CN107698484B - Preparation method and application of lenalidomide derivative - Google Patents

Abstract

The invention discloses a lenalidomide derivative and a preparation method and application thereof. The structural formula of the lenalidomide derivative provided by the invention is shown as a formula I. The lenalidomide derivative shown in the formula I provided by the invention has an obvious anti-inflammatory effect, has no neurotoxicity, and has a good application prospect in the field of design and research of anti-inflammatory drugs. Meanwhile, the compound also has obvious anticancer effect and good application prospect in the research and development field of anticancer drugs.

Description

Preparation method and application of lenalidomide derivative

Technical Field

The invention belongs to the field of medicinal chemistry, and particularly relates to a preparation method and application of a lenalidomide derivative.

Background

Lenalidomide is an anti-tumor drug developed by Celgene biopharmaceutical corporation in America, has a chemical structure similar to that of thalidomide, and has multiple effects of resisting tumor, regulating immunity, resisting angiogenesis and the like. In vitro experiments showed that lenalidomide can inhibit proliferation of certain cell lines such as Namalwa cells. Lenalidomide inhibits the growth of multiple myeloma and MM1S cells in a patient. In addition, lenalidomide can also inhibit the expression of cyclooxygenase 2(COX-2), but has no effect on COX-1. Recent clinical findings indicate that lenalidomide has certain therapeutic effects on myeloma, leukemia, metastatic renal carcinoma, solid tumors, primary systemic amyloidosis and systemic myelofibrosis with myelopoiesis, in addition to the treatment of MDS and MM.

Disclosure of Invention

An object of the present invention is to provide a lenalidomide derivative or a pharmaceutically acceptable salt thereof.

The structural general formula of the lenalidomide derivative provided by the invention is shown as a formula I:

in formula I, R1 is: h OR-C ═ OR5, wherein R5 is the following substituted OR unsubstituted group: linear OR branched C1-C20 alkyl, aryl, heteroaryl, cycloalkyl, arylalkyl, heterocyclylalkyl, OR C1-C8 alkyl-OR 6; r6 is the following substituted or unsubstituted group: C1-C20 alkyl, arylalkyl, or heterocyclylalkyl;

r2 is: substituted or unsubstituted straight or branched C1-C20 alkyl, substituted or unsubstituted arylalkyl, substituted or unsubstituted heterocyclylalkyl;

r3 is: substituted or unsubstituted, straight or branched C1-C20 alkyl;

r4 is: substituted or unsubstituted straight or branched C1-C20 alkyl.

Wherein the substituents in each of the substituted alkyl, aryl, heteroaryl, cycloalkyl, arylalkyl, and heterocyclylalkyl groups are independently optionally selected from the group consisting of: halogen, hydroxyl, amino and nitro. The halogen refers to F, Cl, Br or I.

The above-mentioned substituent-containing group may have a substituent substituted at each substitutable position of the group. When more than one position in a given formula can be substituted with one or more substituents selected from a particular group, the substituents may be substituted at each position, identically or differently.

Further, the cycloalkyl group may be a C3-C20 cycloalkyl group, the arylalkyl group may be a C6-C20 arylalkyl group, and the heterocyclylalkyl group may be a C3-C20 heterocyclylalkyl group.

In some of the embodiments of the present invention,

in formula I, R1 is: h OR-C ═ OR5, R5 is substituted OR unsubstituted C1-C20 alkyl, OR substituted OR unsubstituted phenyl (C1-C5) alkyl;

r2 is: substituted or unsubstituted C1-C5 alkyl, substituted or unsubstituted phenyl (C1-C5) alkyl, or substituted or unsubstituted morpholinyl (C1-C5) alkyl;

r3 is: substituted or unsubstituted C1-C5 alkyl;

r4 is: substituted or unsubstituted C1-C5 alkyl.

In some of the embodiments of the present invention,

in formula I, R1 is: h OR-C ═ OR5, R5 is C1-C20 alkyl OR benzyl;

r2 is: C1-C5 alkyl, benzyl or 2-morpholinoethyl;

r3 is: C1-C5 alkyl or halogen substituted C1-C5 alkyl.

R4 is: C1-C5 alkyl or halogen substituted C1-C5 alkyl.

In some embodiments, the lenalidomide derivatives of the present invention may be exemplified by, but are not limited to, the structures shown below:

the term "alkyl" as used herein refers to a group consisting of only carbon and hydrogen atoms, and having no unsaturation (e.g., double bonds, triple bonds, or rings), which encompasses a wide variety of possible geometric and stereoisomeric groups. This group is connected to the rest of the molecule by a single bond. By way of non-limiting examples of alkyl groups, mention may be made of the following linear or branched groups: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl and seven further isomers thereof, n-hexyl and sixteen further isomers thereof, n-heptyl and various isomers thereof, n-octyl and various isomers thereof, n-nonyl and various isomers thereof, and n-decyl and various isomers thereof.

The term "cycloalkyl" as used herein refers to a saturated non-aromatic ring system consisting of at least 3 carbon atoms, which may be monocyclic, bicyclic, polycyclic, fused, bridged, or spiro. As non-limiting examples of cycloalkyl groups, the following groups may be cited: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl; and fused, bridged or spiro ring groups formed from two or more of the above-mentioned monocyclic rings via a common side and a common carbon atom.

The term "aryl" as used herein, alone or as part of an "arylalkyl" refers to monocyclic, bicyclic, and tricyclic carbocyclic ring systems containing a total of 6 to 14 membered rings, wherein at least one ring system is aromatic, wherein each ring system contains 3 to 7 membered rings and only one attachment point is attached to the rest of the molecule. The term "aryl" may be used interchangeably with the term "aromatic ring", e.g., aromatic rings may include phenyl, naphthyl and anthracenyl.

The term "heteroaryl" as used herein refers to a 5-14 membered aromatic heterocyclic ring system having one or more heteroatoms independently selected from N, O or S, which may be monocyclic, bicyclic, polycyclic, wherein bicyclic and polycyclic rings may be formed from a single ring by single bond linkages or fused. As non-limiting examples of heteroaryl groups, the following groups may be cited: oxazolyl, isoxazolyl, imidazolyl, furyl, indolyl, isoindolyl, pyrrolyl, triazolyl, triazinyl, tetrazolyl, thienyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, benzofuryl, benzothiazolyl, benzoxazolyl, benzimidazolyl, benzothienyl, benzopyranyl, carbazolyl, quinolyl, isoquinolyl, quinazolinyl, cinnolinyl, naphthyridinyl, pteridinyl, purinyl, quinoxalinyl, thiadiazolyl, indolizinyl, acridinyl, phenazinyl, phthalazinyl, coumarinyl, pyrazolopyridinyl, pyridopyridazinyl, pyrrolopyridyl, imidazopyridinyl, pyrazolopyridazinyl; and a group formed by the above-mentioned heteroaryl group by a single bond connection or a fusion connection.

The term "arylalkyl" as used herein, refers to an alkyl group having one or more hydrogen atoms independently replaced by an aryl group, wherein the aryl and alkyl groups are as defined above. Examples include benzyl, phenylethyl, and the like.

The term "heteroarylalkyl" as used herein refers to an alkyl group wherein one or more hydrogen atoms are independently replaced by a heteroaryl group, wherein the heteroaryl and alkyl groups are as defined above. Examples include 2-morpholinoethyl and the like.

The compounds of the present invention may also be used in the form of their pharmaceutically acceptable salts or solvates. Physiologically acceptable salts of the compounds of formula I include conventional salts formed with pharmaceutically acceptable inorganic or organic acids or bases and acid addition salts of quaternary amines. More specific examples of suitable acid salts include salts of hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, perchloric acid, fumaric acid, acetic acid, propionic acid, succinic acid, glycolic acid, formic acid, lactic acid, maleic acid, tartaric acid, citric acid, pamoic acid, malonic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, hydroxynaphthoic acid, hydroiodic acid, malic acid, steroic acid, tannic acid, and the like. More specific examples of suitable base salts include sodium, lithium, potassium, magnesium, aluminum, calcium, zinc, N' -dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, N-methylglucamine, and procaine salts.

It is a further object of the present invention to provide a process for the preparation of the compounds of formula I as described above.

The invention provides a preparation method of a compound of formula I, wherein R1 is H, which comprises the following steps:

1) reacting the compound shown in the formula II with 2-aminophenylacetamide to form a Schiff base compound shown in the formula III;

wherein R3 and R4 in the formula II and the formula III are defined as R3 and R4 in the formula I;

2) carrying out catalytic addition reaction on bromoacetate shown in a formula IV and Schiff base compound shown in a formula III to obtain a compound shown in a formula V;

wherein R2 in the formula IV is defined as R2 in the formula I; in the formula V, R2, R3 and R4 are as defined in the formula I;

3) carrying out hydrogenation reduction on the compound shown in the formula V to obtain an amino compound shown in a formula VI;

r2, R3 and R4 in the formula VI are defined as in the formula I;

4) carrying out ring closure reaction on 2-bromomethyl-3-nitrobenzoic acid methyl ester and an amino compound shown as a formula VI to obtain a compound shown as VII;

in the formula VII, R2, R3 and R4 are defined as formula I;

5) reduction of the nitro group in the compound of formula vii affords a compound of formula I wherein R1 ═ H.

Wherein, the reaction conditions of the reaction in the step 1) are as follows: the reaction temperature is-10 ℃ to 30 ℃, the reaction time is 8 hours to 20 hours, and the molar ratio of the compound shown as the formula II to the 2-aminophenylacetamide is 1: 1.0-1.5; the reaction is carried out in a solvent, preferably methanol.

The reaction conditions of the catalytic addition reaction in the step 2) are as follows: the reaction temperature is 0 to 50 ℃ and the reaction time is 2 to 8 hours; the molar ratio of the Schiff base compound shown in the formula III to the bromoacetate shown in the formula IV is 1: 1-1.5; the catalyst used for the catalytic addition reaction is selected from at least one of the following: diethyl zinc and nickel diacetylacetonate, and the addition amount of the catalyst is 0.05-1 equivalent. The reaction is carried out in a solvent, preferably dichloromethane.

The reaction conditions of the hydrogenation reduction in the step 3) are as follows: the reaction temperature is 20-80 ℃, the reaction time is 3-10 hours, and the reaction pressure is 1-10bar hydrogen pressure; the hydrogenation is carried out in a solvent, preferably methanol or ethanol.

The ring closing reaction in the step 4) is carried out in a solvent, and the solvent is preferably toluene; the reaction conditions of the ring-closing reaction are as follows: heating for 3-10 hours under the reflux condition; the molar ratio of the 2-bromomethyl-3-nitrobenzoic acid methyl ester to the amino compound shown in the formula VI is 1.0-1.5: 1. The ring closing reaction is carried out under the action of alkaline conditions, the used base can be triethylamine, diisopropylethylamine, N-methylmorpholine, pyridine, dimethylaminopyridine or sodium acetate and the like, and triethylamine is preferred. The molar ratio of the base to the amino compound represented by formula VI is 1-5: 1.

the reaction conditions for nitro reduction in the step 5) are as follows: the reaction temperature is 20-80 ℃, the reaction time is 3-10 hours, and the reaction pressure is 1-10bar hydrogen pressure; the hydrogenation is carried out in a solvent, preferably methanol or ethanol.

The invention provides a preparation method of a compound of formula I with R1 ═ C ═ OR5, which comprises the following steps:

acylation of the amino group in the compound of formula I where R1 ═ H affords a compound of formula I where R1 ═ C ═ OR 5.

Another object of the present invention is to provide the use of the compounds of formula I as defined above.

The application of the compound of formula I provided by the invention comprises the following aspects: 1) the use of a compound of formula I for the preparation of a medicament for the prevention and/or treatment of inflammation; 2) the use of a compound of formula I for the preparation of a medicament for the prevention and/or treatment of cancer; 3) the use of a compound of formula I in the manufacture of a medicament for inhibiting cancer cell proliferation.

The inflammation includes acute inflammation, subacute inflammation and immune inflammation.

Such cancers include various cancers known in the art (solid or non-solid), including but not limited to: lymph cancer, myeloma, hepatocarcinoma, cervical cancer, colon cancer, non-small cell lung cancer, breast cancer, esophageal cancer, and leukemia.

The cancer cells comprise lymphoma cells, myeloma cells, liver cancer cells, cervical cancer cells, colon cancer cells, non-small cell lung cancer cells, breast cancer cells, esophageal cancer cells and leukemia cells.

The lymphoma cell can be lymphoma cell P388D1 or non-Hodgkin lymphoma cell Raji

The myeloma cell may specifically be a multiple myeloma cell ARH 77.

The invention also discloses a medicament for preventing and/or treating inflammation and a medicament for preventing and/or treating cancer, which are prepared by using the lenalidomide derivative shown in the formula I as an active ingredient.

The agent for preventing and/or treating inflammation and the agent for preventing and/or treating cancer may be introduced into the body such as muscle, intradermal, subcutaneous, intravenous, mucosal tissue by injection, spray, nasal drop, eye drop, penetration, absorption, physical or chemical mediated method; or mixed or coated with other materials and introduced into body.

If necessary, one or more pharmaceutically acceptable carriers can be added into the medicine. The carrier includes diluent, excipient, filler, binder, wetting agent, disintegrating agent, absorption enhancer, surfactant, adsorption carrier, lubricant, etc. which are conventional in the pharmaceutical field.

The above medicine can be made into various forms such as injection, tablet, powder, granule, capsule, oral liquid, paste, cream, etc. The medicaments in various dosage forms can be prepared according to the conventional method in the pharmaceutical field.

The lenalidomide derivative shown in the formula I provided by the invention has an obvious anti-inflammatory effect, has no neurotoxicity, and has a good application prospect in the field of design and research of anti-inflammatory drugs. The lenalidomide derivative shown in the formula I has an obvious anticancer effect and also has a good application prospect in the research and development field of anticancer drugs.

Drawings

FIG. 1 is a reaction scheme of Compound 6 prepared in example 1.

Detailed Description

The method of the present invention is illustrated by the following specific examples, but the present invention is not limited thereto, and any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included within the scope of the present invention.

The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.

Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.

Synthesis part examples

Example 1: 3- (4-amino-1-isoindolin-2-yl) -3- (3-ethoxy-4-methoxyphenyl) propionic acid methyl ester (Compound 6)

1.1 preparation of step 1, (E) -2- ((3-ethoxy-4-methoxybenzylidene) amino) -2-phenylacetamide (Compound 2)

3-ethoxy-4-methoxybenzaldehyde (compound 1, 31.0g,172mmol) was dissolved in 200mL of methanol, and 2-aminophenylacetamide (25.8g, 172mmol) was added thereto, followed by stirring at room temperature for 18 hours. After the reaction was completed, the reaction solution was concentrated, and ethanol was recrystallized to obtain (E) -2- ((3-ethoxy-4-methoxybenzylidene) amino) -2-phenylacetamide (23.6g, 44.0%) as a white solid.¹H NMR(DMSO-d6,400MHz)δ8.74(s,1H),7.58-6.95(m,8H),5.36(s,1H),4.09(m,2H),3.83(s,3H),1.32(m,3H).ESI-MS m/z:313.4[M+H]⁺.

1.2 step 2 preparation of methyl 3- ((2-amino-2-oxo-1-phenylacetylamino) -3- (3-ethoxy-4-methoxyphenyl) propanoate (Compound 3)

Dissolving (E) -2- ((3-ethoxy-4-methoxybenzylidene) amino) -2-phenylacetamide (compound 2, 23.6g, 75.6mmol) in 150mL of dichloromethane, cooling to-30 ℃, adding diethyl zinc (121mL, 21mmol, 1M n-hexane solution), nickel diacetoacetate (0.646g, 2.52mmol) and ethyl bromoacetate (13.8g, 90.7mmol) to the reaction solution, stirring at constant temperature for 3And (4) hours. After the reaction, ice water was added to quench the reaction, the organic phase was concentrated, and the residual liquid was subjected to column chromatography to give methyl 3- ((2-amino-2-oxo-1-phenylacetylamino) -3- (3-ethoxy-4-methoxyphenyl) propionate (12.0g, 41.3%) as a transparent yellow liquid.¹H NMR(DMSO-d6,400MHz)δ7.33-6.83(m,8H),4.85(s,1H),4.54(m,1H),4.09(m,2H),3.83(s,3H),3.68(s,3H),2.75(m,2H),1.33(m,3H).ESI-MS m/z:387.4[M+H]⁺.

1.3 step 3, preparation of methyl 3-amino-3- (3-ethoxy-4-methoxyphenyl) propanoate (Compound 4)

Methyl 3- ((2-amino-2-oxo-1-phenylacetamido) -3- (3-ethoxy-4-methoxyphenyl) propanoate (compound 3, 12.0g, 31.0mmol) was dissolved in 80mL methanol, 10% palladium on carbon (1.90g, H) was added₂0 w/w-50%), and the mixture was stirred under 9bar of hydrogen at high pressure for 16 hours. The reaction solution was filtered, and the mother liquor was concentrated under reduced pressure. The residual oil was recrystallized to give methyl 3-amino-3- (3-ethoxy-4-methoxyphenyl) propionate (5.00g, 64.1%) as a pale yellow solid.¹H NMR(DMSO-d6,400MHz)δ6.96-6.83(m,3H),4.54(m,1H),4.09(m,2H),3.83(s,3H),3.68(s,3H),2.88(m,2H),1.32(m,3H).ESI-MS m/z:276.3[M+Na]⁺.

1.4 preparation of methyl 3- (3-ethoxy-4-methoxyphenyl) -3- (4-nitro-1-isoindolin-2-yl) propionate (Compound 5), step 4

Methyl 3-amino-3- (3-ethoxy-4-methoxyphenyl) propionate (compound 4, 5.00g, 19.7mmol) was dissolved in 80mL of toluene, triethylamine (2.00g, 19.7mmol) and methyl 2-bromomethyl-3-nitrobenzoate (4.83g, 17.8mmol) were added, respectively, and the mixture was refluxed at 110 ℃ for 5 hours. The reaction mixture was concentrated under reduced pressure, and column chromatography of the residue gave methyl 3- (3-ethoxy-4-methoxyphenyl) -3- (4-nitro-1-isoindolin-2-yl) propionate (4.00g, 49.0%) as a yellow solid.¹H NMR(DMSO-d6,400MHz)δ8.33-6.83(m,6H),5.54(m,1H),4.22(s,2H),4.09(m,2H),3.83(s,3H),3.68(s,3H),2.92(m,2H),1.32(m,3H).ESI-MS m/z:415.3[M+H]⁺.

1.5 preparation of methyl 3- (4-amino-1-isoindolin-2-yl) -3- (3-ethoxy-4-methoxyphenyl) propionate (Compound 6), step 5

Will 3-(3-ethoxy-4-methoxyphenyl) -3- (4-nitro-1-isoindolin-2-yl) propionic acid methyl ester (compound 5, 4.00g, 9.66mmol) was dissolved in 80mL tetrahydrofuran, and 10% palladium on carbon (1.0g, H)₂0 w/w-50%), the reaction was stirred under 9bar of hydrogen at high pressure for 16 hours. The reaction mixture was filtered, the mother liquor was concentrated under reduced pressure, and the residue was recrystallized to give methyl 3- (4-amino-1-isoindolinone-2-yl) -3- (3-ethoxy-4-methoxyphenyl) propionate (2.20g, 59.2%) as a white solid.¹H NMR(DMSO-d6,400MHz)δ7.27-6.72(m,6H),6.27(s,2H),5.54(m,1H),4.22(S,2H),4.09(m,2H),3.83(s,3H),3.68(s,3H),2.92(m,2H),1.32(m,3H).ESI-MS m/z:385.3[M+H]⁺.

Example 2: 3- (4-acetylamino-1-isoindolin-2-yl) -3- (3-ethoxy-4-methoxyphenyl) propionic acid methyl ester (Compound 7)

Methyl 3- (4-amino-1-isoindolinone-2-yl) -3- (3-ethoxy-4-methoxyphenyl) propionate (1.0g, 2.6mmol) was dissolved in 10mL of tetrahydrofuran, and dicyclohexylcarbodiimide (0.54g, 2.6mmol), 4-dimethylaminopyridine (0.32g, 2.6mmol) and acetic acid (0.16g, 2.6mmol) were added to the reaction, respectively, and the mixture was stirred at room temperature for 10 hours. The reaction mixture was filtered, concentrated under reduced pressure, and subjected to column chromatography to give methyl 3- (4-acetylamino-1-isoindolinone-2-yl) -3- (3-ethoxy-4-methoxyphenyl) propionate (0.97g, 88.2%) as a white solid.¹H NMR(DMSO-d6,400MHz)δ7.65-6.74(m,6H),5.51(m,1H),4.22(S,2H),4.09(m,2H),3.83(s,3H),3.68(s,3H),2.92(m,2H),2.04(s,2H),1.32(m,3H).ESI-MS m/z:449.5[M+Na]⁺.

Example 3: 3- (1-oxo-4-valerylaminoisoindolinone-2-yl) -3- (3-ethoxy-4-methoxyphenyl) propionic acid methyl ester (Compound 8)

The procedure is as in example 2, starting from methyl 3- (4-amino-1-isoindolin-2-yl) -3- (3-ethoxy-4-methoxyphenyl) propionateAnd valeric acid to give a white solid powder, yield: 92.5 percent.¹H NMR(DMSO-d6,400MHz)δ7.65-6.74(m,6H),5.51(m,1H),4.22(S,2H),4.09(m,2H),3.83(s,3H),3.68(s,3H),2.92(m,2H),2.39(m,2H),1.63(m,2H),1.32(m,2H),0.90(m,3H).ESI-MS m/z:469.5[M+H]⁺.

Example 4: 3- (4-Decanoylamino-1-isoindolin-2-yl) -3- (3-ethoxy-4-methoxyphenyl) propionic acid methyl ester (Compound 9)

The procedure is as in example 2, starting from methyl 3- (4-amino-1-isoindolinone-2-yl) -3- (3-ethoxy-4-methoxyphenyl) propionate and decanoic acid, giving a white solid in powder form, yield: 85.6 percent.¹H NMR(DMSO-d6,400MHz)δ7.65-6.74(m,6H),5.51(m,1H),4.22(S,2H),4.09(m,2H),3.83(s,3H),3.68(s,3H),2.92(m,2H),2.39(m,2H),1.63-1.29(m,14H),0.88(m,3H).ESI-MS m/z:539.5[M+H]⁺.

Example 5: 3- (4-stearoylamino-1-isoindolin-2-yl) -3- (3-ethoxy-4-methoxyphenyl) propionic acid methyl ester (Compound 10)

The procedure is as in example 2, starting from methyl 3- (4-amino-1-isoindolinone-2-yl) -3- (3-ethoxy-4-methoxyphenyl) propionate and octadecanoic acid to give a white solid powder in yield: 80.6 percent.¹H NMR(DMSO-d6,400MHz)δ7.65-6.74(m,6H),5.51(m,1H),4.22(S,2H),4.09(m,2H),3.83(s,3H),3.68(s,3H),2.92(m,2H),2.39(m,2H),1.63-1.26(m,30H),0.88(m,3H).ESI-MS m/z:673.7[M+Na]⁺.

Example 6: 3- (4-Phenylacetamido-1-isoindolin-2-yl) -3- (3-ethoxy-4-methoxyphenyl) propionic acid methyl ester (Compound 11)

The procedure is as in example 2, starting from methyl 3- (4-amino-1-isoindolinone-2-yl) -3- (3-ethoxy-4-methoxyphenyl) propionate and phenylacetic acid to give a white solid in powder form, yield: 80.6 percent.¹H NMR(DMSO-d6,400MHz)δ7.65-6.74(m,11H),5.51(m,1H),4.22(s,2H),4.09(m,2H),3.90(s,2H),3.83(s,3H),3.68(s,3H),2.92(m,2H),1.32(m,3H).ESI-MS m/z:503.5[M+H]⁺.

Example 7: 3- (4-amino-1-isoindolin-2-yl) -3- (3- (2,2, 2-trifluoroethoxy) -4-methoxyphenyl) propionic acid methyl ester (Compound 12)

The procedure of example 1 was repeated except that 3-ethoxy-4-methoxybenzaldehyde, which was the starting material in step 1.1, was replaced with 4-methoxy-3- (2,2, 2-trifluoroethoxy) benzaldehyde to obtain a white solid powder in total yield: 4.25 percent.¹H NMR(DMSO-d6,400MHz)δ7.27-6.74(m,6H),6.27(s,2H),5.51(m,1H),4.46(s,2H),3.83(s,3H),3.68(s,3H),2.92(m,2H).ESI-MS m/z:439.5[M+H]⁺.

Example 8: 3- (4-amino-1-isoindolin-2-yl) -3- (3-ethoxy-4-propoxyphenyl) propionic acid methyl ester (Compound 14)

The procedure of example 1 was followed except that 3-ethoxy-4-methoxybenzaldehyde, the starting material in step 1.1, was replaced with 3-ethoxy-4-propoxybenzaldehyde to give a white solid powder in total yield: 5.13 percent.¹H NMR(DMSO-d6,400MHz)δ7.27-6.74(m,6H),6.27(s,2H),5.51(m,1H),4.42(s,2H),4.09(m,4H),3.68(s,3H),2.92(m,2H),1.74(m,2H),1.32(m,3H),0.90(m,3H).ESI-MS m/z:413.5[M+H]⁺.

Example 9: 3- (4-amino-1-oxoisoindolin-2-yl) -3- (3-ethoxy-4-methoxyphenyl) propionic acid benzyl ester (Compound 19)

The method is the same as example 1, only the raw material ethyl bromoacetate in step 1.2 is replaced by benzyl bromoacetate, white solid powder is obtained, and the total yield is as follows: 5.33 percent. 1H NMR (DMSO-d6,400MHz) delta 7.47-6.74(M,11H),6.27(s,2H),5.51(M,1H),5.20(s,2H),4.22(s,2H),4.09(M,2H),3.83(s,3H),2.92(M,2H),1.32(M,3H). ESI-MS M/z 483.5[ M + Na ] +.

Example 10: 3- (4-amino-1-oxoisoindolin-2-yl) -3- (3-ethoxy-4-methoxyphenyl) propionic acid 2-morpholineethyl ester (Compound 20)

The method is the same as example 1, only the raw material ethyl bromoacetate in step 1.2 is replaced by ethyl bromoacetate-2-morpholine, white solid powder is obtained, and the total yield is as follows: 3.82 percent. 1H NMR (DMSO-d6,400MHz) delta 7.27-6.74(M,6H),6.27(s,2H),5.51(M,1H),4.35(M,2H),4.22(s,2H),4.09(M,2H),3.83(s,3H),3.65(M,4H),2.97(M,2H),2.67(M,2H),2.36(M,4H),1.32(M,3H), ESI-MS M/z 506.5[ M + Na ] +.

In light of the above examples, one skilled in the art can make appropriate choices for starting materials to synthesize other compounds of similar structure as claimed in the present invention.

Comparative example 1: 3- (5-amino-1-isoindolin-2-yl) -3- (3-ethoxy-4-methoxyphenyl) propionic acid methyl ester (Compound 15)

The same as example 1, the procedure was repeated except that the starting material, methyl 2-bromomethyl-3-nitrobenzoate in step 1.4, was replaced with methyl 4-nitro-2-bromomethylbenzoate to give a white solid powder with a total yield: 4.68 percent.¹H NMR(DMSO-d6,400MHz)δ7.66-6.35(m,6H),6.27(s,2H),5.51(m,1H),4.42(s,2H),4.09(m,2H),3.83(s,3H),3.68(s,3H),2.92(m,2H),1.32(m,3H).ESI-MS m/z:407.5[M+Na]⁺.

Comparative example 2: 3- (4-amino-1, 3-dioxoisoindolin-2-yl) -3- (3-ethoxy-4-methoxyphenyl) propionic acid methyl ester (Compound 16)

The preparation method can be carried out by referring to the known method of similar compounds.

Examples of the drug effect

Example 11: test for Effect of anti-inflammatory action

130 Kunming mice, SPF grade, male, 15-17g are selected. And 3 days of quarantine, randomly dividing the blank into 13 groups according to the weight after the quarantine is finished, wherein each group comprises 10 animals, namely: the model group, the experimental components of the example compounds were 10 groups, and the experimental components of the comparative example compounds were 2 groups. In the experiment, xylene (0.03 mL/mouse) is coated on the right auricle of each mouse to cause auricle swelling, the left auricle is used as a control, the corresponding medicine is administered to the tail vein 30min after the xylene coating, 10mg/kg body weight is administered, and the model group is administered with physiological saline with equal volume. The cervical vertebrae were removed 1 hour after administration to kill the mice, the same part of both ears was cut off in an equal area by a punch having a diameter of 8mm, and the swelling degree was determined by the difference between the weight of the right ear piece and the weight of the left ear piece.

TABLE 1 Effect of paraxylene on ear swelling in Kunming mice

Comparison with model groups: p <0.05, p < 0.01.

Example 12: comparison of neurotoxicity

130 healthy and infertile NIH mice are taken, the weight of the mice is 18-22 g, and the mice are half female and half male. During the test, mice are randomly divided into 3 groups according to sex and weight, wherein each group contains l0 mice, and the mice are half male and female. On the test day, the compounds are divided into 12 groups (10mg/kg), a negative control group is subjected to intragastric administration and is given with an equal volume of sodium chloride injection, the toxic reaction condition of the mice is observed immediately after administration, and the reaction and the duration time of the mice after administration are recorded; after dosing, observations were continued for 14 d. The behavior of the mice in the negative control group and the compound group after administration was recorded in various ways, and the results are shown in table 2.

Compound 15 and compound 16 are neurotoxic, with compound 16 being more neurotoxic; no neurotoxicity was observed with the compounds of examples 1-10.

Example 13 antitumor Effect test

Selection of mouse lymphoma cells P388D1, non-Hodgkin lymphoma cells Raji, multiple myeloma cells ARH77 tumor cells IC of the Compounds of the examples₅₀。

Preparing medicines and reagents: one bag of RPMI1640 medium was supplemented with one liter of water, 2g of sodium bicarbonate, 10 million units of penicillin and 100mg of streptomycin, the pH was adjusted to 7.4, and the mixture was sterilized by filtration through a 0.22 μm sterile filter. The complete culture solution is obtained by adding 5ml of inactivated newborn calf serum into 95ml of culture medium. Preparing trypsin into 0.25% solution with D-hanks buffer solution, filtering, sterilizing, and storing at 4 deg.C.

The compounds of the examples and lenalidomide were weighed to 100mg each, and added to a sterilized 1.5ml centrifuge tube, and 1ml of DMSO was added to prepare a stock solution of 100mg/ml, and the stock solution was stored at-20 ℃ under refrigeration. Before use, the mixture is thawed and diluted with complete culture solution to corresponding concentration for use.

Cell culture and passage: all cells were cultured in a cell culture flask containing 10ml of complete medium in a adherent manner at 37 ℃ with 5% CO₂And culturing under saturated humidity. After the cells are full of the bottom of the bottle, washing the bottle twice by using a sterilized D-hanks solution, adding 0.25% trypsin to digest the cells for 2 minutes, pouring off the trypsin, after the cells can completely shed by shaking lightly, adding 30ml of complete culture solution, blowing the cells by using a pipette, subpackaging the cells in 3 new cell culture bottles, and continuing to culture.

And (3) drug treatment: taking a bottle of freshly grown cells, collecting the cells after trypsinization, and blowing and beating the cells uniformly by a pipetteTwo drops of cell suspension are stained with trypan blue, the number of viable cells is counted under a microscope, and the number of cells is adjusted to 1X 10 with the use of the whole culture medium⁵Individual cells/ml. Add 100. mu.l of cell suspension to each well of a 96-well cell culture plate, place the plate in CO₂Culturing for 12 hours in an incubator, taking out a culture plate, adding 100 mu l of complete culture solution containing experimental drugs with different concentrations into each hole, setting 4 parallel holes for each concentration, adding complete culture solution without drugs into cells of 4 holes as negative control holes, adding complete culture solution containing vincristine into cells of 4 holes as positive control, and setting the final concentration of the vincristine to be 5 mu g/ml. Adding the medicine, vibrating the culture plate on a microplate oscillator, mixing, and placing in CO₂The cultivation was continued in the incubator for 48 hours. And (3) taking out the culture plate, adding 10 mu l of 5mg/ml MTT solution into each hole, uniformly mixing by oscillation, continuously culturing for 4 hours, discarding the original culture solution, adding 150 mu l of DMSO into each hole, fully oscillating to dissolve the blue-violet crystal, measuring the light absorption of each hole on a Bio-Rad 550 microplate reader, and measuring the wavelength of 570nm and the reference wavelength of 630 nm.

And (3) calculating the inhibition rate of the drugs on cell proliferation according to the OD value of each well:

performing linear regression according to the inhibition rate corresponding to logarithm of drug concentration to obtain a linear equation, and calculating the drug concentration corresponding to 50% of the inhibition rate to obtain the half-Inhibitory Concentration (IC) of the experimental drug on tumor cells₅₀)。

Half Inhibitory Concentration (IC) of compounds against different tumor cell lines₅₀，μg/ml)

The compounds 6-14 and 19 of the examples have better inhibiting effect on the above tumor cells P388D1 and Raji than the compounds 15 and 16 and lenalidomide, and the compounds 6-14 of the examples have better inhibiting effect on ARH77 tumor cells than the compounds 15 and 16 and are similar to the lenalidomide.

Claims (10)

1. A compound represented by formula I or a pharmaceutically acceptable salt thereof:

in formula I, R1 is: h OR-C ═ OR5, R5 is C1-C20 alkyl OR benzyl;

r2 is: C1-C5 alkyl, benzyl or 2-morpholinoethyl;

r3 is: C1-C5 alkyl;

r4 is: C1-C5 alkyl.

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: the compound shown in the formula I is a compound shown in any one of the following formulas:

3. a process for the preparation of a compound of formula I as claimed in claim 1, wherein R1 ═ H, comprising the steps of:

1) reacting the compound shown in the formula II with 2-aminophenylacetamide to form a Schiff base compound shown in the formula III;

wherein R3 and R4 in the formula II and the formula III are defined as R3 and R4 in the formula I;

2) carrying out catalytic addition reaction on bromoacetate shown in a formula IV and Schiff base compound shown in a formula III to obtain a compound shown in a formula V;

wherein R2 in the formula IV is defined as R2 in the formula I; in the formula V, R2, R3 and R4 are as defined in the formula I;

3) carrying out hydrogenation reduction on the compound shown in the formula V to obtain an amino compound shown in a formula VI;

r2, R3 and R4 in the formula VI are defined as in the formula I;

4) carrying out ring closure reaction on 2-bromomethyl-3-nitrobenzoic acid methyl ester and an amino compound shown as a formula VI to obtain a compound shown as VII;

in the formula VII, R2, R3 and R4 are defined as formula I;

5) reduction of the nitro group in the compound of formula vii affords a compound of formula I wherein R1 ═ H.

4. A process for the preparation of a compound of formula I as claimed in claim 1, wherein R1 ═ C ═ OR5, comprising the steps of:

acylation of the amino group in the compound of formula I where R1 ═ H affords a compound of formula I where R1 ═ C ═ OR 5.

5. The use of a compound of formula I, or a pharmaceutically acceptable salt thereof, as claimed in any one of claims 1-2, in the manufacture of a medicament for: 1) a medicament for preventing and/or treating inflammation; 2) a preventive and/or therapeutic agent for cancer; 3) a medicine for inhibiting cancer cell proliferation.

6. Use according to claim 5, characterized in that: the inflammation is acute inflammation, subacute inflammation and immune inflammation;

the cancer is a solid or non-solid cancer;

the cancer cells are: lymphoma cells, myeloma cells, liver cancer cells, cervical cancer cells, colon cancer cells, non-small cell lung cancer cells, breast cancer cells, esophageal cancer cells, or leukemia cells.

7. Use according to claim 6, characterized in that:

the cancer is lymphoma, myeloma, liver cancer, cervical cancer, colon cancer, non-small cell lung cancer, breast cancer, esophageal cancer or leukemia.

8. A medicament, the active ingredient of which is a compound of formula I according to any one of claims 1 to 2 or a pharmaceutically acceptable salt thereof; the medicine is as follows: 1) a medicament for preventing and/or treating inflammation; 2) a preventive and/or therapeutic agent for cancer; 3) a medicine for inhibiting cancer cell proliferation.

9. The medicament of claim 8, wherein: the inflammation is acute inflammation, subacute inflammation and immune inflammation;

the cancer is a solid or non-solid cancer;

the cancer cells are: lymphoma cells, myeloma cells, liver cancer cells, cervical cancer cells, colon cancer cells, non-small cell lung cancer cells, breast cancer cells, esophageal cancer cells, or leukemia cells.

10. The medicament of claim 9, wherein:

the cancer is lymphoma, myeloma, liver cancer, cervical cancer, colon cancer, non-small cell lung cancer, breast cancer, esophageal cancer or leukemia.

Images