30

The procedure used in example 13 was repeated except for using 1, 3-benzodioxol-4-alkylmethanol instead of 2-chlorobenzyl alcohol to give 92mg of a white solid in a yield of 57%. Melting point: 102.5-103.8 ℃. ¹ H NMR(400MHz,CD ₃ OD)δ7.10(d,J＝8.0Hz,2H),6.90(t,J＝10.0Hz,3H),6.85–6.75(m,2H),5.97(s,2H),5.00(s,2H),3.26(d,J＝6.8Hz,1H),2.71–2.41(m,4H),1.80(d,J＝6.9Hz,2H),1.28(d,J＝6.7Hz,3H).LCMS(M+H) ⁺ :356.9.HRMS(ESI,m/z):calculated for C ₂₀ H ₂₄ N ₂ O ₄ (M+H) ⁺ 357.1809,found 357.1821.

EXAMPLE 31 preparation of Compound I-31

The procedure of example 13 was followed using 2, 3-dihydro-1, 4-benzodioxan-5-methyl alcohol instead of 2-chlorobenzyl alcohol to give 63mg of a pale yellow solid in 38% yield. Melting point: 116.0-117.1 ℃. ¹ H NMR(400MHz,CD ₃ OD)δ7.05(d,J＝4.7Hz,2H),6.87(d,J＝24.7Hz,3H),6.76(s,2H),4.95(s,2H),4.18(d,J＝12.4Hz,4H),3.30(s,1H),2.55(s,4H),1.78(s,2H),1.28(s,3H).LCMS(M+H) ⁺ :371.2.HRMS(ESI,m/z):calculated for C ₂₁ H ₂₆ N ₂ O ₄ (M+H) ⁺ 371.1965,found 371.1971.

EXAMPLE 32 preparation of Compound I-32

The preparation method used in example 13 was the same as that of example 13 except that 1-adamantanemethanol was used instead of 2-chlorobenzyl alcohol, whereby 18mg of a white solid was obtained in a yield of 11%. ¹ H NMR(400MHz,CD ₃ OD)δ7.08(s,2H),6.80(s,2H),3.45(s,2H),3.32(d,J＝6.8Hz,1H),2.59(s,4H),1.99(s,3H),1.75(d,J＝17.4Hz,8H),1.67(s,6H),1.31(s,3H).LCMS(M+H) ⁺ :371.3.

EXAMPLE 33 preparation of Compound I-33

The same procedures used in example 13 were repeated except for using (S) -1- (2-fluorophenyl) ethanol instead of 2-chlorobenzyl alcohol to give 54mg of a white solid in 35% yield. Melting point: 64.0-65.6 ℃. ¹ H NMR(400MHz,CD ₃ OD)δ7.51–7.32(m,1H),7.22(d,J＝5.4Hz,1H),7.12–6.94(m,4H),6.74(t,J＝11.9Hz,2H),5.71–5.47(m,1H),3.29–3.16(m,1H),2.54(dd,J＝13.6,6.6Hz,4H),1.80–1.67(m,2H),1.60–1.51(m,3H),1.28–1.20(m,3H).LCMS(M+H) ⁺ :345.3.HRMS(ESI,m/z):calculated for C ₂₀ H ₂₅ FN ₂ O ₂ (M+H) ⁺ 345.1973,found345.1980.

EXAMPLE 34 preparation of Compound I-34

The same procedures used in example 14 were repeated except for using (R) -1- (2-fluorophenyl) ethanol instead of 2-chlorobenzyl alcohol to give 73mg of a white solid in 47% yield. Melting point: 90.6-97.4 ℃. ¹ H NMR(400MHz,CD ₃ OD)δ7.49–7.36(m,1H),7.24(s,1H),7.14–6.94(m,4H),6.74(d,J＝8.4Hz,2H),5.69–5.54(m,1H),3.21(dd,J＝13.1,6.3Hz,1H),2.52(d,J＝6.0Hz,4H),1.74(d,J＝5.8Hz,2H),1.57(d,J＝6.4Hz,3H),1.25(s,3H).LCMS(M+H) ⁺ :345.2.HRMS(ESI,m/z):calculated for C ₂₀ H ₂₅ FN ₂ O ₂ (M+H) ⁺ 345.1973,found345.1981.

EXAMPLE 35 preparation of Compound I-35

Replacing methyl p-hydroxyphenylpropionate with methyl p-hydroxybenzoate to obtain 4- (4- ((2-fluorobenzyl) oxy)Phenyl) butyraldehyde, which was used instead of 3- (4- ((2-fluorobenzyl) oxy) phenyl) propionaldehyde, was prepared by the same method as in example 1, giving a yield of 62% in 213mg of white solid. Melting point: 113.0-113.5 ℃. ¹ H NMR(400MHz,CD ₃ OD)δ7.50(s,1H),7.35(s,1H),7.25–7.06(m,4H),6.92(d,J＝14.1Hz,2H),5.11(d,J＝22.3Hz,2H),3.19(s,1H),2.56(s,4H),1.56(d,J＝39.8Hz,4H),1.27(d,J＝15.9Hz,3H).LCMS(M+H) ⁺ :345.2.HRMS(ESI,m/z):calculated for C ₂₀ H ₂₅ FN ₂ O ₂ (M+H) ⁺ 345.1973,found 345.1978.

Performance testing

Test example 1: electrophysiological assay

Culture and passage of stably transfected cell lines

HEK293 cells stably expressing Nav1.7 and the sodium channel subtype to be determined, using DMEM medium containing 10% Fetal Bovine Serum (FBS), 1% Penicillin/Streptomyces (P/S) and 500. Mu.g/ml G418 at 37 ℃ 5% CO ₂ Culturing in an incubator with concentration. When the cell fusion degree reaches about 90%, 0.25% pancreatin is digested and passaged at 1 × 10 ⁵ Mu.l of single cell suspension was inoculated at a density of/L onto 8mm cell slides and used 24h later for patch clamp experiments.

Recording of electric current

The test compound was dissolved in DMSO to prepare a 50mM stock solution, which was dispensed and frozen at-20 ℃. Test compounds were diluted to the corresponding working concentration (DMSO)<0.3%) was subjected to patch clamp electrophysiological activity test. Placing the slide inoculated with cells under an inverted microscope, perfusing extracellular fluid (flow rate about 1mL/min, formula (mM): 140NaCl, 1.8 CaCl) ₂ 、1MgCl ₂ 10Glucose, 4KCl, 10HEPES, naOH to pH 7.4). Capillary glass tubes were drawn into recording electrodes by a multi-step procedure using a horizontal drawing instrument (P-97, sutter Instrument, USA), and filled with electrode solutions (formulation (mM): 65CsCl, 75CsF, 5EGTA, 2.5 MgCl) under an inverted microscope ₂ 10HEPES, csOH to regulate pH to 7.3) to the cell, negative pressure suction is applied to form high-resistance seal, negative pressure is applied again to break cell membrane to form whole-cell markerRecording mode.

All current recordings were performed at room temperature (20-25 deg.C) using an Axon patch 700B patch clamp amplifier and signal acquisition using pClamp 10.5 software (Axon Instruments, CA) with 10KHz filtering. The Nav1.7 current recording used a stimulation program of: cells were first clamped at-40 mV to almost completely inactivate the Nav1.7 current, then hyperpolarized to-150 mV for 20ms to reactivate a portion of the channels, depolarized to 0mV to induce a reactivated portion of the Nav1.7 current, and finally returned to-40 mV, with the entire stimulation program run once in 1 s. Repeating for several times to deactivate the current by 50%, and allowing the current to stabilize (the current is I) ₀ ) The effect of the compound to be tested on the Nav1.7 current is observed by perfusion, and the current after administration is recorded as I _Medicine . In this test, all compounds were prototype compounds.

Data analysis

Data analysis and image processing were performed using OriginPro 8.0 (Origin Lab, usa) and Adobe Illustrator, etc. software. All experimental data are expressed as mean ± standard error (mean ± s.e.m.). The inhibition rate was calculated as (I) ₀ -I _Medicine )/I ₀ . The dose-effect relationship curve is fitted by the logistic equation as follows: y = A ₂ +(A ₁ -A ₂ )/[1+(x/x ₀ ) ^p ]Wherein y represents the effect, A ₁ And A ₂ Representing maximum and minimum effects, respectively, x represents drug concentration and p is the Hill coefficient. Statistical analysis of the data was performed using unpaired t-test (Student's unpaired t-test), and differences between the two groups were considered statistically significant when P ≦ 0.05.

Results of the experiment

The results of the experiment are shown in tables 1 to 2.

TABLE 1 Nav1.7 inhibition rates of Compounds I-1 to 35 at a concentration of 10. Mu.M

Table 2 partial compounds Nav1.7 Activity test results (IC) ₅₀ )

Number of	IC 50 (μM)	Number of	IC 50 (μM)	Numbering	IC 50 (μM)
						I-1	5.00±0.89	I-13	9.86±1.90	I-31	7.53±1.22
I-2	10.06±2.06	I-15	11.65±2.03	I-33	12.08±3.28
						I-3	15.29±2.01	I-16	10.18±2.10	I-34	15.39±2.51
I-4	9.34±2.53	I-21	12.00±2.58	I-35	9.89±1.43
						I-5	21.44±5.18	I-24	14.02±0.56	Ralfinamide	35.24±2.80
I-10	18.36±4.19	I-30	2.93±0.80	-	-

As can be seen from tables 1-2, the compounds of the examples of the present invention have better inhibitory action on sodium ion channel Nav1.7, and the inhibitory activity is better than Ralfinamide.

Test example 2: mouse sciatic nerve branch selective injury pain model test

Construction of mouse sciatic Nerve Branch Selective Injury (SNI) model

Before the experiment, surgical instruments are disinfected and then soaked in normal saline, 8% chloral hydrate is injected into the abdominal cavity of a mouse for anesthesia, and the hair near the hind limb of the left leg of the mouse is removed by an automatic hair remover and is placed on an operation table. The skin surface of the mouse is disinfected by dipping the iodophor in cotton, and then the mouse is wiped clean by a cotton ball dipped with physiological saline. The skin of the left hind limb was incised with scissors and the biceps brachii muscle of the hind limb was gently peeled off with a glass needle, exposing the sciatic nerve and three branches distal to it: sural nerve, common peroneal nerve, tibial nerve. The common peroneal nerve and the tibial nerve of 1-2mm are cut off by scissors respectively to prevent the nerves from growing and combining again after suturing, and the sural nerve is prevented from being touched in the process. The wound was then sutured and the mice were placed on a heat blanket to maintain the body temperature of the mice, helping the mice to recover from anesthesia as quickly as possible. The mice in the sham operation group are not cut after the three nerves are separated, and the other operations are the same as the operation group.

Mechanical paw withdrawal pain threshold measurement in mice

The mouse adapts to an experimental environment for two days before an experiment, and the basal foot-contraction pain threshold of the mouse is measured on the third day, wherein the specific measurement steps are as follows: the mice were placed in a box with a metal grid in advance, acclimated for 15-30min until they remained familiar and quiet to the strange environment (the mice did not walk around in the box, scurry up, lick themselves). The Von Frey hair measured was chosen to be 0.008, 0.02, 0.04, 0.07, 0.16, 0.40, 0.60, 1.0, 1.4, 2.0, (unit: g) and the corresponding log units were 1.65, 2.36, 2.44, 2.83, 3.22, 3.61, 3.84, 4.08, 4.17, 4.31. During measurement, 0.40g of the middle value is selected as the starting point, the testing fiber is vertically contacted with the sole of the foot at the hairless position of the sole of the left side, when the fiber is bent in an S shape and lasts for more than 5S, the reaction of the mouse is observed, if the mouse is rapidly retracted, is afraid of contracting, is lifted or licks and bites the left hind paw, the positive reaction is obtained, the fiber with the lower force is replaced for measurement, otherwise, the fiber with the higher force is replaced, and the interval between adjacent stimulations is at least 7 seconds. The positive reaction is needed to be continuously measured for four times for the first time, and the log unit value corresponding to the Von Frey hair measured for the last time is substituted into the formula for calculation. In the stimulation process, the mice walk back and forth or just lift feet to be positive suspicions just after contacting with stimulation, and the measurement needs to be carried out when the mice are quiet. Positive results are marked as X, negative is O. The mechanical foot-contraction pain threshold of the mice was calculated by the up-and-down method.

Animal grouping and dosing

The above surgical mice were divided into 6 groups: solvent control group (Vehicle), 10 mg. Kg ^-1 Ralfinamide group, 5 mg.kg ^-1 Morphine group and compound I-1, 2 mg/kg ^-1 、5mg·kg ^-1 、10mg·kg ^-1 Or compound I-30 in three dose groups of 5 mg/kg ^-1 、10mg·kg ^-1 、20mg·kg ^-1 . In this test, I-1 and Ralfinamide were prototypes of drugs and morphine was the hydrochloride salt.

And (4) measuring the pain threshold of the mice for two consecutive days from the fifth day after four days of postoperative recovery of the mice, and judging whether the mice are successfully modeled. The mice were evaluated for mechanical paw withdrawal pain threshold on days 1,3, 5, 7 and 10 following the procedure, i.e., intraperitoneal injections of the drug once a day, starting on day 8 after the operation.

Analgesic Effect of Compound I-1 in the mouse SNI model

The operation of the SNI neuropathological pain model resulted in a significant and sustained increase in mechanical sensitivity in mice. On day 6 after surgery, the mechanical paw withdrawal pain threshold of the mouse on the operative side was 0.04 ± 0.009g, which is significantly lower than the mechanical withdrawal pain threshold of 1.32 ± 0.11g in the sham group mice, indicating that the SNI model was successfully prepared (table 3, fig. 1). Injections 2,5 and 10mg/kg ^-1 I-1 and 10mg kg ^-1 The mean mechanical paw withdrawal thresholds were raised to 0.25. + -. 0.02g, 0.45. + -. 0.03g, 0.98. + -. 0.04g and 0.52. + -. 0.04g respectively after Ralfinamide. The experimental result shows that the compound I-1 dose-dependently relieves the neuropathic pain, the analgesic effect is better than Ralfinamide under the dose of 10mg/kg, and 10mg/kg is taken in the tenth day after the administration ^-1 The mechanical foot pain threshold of mice in group I-1 is 0.98 + -0.11 g, and is compared with morphine (5 mg. Kg) in positive control group ^-1 ) 1.19 + -0.07 g has equivalent analgesic effect. Each data point is expressed as the mean ± sem, ^*** P<0.001vs Sham， ^# P<0.05， ^### P<0.01， ^### P<0.001vs Vehicle。

TABLE 3 SNI test results for Compound I-1

Each data point is expressed as the mean ± sem, ^*** P<0.001vs Sham， ^# P<0.05， ^### P<0.01， ^### P<0.001vs Vehicle。

analgesic Effect of Compound I-30 in the mouse SNI model

The operation of the SNI neuropathological pain model resulted in a significant and sustained increase in mechanical sensitivity in mice. On the 6 th day after operation, the mechanical foot-contracting pain threshold of the mouse on the postoperative side hind paw is 0.03 +/-0.002 g, which is obviously lower than that of the mouse in the sham operation group which is 1.03 +/-0.07 g, which indicates that the SNI model is successfully prepared (Table 4 and figure 2). Injections 5, 10 and 20 mg/kg ^-1 I-30 and 10 mg.kg of ^-1 After Ralfinamide, the mean mechanical foot pain threshold rises to 0.41 + -0.03 g, 0.75 + -0.03 g, 0.97 + -0.06 g and 0.62 + -0.02 g, respectively. The experimental result shows that the compound I-30 can reduce neuropathic pain in a dose-dependent manner, the analgesic effect is better than Ralfinamide under the dose of 10mg/kg, and 20 mg/kg is taken in the tenth day after administration ^-1 The threshold value of mechanical foot-contracting pain of the mice in the group I-30 is 1.06 +/-0.07 g, and the effect is equivalent to that of the mice in the sham operation group 1.17 +/-0.05 g. Each data point is expressed as the mean ± sem, ^*** P<0.001vs Sham， ^# P<0.05， ^### P<0.01， ^### P<0.001vs vessel. In this test, ralfinamide was the drug prototype and I-30 and morphine were the hydrochloride salts.

TABLE 4 SNI test results for Compound I-30

Each data point is expressed as mean ± sem, ^*** P<0.001vs Sham， ^# P<0.05， ^### P<0.01， ^### P<0.001vs Vehicle。

test example 3: formalin pain model test in mice

Construction of mouse formalin pain model

Before the experiment is started, the mice are placed in an observation box to adapt for 20min, so that the mice are prevented from causing a nervous and panic emotion in an unfamiliar environment, the drugs are respectively injected into the abdominal cavity in advance according to the detection time of the detected drugs, the needle is inserted into the skin at the center of the left sole of each mouse, the needle is slowly pushed into 20 mu L of a prepared 5% formalin solution after being inserted into the skin, the needle is taken out while rotating when being pulled out, and the administration effect is prevented from being influenced by the seepage of the formalin solution from the sole along with the needle. Then, the mice were quickly placed in an observation box for recording and observation, and the control group was subjected to intraperitoneal injection of physiological saline for observation. In a formalin-induced mouse inflammatory pain model experiment, formalin can enable a mouse to have pain reactions such as paw licking, foot lifting, leg contraction and the like, the phase I reaction recording time is within 0-5min after formalin injection, the phase II pain reaction recording time is within 6-45min, and the total time of the mouse for licking, biting, lifting and contracting the left hind paw in the two-phase pain reaction time is calculated respectively.

Animal grouping and dosing

The above surgical mice were divided into 6 groups: solvent control group (Vehicle), 10 mg. Kg ^-1 Ralfinamide group, 10 mg. Kg ^-1 Morphine group and Compound I-1 in three dose groups, each 5mg kg ^-1 、10mg·kg ^-1 And 20mg kg ^-1 . In this test, I-1 and Ralfinamide were prototypes of drugs and morphine was the hydrochloride salt.

Analgesic Effect of Compound I-1 in formalin pain model

Compound I-1 was effective in inhibiting inflammatory pain in the evaluation of the formalin-induced mouse pain model (table 5, fig. 3). The injection is administered at 5, 10, 20 mg/kg ^-1 After I-1, the total paw Licking times (Licking time) of the I-phase reaction are respectively 27.3 +/-6.02 s, 13.58 +/-4.86 s and 16.3 +/-5.57 s, and no obvious analgesic effect is observed compared with 26.25 +/-5.71 s of a solvent control group; the total number of times of paw licking in II phase reaction is 291.00 +/-46.19 s, 201.2 +/-30.29 s and 164.80 +/-26.04 s respectively, and compared with 392.75 +/-42.26 s of solvent control group, the compound has dose-dependent analgesic effect. Intraperitoneal injection of 10mg/kg ^-1 Ralfinamide, the total number of paw licks in phase I reaction is 18.20 + -3.96s, and the total number of paw licks in phase II reaction is 294.90 + -51.94 s, which causes 10mg/kg pain in phase I and phase II ^-1 None of the Ralfinamides showed a more pronounced analgesic effect than I-1. Each data point is expressed as the mean ± sem, ^** P<0.01， ^*** P<0.001vs Vehicle。

TABLE 5 formalin test results for Compound I-1

Note: each data point is expressed as the mean ± sem, ^** P<0.01， ^*** P<0.001vs Vehicle。

test example 4: metabolic stability testing

Test compounds: compounds I-1 and I-30.

Positive control compound: ketanserin, the stock concentrations of the above compounds are all 10mM.

Buffer solution: 100mM potassium phosphate buffer, pH 7.4.

Acetonitrile solution with drug concentration 500 μ M: mu.L of stock solution (10 mM) of test compound or positive control compound was diluted with 95. Mu.L of acetonitrile.

Liver microsome solution with drug concentration of 1.5 μ M: mu.L of a 500. Mu.M acetonitrile solution having a drug concentration of 500. Mu.M and 18.75. Mu.L of 20mg/mL liver particles were taken into 479.75. Mu.L of 100mM potassium phosphate buffer.

3 parts NADPH stock (6 mM,5 mg/mL): the NADPH is dissolved in a buffer solution to prepare the compound.

And (3) testing: 30 μ L of liver microsome solution at a drug concentration of 1.5 μ M was dispensed to assay plates designated for different time points (0-, 5-,15-,30-, 45-min). The reaction stop solution was added at 0min to the 0min assay plate, followed by 15. Mu.L of NADPH stock. The assay plate incubated the mixture of microsomal solution and compound at 37 ℃ for about 5 minutes. Add 15. Mu.L of NADPH stock and start the reaction and time. Adding reaction stop solution at 5-min,15-min,30-min and 45-min, and stopping reaction. The sample plate was shaken for about 10 minutes and the sample was centrifuged at 600rpm for 10 minutes and 6000rpm for 15 minutes. After centrifugation, 80. Mu.L of the supernatant and 120. Mu.L of HPLC water were added to an 8X 96 well plate, mixed and detected by LCMS/MS.

TABLE 6I-1 stability test results in human liver microsomes

TABLE 7 stability test results of I-1 in liver microsomes of mice

TABLE 8I-30 stability test results in human liver microsomes

TABLE 9 stability test results of I-30 in liver microsomes of mice

As seen from tables 6-9, the half-lives of compounds I-1 and I-30 in human liver microsomes were both greater than 120 minutes and in mouse liver microsomes were 44.57 and 36.94 minutes, respectively, so that compounds I-1 and I-30 had better metabolic stability. In this test, I-1 and I-30 are the hydrochloride salts.

Experimental procedures without specifying specific conditions in the following examples were selected in accordance with conventional procedures and conditions, or in accordance with commercial instructions. The reagents and starting materials used in the present invention are commercially available.

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.

Claims (10)

1. An α -aminoamide compound represented by the formula (i) below, a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing the same as an active ingredient, wherein:

characterized in that the formula (I) is selected from:

each R ₁ Are respectively and independently selected from H, F, cl, br, I and C _1- C ₄ Alkyl of (C) _1- C ₄ Alkoxy of (a), the C _1- C ₄ Alkyl and C _1- C ₄ Is optionally substituted with 1, 2 or 3R; wherein m =1-5;

R ₂ selected from H, CH ₃ And CF ₃ ；

R _3- R ₇ 、R _11- R ₁₂ Is selected from H;

n is 1 or 2;

R ₈ selected from H, C ₁ ～C ₄ Alkyl and benzyl of (a); said C is ₁ ～C ₄ Is optionally substituted with 1, 2 or 3R;

R ₉ and R ₁₀ Each independently selected from H, C ₁ ～C ₄ Or R is ₉ And R ₁₀ Are linked to the carbon atom to which they are attached to form a cyclopentyl group, said C ₁ ～C ₄ Is optionally substituted with 1, 2 or 3R;

each R is independently selected from H, F, cl, br, I, CN, OH and NH ₂ 。

2. The α -aminoamide compound according to claim 1, wherein R is a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing the same as an active ingredient ₁ Selected from H, F, cl, br, I, CH ₃ 、CF ₃ 、OCH ₃ And OCF ₃ ；R ₂ Is selected from H; r is ₈ Selected from H, CH ₃ 、CH ₂ CH ₃ And

R ₉ and R ₁₀ Are respectively and independently selected from H and CH ₃ 、CH ₂ OH、CH ₂ CH ₃ 、CH(OH)CH ₃ 、CH(CH ₃ ) ₂ And CH ₂ CH(CH ₃ ) ₂ (ii) a Or R ₉ And R ₁₀ To the carbon atom to which they are attached to form a cyclopentyl group.

3. The α -aminoamide compounds according to claim 1, pharmaceutically acceptable salts thereof or pharmaceutical compositions containing these compounds as an active ingredient, characterized in that the structural unit in the formula (i)

Selected from:

structural unit in formula (I)

Selected from:

4. the α -aminoamide compounds according to claim 1, which have a structure of formula (i) selected from the group consisting of:

5. use of the α -aminoamide compounds according to any one of claims 1 to 4, pharmaceutically acceptable salts thereof or pharmaceutical compositions containing these compounds as active ingredients for the preparation of analgesic drugs.

6. Use according to claim 5, wherein the pain comprises the treatment or alleviation of pain during surgery, chronic pain, neuropathic pain, cancer pain.

7. The process for the preparation of α -aminoamides, pharmaceutically acceptable salts thereof or pharmaceutical compositions containing these compounds as active ingredient according to any of claims 1 to 4, characterized by the following synthetic route:

wherein the substituent group X is selected from O and the other substituent groups are as defined in any one of claims 1 to 6.

8. The method of claim 7, comprising the steps of:

(1) The synthesis of the target compound shown in the formula I directly uses 2-fluorobenzyl bromide as a raw material, potassium carbonate as alkali and a compound b to perform nucleophilic substitution reaction to obtain a compound c, then uses sodium hydroxide as alkali to perform ester hydrolysis to obtain a compound d, reduces the compound d by lithium aluminum hydride to obtain a compound e, then oxidizes the compound d by a desselin oxidizer to obtain a corresponding aldehyde to obtain a compound f, and performs reductive amination reaction on the compound f by sodium cyanoborohydride and corresponding alpha-aminoamide to obtain a series of target compounds g;

(2) Removing benzyl protection from the compound g through palladium-carbon hydrogenation to obtain a compound h, and then reacting to obtain a series of target compounds i;

(3) Carrying out reductive amination reaction or substitution reaction on the compound i to obtain a series of target compounds j;

(4) Similar to the synthesis, a series of target products l are obtained by the reductive amination reaction of ketone compounds k;

(5) Removing benzyl protection from the compound l through palladium-carbon hydrogenation to obtain a compound m, and reacting to obtain a series of target compounds n;

(6) And carrying out reductive amination reaction or substitution reaction on the compound n to obtain a series of target compounds I.

9. The α -aminoamide compounds according to claim 1, pharmaceutically acceptable salts thereof or pharmaceutical compositions containing these compounds as an active ingredient, wherein the pharmaceutical composition is in the form of: tablet, capsule, injection, and solution.

10. The α -aminoamide compound according to claim 1, a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing the same as an active ingredient, characterized in that it is administered by injection route or orally.

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