TRPV1 AGONIST AND PREPARATION METHOD THEREFOR AND USE THEREOF
20220371988 · 2022-11-24
Inventors
Cpc classification
C07C233/09
CHEMISTRY; METALLURGY
A61P29/00
HUMAN NECESSITIES
C07D207/06
CHEMISTRY; METALLURGY
A61P21/00
HUMAN NECESSITIES
C07C233/20
CHEMISTRY; METALLURGY
C07D211/14
CHEMISTRY; METALLURGY
A61P15/00
HUMAN NECESSITIES
International classification
C07C233/09
CHEMISTRY; METALLURGY
C07C233/20
CHEMISTRY; METALLURGY
C07D207/06
CHEMISTRY; METALLURGY
C07D211/14
CHEMISTRY; METALLURGY
Abstract
A compound is represented by formula I. A stereoisomer, tautomer, solvate, polymorph of the compound or a pharmaceutically acceptable salt of the compound, a pharmaceutical composition containing the compound, a preparation method of the compound, and the medical use of the compound are provided.
##STR00001##
Claims
1-10. (canceled)
11. A compound of formula I, or a stereoisomer, a tautomer, a solvate, a polymorph or a pharmaceutically acceptable salt thereof: ##STR00031## wherein, R.sub.1 is selected from H, halogen, hydroxyl, amino, and the following groups unsubstituted or optionally substituted with one, two or more R: a (C.sub.1-C.sub.12) aliphatic hydrocarbyl, a (C.sub.1-C.sub.12) aliphatic hydrocarbyl optionally comprising one, two or more heteroatoms, a C.sub.3-12 cycloalkyl, a 3-12 membered heterocycloalkyl, a C.sub.6-20 aryl, a 5-14 membered heteroaryl, S(═O).sub.2NH.sub.2, —S(═O).sub.2NH—(C.sub.1-C.sub.12)aliphatic hydrocarbyl, and —S(═O).sub.2N((C.sub.1-C.sub.12)aliphatic hydrocarbyl).sub.2; R.sub.2 and R.sub.3 are each independently selected from H, halogen, hydroxyl, amino, and the following groups unsubstituted or optionally substituted with one, two or more R: a (C.sub.1-C.sub.12)aliphatic hydrocarbyl, a (C.sub.1-C.sub.12)aliphatic hydrocarbyl optionally comprising one, two or more heteroatoms, a C.sub.3-12 cycloalkyl, a 3-12 membered heterocycloalkyl, a C.sub.6-20 aryl, a 5-14 membered heteroaryl, S(═O).sub.2NH.sub.2, —S(═O).sub.2NH—(C.sub.1-C.sub.12)aliphatic hydrocarbyl, and —S(═O).sub.2N((C.sub.1-C.sub.12)aliphatic hydrocarbyl).sub.2, and at least one of R.sub.2 and R.sub.3 is not H; or R.sub.2 and R.sub.3, together with an N atom connected thereto, form an N-containing 3-12 membered heterocycloalkyl or an N-containing 5-14 membered heteroaryl unsubstituted or optionally substituted with one, two or more R; the R is selected from halogen, CN, OH, NH.sub.2, COOH, ═O, a (C.sub.1-C.sub.12)aliphatic hydrocarbyl, a (C.sub.1-C.sub.12)aliphatic hydrocarbyl optionally comprising one, two or more heteroatoms, a C.sub.3-12 cycloalkyl, a 3-12 membered heterocycloalkyl, a C.sub.6-20 aryl, and a 5-14 membered heteroaryl.
12. The compound of formula I, or the stereoisomer, the tautomer, the solvate, the polymorph or the pharmaceutically acceptable salt thereof according to claim 11, wherein the N-containing 3-12 membered heterocycloalkyl or the N-containing 5-14 membered heteroaryl may be selected from an N-containing 5-6 membered heterocycloalkyl and an N-containing 5-6 membered heteroaryl; R.sub.2 and R.sub.3 are each independently selected from the following groups unsubstituted or optionally substituted with one, two or more R: a (C.sub.1-C.sub.12)aliphatic hydrocarbyl, and a (C.sub.1-C.sub.12)aliphatic hydrocarbyl optionally comprising one, two or more heteroatoms; the halogen is selected from F, Cl, Br and I; the (C.sub.1-C.sub.12)aliphatic hydrocarbyl is selected from a C.sub.1-C.sub.12 alkyl, a C.sub.2-C.sub.12 alkenyl and a C.sub.2-C.sub.12 alkynyl.
13. The compound of formula I, or the stereoisomer, the tautomer, the solvate, the polymorph or the pharmaceutically acceptable salt thereof according to claim 11, wherein the “(C.sub.1-C.sub.12) aliphatic hydrocarbyl optionally comprising one, two or more heteroatoms” may be selected from a (C.sub.1-C.sub.6)aliphatic hydrocarbyloxy, a (C.sub.1-C.sub.6)aliphatic hydrocarbylthio, a (C.sub.1-C.sub.6)aliphatic hydrocarbyloxy(C.sub.1-C.sub.6)aliphatic hydrocarbyl, a (C.sub.1-C.sub.6)aliphatic hydrocarbylthio(C.sub.1-C.sub.6)aliphatic hydrocarbyl, an N—(C.sub.1-C.sub.3)aliphatic hydrocarbylamino(C.sub.1-C.sub.6)aliphatic hydrocarbyl, an N,N-di-(C.sub.1-C.sub.3)aliphatic hydrocarbylamino(C.sub.1-C.sub.6)aliphatic hydrocarbyl, a (C.sub.1-C.sub.6)aliphatic hydrocarbyl-NH—, and an N((C.sub.1-C.sub.6)aliphatic hydrocarbyl).sub.2; preferably, the (C.sub.1-C.sub.12)aliphatic hydrocarbyl substituted with one, two or more R and optionally containing one, two or more heteroatoms may be selected from a —(C.sub.1-C.sub.6)aliphatic hydrocarbyl OC(═O)NH.sub.2, a —(C.sub.1-C.sub.6)aliphatic hydrocarbyl OC(═O)NH(C.sub.1-C.sub.6)aliphatic hydrocarbyl, a —(C.sub.1-C.sub.6)aliphatic hydrocarbyl OC(═O)N((C.sub.1-C.sub.6)aliphatic hydrocarbyl).sub.2, a —(C.sub.1-C.sub.6)aliphatic hydrocarbyl OC(═O)(C.sub.1-C.sub.6)aliphatic hydrocarbyl, and a —(C.sub.1-C.sub.6)aliphatic hydrocarbyl C(═O)O(C.sub.1-C.sub.6)aliphatic hydrocarbyl.
14. The compound of formula I, or the stereoisomer, the tautomer, the solvate, the polymorph or the pharmaceutically acceptable salt thereof according to claim 11, wherein R.sub.1 may be selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, ethenyl, 1-propenyl, 2-propenyl, 1-methylethenyl, 1-butenyl, 1-ethylethenyl, 1-methyl-2-propenyl, 2-butenyl, 3-butenyl, 2-methyl-1-propenyl, 2-methyl-2-propenyl, 1-pentenyl, 1-hexenyl, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 1-methyl-2-propynyl, 3-butynyl, 1-pentynyl, 1-hexynyl, phenyl, thienyl, furyl, pyrrolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, and triazinyl; R.sub.2 and R.sub.3 may be each independently selected from the following groups unsubstituted or optionally substituted with one, two or more R: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, ethenyl, 1-propenyl, 2-propenyl, 1-methylethenyl, 1-butenyl, 1-ethylethenyl, 1-methyl-2-propenyl, 2-butenyl, 3-butenyl, 2-methyl-1-propenyl, 2-methyl-2-propenyl, 1-pentenyl, 1-hexenyl, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 1-methyl-2-propynyl, 3-butynyl, 1-pentynyl, and 1-hexynyl; or R.sub.2 and R.sub.3, together with an N atom connected thereto, form the following groups optionally substituted with one, two or more R: tetrahydropyrrolyl, piperidinyl, and ##STR00032## for example, may be selected from ##STR00033##
15. The compound of formula I, or the stereoisomer, the tautomer, the solvate, the polymorph or the pharmaceutically acceptable salt thereof according to claim 11, wherein the compound of formula I may be specifically selected from the following structures: ##STR00034## ##STR00035## ##STR00036##
16. The compound of formula I, or the stereoisomer, the tautomer, the solvate, the polymorph or the pharmaceutically acceptable salt thereof according to claim 11, wherein the pharmaceutically acceptable salt of the compound of formula I may be selected from inorganic acid salts and organic acid salts, such as hydrochloride, oxalate, formate, and acetate.
17. The preparation method for the compound of formula I, or the stereoisomer, the tautomer, the solvate, the polymorph or the pharmaceutically acceptable salt thereof according to claim 11, comprising the following steps: (1) reacting ##STR00037## with chloroacetic acid to give ##STR00038## R.sub.1, R.sub.2 and R.sub.3 are defined as those of the compound of formula I above; (2) reacting the M-2 with trans-8-methyl-N-vanillyl-6-nonenamide to give the compound of formula I.
18. A pharmaceutical composition comprising the compound of formula I, or the stereoisomer, the tautomer, the solvate, the polymorph or the pharmaceutically acceptable salt thereof according to claim 11.
19. Use of the compound of formula I, or the stereoisomer, the tautomer, the solvate, the polymorph or the pharmaceutically acceptable salt thereof according to claim 11.
20. Use of the pharmaceutical composition according to claim 18 in preparing a TRPV1 agonist.
21. Use of the compound of formula I, or the stereoisomer, the tautomer, the solvate, the polymorph or the pharmaceutically acceptable salt thereof according to claim 11 in preparing a medicament for treating diseases or conditions responsive to trans-8-methyl-N-vanillyl-6-nonenamide.
22. Use of the pharmaceutical composition according to claim 18 in preparing a medicament for treating diseases or conditions responsive to trans-8-methyl-N-vanillyl-6-nonenamide.
23. The use according to claim 20, wherein the disease or condition is selected from post-operative pain, neuropathic pain, post-herpetic neuralgia, diabetic neuropathy, HIV-associated neuropathy, complex regional pain syndrome, cancer, nerve injury, cancer chemotherapy, vulvodynia, trauma, surgery, chronic musculoskeletal pain, lower back pain, osteoarthritis and rheumatoid arthritis-related conditions.
24. A method for treating diseases or conditions responsive to trans-8-methyl-N-vanillyl-6-nonenamide comprising administering to the subject a therapeutically effective amount of the compound of formula I, or the stereoisomer, the tautomer, the solvate, the polymorph or the pharmaceutically acceptable salt thereof according to claim 11.
25. A method for treating diseases or conditions associated with the modulation of TRPV1 activity comprising administering to the subject a therapeutically effective amount of the compound of formula I, or the stereoisomer, the tautomer, the solvate, the polymorph or the pharmaceutically acceptable salt thereof according to claim 11.
26. The method according to claim 24, wherein the disease or condition is selected from post-operative pain, neuropathic pain, post-herpetic neuralgia, diabetic neuropathy, HIV-associated neuropathy, complex regional pain syndrome, cancer, nerve injury, cancer chemotherapy, vulvodynia, trauma, surgery, chronic musculoskeletal pain, lower back pain, osteoarthritis and rheumatoid arthritis-related conditions.
27. The method according to claim 25, wherein the disease or condition is selected from post-operative pain, neuropathic pain, post-herpetic neuralgia, diabetic neuropathy, HIV-associated neuropathy, complex regional pain syndrome, cancer, nerve injury, cancer chemotherapy, vulvodynia, trauma, surgery, chronic musculoskeletal pain, lower back pain, osteoarthritis and rheumatoid arthritis-related conditions.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
[0057]
[0058]
[0059]
DETAILED DESCRIPTION
[0060] The technical solution of the present invention will be further illustrated in detail with reference to the following specific examples. It should be understood that the following examples are merely exemplary illustration and explanation of the present invention, and should not be construed as limiting the scope of protection of the present invention. All techniques implemented based on the aforementioned contents of the present invention are encompassed within the scope of protection of the present invention.
[0061] Unless otherwise stated, the starting materials and reagents used in the following examples are all commercially available or can be prepared by known methods.
[0062] Unless otherwise stated, it should be understood by those skilled in the art that pharmaceutically acceptable salts can be prepared from the compound using conventional acid or base addition procedures, and the salts of the compound can be converted into free products under acid or base addition conditions.
Example 1. Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido)methyl)phenyldiethylglycine hydrochloride (Compound A-1)
[0063] ##STR00013##
Synthesis of Ethylglycine (Compound 3)
[0064] Compound 1 (10 mL, 7 g, 95.7 mmol) was added to a reaction flask and cooled to 5° C. in an ice bath. The reaction system was slowly added with compound 2 (129 g, 13.6 mmol) in portions, warmed to 30° C., and stirred for 6 h until the reaction system turned yellow and finally reddish brown. The reaction solution was filtered in vacuum, and the filter cake was washed with ethyl acetate. The filtrate was concentrated by rotary evaporation to give a reddish-brown oil (1.6 g, 89.3% yield).
Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido) methyl) phenyldiethylglycine (Compound 4)
[0065] To a solution of compound 3 (1.28 g, 9.82 mmol) in dichloromethane were added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.03 g, 5.401 mmol) and 4-dimethylaminopyridine (0.059 g, 0.491 mmol). The reaction system was stirred for 30 min, added with trans-8-methyl-N-vanillyl-6-nonenamide (1.5 g, 4.91 mmol), and stirred at room temperature for 3 h. Then the reaction system was washed with hydrochloric acid (2 N) three times, washed with water three times, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography to give an oil (about 0.89 g, 43.6% yield).
Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido) methyl) phenyldiethylglycine hydrochloride (Compound A-1)
[0066] Compound 4 (0.89 g, 2.127 mmol) was dissolved in a certain amount of ethyl acetate. The reaction system was purged with dry HCl gas generated by the reaction of sodium chloride with concentrated sulfuric acid, with the pH maintained at 3-4, reacted at room temperature for 1 h, and filtered in vacuum to give a white solid (0.92 g, 95.3% yield). .sup.1H NMR (300 MHz, DMSO-d.sub.6): δ 10.62 (s, 1H), 8.47 (t, J=5.9 Hz, 1H), 7.10 (dd, J=16.3, 4.9 Hz, 2H), 6.86 (dd, J=8.2, 1.7 Hz, 1H), 5.42-5.25 (m, 2H), 4.52 (d, J=3.9 Hz, 2H), 4.26 (d, J=5.9 Hz, 2H), 3.76 (s, 3H), 3.27 (dd, J=7.0, 3.9 Hz, 4H), 2.24-2.10 (m, 3H), 1.94 (dd, J=12.9, 6.9 Hz, 2H), 1.57-1.45 (m, 2H), 1.27 (t, J=7.2 Hz, 8H), 0.92 (d, J=6.7 Hz, 6H). HRMS (ESI): m/z, calcd for C.sub.24H.sub.39N.sub.2O.sub.4 [M+H].sup.+, 419.2904; found: 419.2886.
Example 2. Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamidoyl)methyl)phenyl N-ethyl-N-isopropylglycine hydrochloride (Compound A-2)
[0067] ##STR00014##
Synthesis of N-ethyl-N-isopropylglycine (Compound 6)
[0068] Compound 5 (2.3 g, 26.47 mmol) was dissolved in a certain amount of dichloromethane. The reaction system was cooled to 5° C. in an ice bath, slowly added with chloroacetic acid (0.5 g, 5.29 mmol), and reacted at room temperature for 3 h. Then the reaction system was concentrated under reduced pressure to remove dichloromethane, and the residue was added with a certain amount of ethyl acetate, and filtered in vacuum to remove the precipitated salts. The filtrate was concentrated by rotary evaporation to give an oil (compound 6).
Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamidoyl) methyl) phenyl N-ethyl-N-isopropylglycine hydrochloride (Compound A-2)
[0069] Compound 6 (0.48 g, 3.274 mmol) was dissolved in a certain amount of dichloromethane. The reaction system was cooled to 5° C. in an ice bath, slowly added with trans-8-methyl-N-vanillyl-6-nonenamide (0.5 g, 1.64 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.48 g, 2.45 mmol) and 4-dimethylaminopyridine (0.04 g, 0.75 mmol), and stirred at room temperature for 2 h. Then the reaction system was concentrated under reduced to remove the solvent, and the residue was added with a certain amount of ethyl acetate, washed with hydrochloric acid (2 N) three times, washed with water twice, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to give an oil. The oil was dissolved in a certain amount of ethyl acetate. The reaction system was purged with HCl gas generated by the reaction of sodium chloride with concentrated sulfuric acid until the pH was adjusted to 3-4, stirred at room temperature for 1 h, and filtered in vacuum to give a white solid (compound A-2). HRMS (ESI): m/z, calcd for C.sub.25H.sub.41N.sub.2O.sub.4 [M+H].sup.+, 433.3061; found: 433.3057.
Example 3. Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido) methyl) phenyl N-ethyl-N-propylglycine hydrochloride (Compound A-3)
[0070] ##STR00015##
Synthesis of N-ethyl-N-propylglycine (Compound 8)
[0071] Compound 7 (3.9 g, 26.9 mmol) was dissolved in a certain amount of dichloromethane. The reaction system was cooled to 5° C. in an ice bath, slowly added with chloroacetic acid (0.5 g, 5.29 mmol), and reacted at room temperature for 3 h. Then the reaction system was concentrated under reduced pressure to remove dichloromethane, and the residue was added with a certain amount of ethyl acetate, and filtered in vacuum to remove the precipitated salts. The filtrate was concentrated by rotary evaporation to give an oil (compound 8).
Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido) methyl) phenyl N-ethyl-N-propylglycinate hydrochloride (Compound A-3)
[0072] Compound 8 (0.11 g, 0.79 mmol) was dissolved in a certain amount of dichloromethane. The reaction system was cooled to 5° C. in an ice bath, slowly added with trans-8-methyl-N-vanillyl-6-nonenamide (0.2 g, 0.65 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.19 g, 0.98 mmol) and 4-dimethylaminopyridine (0.016 g, 0.13 mmol), and stirred at room temperature for 2 h. Then the reaction system was concentrated under reduced to remove the solvent, and the residue was added with a certain amount of ethyl acetate, washed with hydrochloric acid (2 N) three times, washed with water twice, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to give an oil. The oil was dissolved in a certain amount of ethyl acetate. The reaction system was purged with HCl gas generated by the reaction of sodium chloride with concentrated sulfuric acid until the pH was adjusted to 3-4, stirred at room temperature for 1 h, and filtered in vacuum to give a white solid (compound A-3). HRMS (ESI): m/z calcd for C.sub.25H.sub.41N.sub.2O.sub.4 [M+H].sup.+, 433.3061; found: 433.3050.
Example 4. Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido) methyl) phenyl N-butyl-N-ethylglycine hydrochloride (Compound A-4)
[0073] ##STR00016##
Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido) methyl) phenyl 2-chloroacetate (Compound 10)
[0074] Trans-8-methyl-N-vanillyl-6-nonenamide (1 g, 3.274 mmol) was dissolved in a certain amount of dichloromethane. The reaction system was cooled to 0° C. in an ice bath, added with triethylamine (0.66 g, 6.548 mmol), slowly added dropwise with chloroacetyl chloride (0.55 g, 4.911 mmol), and reacted at room temperature for 1 h. Then the reaction system was concentrated under reduced pressure to remove excess dichloromethane to give an oil (compound 10).
Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido) methyl) phenyl N-butyl-N-ethylglycinate hydrochloride (Compound A-4)
[0075] Compound 10 (0.3 g, 0.79 mmol) was dissolved in a certain amount of acetonitrile. The reaction system was cooled to 5° C. in an ice bath, slowly added with N-ethyl butylamine (0.24 g, 2.36 mmol), and stirred at room temperature for 3 h. Then the reaction system was concentrated under reduced pressure to removed acetonitrile to give an oil. The oil was dissolved in a certain amount of ethyl acetate. The reaction system was purged with HCl gas generated by the reaction of sodium chloride with concentrated sulfuric acid until the pH was adjusted to 3-4, stirred at room temperature for 1 h, and filtered in vacuum to give a white solid (compound A-4). HRMS (ESI): m/z, calcd for C.sub.26H.sub.43N.sub.2O.sub.4 [M+H].sup.+, 447.3217, found: 433.3222.
Example 5. Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido)methyl)phenyldimethylglycine hydrochloride (Compound A-5)
[0076] ##STR00017##
Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido)methyl)phenyldimethylglycine hydrochloride (Compound A-5)
[0077] Compound 10 (0.3 g, 0.79 mmol) was dissolved in a certain amount of acetonitrile. The reaction system was cooled to 5° C. in an ice bath, slowly added with dimethylamine (0.11 g, 2.36 mmol), and stirred at room temperature for 3 h. Then the reaction system was concentrated under reduced pressure to removed acetonitrile to give an oil. The oil was dissolved in a certain amount of ethyl acetate. The reaction system was purged with HCl gas generated by the reaction of sodium chloride with concentrated sulfuric acid until the pH was adjusted to 3-4, stirred at room temperature for 1 h, and filtered in vacuum to give a white solid (compound A-5). HRMS (ESI): m/z, calcd for C.sub.22H.sub.35N.sub.2O.sub.4 [M+H].sup.+, 391.2591; found: 351.2592.
Example 6. Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido)methyl)phenyl 2-(pyrrolidin-1-yl)acetic acid hydrochloride (Compound A-6)
[0078] ##STR00018##
Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido)methyl)phenyl 2-(pyrrolidin-1-yl)acetic acid hydrochloride (Compound A-6)
[0079] Compound 10 (0.3 g, 0.79 mmol) was dissolved in a certain amount of acetonitrile. The reaction system was cooled to 5° C. in an ice bath, slowly added with pyrrolidine (0.17 g, 2.36 mmol), and stirred at room temperature for 3 h. Then the reaction system was concentrated under reduced pressure to removed acetonitrile to give an oil. The oil was dissolved in a certain amount of ethyl acetate. The reaction system was purged with HCl gas generated by the reaction of sodium chloride with concentrated sulfuric acid until the pH was adjusted to 3-4, stirred at room temperature for 1 h, and filtered in vacuum to give a white solid (compound A-6). HRMS (ESI): m/z, calcd for C.sub.24H.sub.37N.sub.2O.sub.4 [M+H].sup.+, 417.2748; found: 417.2740.
Example 7. Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido)methyl)phenyl 2-(methyl-1-azanaphthyl)propionic acid hydrochloride (Compound A-7)
[0080] ##STR00019##
Synthesis of N,N-diethylalanine (Compound 12)
[0081] Compound 11 (1.97 g, 26.9 mmol) was dissolved in a certain amount of dichloromethane. The reaction system was cooled to 5° C. in an ice bath, slowly added with chloroacetic acid (0.5 g, 5.29 mmol), and reacted at room temperature for 3 h. Then the reaction system was concentrated under reduced pressure to remove dichloromethane, and the residue was added with a certain amount of ethyl acetate, and filtered in vacuum to remove the precipitated salts. The filtrate was concentrated by rotary evaporation to give an oil (compound 12).
Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido)methyl)phenyldiethylalanine hydrochloride (Compound A-7)
[0082] Compound 12 (0.11 g, 0.79 mmol) was dissolved in a certain amount of dichloromethane. The reaction system was cooled to 5° C. in an ice bath, slowly added with trans-8-methyl-N-vanillyl-6-nonenamide (0.2 g, 0.65 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.19 g, 0.98 mmol) and 4-dimethylaminopyridine (0.016 g, 0.13 mmol), and stirred at room temperature for 2 h. Then the reaction system was concentrated under reduced to remove the solvent, and the residue was added with a certain amount of ethyl acetate, washed with hydrochloric acid (2 N) three times, washed with water twice, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to give an oil. The oil was dissolved in a certain amount of ethyl acetate. The reaction system was purged with HCl gas generated by the reaction of sodium chloride with concentrated sulfuric acid until the pH was adjusted to 3-4, stirred at room temperature for 1 h, and filtered in vacuum to give a white solid (compound A-7). HRMS (ESI): m/z, calcd for C.sub.25H.sub.41N.sub.2O.sub.4 [M+H].sup.+, 433.3061; found: 433.3057.
Example 8. Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido)methyl)phenyldiethyl-L-valine hydrochloride (Compound A-8)
[0083] ##STR00020##
Synthesis of N,N-diethyl-L-valine (Compound 14)
[0084] L-valine (0.35 g, 3 mmol) was dissolved in a certain amount of mixed solution of water and methanol. The reaction system was cooled to 0° C. in an ice bath, slowly added with sodium cyanoborohydride (0.48 g, 7.3 mmol), slowly added dropwise with an aqueous acetaldehyde solution (1.44 mL, 14.26 mmol) after the solution was clear, and stirred at room temperature for 5 h. Then the reaction system was added with a certain amount of concentrated hydrochloric acid to adjust the pH to about 1.5, and concentrated under reduced pressure to remove the solvent to give an oil (compound 14), which was directly used in the next step.
Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido)methyl)phenyldiethyl-L-valine hydrochloride (Compound A-8)
[0085] Compound 14 (0.14 g, 0.79 mmol) was dissolved in a certain amount of dichloromethane. The reaction system was cooled to 5° C. in an ice bath, slowly added with trans-8-methyl-N-vanillyl-6-nonenamide (0.2 g, 0.65 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.19 g, 0.98 mmol) and 4-dimethylaminopyridine (0.016 g, 0.13 mmol), and stirred at room temperature for 2 h. Then the reaction system was concentrated under reduced to remove the solvent, and the residue was added with a certain amount of ethyl acetate, washed with hydrochloric acid (2 N) three times, washed with water twice, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to give an oil. The oil was dissolved in a certain amount of ethyl acetate. The reaction system was purged with HCl gas generated by the reaction of sodium chloride with concentrated sulfuric acid until the pH was adjusted to 3-4, stirred at room temperature for 1 h, and filtered in vacuum to give a white solid (compound A-8). HRMS (ESI): m/z, calcd for C.sub.27H.sub.45N.sub.2O.sub.4 [M+H].sup.+, 461.3374; found: 461.3371.
Example 9. Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido)methyl)phenyldiethyl-L-leucine hydrochloride (Compound A-9)
[0086] ##STR00021##
Synthesis of N,N-diethyl-L-leucine (Compound 16)
[0087] L-leucine (0.39 g, 3 mmol) was dissolved in a certain amount of mixed solution of water and methanol. The reaction system was cooled to 0° C. in an ice bath, slowly added with sodium cyanoborohydride (0.48 g, 7.3 mmol), slowly added drop/vise with an aqueous acetaldehyde solution (1.44 mL, 14.26 mmol) after the solution was clear, and stirred at room temperature for 5 h. Then the reaction system was added with a certain amount of concentrated hydrochloric acid to adjust the pH to about 1.5, and concentrated under reduced pressure to remove the solvent to give an oil (compound 16), which was directly used in the next step.
Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido)methyl)phenyldiethyl-L-leucine hydrochloride (Compound A-9)
[0088] Compound 16 (0.15 g, 0.79 mmol) was dissolved in a certain amount of dichloromethane. The reaction system was cooled to 5° C. in an ice bath, slowly added with trans-8-methyl-N-vanillyl-6-nonenamide (0.2 g, 0.65 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.19 g, 0.98 mmol) and 4-dimethylaminopyridine (0.016 g, 0.13 mmol), and stirred at room temperature for 2 h. Then the reaction system was concentrated under reduced to remove the solvent, and the residue was added with a certain amount of ethyl acetate, washed with hydrochloric acid (2 N) three times, washed with water twice, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to give an oil. The oil was dissolved in a certain amount of ethyl acetate. The reaction system was purged with HCl gas generated by the reaction of sodium chloride with concentrated sulfuric acid until the pH was adjusted to 3-4, stirred at room temperature for 1 h, and filtered in vacuum to give a white solid (compound A-9). HRMS (ESI): m/z, calcd for C.sub.28H.sub.47N.sub.2O.sub.4 [M+H].sup.+, 475.3530; found: 475.3497.
Example 10. Synthesis of 2-methoxy-4-((8-methylnon-6-enamido)methyl)phenyldiethyl-L-isoleucine hydrochloride (Compound A-10)
[0089] ##STR00022##
Synthesis of N,N-diethyl-L-isoleucine (Compound 18)
[0090] L-leucine (0.39 g, 3 mmol) was dissolved in a certain amount of mixed solution of water and methanol. The reaction system was cooled to 0° C. in an ice bath, slowly added with sodium cyanoborohydride (0.48 g, 7.3 mmol), slowly added dropwise with an aqueous acetaldehyde solution (1.44 mL, 14.26 mmol) after the solution was clear, and stirred at room temperature for 5 h. Then the reaction system was added with a certain amount of concentrated hydrochloric acid to adjust the pH to about 1.5, and concentrated under reduced pressure to remove the solvent to give an oil (compound 18), which was directly used in the next step.
Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido)methyl)phenyldiethyl-L-leucine hydrochloride (Compound A-10)
[0091] Compound 18 (0.15 g, 0.79 mmol) was dissolved in a certain amount of dichloromethane. The reaction system was cooled to 5° C. in an ice bath, slowly added with trans-8-methyl-N-vanillyl-6-nonenamide (0.2 g, 0.65 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.19 g, 0.98 mmol) and 4-dimethylaminopyridine (0.016 g, 0.13 mmol), and stirred at room temperature for 2 h. Then the reaction system was concentrated under reduced to remove the solvent, and the residue was added with a certain amount of ethyl acetate, washed with hydrochloric acid (2 N) three times, washed with water twice, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to give an oil. The oil was dissolved in a certain amount of ethyl acetate. The reaction system was purged with HCl gas generated by the reaction of sodium chloride with concentrated sulfuric acid until the pH was adjusted to 3-4, stirred at room temperature for 1 h, and filtered in vacuum to give a white solid (compound A-10). HRMS (ESI): m/z, calcd for C.sub.28H.sub.47N.sub.2O.sub.4 [M+H].sup.+, 475.3530; found: 475.3502.
Example 11. Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido)methyl)phenyl N-ethyl-N-methylalanine hydrochloride (Compound A-11)
[0092] ##STR00023##
Synthesis of N-ethyl methyl alanine (Compound 20)
[0093] Compound 19 (1.59 g, 26.9 mmol) was dissolved in a certain amount of dichloromethane. The reaction system was cooled to 5° C. in an ice bath, slowly added with chloroacetic acid (0.5 g, 5.29 mmol), and reacted at room temperature for 3 h. Then the reaction system was concentrated under reduced pressure to remove dichloromethane, and the residue was added with a certain amount of ethyl acetate, and filtered in vacuum to remove the precipitated salts. The filtrate was concentrated by rotary evaporation to give an oil (compound 20).
Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido)methyl)phenyl N-ethyl-N-methylalanine hydrochloride (Compound A-11)
[0094] Compound 20 (0.10 g, 0.79 mmol) was dissolved in a certain amount of dichloromethane. The reaction system was cooled to 5° C. in an ice bath, slowly added with trans-8-methyl-N-vanillyl-6-nonenamide (0.2 g, 0.65 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.19 g, 0.98 mmol) and 4-dimethylaminopyridine (0.016 g, 0.13 mmol), and stirred at room temperature for 2 h. Then the reaction system was concentrated under reduced to remove the solvent, and the residue was added with a certain amount of ethyl acetate, washed with hydrochloric acid (2 N) three times, washed with water twice, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to give an oil. The oil was dissolved in a certain amount of ethyl acetate. The reaction system was purged with HCl gas generated by the reaction of sodium chloride with concentrated sulfuric acid until the pH was adjusted to 3-4, stirred at room temperature for 1 h, and filtered in vacuum to give a white solid (compound A-11). HRMS (ESI): m/z, calcd for C.sub.24H.sub.39N.sub.2O.sub.4 [M+H].sup.+, 419.2904: found: 419.2901.
Example 12. Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido)methyl)phenyl N-ethyl-N-methylglycine hydrochloride (Compound A-12)
[0095] ##STR00024##
Synthesis of N-ethyl-N-methylglycine (Compound 22)
[0096] Compound 21 (3.18 g, 53.80 mmol) was dissolved in a certain amount of dichloromethane. The reaction system was cooled to 5° C. in an ice bath, slowly added with chloroacetic acid (1 g, 10.58 mmol), and reacted at room temperature for 3 h. Then the reaction system was concentrated under reduced pressure to remove dichloromethane, and the residue was added with a certain amount of ethyl acetate, and filtered in vacuum to remove the precipitated salts. The filtrate was concentrated by rotary evaporation to give an oil (compound 22).
Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido)methyl)phenyl N-ethyl-N-methylglycinate hydrochloride (Compound A-12)
[0097] Compound 22 (0.09 g, 0.79 mmol) was dissolved in a certain amount of dichloromethane. The reaction system was cooled to 5° C. in an ice bath, slowly added with trans-8-methyl-N-vanillyl-6-nonenamide (0.2 g, 0.65 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.19 g, 0.98 mmol) and 4-dimethylaminopyridine (0.016 g, 0.13 mmol), and stirred at room temperature for 2 h. Then the reaction system was concentrated under reduced to remove the solvent, and the residue was added with a certain amount of ethyl acetate, washed with hydrochloric acid (2 N) three times, washed with water twice, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to give an oil. The oil was dissolved in a certain amount of ethyl acetate. The reaction system was purged with HCl gas generated by the reaction of sodium chloride with concentrated sulfuric acid until the pH was adjusted to 3-4, stirred at room temperature for 1 h, and filtered in vacuum to give a white solid (compound A-12). HRMS (ESI): m/z, calcd for C23H37N2O4 [M+H].sup.+, 405.2748; found: 405.2742.
Example 13. Synthesis of (E)-2-methoxy-4-(8-methylnonanamide)phenylglycine hydrochloride (Compound A-13)
[0098] ##STR00025##
Synthesis of (tert-butoxycarbonyl)glycine (Compound 24)
[0099] Compound 23 (2 g, 26.64 mmol) was dissolved in a certain amount of water. The reaction system was slowly added with di-tert-butyl methyl dicarbonate (8.73 g, 39.96 mmol) and triethylamine (8.09 g, 79.92 mmol), and reacted at room temperature for 1 h. Then the reaction system was adjusted to pH 4-5 with hydrochloric acid (2 M), and extracted with ethyl acetate to give a pale yellow oil (compound 24, 3.74 g).
Synthesis of (E)-2-methoxy-4-((8-methylnon-6-yl)methyl)phenyl(tert-butoxycarbonyl)glycine (Compound 25)
[0100] Compound 24 (0.31 g, 1.639 mmol) was dissolved in a certain amount of dichloromethane. The reaction system was cooled to 5° C. in an ice bath, slowly added with trans-8-methyl-N-vanillyl-6-nonenamide (0.5 g, 1.637 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.48 g, 2.45 mmol) and 4-dimethylaminopyridine (0.04 g, 0.325 mmol), and stirred at room temperature for 2 h. Then the reaction system was concentrated under reduced to remove the solvent, and the residue was added with a certain amount of ethyl acetate, washed with hydrochloric acid (2 N) three times, washed with water twice, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to give an oil (compound 25).
Synthesis of (E)-2-methoxy-4-(8-methylnonanamide)phenylglycine hydrochloride (Compound A-13)
[0101] The oil (compound 25) was dissolved in a certain amount of ethyl acetate. The reaction system was purged with HCl gas generated by the reaction of sodium chloride with concentrated sulfuric acid until the pH was adjusted to 3-4, stirred at room temperature for 0.5 h, and filtered in vacuum to give a white solid (compound A-13), 1H NMR (300 MHz, MeOD) δ 7.07 (d, J=8.0 Hz, 2H), 6.90 (dd, J=8.2, 1.6 Hz, 1H), 5.43-5.28 (m, 2H), 4.37 (s, 2H), 4.15 (s, 2H), 3.83 (d, J=4.5 Hz, 3H), 2.23 (dt, J=12.6, 6.3 Hz, 3H), 2.00 (dd, J=12.5, 7.2 Hz, 2H), 1.70-1.58 (m, 2H), 1.43-1.31 (m, 2H), 0.96 (d, J=6.7 Hz, 6H). HRMS (ESI): m/z, calcd for C.sub.20H.sub.30N.sub.2O.sub.4 [M+H].sup.+, 363.2278; found: 363.2273.
Example 14. Synthesis of (E)-2-methoxy-4-(8-methylnonanamide)phenyl 2-(piperidin-1-yl)acetic acid hydrochloride (Compound A-14)
[0102] ##STR00026##
Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido)methyl)phenyl 2-chloroacetate (Compound 26)
[0103] Trans-8-methyl-N-vanillyl-6-nonenamide (1 g, 3.274 mmol) was dissolved in a certain amount of dichloromethane. The reaction system was cooled to 0° C. in an ice bath, added with triethylamine (0.66 g, 6.548 mmol), slowly added dropwise with chloroacetyl chloride (0.55 g, 4.911 mmol), and reacted at room temperature for 1 h. Then the reaction system was concentrated under reduced pressure to remove excess dichloromethane to give an oil (compound 26).
Synthesis of (E)-2-methoxy-4-(8-methylnonanamide)phenyl 2-(piperidin-1-yl)acetic acid hydrochloride (Compound A-14)
[0104] Compound 26 (0.3 g, 0.79 mmol) was dissolved in a certain amount of DCM. The reaction system was cooled to 5° C. in an ice bath, slowly added with piperidine (0.17 g, 2.36 mmol), and stirred at room temperature for 3 h. Then the reaction system was concentrated under reduced pressure to removed acetonitrile to give an oil. The oil was dissolved in a certain amount of ethyl acetate. The reaction system was purged with HCl gas generated by the reaction of sodium chloride with concentrated sulfuric acid until the pH was adjusted to 3-4, stirred at room temperature for 1 h, and filtered in vacuum to give a white solid (compound A-14). 1H NMR (300 MHz, DMSO) δ 10.64 (s, 1H), 8.47 (t, J=5.9 Hz, 1H), 7.18-7.02 (m, 2H), 6.86 (d, J=8.4 Hz, 1H), 5.42-5.25 (m, 2H), 4.53 (s, 2H), 4.27 (d, J=5.9 Hz, 2H), 3.77 (s, 3H), 3.55 (s, 2H), 3.08 (s, 2H), 2.19 (dt, J=19.9, 7.1 Hz, 3H), 1.94 (dd, J=12.9, 6.7 Hz, 2H), 1.88-1.66 (m, 5H), 1.57-1.46 (m, 2H), 1.30 (dt, J=19.2, 9.8 Hz, 3H), 0.93 (d, J=6.7 Hz, 6H). HRMS (ESI): m/z, calcd for C25H38N2O4 [M+H].sup.+, 431.2904; found: 431.2464.
Example 15. Synthesis of (E)-2-methoxy-4-(8-methylnonanamide)phenyl 2-(dimethylamino)-2-phenylacetic acid hydrochloride (Compound A-15)
[0105] ##STR00027##
Synthesis of 2-(dimethylamino)-2-phenylacetic acid hydrochloride (Compound 28)
[0106] Compound 27 (2 g, 9.3 mmol) was dissolved in a certain amount of acetone solution. The reaction system was cooled to 5° C. in an ice bath, slowly added with dimethylamine (0.84 g, 18.6 mmol) and stirred at room temperature for 1 h. Then the reaction system was added with hydrochloric acid to adjust the pH to 3-4, and concentrated by rotary evaporation to remove the solvent. The residue was re-crystallized from ethanol to give a white solid (compound 28).
Synthesis of (E)-2-methoxy-4-(8-methylnonanamide)phenyl 2-(dimethylamino)-2-phenylacetic acid hydrochloride (Compound A-15)
[0107] Compound 28 (0.17 g, 0.79 mmol) was dissolved in a certain amount of dichloromethane. The reaction system was cooled to 5° C. in an ice bath, slowly added with trans-8-methyl-N-vanillyl-6-nonenamide (0.2 g, 0.65 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.19 g, 0.98 mmol) and 4-dimethylaminopyridine (0.016 g, 0.13 mmol), and stirred at room temperature for 2 h. Then the reaction system was concentrated under reduced to remove the solvent, and the residue was added with a certain amount of ethyl acetate, washed with hydrochloric acid (2 N) three times, washed with water twice, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to give an oil. The oil was dissolved in a certain amount of ethyl acetate. The reaction system was purged with HCl gas generated by the reaction of sodium chloride with concentrated sulfuric acid until the pH was adjusted to 3-4, stirred at room temperature for 1 h, and filtered in vacuum to give a white solid (compound A-15). 1H NMR (300 MHz, DMSO) δ 8.43 (t, J=5.9 Hz, 1H), 7.69 (s, 2H), 7.58 (d, J= 2.9 Hz, 2H), 7.52 (d, J=3.9 Hz, 1H), 7.00 (d, J=8.2 Hz, 2H), 6.82 (d, J=7.8 Hz, 1H), 5.85 (s, 1H), 5.41-5.24 (m, 2H), 4.24 (d, J=5.8 Hz, 2H), 3.64 (s, 3H), 3.00 (s, 2H), 2.89 (d, J=6.5 Hz, 2H), 2.59 (s, 2H), 2.16 (dt, J=14.6, 7.0 Hz, 3H), 1.93 (dd, J=12.6, 7.1 Hz, 2H), 1.49 (dd, J=15.0, 7.2 Hz, 2H), 1.27 (dd, J=14.7, 7.2 Hz, 2H), 0.91 (d, J=6.7 Hz, 6H). HRMS (ESI): m/z, calcd for C28H38N2O4 [M+H].sup.+, 467.2904; found: 467.2895.
Example 16. Synthesis of (E)-2-methoxy-4-(8-methylnonanamide)phenyl 2-([[1,4′-bipiperidin]-1′-yl) propanoate hydrochloride (Compound A-16)
[0108] ##STR00028##
Synthesis of 2-([1,4′-bipiperidin]-1′-yl)propionic acid hydrochloride (Compound 30)
[0109] Compound 29 (2 g, 11.89 mmol) was dissolved in a certain amount of dichloromethane. The reaction system was cooled to 5° C. in an ice bath, slowly added with 2-chloropropionic acid (0.65 g, 5.95 mmol), and reacted at room temperature for 3 h. Then the reaction system was concentrated under reduced pressure to remove dichloromethane, and the residue was added with a certain amount of ethyl acetate, and filtered in vacuum to remove the precipitated salts. The filtrate was added with hydrochloric acid to adjust the pH to 3-4, concentrated by rotary evaporation to remove the solvent, and re-crystallized from ethanol to give a white solid (compound 30).
Synthesis of (E)-2-methoxy-4-(8-methylnonanamide)phenyl 2-([[1,4′-bispiperidin]-1′-yl)propionate hydrochloride (Compound A-16)
[0110] Compound 30 (0.22 g, 0.79 mmol) was dissolved in a certain amount of dichloromethane. The reaction system was cooled to 5° C. in an ice bath, slowly added with trans-8-methyl-N-vanillyl-6-nonenamide (0.2 g, 0.65 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.19 g, 0.98 mmol) and 4-dimethylaminopyridine (0.016 g, 0.13 mmol), and stirred at room temperature for 2 h. Then the reaction system was concentrated under reduced to remove the solvent, and the residue was added with a certain amount of ethyl acetate, washed with hydrochloric acid (2 N) three times, washed with water twice, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to give an oil. The oil was dissolved in a certain amount of ethyl acetate. The reaction system was purged with HCl gas generated by the reaction of sodium chloride with concentrated sulfuric acid until the pH was adjusted to 3-4, stirred at room temperature for 1 h, and filtered in vacuum to give a white solid (compound A-16). 1H NMR (300 MHz, MeOD) δ 7.16-7.05 (m, 2H), 6.91 (d, J=8.1 Hz, 1H), 5.42-5.29 (m, 2H), 4.63 (d, J=6.9 Hz, 2H), 4.36 (s, 2H), 3.83 (s, 3H), 3.57 (dd, J=27.6, 15.3 HZ, 5H), 3.16-3.00 (m, 3H), 2.52 (d, J=12.2 Hz, 2H), 2.22 (dt, J=12,2, 6.4 HZ, 5H), 1.96 (t, J=11.3 Hz, 5H), 1.83 (t, J=9.0 Hz, 4H), 1.62 (dt, J=15.3, 7.6 Hz, 3H), 1.36 (dt, J=14.9, 7.6 Hz, 3H), 0.94 (d, J=6.7 Hz, 6H). HRMS (ESI): m/z, calcd for C31H49N3O4 [M+H].sup.+, 528.3796; found: 528.3796,
Example 17. Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido)methyl)phenyldiethylglycine oxalate (Compound A-17)
[0111] ##STR00029##
Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido)methyl)phenyldiethylglycine oxalate (Compound A-17)
[0112] Compound 10 (4.65 g, 12.31 mmol) and diethylamine (3.56 g, 48.67 mmol) were dissolved in a certain amount of ethyl acetate. The reaction system was heated to 60° C. and reacted for 3 h. Then the reaction system was added with a certain amount of water followed by liquid separation. The organic phase was added with dilute hydrochloric acid to adjust the pH to 1-2, and the aqueous phase was added with a certain amount of ethyl acetate and adjusted to pH 8-9 followed by liquid separation. The organic phase was concentrated to give an oil. The oil was dissolved in a certain amount of ethyl acetate, added with oxalic acid dihydrate (1.16 g, 9.10 mmol), and reacted at 40° C. for 15 min. Then the reaction system was cooled to room temperature, stirred for 1 h, and filtered in vacuum to give an off-white solid (3.62 g, 62.5%). .sup.1H NMR (300 MHz, DMSO-d.sub.6): 8.35 (t, J=5.9 Hz, 1H), 7.06 (dd, J=16.3, 4.9 Hz, 2H), 6.84 (dd, J=8.2, 1.7 Hz, 1H), 5.36-5.32 (m, 2H), 4.26 (d, J=3.9 Hz, 2H), 3.98 (s, 2H), 3.75 (s, 3H), 2.95 (dd, J=7.0, 3.9 Hz, 4H), 2.24-2.10 (m, 3H), 1.94 (dd, J=12.9, 6.9 Hz, 2H), 1.57-1.45 (m, 2H), 1.34-1.28 (m, 2H), 1.14 (t, J=7.2 Hz, 6H), 0.92 (d, J=6.7 Hz), 6H), MS (ESI): m/z, [M+H].sup.+, 419.45.
Example 18. Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido)methyl)phenyldimethylglycine oxalate (Compound A-18)
[0113] ##STR00030##
Synthesis of (E)-2-methoxy-4-((8-methylnon-6-enamido)methyl)phenyldimethylglycine oxalate (Compound A-18)
[0114] Compound 10 (21 g, 54.99 mmol), dimethylamine hydrochloride (27.9 g, 342 mmol) and triethylamine (33.3 g, 333 mmol) were dissolved in a certain amount of ethyl acetate. The reaction system was heated to 60° C. and reacted for 4 h. Then the reaction system was added with a certain amount of water followed by liquid separation. The organic phase was added with dilute hydrochloric acid to adjust the pH to 1-2, and the aqueous phase was added with a certain amount of ethyl acetate and adjusted to pH 8-9 followed by liquid separation. The organic phase was concentrated to give an oil. The oil was dissolved in a certain amount of ethyl acetate, added with oxalic acid dihydrate (1.60 g, 12.70 mmol), and reacted at 40° C. for 15 min. Then the reaction system was cooled to room temperature, stirred for 1 h, and filtered in vacuum to give an off-white solid (6.2 g, 23.5%). .sup.1H NMR (300 MHz, DMSO-d.sub.6): 8.35 (t, J=5.9 Hz, 1H), 7.07 (dd, J=16.3, 4.9 Hz, 2H), 6.85 (dd, J=8.2, 1.7 Hz, 1H), 5.40-5.28 (m, 2H), 4.26 (d, J=3.9 Hz, 2H), 3.93 (s, 2H), 3.75 (s, 3H), 2.59 (s, 6H), 2.22-2.12 (m, 3H), 1.97 (dd, J=12.9, 6.9 Hz, 2H), 1.57-1.45 (m, 2H), 1.34-1.27 (m, 2H), 0.92 (d, J=6.7 Hz, 6H). MS (ESI): m/z, [M+H].sup.+, 391.25.
Example 19. Determination of Solubility of Compounds Disclosed Herein
[0115] To determine the solubility of the compounds disclosed herein in water, 10 mg of each of the compounds according to the examples of the present invention was added to a colorimetric tube, followed by the addition of 0.1 mL of water. The reaction system was sonicated for 5 min, and if the compound was not dissolved, 0.1 mL of water was supplemented until no undissolved material was observed. After the solubility measurement, it was found that the solubility of the compounds according to the examples of the present invention in water was much greater than that of the parent drug trans-8-methyl-N-vanillyl-6-nonenamide (solubility: 0.00841 mg/mL). The results are shown in Table 19-1.
TABLE-US-00001 TABLE 19-1 Solubility of compounds disclosed herein Examples Solubility Compound A-1 244 mg/mL Compound A-14 164 mg/mL Compound A-15 105 mg/mL Compound A-16 101 mg/mL Compound A-17 175 mg/mL Compound A-18 70 mg/mL Trans-8-methyl-N-vanillyl-6-nonenamide 0.00841 mg/mL
Example 20. In Vitro pH Stability of Compounds Disclosed Herein
[0116] The compounds according to the examples of the present invention were incubated in the different buffers specified. The incubation was performed at room temperature, and samples were collected at specific time points. The formation of trans-8-methyl-N-vanillyl-6-nonenamide and the consumption of the starting compound were analyzed by HPLC, and the release half-life was calculated. The experiment demonstrated that the compounds disclosed herein have a certain stability at pH levels suitable for the preparation of pharmaceutical preparations, and can be stored, processed under appropriate conditions and prepared as medicaments. The results are shown in Table 20-1.
TABLE-US-00002 TABLE 20-1 Release half-life of compounds disclosed herein at different pH Release half-life t.sub.1/2 Medium A-1 A-16 A-17 A-18 0.1 mol HCl 68.6 h About 82 h About 104 h About 59 h pH4.5 About 24.7 h About 73 h About 48 h About 31 h pH7.4 About 45.7 min About 157 h About l h About 3 h
Example 21. In Vitro Simulation of In Vivo Drug Release
[0117] 0.1 mg of each of the compounds according to the examples of the present invention was incubated with the plasma sample at physiological temperature. Samples were collected at different time points. 100 μL of drug-containing plasma was precisely measured out, added with 1 mL of methanol to precipitate protein, and centrifuged. The supernatant was collected, and injected for analysis. The percentage of trans-8-methyl-N-vanillyl-6-nonenamide released by the compound disclosed herein was measured, and the release half-life t.sub.1/2 was calculated. The experiments demonstrated that about 50% of the compound disclosed herein was converted to the parent drug in a relatively short time. The test results are shown in Table 21-1.
TABLE-US-00003 TABLE 21-1 Release half-life of compounds disclosed herein Compound Release half-life t.sub.1/2 Compound A-1 <1 min Compound A-16 2.5 h Compound A-17 <1 min Compound A-18 <1 min
Example 22. Irritation Test (1)
[0118] The equal amount of the compounds according to the examples of the present invention were compared with capsaicine in odor. The results showed that the compounds according to the examples of the present invention were all white and tasteless, and had significantly less irritation.
Example 23. Irritation Test (2)
[0119] The experiment was performed to demonstrate that the irritation of the compounds disclosed herein to the nasal mucosa and perinasal skin of the rats is less than that of trans-8-methyl-N-vanillyl-6-nonenamide.
23.1 Test Drugs
[0120] Sample: compound A-17 solution (16.67 mg/mL, equimolar with trans-8-methyl-N-vanillyl-6-nonenamide). Positive control: trans-8-methyl-N-vanillyl-6-nonenamide solution (10 mg/mL). Negative control solution: normal saline and blank solvent.
23.2 Laboratory Animals
[0121] 12 SPF male SD rats weighed 180-220 g, room temperature of 22±2° C., humidity of about 50%, ammonia concentration below 20 ppm, and regular ventilation. All rats were fed with complete nutritional feed and housed in IVC cage boxes, 4 rats in each boxes, with free access to food and water.
23.3 Test Methods
23.3.1 Grouping
[0122] Healthy SD rats were randomly divided into 4 groups, i.e., a treatment group (i.e., compound A-17 group), a positive control group, negative control groups (i.e., normal saline group and blank solvent group); 4 rats in each of the treatment group and the positive control group, and 2 rats in each of the negative control groups.
23.12 Administration
[0123] During the test, 10 μL of the sample solution or control solution was quantitatively pipetted and carefully dropped into the left nostril of the rats; the rats were observed for 30 min immediately after the administration and behavioral changes were recorded.
23.3.3 Observation Index
[0124] For general states: the eye closure and the hypokinesia symptoms, and the beginning and ending time and the severity of the symptoms of the rats within 30 min after the administration;
[0125] For local irritative symptoms: the symptoms such as the congestion and redness and swelling of the nasal cavity and periphery and nasal secretion, and the times of sneezing and nasal scratching symptoms of the rats within 10 min after the administration and within 10-30 min after the administration.
TABLE-US-00004 TABLE 23-1 Scoring standard of congestion and redness and swelling of nasal cavity and periphery of rats Irritative symptoms Score Normal 0 Mild hyperemia 1 Obvious congestion and redness 2 and swelling Severe redness and swelling with 3 secretion
[0126] The calculation method for the times of nasal scratching: according to continuous nasal scratching of the rats after irritation to nasal mucosa; a continuous nasal scratching of no more than 10 s is counted as once; a continuous scratching of more than 10 s is counted as twice or more times, (for example, a continuous scratching of 30 s is counted as 3 times).
23.4 Results
[0127] Data were processed using the software SPSS19 for non-parametric tests, P<0.05 showed a statistically significant difference.
23.4.1 General State Observation
[0128] Negative control group: no eye closure or hypokinesia symptom was observed in the rats after the nasal administration of normal saline or blank solvent.
[0129] Positive control group: after the rats were administered trans-8-methyl-N-vanillyl-6-nonenamide, eye closure symptom that lasts for about 1.4 min was observed in 1 rat about 2 min later after the administration, and no eye closure symptom was observed in the other rats. Hypokinesia symptom that lasts for 3-12 min was observed in 3 of the 4 rats about 2 min later after the administration, with prostration and deep-breathing symptoms observed in 1 of the 3 rats.
[0130] Treatment group: no eye closure or hypokinesia was observed in the rats after the administration of compound A-17 during the observation.
23.4.2 Local Irritation Symptom Observation
[0131] Negative control group: only transient and accidental sneezing or nasal scratching symptoms (less than 3 times) were observed in the rats after the nasal administration of normal saline or blank solvent, and the other symptoms such as the congestion and redness and swelling of nasal cavity and periphery were not observed.
[0132] Comparison of the local irritation of the compounds disclosed herein and trans-8-methyl-N-vanillyl-6-nonenamide to the nasal cavity of the rats: the severity of the congestion and redness and swelling of the nasal cavity and periphery of the rats induced by the treatment group (compound A-17) was obviously lower than that induced by the positive control group (trans-8-methyl-N-vanillyl-6-nonenamide administration group) (P<0.001). The times of sneezing of the rats within 10 min and 10-30 min after the administration of the treatment group was less than that of the positive group (trans-8-methyl-N-vanillyl-6-nonenamide) (P<0.001), the times of nasal scratching of the rats within 10 min after the administration of the treatment group was more than that of the positive control group (trans-8-methyl-N-vanillyl-6-nonenamide) (P<0.001), but the times of nasal scratching of the rats within 10-30 min after the administration of the treatment group was less than that of the positive group (trans-8-methyl-N-vanillyl-6-nonenamide) (P<0.001). The results are shown in Table 23-2.
TABLE-US-00005 TABLE 23-2 Local irritation symptom statistics (n = 4,
Example 24. Study of Analgesic Effect on Rats as Models of Pain After Incision
24.1 Test Materials and Laboratory Animals
[0133] Gas anesthesia machine for small animals, Shanghai Yuyan Instruments Co., Ltd., model number: ABS
[0134] Plantar tester, IITC Life Science, USA, model number: Series 8 Model 390
[0135] Von-Frey filaments, North Coast Medical Inc., model number: NC12775-99
[0136] Sterile syringe (1 mL), Shanghai Kindly Enterprise Development Group Co., Ltd.
[0137] Laboratory animals
[0138] Species: SD rat Sex: male
[0139] Week age: 5-6 weeks body weight: 180-200 g
[0140] Quantity: 36
24.2 Preparation of Rats as Models of Pain After Incision
[0141] After 3-5 days of adaptive feeding, all rats were anesthetized with 3.5%-4% isoflurane/oxygen, and then were maintained with 1.5%-2% isoflurane. The drug (150 μL) was infiltrated by intraplantar injection shortly before the incision (1-2 min). Under sterile conditions, each rat was made a 1 cm longitudinal incision forward with a No. 11 scalpel blade at 0.5 cm posterior to the metatarsals, the fascia and muscle were separated, the flexor was raised, and a longitudinal incision was made via blunt dissection, leaving the muscle source and insertion intact. After the hemostasis by light pressure, the drug was injected into the flexor (30 μL) before the wound was closed, and the skin was closed with two interrupted stitches at the incision site with a No. 5 nylon thread.
24.3 Grouping and Administration
[0142] The modeled rats were randomly divided into 6 groups of 6 animals each: the high-dose (HD), medium-dose (MD) and low-dose (LD) groups of compound A-17, the LD and HD groups of trans-8-methyl-N-vanillyl-6-nonenamide, and a blank preparation group. Mode of administration: sample or control (150 μL) was infiltrated by intraplantar injection 1 min before the incision, and injected into the flexor (30 μL) before the wound was stitched. The specific grouping and administration design are shown in Table 24-1.
TABLE-US-00006 TABLE 24-1 Administration design Admin- Admin- Admin- istration istration istration concen- volume dose Number Mode of tration (mL/ (mg/ of admin- Grouping (mg/mL) animal) animal) Animals* istration A-17 (LD) 0.83 0.18 0.15 6 150 μL of the A-17 (MD) 1.665 0.18 0.3 6 compound A-17 (HD) 3.33 0.18 0.6 6 was injected Trans-8- 0.5 0.18 0.09 6 subcutaneously methyl-N- in foot 1-2 min vanillyl-6- before the nonenamide incision, and (LD) 30 μL of the Trans-8- 2 0.18 0.36 6 compound was methyl-N- injected in vanillyl-6- the flexor nonenamide before the (HD) incision Blank / 0.18 / 6 suture preparation *each animal received two tests after the administration, i.e., mechanica allodynia test and thermal hyperalgesia test.
24.4 Test Results
Mechanical Allodynia Test in Rats—Von Frey Method
[0143] The mechanical paw withdrawal threshold (PWT) of rats was tested as the pain threshold by irritating the middle plantar of hind limbs with Von Frey monofilaments at different time points after the administration. The Von Frey filaments were in 7 strengths, and the irritation intensity was increased approximately logarithmically (equivalent to 2 g, 4 g, 6 g, 8 g, 10 g, 15 g, 26 g). The rats was placed in a customized plexiglass box with a metal grid laid on the bottom, and acclimatized for at least 15 min before the test. The plantar surface of the hind paw of rats was contacted with the filaments, and certain pressure was applied to slightly bend the filaments. The filament strength was gradually increased with each filament tested 3 times on each plantar surface, and the pain threshold was defined as the lowest strength at which at least two paw withdrawals occurred over 3 consecutive tests. The test was performed at 1 h, 4 h, 24 h, 48 h, 72 h, 96 h and 168 h after the administration.
Thermal Hyperalgesia Test in Rats—Hargreaves Method
[0144] The thermal withdrawal latency (TWL) of rats was measured using a IITC 390 thermal stimulation pain tester (Life science, USA). A 15 cm×15 cm×15 cm transparent plexiglass box was placed on the glass plate of the instrument, and the rats in the box were acclimatized for 10 min to rest before the test. When using the thermal radiation stimulator, the guide light source from the test head below the paw was first moved to the bottom of the paw of the rats, with the position determined by the reflector, and the heating light beam was then started to irradiate the bottom of the paw of the rats. The time from the start of irradiation to the occurrence of paw withdrawal in the rats was TWL. The light source irritation intensity was constant (90 W), and the automatic cut-off time was set to be 15 s to prevent the tissue from burning. The left and right feet of the rats were each tested for 5 times, with each interval of no less than 20 s. The maximum value and the minimum value of the 5 tests were discarded, only the average of the 3 middle values was taken as the test result of a certain specific time point, and then the average was taken. The average normal latency was about 8-10 s, and the rat whose measurements was less than 6 s or more than 12 s was considered to have a disqualified basal threshold, and cannot be given administration and subsequent tests. The test was performed at 1 h, 4 h, 24 h, 48 h, 72 h, 96 h and 168 h after the administration.
[0145] The test results were tested for Two-Way ANOVA using the statistical software Graphpad Prism 6, and P<0.05 was considered a significant difference.
Results
(1) Effect of A-17 on Thermal Hyperalgesia in Rats as Models of Pain After Incision
[0146] The effect of A-17-HD, -MD and -LD administered by subcutaneous injection at the incision site on the thermal hyperalgesia in the rats as models of pain after incision was evaluated, with the blank preparation (Vehicle) as a negative control and trans-8-methyl-N-vanillyl-6-nonenamide-HD and -LD as positive controls. The results showed that A-17 relieved the thermal hyperalgesia in rats as models of pain after incision in a dose-dependent manner and a time-dependent manner, as shown in
[0147] The area under the time-TWL value curve (AUC) of each group within 1-168 h after the surgery in
(2) Effect of A-17 on Mechanical Allodynia in Rats as Models of Pain After Incision
[0148] The relieving effect of A-17-HD, -MD and -LD administered by subcutaneous injection at the incision site on the mechanical allodynia in the rats as models of pain after incision was evaluated, with the blank preparation as a negative control and trans-8-methyl-N-vanillyl-6-nonenamide-HD and -LD as positive controls. The results are shown in
[0149] The above test results suggest that: the A-17-HD group has a strong and lasting analgesic effect on thermal hyperalgesia induced by the incision pain of rats after the surgery, which can last for 4 days (1-96 h), and is superior to the trans-8-methyl-N-vanillyl-6-nonenamide-HD group (3 days), and the equal dose of trans-8-methyl-N-vanillyl-6-nonenamide; the A-17-MD group also has a strong analgesic effect, which lasts for 3 days (1-72 h); and the A-17-LD group exerts an analgesic effect only within 1 day (1-24 h). Furthermore, in terms of the intensity of the analgesic effect (TWL values at all time points and overall thermal hyperalgesia within 7 days), the analgesic effect of the A-17-HD group is significantly better than that of the A-17-MD group and that of the A-17-LD group. Therefore, A-17 relieve the thermal hyperalgesia induced by the incision pain after the surgery in a dose-dependent manner and a time-dependent manner, with the intensity and the duration of the analgesic effect better than those of the trans-8-methyl-N-vanillyl-6-nonenamide at the same molar dose.
[0150] The A-17-HD group has a certain analgesic effect on the mechanical allodynia induced by the incision pain after the surgery, which can last for 2 days (1-4 h, 48 h); the A-17-MD group also has a certain analgesic effect, which starts at 24 h after the surgery and lasts for 1 day (24-48 h); and the A-17-LD group exerts an analgesic effect only at 1 h after the surgery. There is no significant difference among the A-17 dose groups. The trans-8-methyl-N-vanillyl-6-nonenamide-HD group has a certain analgesic effect only at 4 h after the surgery; and the trans-8-methyl-N-vanillyl-6-nonenamide-LD group has no analgesic effect. There is no significant difference between the A-17-HD group and the trans-8-methyl-N-vanillyl-6-nonenamide-HD group.
Example 25. Pharmacokinetics of Compounds Disclosed Herein In Vivo
Example 25. Clinical Compatibility Stability
[0151] 10 mg of each of the compounds according to the examples of the present invention was added with 10 mL of 0.9% sodium chloride and glucose injection. The reaction system was shaken to mix well, and the test tubes were left, to stand at room temperature and sampled at 1 h, 2 h, 4 h, 6 h and 8 h, The appearance of the sample was observed, and the related substances and pH value were determined. It suggests that the compounds disclosed herein have no obvious pH change within 6 h and have an impurity change of less than 10% within 8 h, feature good compatibility stability and are suitable for clinical application. The clinical preparation data for compound A-1 are shown in the following table.
TABLE-US-00007 TABLE 25-1 Clinical compatibility stability 0.9% sodium chloride injection (25° C.) Investigation time (h) 0 1 2 4 6 8 Character Clear liquid pH 3.39 3.38 3.37 3.34 3.31 / Related Capsaicine 2.57 3.22 3.84 4.95 6.1 7.22 substances Total impurities 6.25 6.9 7.52 8.59 9.77 10.89 Glucose injection (25° C.) Investigation time (h) 0 1 2 4 6 8 Character Clear liquid pH 3.30 3.31 3.28 3.26 3.21 / Related Capsaicine 2.69 3.64 4.24 5.62 6.94 8.32 substances Total impurities 6.46 7.41 8.00 9.38 10.7 12.02
[0152] The examples of the present invention have been described above. However, the present invention is not limited to the above examples. Any modification, equivalent, improvement and the like made without departing from the spirit and principle of the present invention shall fall within the protection scope of the present invention.