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

Abstract

The invention discloses the use of α-aminoamide compounds as sodium ion channel (especially Nav1.7) inhibitors in pain treatment. Specifically, the present invention discloses α-aminoamide compounds, stereoisomers, pharmaceutically acceptable salts, solvates, prodrugs or pharmaceutical compositions containing these compounds as active ingredients represented by structural formula I as sodium Use of ion channel (especially Nav1.7) inhibitor analgesic drugs, and preparation method of α-aminoamide compounds.

Description

α-Amino amide compound and its preparation method and application

technical field

The invention belongs to the field of medicinal chemistry, and relates to α-aminoamide compounds, stereoisomers, pharmaceutically acceptable salts, solvates, prodrugs and pharmaceutical compositions containing these compounds as active ingredients, and preparation of the compounds The method and its use as an analgesic drug for sodium ion channel (especially Nav1.7) inhibitor.

Background technique

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 the body's self-protection, and it is also a major health problem. It greatly reduces the quality of life and brings high health costs and economic losses to the society. Although a considerable number of analgesic drugs have been used in clinical pain treatment, due to the complex pain mechanism, the drugs have strong side effects or low efficacy, the existing pain treatment drugs are still far from satisfying clinical pain treatment. Therefore, the development of new mechanisms of analgesic drugs with strong analgesic activity and good safety has great research significance and application value.

Ralfinamide is the only Nav1.7 inhibitor that has entered clinical phase III. It has a certain therapeutic effect in the clinical phase II of moderate to severe neuropathic pain, but the clinical phase III of chronic neuropathic low back pain failed to reach the primary endpoint. Therefore, the inventor of the present patent has designed and synthesized a series of α-aminoamide compounds to screen them by activity test, which is beneficial to develop the application of analgesic active compounds and clinical drug resources, and there is no relevant report at present.

SUMMARY OF THE INVENTION

On the basis of conforming to common knowledge in the art, the above preferred conditions can be combined arbitrarily without departing from the concept and protection scope of the present invention.

In order to develop and utilize the existing clinical medicine resources, the purpose of the present invention is to provide a class of α-aminoamide compounds, stereoisomers, pharmaceutically acceptable salts, solvent compounds, prodrugs and containing these compounds as active ingredients secondly, the purpose of the present invention is to provide its preparation method, thereby opening up a new way for finding new analgesic active compounds; another purpose of the present invention is to provide it as a sodium ion channel (especially Nav1.7) The use of depressant analgesics.

The present invention provides an α-aminoamide compound represented by formula (I), a stereoisomer, a pharmaceutically acceptable salt thereof, a solvent compound, a prodrug or a pharmaceutical composition containing these compounds as active ingredients, The structural formula of 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, C _6- C ₁₀ cycloalkyl, C _6- C ₁₀ heteroaryl and C _6- C ₁₀ heterocyclic group, the C _6- C ₁₀ aryl group, C _6- C ₁₀ cycloalkyl group, C _6- C ₁₀ heteroaryl group and C _6- C ₁₀ heterocyclic group are optionally 1, 2, 3, 4 or 5 R ₁ substitutions; each R ₁ is independently selected from H, F, Cl, Br, I, CN, OH, NH ₂ , C _1- C ₄ alkyl, C _{1 -} C ₄ alkoxy group, C _1- C ₄ epoxy group, said C _1- C ₄ alkyl group and C _1- C ₄ alkoxy group are optionally substituted by 1, 2 or 3 R; R ₂ is selected from H, C ₁ -C ₄ alkyl, C ₃ -C ₆ cycloalkyl and C ₃ -C ₆ heterocycloalkyl, the C ₁ -C ₄ alkyl, C ₃ -C C ₆ cycloalkyl and C ₃ -C ₆ heterocycloalkyl are optionally substituted by 1, 2 or 3 R; R ₃ , R ₄ , R ₅ and R ₆ are independently selected from H, F, Cl, Br, I, CN, OH, NH ₂ , C _1- C ₄ alkyl and C _1- C ₄ alkoxy, the C _1- C ₄ alkyl and C _1- C ₄ alkoxy Optionally substituted by 1, 2 or 3 R; n is 1 or 2; R ₇ is selected from H, C ₁ -C ₄ alkyl, C ₃ -C ₆ cycloalkyl and C ₃ -C ₆ heterocycle Alkyl, the C ₁ -C ₄ alkyl, C ₃ -C ₆ cycloalkyl and C ₃ -C ₆ heterocycloalkyl are optionally substituted by 1, 2 or 3 R; R ₈ is selected from H , C ₁ -C ₄ alkyl, C ₃ -C ₆ cycloalkyl, C ₃ -C ₆ heterocycloalkyl, aromatic, heteroaryl and benzyl, the C ₁ -C ₄ alkyl, C ₃ -C ₆ cycloalkyl or heterocycloalkyl, aromatic, heteroaryl and benzyl are optionally substituted by 1, 2 or 3 R; R ₉ and R ₁₀ are independently selected from H, C ₁ - C ₄ alkyl, C ₃ -C ₆ cycloalkyl and C ₃ -C ₆ heterocycloalkyl, the C ₁ -C ₄ alkyl, C ₃ -C ₆ cycloalkyl and C ₃ -C C ₆ heterocycloalkyl is optionally substituted with 1, 2 or 3 R; or R ₉ and R ₁₀ are connected with the carbon atoms to which they are attached to form a C ₃ -C ₆ cycloalkyl, the C ₃ -C ₆ cycloalkyl Cycloalkyl is optionally substituted by 1, 2 or 3 R; R ₁₁ and R ₁₂ are independently selected from H, C ₁ -C ₄ alkyl, C ₃ -C ₆ cycloalkyl and C ₃ -C ₆ heterocycloalkyl, the C ₁ -C ₄ alkyl, C ₃ -C ₆ cycloalkyl and C ₃ -C ₆ heterocycloalkyl are optionally substituted by 1, 2 or 3 R; or R ₁₁ and R ₁₂ are connected to the carbon atoms to which they are attached to form a C ₃ -C ₆ cycloalkane group, the C ₃ -C ₆ cycloalkyl is optionally substituted by 1, 2 or 3 R; each R is independently selected from H, F, Cl, Br, I, CN, OH and NH ₂ .

As a preferred technical solution, the ring A in the present invention is selected from C _6- C ₁₀ aryl groups, C _6- C ₁₀ heteroaryl groups; the C _6- C ₁₀ aryl groups, C ₆ -C 10 aryl groups C ₁₀ heteroaryl is optionally substituted with 1, 2, 3, 4 or 5 R ₁ ; each R ₁ is independently selected from H, F, Cl, Br, I, C _1- C ₄ alkoxy , C _1- C ₄ epoxy group, the C _1- C ₄ alkyl group and C _1- C ₄ alkoxy group are optionally substituted by 1, 2 or 3 R; R ₂ is selected from H, C ₁ -C ₄ alkyl group; the C ₁ -C ₄ alkyl group is optionally substituted by 1, 2 or 3 R; R ₃ , R ₄ , R ₅ and R ₆ are independently selected from H, F, Cl, Br, I, C _1- C ₄ alkyl group and C _1- C ₄ alkoxy group, the C _1- C ₄ alkyl group and C _1- C ₄ alkoxy group are optionally replaced by 1, 2 or 3 R substitutions; n is 1; R ₇ is selected from H, C ₁ -C ₄ alkyl; the C ₁ -C ₄ alkyl is optionally substituted by 1, 2 or 3 R; R ₈ is selected from H, C ₁ -C ₄ alkyl; the C ₁ -C ₄ alkyl is optionally substituted by 1, 2 or 3 R; R ₉ and R ₁₀ are independently selected from H, C ₁ -C C ₄ alkyl; the C ₁ -C ₄ alkyl is optionally substituted with 1, 2 or 3 R; or R ₉ and R ₁₀ are linked to the carbon atoms to which they are attached to form a C ₃ -C ₆ cycloalkane base, the C ₃ -C ₆ cycloalkyl is optionally substituted by 1, 2 or 3 R; R ₁₁ and R ₁₂ are independently selected from H, C ₁ -C ₄ alkyl, the C ₁ -C The _C4 alkyl group is optionally substituted with 1, 2 or 3 Rs; each R is independently selected from H, F, Cl, Br, I, CN, OH and _NH2 .

As a preferred technical scheme, the α-aminoamide compounds, stereoisomers, pharmaceutically acceptable salts, solvates, prodrugs or pharmaceutical compositions containing these compounds as active ingredients, the formula ( Ⅰ) The structural formula is selected from:

Wherein, each R ₁ is independently selected from H, F, Cl, Br, I, C _1- C ₄ alkoxy group, C _1- C ₄ epoxy group, said C _1- C ₄ alkyl group and C _1- C ₄ alkoxy is optionally substituted by 1, 2 or 3 R; R ₂ is selected from H, CH ₃ , CF ₃ ; R ₈ is selected from H, C ₁ -C ₄ alkyl; The C ₁ -C ₄ alkyl is optionally substituted by 1, 2 or 3 R; R ₉ and R ₁₀ are independently selected from H, C ₁ -C ₄ alkyl; the C ₁ -C ₄ alkyl Alkyl is optionally substituted with 1, 2 or 3 Rs; each R is independently selected from H, F, Cl, Br, I, CN, OH and _NH2 .

R₉和R₁₀分别独立地选自H、CH₃、CH₂OH、CH₂CH₃、CH(OH)CH₃、CH(CH₃)₂和CH₂CH(CH₃)₂；或者R₉和R₁₀与它们相连的碳原子连接形成环戊基。As a preferred technical solution, the α-aminoamide compounds, stereoisomers, pharmaceutically acceptable salts, solvates, prodrugs or pharmaceutical compositions containing these compounds as active ingredients described in the present invention , wherein, R ₁ is 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 is selected from H, CH ₃ , CH ₂ CH ₃ and

R ₉ and R ₁₀ are each independently selected from H, CH ₃ , CH ₂ OH, CH ₂ CH ₃ , CH(OH)CH ₃ , CH(CH ₃ ) ₂ and CH ₂ CH(CH ₃ ) ₂ ; or R ₉ and R ₁₀ are attached to the carbon atom to which they are attached to form a cyclopentyl group.

选自：As a preferred technical solution, the α-aminoamide compounds, stereoisomers, pharmaceutically acceptable salts, solvates, prodrugs or pharmaceutical compositions containing these compounds as active ingredients described in the present invention , the structural unit in formula (I)

Selected from:

选自：in formula (I)

Selected from:

As a preferred technical solution, the α-aminoamide compounds, stereoisomers, pharmaceutically acceptable salts, solvates, prodrugs or pharmaceutical compositions containing these compounds as active ingredients described in the present invention, The structure of formula (I) is selected from:

The second aspect of the present invention provides the α-aminoamide compounds, stereoisomers, pharmaceutically acceptable salts, solvates, prodrugs or pharmaceutical compositions containing these compounds as active ingredients Use as analgesic.

As a preferred technical solution, it is used to treat pain; the pain includes treatment or relief of pain during surgery, chronic pain, neuropathic pain, and cancer pain.

The third aspect of the present invention provides the α-aminoamide compounds, stereoisomers, pharmaceutically acceptable salts, solvates, prodrugs or pharmaceutical compositions containing these compounds as active ingredients preparation method,

The following synthetic route was used:

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

As a kind of preferred technical scheme, described preparation method, its preparation method step comprises:

(1) The synthesis of the target compound shown in formula I directly takes 2-fluorobenzyl bromide as a raw material and potassium carbonate as a base and compound b through nucleophilic substitution reaction to obtain compound c, then with sodium hydroxide as an alkali, through ester hydrolysis to obtain compound d. Compound d is reduced by lithium aluminum hydride to obtain compound e, which is then oxidized to the corresponding aldehyde by Dess-Martin oxidant to obtain compound f. Compound f was subjected to reductive amination of sodium cyanoborohydride with the corresponding α-aminoamide to obtain a series of target compounds g.

(2) Compound g is deprotected by palladium-carbon hydrogenation to obtain compound h, which is then reacted to obtain a series of target compounds i.

(3) Compound i is subjected to reductive amination reaction or substitution reaction to obtain a series of target compounds j.

(4) Similar to the above synthesis, a series of target products l were obtained by reductive amination reaction with ketone compound k.

(5) Compound 1 is deprotected by palladium-carbon hydrogenation to obtain compound m, which is then reacted to obtain a series of target compounds n.

(6) Compound n can obtain a series of target compounds I through reductive amination reaction or substitution reaction.

The fourth aspect of the present invention provides the α-aminoamide compounds, stereoisomers, pharmaceutically acceptable salts, solvates, prodrugs or pharmaceutical compositions containing these compounds as active ingredients The dosage form is: solvent tablet, tablet, capsule, injection, solution.

The fifth aspect of the present invention provides the α-aminoamide compounds, stereoisomers, pharmaceutically acceptable salts, solvates, prodrugs or pharmaceutical compositions containing these compounds as active ingredients The method of administration is by injection or oral administration.

Beneficial effects:

The α-aminoamide compound of the present invention has a good inhibitory effect on the sodium ion channel Nav1.7, and can be applied to the treatment or alleviation of pain. The inhibitory activity is better than that of Ralfinamide. In preliminary druggability studies, especially, compounds I-1 and I-30 have good metabolic stability, and in preliminary in vivo analgesia experiments, and compounds I-1 and I-30 are selected in the sciatic nerve branch of mice Sexual injury experiment (SNI model) showed good analgesic effect, meanwhile, compound I-1 also showed good analgesic effect in mouse formalin model.

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

Unless otherwise specified or indicated, the term " _C6 - _C10 aryl" refers to an all-carbon monocyclic or fused polycyclic group having a conjugated pi-electron system, and refers to an aryl group containing 6 to 10 carbon atoms , preferably phenyl and naphthyl.

Unless otherwise specified or indicated, the term "C6 _{- C10} cycloalkyl" refers to saturated or partially unsaturated cyclic alkyl, cyclic alkyl containing 6 to 10 carbon atoms, partially unsaturated means containing One or more unsaturated bonds, but not a fully conjugated pi electron system.

Unless otherwise specified or indicated, the term " _C6-10 heteroaryl" refers to an aryl group containing from 6 to 10 carbon atoms, wherein one or more atoms are selected from nitrogen, oxygen, and sulfur.

Unless otherwise specified or indicated, the term " _C6-10 heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic alkyl group, and wherein one or more atoms are selected from nitrogen, oxygen and sulfur, moiety Unsaturation is defined as above.

Unless otherwise specified or indicated, the term "C ₁ -C ₄ alkyl" refers to a straight or branched hydrocarbon chain containing 1 to 4 carbon atoms, including methyl, ethyl, propyl, isopropyl butyl, butyl, isobutyl, sec-butyl and tert-butyl.

Unless otherwise stated or indicated, the term C3 _{- C6} cycloalkyl refers to saturated or partially unsaturated cyclic alkyl groups, preferably cyclopropyl, cyclobutyl, cyclopentyl and cyclohexane.

Unless otherwise specified or indicated, the term C _1-4 alkoxy refers to OC _1-4 alkyl, including methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy group, sec-butoxy and tert-butoxy.

Unless otherwise specified or indicated, the α-aminoamide compounds, stereoisomers, pharmaceutically acceptable salts, solvates, prodrugs or pharmaceutical compositions containing these compounds as active ingredients of the general formula I, It is characterized in that, the stereoisomer can be left-handed body, dextrorotatory body, racemate of chiral compounds, and mixtures thereof in any proportion;

Unless otherwise specified or indicated, the α-aminoamide compounds, stereoisomers, pharmaceutically acceptable salts, solvates, prodrugs or pharmaceutical compositions containing these compounds as active ingredients of the general formula I, It is characterized in that, the salts are pharmaceutically acceptable acid addition salts formed by the α-aminoamide compounds and inorganic or organic acids.

Unless otherwise specified or indicated, the α-aminoamide compounds, stereoisomers, pharmaceutically acceptable salts, solvates, prodrugs or pharmaceutical compositions containing these compounds as active ingredients of the general formula I, It is characterized in that, described inorganic acid is one or any two or more compositions in hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, sulfuric acid, phosphoric acid; Described organic acid is 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, or a combination of any two or more.

Description of drawings

Figure 1. Effects of Compound 1-1 on nerve injury (SNI)-induced neuropathic pain.

Figure 2. Effects of Compound 1-30 on nerve injury (SNI)-induced neuropathic pain.

Figure 3. Effects of Compound 1-1 on formalin-induced inflammatory pain.

Detailed ways

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

Example 1 Preparation of Compound I-1

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

Take 5.41g (30.02mmol) methylparaben in a 250mL single-necked flask, add 150mL acetonitrile and stir, add 4.98g (36.03mmol) potassium carbonate and 6.24g (33.01mmol) 2-fluorobenzyl bromide successively, slowly heat up Reaction at 90°C for 4h. The reaction solution was cooled to room temperature, filtered with suction, washed twice with acetonitrile, the organic phase was concentrated, 250 mL of water was added, extracted three times with 250 mL of dichloromethane, the organic phases were combined and washed twice with 50 mL of saturated brine. Dry over anhydrous sodium sulfate, filter, concentrate under reduced pressure, and purify by silica gel column chromatography to obtain 6.09 g of a white solid with a yield of 70%.

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

Take the above-mentioned 6.09g (21.12mmol) methyl 3-(4-((2-fluorobenzyl)oxy)phenyl)propanoate in a 250mL single-neck flask, add 50mL methanol and 50mL tetrahydrofuran, stir, add 0.62M hydrogen 51 mL of sodium oxide aqueous solution was slowly heated to 60°C. After the reaction for 3 h, the reaction solution was cooled to room temperature and concentrated under reduced pressure. The concentrate was dissolved in 300 mL of dichloromethane and 200 mL of water to adjust pH=6. The aqueous phase was treated with 250 mL of dichloromethane. It was extracted twice with methyl chloride, 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 obtain 5.39 g of a white solid with a yield of 93%.

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

Take the above 5.39g (19.65mmol) 3-(4-((2-fluorobenzyl)oxy)phenyl)propionic acid in a 250mL there-necked flask, add 100mL anhydrous tetrahydrofuran and stir, under argon protection, under ice bath conditions 39 mL (39 mmol) of 1M lithium aluminum hydride solution in tetrahydrofuran was slowly added dropwise, the temperature was slowly raised to 80° C., and the reaction solution was cooled to room temperature after 2 h of reaction. Under ice bath conditions, slowly add saturated ammonium chloride solution dropwise until no bubbles are generated, filter, and concentrate under reduced pressure, the concentrate is dissolved in 300 mL of dichloromethane, washed with 200 mL of water, and the aqueous phase is extracted twice with 100 mL of dichloromethane , 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 obtain 3.05 g of a colorless oil with a yield of 60%.

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

Take the above 1.54g (5.92mmol) of 3-(4-((2-fluorobenzyl)oxy)phenyl)propanol in a 100mL single-neck flask, add 60mL of dichloromethane and stir, add 3.02g (7.12mmol) of S-Martin reagent, react at room temperature for 4h. 60 mL of water was added to wash, the aqueous phase was extracted twice with 60 mL of dichloromethane, the organic phases were combined, the organic layers were dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain 1.23 g of yellow oil, Yield 80%.

1.5 Synthesis of compound I-1

Take the above 2.99g (11.58mmol) 3-(4-((2-fluorobenzyl)oxy)phenyl)propanal in a 100mL single-necked flask, add 60mL methanol and stir, then add 1.73g (13.89mmol) L- Alaninamide hydrochloride, 0.86g (13.91mmol) sodium cyanoborohydride, 6mL of glacial acetic acid, warmed to 60°C, and reacted for 4h. Cooled to room temperature, concentrated under reduced pressure, the concentrate was dissolved in 300 mL of ethyl acetate and 200 mL of water, adjusted to pH=9 with ammonia water, the aqueous phase was extracted three times with 50 mL of ethyl acetate, the organic phases were combined, and the organic layer was dried over anhydrous sodium sulfate , filtered, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain 2.63 g of a white solid with a yield of 69%. Melting point: 113.6-115.9℃. ¹ H NMR (400MHz, CD ₃ OD) δ 7.50(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

Take the above 103mg (0.40mmol) 3-(4-((2-fluorobenzyl)oxy)phenyl)propanal in a 50mL single-necked flask, add 10mL methanol and stir, then add 44mg (0.40mmol) glycinamide salt acid salt, 50 mg (0.80 mmol) of sodium cyanoborohydride, 0.20 mL of glacial acetic acid, stirred at room temperature overnight, concentrated under reduced pressure, the concentrate was dissolved in 10 mL of ethyl acetate and 10 mL of water, adjusted to pH=9 with ammonia, and 10 mL of water was used for the aqueous phase. Ethyl acetate was extracted three times, 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 obtain 56 mg of white solid with a yield of 44%. Melting point: 61.5-62.3℃. ¹ H NMR (400MHz, CD ₃ OD) δ 7.49(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,found317.1668.

Example 3 Preparation of Compound I-3

The glycinamide hydrochloride was replaced with 2-methylalaninamide, and the preparation method was the same as that of Example 2, and 98 mg of white solid was obtained with a yield of 71%. Melting point: 87.4-88.3°C. ¹ H NMR (400MHz, CD ₃ OD) δ 7.48 (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 glycinamide hydrochloride was replaced with L-2-aminobutanamide hydrochloride, the preparation method was the same as that of Example 2, and 76 mg of white solid was obtained with a yield of 55%. Melting point: 102.1-101.2°C. ¹ H NMR (400MHz, CD ₃ OD) δ 7.48 (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

L-valineamide hydrochloride was used instead of glycinamide hydrochloride, the preparation method was the same as that of Example 2, and 79 mg of white solid was obtained with a yield of 55%. Melting point: 98.1-100.2℃. ¹ H NMR (400MHz, CD ₃ OD) δ 7.48(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

L-leucinamide hydrochloride was used instead of glycinamide hydrochloride, and the preparation method was the same as that of Example 2, and 118 mg of white solid was obtained with a yield of 79%. ¹ H NMR (400MHz, CD ₃ OD) δ 7.49(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

L-serinamide hydrochloride was used instead of glycinamide hydrochloride, the preparation method was the same as that of Example 2, and 110 mg of white solid was obtained with a yield of 79%. ¹ H NMR (400MHz, CD ₃ OD) δ 7.47(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 glycinamide hydrochloride was replaced with threonine amide hydrochloride, and the preparation method was the same as that of Example 2, and 106 mg of white solid was obtained with a yield of 74%. ¹ H NMR (400 MHz, CD ₃ OD) δ 7.46 (t, J=6.7 Hz, 1H), 7.31 (s, 1H), 7.17-7.01 (m, 4H), 6.88 (d, J=7.6 Hz, 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.

Preparation of Example 9 Compound I-9

The glycinamide hydrochloride was replaced with 1-amino-1-cyclopentanecarboxamide, the preparation method was the same as that of Example 2, and 133 mg of white solid was obtained with a yield of 90%. ¹ H NMR (400MHz, CD ₃ OD) δ 7.46(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

Take the above-mentioned 99mg (0.30mmol) (S)-2-((3-(4-((2-fluorobenzyl)oxy)phenyl)propyl)amino)propanamide in a 50mL single-neck flask, add 10mL methanol Stir, successively add 180 mg (6 mmol) formaldehyde, 378 mg (6 mmol) sodium cyanoborohydride, 0.30 mL glacial acetic acid, stir at room temperature overnight, concentrate under reduced pressure, dissolve the concentrate in 10 mL ethyl acetate and 10 mL water, and adjust pH= 9. The aqueous phase was extracted three times with 10 mL of ethyl acetate, the organic phases were combined, the organic layers were dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain 91 mg of 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,found345.1979.

Example 11 Synthesis of Compound I-11

The formaldehyde was replaced with acetaldehyde, and the preparation method was the same as that of Example 10, and 78 mg of light yellow oil was obtained, with a yield of 73%. ¹ H NMR (400 MHz, CD ₃ OD) δ 7.23 (dd, J=13.2, 7.1 Hz, 1H), 7.08 (d, J=5.8 Hz, 1H), 6.85 (d, J=16.6 Hz, 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

Substitute formaldehyde with benzaldehyde, and the preparation method is the same as that in Example 10, to obtain 31 mg of white solid with a yield of 25%. ¹ H NMR (400MHz, CD ₃ OD) δ 7.48 (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

Take the above-mentioned 330mg (1mmol) (S)-2-((3-(4-((2-fluorobenzyl)oxy)phenyl)propyl)amino)propanamide in a 50mL single-neck flask, add 25mL ethanol and stir , 320 mg (0.15 mmol) of 5% palladium on carbon was added, replaced by hydrogen for 5 times, and the temperature was slowly raised to 45° C. After overnight reaction, the reaction solution was cooled to room temperature. It was filtered through celite, washed twice with 10 mL of ethanol, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain 200 mg of a colorless oil with a yield of 90%.

13.2 Preparation of compound I-13

Take 142 mg (0.54 mmol) of triphenylphosphine and 124 mg (0.54 mmol) of di-tert-butyl azodicarboxylate in a 10 mL sealed tube, add 2 mL of tetrahydrofuran and stir until the solution is clear, add 77 mg (0.54 mmol) of 2-chlorobenzyl alcohol and stir To dissolve, add 222mg (0.45mmol) (S)-2-((3-(4-hydroxyphenyl)propyl)amino)propanamide, slowly warm up to 90°C, react overnight, concentrate under reduced pressure, pass through Purification by silica gel column chromatography gave 51 mg of white solid in 33% yield. 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

Substitute 2-bromobenzyl alcohol for 2-chlorobenzyl alcohol, the preparation method is the same as that of Example 13, and 41 mg of white solid is obtained, and the yield is 23%. ¹ H NMR (400 MHz, CD ₃ OD) δ 7.71–7.47 (m, 2H), 7.31 (dd, J=51.9, 5.9 Hz, 2H), 7.14 (d, J=6.1 Hz, 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 2-chlorobenzyl alcohol was replaced by 2-methoxybenzyl alcohol, the preparation method was the same as that of Example 13, and 51 mg of white solid was obtained with a yield of 33%. Melting point: 81.8-83.6°C. ¹ H NMR (400MHz, CD ₃ OD) δ 7.37 (d, J=7.1 Hz, 1H), 7.26 (d, J=7.6 Hz, 1H), 7.08 (d, J=7.8 Hz, 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 forC ₂₀ H ₂₆ N ₂ O ₃ (M+H) ⁺ 343.2016,found 343.2023.

Example 16 Preparation of compound I-16

Substitute 2-methylbenzyl alcohol for 2-chlorobenzyl alcohol, and the preparation method is the same as that in Example 13, to obtain 37 mg of white solid, with 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 compound I-17

Substitute 2-trifluoromethoxybenzyl alcohol for 2-chlorobenzyl alcohol, the preparation method is the same as that of Example 13, and 70 mg of light white solid is obtained, and the yield is 39%. ¹ H NMR (400 MHz, CD ₃ OD) δ 7.58 (d, J=5.9 Hz, 1H), 7.40 (d, J=7.3 Hz, 1H), 7.33 (s, 2H), 7.10 (d, J=7.9 Hz, 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

Substitute 2-trifluoromethylbenzyl alcohol for 2-chlorobenzyl alcohol, and the preparation method is the same as that of Example 13, to obtain 60 mg of white solid with a yield of 35%. ¹ H NMR (400MHz, CD ₃ OD) δ 7.80-7.65 (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 compound I-19

Substitute 2,3-difluorobenzyl alcohol for 2-chlorobenzyl alcohol, and the preparation method is the same as that of Example 13, to obtain 72 mg of white solid with a yield of 46%. ¹ H NMR (400 MHz, CD ₃ OD) δ 7.27 (s, 1H), 7.21 (d, J=7.9 Hz, 1H), 7.13 (t, J=11.6 Hz, 3H), 6.89 (d, J=8.5 Hz, 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

Substitute 2,3-methoxybenzyl alcohol for 2-chlorobenzyl alcohol, and the preparation method is the same as that of Example 13, to obtain 80 mg of pale yellow solid with a yield of 46%. ¹ H NMR (400 MHz, CD ₃ OD) δ 7.02 (s, 2H), 7.00-6.86 (m, 3H), 6.83 (d, J=6.6 Hz, 2H), 4.94 (d, J=11.7 Hz, 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

Substitute 2,4-difluorobenzyl alcohol for 2-chlorobenzyl alcohol, and the preparation method is the same as that of Example 13, to obtain 71 mg of white solid with a yield of 45%. Melting point: 105.9-106.2°C. ¹ H NMR (400MHz, CD ₃ OD) δ 7.50 (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

Substitute 2-chlorobenzyl alcohol with 2-fluoro-4-methoxybenzyl alcohol, the preparation method is the same as that of Example 13, and 48 mg of white solid is obtained, and the yield is 30%. ¹ H NMR (400MHz, CD ₃ OD) δ 7.51(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

Substitute 2,5-difluorobenzyl alcohol for 2-chlorobenzyl alcohol, and the preparation method is the same as that of Example 13, to obtain 36 mg of white solid with a yield of 23%. ¹ H NMR (400MHz, CD ₃ OD) δ 7.24 (s, 1H), 7.12 (d, J=6.0 Hz, 4H), 6.90 (d, J=5.5 Hz, 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

Substitute 2,6-difluorobenzyl alcohol for 2-chlorobenzyl alcohol, and the preparation method is the same as that of Example 13, to obtain 28 mg of white solid with a yield of 18%. Melting point: 98.8-99.4°C. ¹ H NMR (400MHz, CD ₃ OD) δ 7.46-7.32 (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

Substitute 2,4,5-trifluorobenzyl alcohol for 2-chlorobenzyl alcohol, the preparation method is the same as that of Example 13, and 36 mg of white solid is obtained, and the yield is 22%. ¹ H NMR (400 MHz, CD ₃ OD) δ 7.45 (d, J=8.2 Hz, 1H), 7.18 (dd, J=22.4, 8.0 Hz, 3H), 6.88 (dd, J=36.7, 7.4 Hz, 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

Substitute 2,4,6-trifluorobenzyl alcohol for 2-chlorobenzyl alcohol, the preparation method is the same as that of Example 13, and 44 mg of white solid is obtained, the yield is 27%. ¹ H NMR (400 MHz, CD ₃ OD) δ 7.11 (d, J=7.7 Hz, 2H), 6.89 (d, J=4.6 Hz, 4H), 5.02 (s, 2H), 3.24 (q, J=6.5 Hz, 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 compound 1-27

Substitute 2,3,4,5,6-pentafluorobenzyl alcohol for 2-chlorobenzyl alcohol, the preparation method is the same as that of Example 13, and 42 mg of white solid is obtained, the yield is 23%. ¹ H NMR (400MHz, CD ₃ OD) δ 7.15(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

Substitute benzyl alcohol for 2-chlorobenzyl alcohol, and the preparation method is the same as that in Example 13, to obtain 45 mg of white solid with a yield of 32%. ¹ H NMR (400MHz, CD ₃ OD) δ 7.36 (dd, J=29.1, 16.1 Hz, 5H), 7.09 (s, 2H), 6.88 (s, 2H), 5.01 (d, J=7.2 Hz, 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 compound I-29

Substitute 2-chlorobenzyl alcohol with naphthalene-1-ylmethanol, the preparation method is the same as that of Example 13, and 49 mg of white solid is obtained, and the yield is 30%. ¹ HNMR (400MHz, CD ₃ OD) δ 8.02(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

Substitute 2-chlorobenzyl alcohol with 1,3-benzodioxolane-4-alkylmethanol, the preparation method is the same as that of Example 13, and 92 mg of white solid is obtained, with 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

Substitute 2,3-dihydro-1,4-benzodioxane-5-methyl alcohol for 2-chlorobenzyl alcohol, the preparation method is the same as that of Example 13, and 63 mg of light yellow solid is obtained, and the yield is 38%. 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

1-adamantane methanol was used instead of 2-chlorobenzyl alcohol, and the preparation method was the same as that in Example 13, and 18 mg of white solid was obtained with a yield of 11%. ¹ H NMR (400MHz, CD ₃ OD) δ 7.08(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

Substitute (S)-1-(2-fluorophenyl)ethanol for 2-chlorobenzyl alcohol, the preparation method is the same as that of Example 13, and 54 mg of white solid is obtained, the yield is 35%. Melting point: 64.0-65.6℃. ¹ H NMR (400MHz, CD ₃ OD) δ 7.51-7.32 (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

Substitute (R)-1-(2-fluorophenyl)ethanol for 2-chlorobenzyl alcohol, and the preparation method is the same as that of Example 14, to obtain 73 mg of white solid with a yield of 47%. Melting point: 90.6-97.4℃. ¹ H NMR (400MHz, CD ₃ OD) δ 7.49-7.36(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

Substitute methyl paraben with methyl paraben to give 4-(4-((2-fluorobenzyl)oxy)phenyl)butanal, which is used instead of 3-(4-(( 2-Fluorobenzyl)oxy)phenyl)propanal, the preparation method was the same as that of Example 1, and 213 mg of white solid was obtained in a yield of 62%. Melting point: 113.0-113.5℃. ¹ H NMR (400MHz, CD ₃ OD) δ 7.50(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 tested were used in DMEM medium containing 10% fetal bovine serum (Fetalbovine Serum, FBS), 1% Penicillin/Streptomycin (P/S) and 500 μg/ml G418, Incubate at 37°C in a 5% CO ₂ incubator. When the cell confluency reached about 90%, 0.25% trypsin was used for passage, and 50 μl of single cell suspension was inoculated on 8 mm cell slides at a density of 1×10 ⁵ /L, and used for patch clamp experiments 24 hours later.

Current recording

The compounds to be tested were dissolved in DMSO to prepare a 50 mM stock solution, which was aliquoted and stored at -20°C. The compounds to be tested were diluted to the corresponding working concentration (DMSO<0.3%) for patch-clamp electrophysiological activity test. Place the slides seeded with cells under an inverted microscope, perfuse extracellular fluid (flow rate is about 1mL/min, formula (mM): 140NaCl, 1.8CaCl ₂ , 1MgCl ₂ , 10Glucose, 4KCl, 10HEPES, NaOH adjusts the pH to 7.4 ). Using a horizontal drawing machine (P-97, Sutter instrument, USA), the capillary glass tube was drawn into a recording electrode through a multi-step procedure. , 2.5MgCl ₂ , 10HEPES, CsOH to adjust the pH to 7.3) the recording electrode (resistance is 2-4MΩ) is in contact with the cell, and the negative pressure suction is given to form a high resistance seal, and the negative pressure is applied again to break the cell membrane to form a whole cell record mode.

All current recordings were performed at room temperature (20°C-25°C) using an Axon patch 700B patch clamp amplifier, and signal acquisition was performed using pClamp 10.5 software (Axon Instruments, CA) with a filter of 10KHz. The stimulation program used for Nav1.7 current recording was as follows: firstly, the cells were clamped at -40mV to almost completely inactivate Nav1.7 current, then hyperpolarized to -150mV for 20ms to revive some channels, and then depolarized to 0mV , induced the partial Nav1.7 current that had been resurrected, and finally recovered to -40mV, and the whole stimulation program was run once for 1s. Repeat several times to finally inactivate about 50% of the current. After the current is stabilized (the current at this time is I ₀ ), the compound to be tested is perfused to observe its effect on Nav1.7 current, and the current after administration is recorded as I _medicine . In this test, all compounds are prototype compounds.

data analysis

Data analysis and image processing were performed using software such as OriginPro 8.0 (Origin Lab, USA) and Adobe Illustrator. All experimental data are expressed as mean ± standard error (mean ± SEM). The inhibition rate was calculated as (I ₀ -I _drug )/I ₀ . The dose-response relationship curve was fitted by the logistic equation, the formula is as follows: y=A ₂ +(A ₁ -A ₂ )/[1+(x/x ₀ ) ^p ], where y represents the effect, A ₁ and A ₂ respectively represents the maximum effect and the minimum effect, x represents the drug concentration, and p is the Hill coefficient. Unpaired t-test (Student's unpaired t-test) was used for statistical analysis of the obtained data, and the difference between the two groups was considered to be statistically significant when P≤0.05.

Experimental results

The experimental results are shown in Tables 1-2.

Table 1 Nav1.7 inhibition rate of compounds I-1～35 at 10 μM concentration

The Nav1.7 activity test results (IC ₅₀ ) of some compounds in Table 2

Numbering IC₅₀(μM) Numbering IC₅₀(μM) Numbering IC₅₀(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 - -

It can be seen from Tables 1-2 that the example compounds of the patent of the present invention have a better inhibitory effect on the sodium ion channel Nav1.7, and the inhibitory activity is better than that of Ralfinamide.

Test Example 2: Mouse Sciatic Nerve Branch Selective Injury Pain Model Test

Construction of a mouse model of selective injury of sciatic nerve branches (Spared Nerve Injury, SNI)

Before the experiment, the surgical instruments were sterilized and soaked in normal saline. The mice were anesthetized by intraperitoneal injection of 8% chloral hydrate, and the hair near the hind limbs of the left leg of the mice was removed with an automatic hair remover and placed on the operating table. Use cotton dipped in iodophor to apply iodine to the surface of mouse skin to disinfect, and then wipe clean with cotton balls dipped in normal saline. The skin of the left hindlimb was incised with scissors, and the biceps muscle of the hindlimb was gently dissected with a glass minute needle to expose the sciatic nerve and its three distant branches: sural nerve, common peroneal nerve, and tibial nerve. The common peroneal nerve and the tibial nerve were cut with scissors of 1-2 mm respectively to prevent the nerves from growing and combining again after suture. Avoid touching the sural nerve during this process. The wounds were then sutured, and the mice were placed on a heating blanket to maintain their body temperature to help the mice wake up from anesthesia as soon as possible. Mice in the sham-operated group were not cut off after the above three nerves were isolated, and the rest of the operations were the same as those in the surgery group.

Measurement of pain threshold of mechanical withdrawal in mice

The mice were acclimated to the experimental environment for two days before the experiment, and the basal withdrawal pain threshold of the mice was measured on the third day. The specific measurement steps were as follows: The mice were placed in a box with a metal grid in advance, and acclimated for 15-30 minutes. , until the mice keep a familiar and quiet state to the unfamiliar environment (the mice do not walk back and forth in the box, run up and lick themselves). Select the measured VonFrey hair as 0.008, 0.02, 0.04, 0.07, 0.16, 0.40, 0.60, 1.0, 1.4, 2.0, (unit: g), the corresponding log unit is 1.65, 2.36, 2.44, 2.83, 3.22, 3.61, 3.84, 4.08, 4.17, 4.31. When measuring, first select the middle value of 0.40g as the starting point, and vertically touch the test fiber to the sole of the foot at the hairless part of the left foot. When the fiber shows an "S" shape and lasts for more than 5s, observe the reaction of the mouse. If the mouse quickly retracts, flinches, lifts, or licks or bites the left hind paw, it is a positive reaction. After a positive reaction, the fiber measurement with a smaller intensity is replaced, otherwise, the fiber with a larger intensity is replaced, and the adjacent stimulation interval at least 7 seconds. When a positive reaction occurs for the first time, it is necessary to continue to measure four times, and the log unit value corresponding to the VonFrey hair measured at the last time is substituted into the formula for calculation. During the stimulation process, if the mouse walks back and forth or lifts its foot just after touching the stimulation, it is suspiciously positive, and the measurement should be performed after the mouse is quiet. Positive results are marked as X, and negative results are marked as O. The up-and-down method was used to calculate the pain threshold of mechanical withdrawal in mice.

Animal groups and doses

The above-mentioned operated mice were divided into 6 groups: vehicle control group (Vehicle), 10 mg·kg ^-1 Ralfinamide group, 5 mg·kg ^-1 morphine group, and compound I-1 three dose groups, 2 mg·kg ^-1 and 5 mg respectively. ·kg ^-1 , 10 mg·kg ^-1 , or three dose groups of compound I-30, 5 mg·kg ^-1 , 10 mg·kg ^-1 , and 20 mg·kg ^-1 , respectively. In this test, I-1 and Ralfinamide are the prototype drugs, and morphine is the hydrochloride salt.

The mice recovered for four days after the operation, and the pain threshold of the mice was measured for two consecutive days from the fifth day to determine whether the mice were successfully modeled. From the 8th day after the operation, the drug was injected intraperitoneally once a day, and on the 1st, 3rd, 5th, 7th and 10th days of administration, the mechanical withdrawal pain threshold of mice was evaluated according to the above operation.

Analgesic effect of compound I-1 in mouse SNI model

Surgical manipulation of the SNI neuropathic pain model resulted in a significant and sustained increase in mechanical sensitivity in mice. On the 6th day after the operation, the pain threshold of mechanical withdrawal in the hind paw of the mouse was 0.04±0.009g, which was significantly lower than the mechanical withdrawal pain threshold of 1.32±0.11g in the sham operation group, indicating that the SNI model was successfully prepared (Table 1). 3. Figure 1). After injection of 2, 5 and 10 mg·kg ^-1 of I-1 and 10 mg·kg ^-1 of Ralfinamide, the mean mechanical withdrawal pain thresholds increased to 0.25±0.02g, 0.45±0.03g, 0.98±0.04g and 0.52±0.52± 0.04g. The experimental results showed that compound I- ¹ dose-dependently reduced neuropathic pain, and the analgesic effect was better than that of Ralfinamide at a dose of 10 mg/kg. The foot pain threshold was 0.98±0.11g, which was equivalent to the analgesic effect of morphine (5mg·kg ^-1 ) 1.19±0.07g in the positive control group. Each data point is expressed as mean ± SE, ^*** P<0.001vs Sham, ^#P <0.05, ^### P<0.01, ^### P<0.001vsVehicle.

Table 3 SNI experimental results of compound I-1

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

Analgesic effect of compound I-30 in mouse SNI model

Surgical manipulation of the SNI neuropathic pain model resulted in a significant and sustained increase in mechanical sensitivity in mice. On the 6th day after the operation, the pain threshold of mechanical withdrawal in the hind paw of the mice was 0.03±0.002g, which was significantly lower than the mechanical withdrawal pain threshold of 1.03±0.07g in the sham-operated group, indicating that the SNI model was successfully prepared (Table 1). 4, Figure 2). After injection of 5, 10 and 20 mg·kg ^-1 of I-30 and 10 mg·kg ^-1 of Ralfinamide, the mean mechanical withdrawal pain threshold increased to 0.41±0.03g, 0.75±0.03g, 0.97±0.06g and 0.62±0.62± 0.02g. The experimental results showed that compound I-30 dose-dependently reduced neuropathic pain, and the analgesic effect was better than that of ^Ralfinamide at a dose of 10 mg/kg. The foot pain threshold was 1.06±0.07g, which was equivalent to the 1.17±0.05g effect of the sham operation group. Each data point is expressed as mean ± standard error, ^*** P<0.001vs Sham, ^#P <0.05, ^### P<0.01, ^### P<0.001vs Vehicle. In this test, Ralfinamide was the prototype drug, and I-30 and morphine were the hydrochloride salts.

Table 4 SNI experimental results of compound I-30

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

Test Example 3: Mouse Formalin Pain Model Test

Construction of a mouse formalin pain model

Before the start of the experiment, the mice were placed in the observation box for 20 minutes to avoid anxiety and panic caused by the mice in an unfamiliar environment. According to the detection time of the tested drugs, the drugs were injected intraperitoneally in advance, and the center of the left foot of each mouse was loose Insert the needle at the place, insert the needle into the skin and slowly push in 20 μL of the 5% formalin solution prepared in advance. When pulling out the needle, rotate the needle to prevent the formalin solution from leaking from the sole of the foot with the needle. affect the effect of drug delivery. Then the mice were quickly put into the observation box to record and observe, and the control group was injected with normal saline for observation. In the formalin-induced inflammatory pain model experiment in mice, formalin can cause pain responses such as paw licking, foot lift, and leg retraction in mice. The first phase response is recorded during the injection of formalin. Within the next 0-5 min, the recording time of the second phase pain response was 6-45 min, and the total time of licking, biting, raising, and contracting the left hind foot of the mice was calculated respectively within the two-phase pain response time.

Animal groups and doses

The above-mentioned operated mice were divided into 6 groups: vehicle control group (Vehicle), 10 mg·kg ^-1 Ralfinamide group, 10 mg·kg ^-1 morphine group and three dose groups of compound I-1, 5 mg·kg ^-1 , 10 mg·kg ^-1 and 20 mg·kg ^-1 . In this test, I-1 and Ralfinamide are the prototype drugs, and morphine is the hydrochloride salt.

Analgesic effect of compound I-1 in formalin pain model

In the evaluation of the formalin-induced pain model in mice, Compound I-1 can effectively inhibit inflammatory pain, (Table 5, Figure 3). After intraperitoneal injection of 5, 10, and 20 mg·kg ^-1 I-1, the total licking times (Licking time) of the phase I reaction were 27.3±6.02s, 13.58±4.86s, 16.3±5.57s, respectively, which were different from those of the solvent control group. Compared with 26.25±5.71s, no obvious analgesic effect was observed; the total number of paw licking in phase II reaction was 291.00±46.19s, 201.2±30.29s, 164.80±26.04s, compared with 392.75±42.26s in the solvent control group Has a dose-dependent analgesic effect. Intraperitoneal injection of 10 mg·kg ^-1 Ralfinamide, the total number of paw licking in phase I was 18.20±3.96s, and the total number of paw licking in phase II was 294.90±51.94s. No matter in phase I or phase II pain, 10 mg·kg ^-1 None of Ralfinamide exhibited more pronounced analgesic effect than I-1. Each data point is expressed as mean ± SE, ^** P<0.01, ^*** P<0.001 vs Vehicle.

Table 5 Formalin test results of compound I-1

Note: Each data point is expressed as mean ± standard error, ^** P<0.01, ^*** P<0.001 vs Vehicle.

Test Example 4: Metabolic Stability Test

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

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

Buffer: 100 mM potassium phosphate buffer, pH 7.4.

Acetonitrile solution with a drug concentration of 500 μM: Take 5 μL of the stock solution (10 mM) of the test compound or positive control compound and dilute with 95 μL of acetonitrile.

Liver microsome solution at 1.5 μM drug concentration: Take 15 μL of 500 μM drug concentration in 500 μM acetonitrile solution and 18.75 μL of 20 mg/mL liver microsomes into 479.75 μL of 100 mM potassium phosphate buffer.

3 NADPH stock solutions (6 mM, 5 mg/mL): Prepared by dissolving NADPH in buffer solution.

Assay: 30 μL of a drug concentration of 1.5 μM liver microsome solution was dispensed to assay plates designated for different time points (0-, 5-, 15-, 30-, 45-min). Add reaction stop solution to assay plate at 0 min, then add 15 μL of NADPH stock solution. The assay plate was incubated with the mixture of microsomal solution and compound for approximately 5 minutes at 37°C. Add 15 μL of NADPH stock solution, start the reaction and time it. Stop the reaction by adding reaction stop solution at 5-min, 15-min, 30-min and 45-min. The sampling plate was shaken for about 10 minutes, and the samples were centrifuged at 600 rpm for 10 minutes and 6000 rpm for 15 minutes. After centrifugation, 80 μL of the supernatant and 120 μL of HPLC water were added to an 8×96-well plate, mixed and detected by LCMS/MS.

Table 6 Stability test results of I-1 in human liver microsomes

Table 7 Stability test results of I-1 in mouse liver microsomes

Table 8 Stability test results of I-30 in human liver microsomes

Table 9 Stability test results of I-30 in mouse liver microsomes

It can be seen from Tables 6-9 that the half-lives of compounds I-1 and I-30 in human liver microsomes are both greater than 120 minutes, and the half-lives in mouse liver microsomes are 44.57 and 36.94 minutes, respectively. Therefore, compound I -1 and I-30 have better metabolic stability. In this test, I-1 and I-30 are the hydrochloride salts.

The experimental methods that do not specify specific conditions in the following examples are selected according to conventional methods and conditions, or according to the product description. The reagents and raw materials used in the present invention are all commercially available.

The above-mentioned embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited by the above-mentioned embodiments, and any other changes, modifications, substitutions, combinations, The simplification should be equivalent replacement manners, which are all included in the protection 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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