CN111925313B

CN111925313B - Compound with PDE4 inhibitory activity, preparation method, composition and application

Info

Publication number: CN111925313B
Application number: CN202010999493.XA
Authority: CN
Inventors: 王永广; 常俊美; 贾冰洁; 苏小庭; 戴信敏
Original assignee: Nalanga Shanghai Biomedical Technology Co ltd
Current assignee: Nalan Jia Shanghai Pharmaceutical Research Co ltd
Priority date: 2020-09-22
Filing date: 2020-09-22
Publication date: 2021-06-25
Anticipated expiration: 2040-09-22
Also published as: CN111925313A

Abstract

The invention belongs to the field of medicines, and particularly relates to a compound with PDE4 inhibitory activity, a preparation method, a composition and application thereof, wherein the compound is a compound with a structure shown in a formula I or a pharmaceutically acceptable salt thereof, and R in the formula₁Represents N, N-dimethylaminoethyl, 3-amino-3-oxopropyl, 3-difluoropropyl, methoxyalkyl; r₂Represents cyano, aminosulfonyl, aminocarbonyl, alkylacylamino. The compound provided by the invention has better PDE4 inhibitory activity, has the advantages of high efficacy and low side effect, and can be used for preparing medicaments for preventing and/or treating allergic diseases, autoimmune diseases or inflammatory diseases.

Description

Compound with PDE4 inhibitory activity, preparation method, composition and application

Technical Field

The invention belongs to the field of medicines, and particularly relates to a compound with PDE4 inhibitory activity, a preparation method, a composition and application thereof.

Background

Chronic inflammation is a multifactorial complication characterized by the activation of various types of inflammatory cells, such as lymphoid lineage (T lymphocytes) and myeloid lineage (including granulocytes, macrophages, and monocytes), which produce proinflammatory mediators, including cytokines, such as Tumor Necrosis Factor (TNF) and interleukin (IL-1). Thus, agents that inhibit the activation of these cells or their production of pro-inflammatory cytokines may be useful in the treatment of inflammatory diseases and other diseases involving elevated levels of cytokines.

Phosphodiesterases (PDEs) are a large family of polygenes that include over 30 types 11 and have different substrate specificities, PDE4, 7, 8 act exclusively on cyclic adenosine monophosphate (cAMP), a second messenger of the organism responsible for mediating cell stimulation to a wide range of extracellular stimuli, adenylate cyclase is activated when appropriate agonists bind to specific cell surface receptors, converting Adenosine Triphosphate (ATP) to cAMP, and inhibition of PDE function prevents conversion of cAMP to the inactive metabolite 5-AMP, thus maintaining high cAMP levels and inhibiting cell activation.

The research shows that PDE4 is related to allergic, autoimmune and inflammatory diseases, but the existing PDE4 inhibitor has side effects of vomit, central nervous system disorder and the like, so that the determination of a PDE4 activity inhibiting substance with low side effect is crucial to the allergic, autoimmune and inflammatory diseases.

Disclosure of Invention

The compound has high efficacy and low side effect, can be applied to the development of medicaments for allergic, autoimmune and inflammatory diseases, and has important significance for the prevention and treatment of the allergic, autoimmune and inflammatory diseases.

In a first aspect, the present invention provides a compound having the structure of formula i:

；

in the formula, R₁Represents N, N-dimethylaminoethyl3-amino-3-oxopropyl, 3-difluoropropyl, methoxyalkyl;

R₂represents cyano, aminosulfonyl, aminocarbonyl, alkylacylamino.

The compound provided by the invention has better PDE4 inhibitory activity, has the advantages of high efficacy and low side effect, can be applied to the development of drugs for allergic, autoimmune and inflammatory diseases, and has important significance for the prevention and treatment of the allergic, autoimmune and inflammatory diseases.

In a second aspect, there is provided a process for the preparation of a compound of the first aspect, or a pharmaceutically acceptable salt thereof, comprising the steps of:

synthesis of intermediate IV:

heating the compound II and the compound III in a first reaction solvent under the action of a first alkaline substance to react to obtain an intermediate IV;

synthesis of intermediate V:

reacting the intermediate IV in a second reaction solvent under the action of a second alkaline substance to obtain an intermediate V;

synthesis of Compound I:

and heating the intermediate V in a third reaction solvent to react under the action of a third alkaline substance to obtain the compound I.

The preparation method provided by the invention takes the compound II and the compound III as starting raw materials, and prepares the compound I by synthesizing the intermediate IV and the intermediate V. In the synthesis of the intermediate IV, the compound II and the compound III are heated to react under the action of a first alkaline substance; when the intermediate V is synthesized, the previously synthesized intermediate IV reacts under the action of a second alkaline substance; in the synthesis of compound I, the previously synthesized intermediate V reacts with compound VI under the action of a third basic substance. The preparation method is simple and convenient to operate, and has low requirements on equipment.

In a third aspect, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound selected from the compounds of the first aspect or a pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable carrier.

In a fourth aspect, the present invention provides a PDE4 inhibitor comprising a therapeutically effective amount of a compound selected from those described in the first aspect or a pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable carrier.

In a fifth aspect, the present invention provides the use of a compound according to the first aspect or a pharmaceutically acceptable salt thereof as a PDE4 inhibitor for the manufacture of a medicament for the prevention and/or treatment of an allergic disease, an autoimmune disease or an inflammatory disease.

Detailed Description

In order to make the technical problems, technical solutions and advantageous effects to be solved by the present invention more clearly apparent, the present invention is further described in detail below with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

In one aspect, this embodiment provides a compound having the structure of formula i:

；

in the formula, R₁Represents N, N-dimethylaminoethyl, 3-amino-3-oxopropyl, 3-difluoropropyl, methoxyalkyl;

R₂represents cyano, aminosulfonyl, aminocarbonyl, alkylacylamino.

The compound provided by the embodiment has better PDE4 inhibitory activity, has the advantages of high efficacy and low side effect, can be applied to the development of drugs for allergic, autoimmune and inflammatory diseases, and has important significance for the prevention and treatment of the allergic, autoimmune and inflammatory diseases.

Further, when said R is₁When represents a methoxyalkyl group, said R₁Is methoxyethyl orAnd (3) methoxypropyl.

Further, when said R is₂When represents an alkylacylamino group, said R₂Is pivaloylamino or acetylamino.

Further, the structure shown in formula I can be, but is not limited to, the following structure:

in a second aspect, a process for the preparation of a compound according to any one of the first aspect, or a pharmaceutically acceptable salt thereof, comprising the steps of:

step S10: synthesis of intermediate IV: heating the compound II and the compound III in a first reaction solvent under the action of a first alkaline substance to react to obtain an intermediate IV;

step S20: synthesis of intermediate V: reacting the intermediate IV in a second reaction solvent under the action of a second alkaline substance to obtain an intermediate V;

step S30: and (3) synthesizing a compound I, namely heating the intermediate V in a third reaction solvent to react under the action of a third alkaline substance to obtain the compound I.

In the preparation method provided in this example, compound I is prepared by synthesizing intermediate IV and intermediate V using compound II and compound III as starting materials. In the synthesis of the intermediate IV, the compound II and the compound III are heated to react under the action of a first alkaline substance; when the intermediate V is synthesized, the previously synthesized intermediate IV reacts under the action of a second alkaline substance; in the synthesis of compound I, the previously synthesized intermediate V reacts with compound VI under the action of a third basic substance. The preparation method is simple and convenient to operate, and has low requirements on equipment.

Further, in step S10, the first basic substance includes at least one of triethylamine, diisopropylethylamine, N-methylmorpholine, 4-Dimethylaminopyridine (DMAP), sodium carbonate, cesium carbonate, potassium carbonate, lithium hydroxide, and sodium hydroxide.

Further, in step S10, the first reaction solvent includes at least one of dichloromethane, tetrahydrofuran, dioxane, acetonitrile, dimethyl sulfoxide (DMSO), N-Dimethylformamide (DMF), and ethylene glycol dimethyl ether.

Further, the reaction temperature is 20 ℃ to 100 ℃, for example, the reaction temperature can be 20 ℃, 40 ℃, 60 ℃, 80 ℃ or 100 ℃.

Further, in step S20, the second basic substance includes at least one of n-butyl lithium, sec-butyl lithium, tert-butyl magnesium chloride, tert-butyl magnesium bromide, and isopropyl magnesium chloride.

Further, in step S20, the second reaction solvent includes at least one of Tetrahydrofuran (THF), diethyl ether, and methyl tert-butyl ether.

Further, in step S20, the reaction temperature includes-90 ℃ to 30 ℃, for example, the reaction temperature may be-90 ℃, 78 ℃, 30 ℃, 0 ℃, 10 ℃, 30 ℃, etc.

Further, in step S30, the third basic substance includes at least one of triethylamine, diisopropylethylamine, pyridine, sodium carbonate, potassium carbonate, lithium hydroxide, sodium hydroxide, and potassium hydroxide.

Further, in step S30, the third reaction solvent includes at least one of dichloromethane, tetrahydrofuran, dioxane, N-dimethylformamide, and N, N-dimethylacetamide.

Further, in step S30, the reaction temperature includes 20 ℃ to 100 ℃, for example, the reaction temperature may be 20 ℃, 25 ℃, 30 ℃, 80 ℃, 100 ℃, and the like.

It is noted that "pharmaceutically acceptable salts" refer to those salts that retain the biological effectiveness and properties of the parent compound. The salt comprises: acid addition salts obtained by reaction of the free base of the parent compound with an inorganic acid or with an organic acid; such as hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, phosphoric acid, sulfuric acid, perchloric acid, and the like; such as acetic acid, oxalic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, tartaric acid, benzenesulfonic acid (benzenesulfonate), benzoic acid, camphorsulfonic acid, citric acid, fumaric acid, gluconic acid, glutamic acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, mucic acid, pamoic acid, pantothenic acid, succinic acid, tartaric acid, malonic acid, or the like; preferably hydrochloric acid or (L) -malic acid; or when the acid proton present in the parent compound is replaced by a metal ion, such as an alkali metal ion, alkaline earth ion or aluminum ion, or coordinated with an organic base, a salt is formed; such as ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine and the like.

In a third aspect, the present embodiment provides a pharmaceutical composition comprising a therapeutically effective amount of a compound selected from the compounds of the first aspect, or a pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable carrier.

The compositions provided in this example can be administered to a patient in need of such treatment by oral, rectal, or parenteral administration. For oral administration, it can be made into conventional solid preparations such as tablet, powder, granule, capsule, etc., or liquid preparations such as water or oil suspension, or other liquid preparations such as syrup, etc.; for parenteral administration, it can be formulated into solution for injection, aqueous or oily suspension, etc.

A therapeutically effective amount refers to an amount of a compound of the present invention sufficient to effect the intended use. The therapeutically effective amount may vary depending on the following factors: the intended application (in vitro or in vivo), or the subject and disease condition being treated, such as the weight and age of the subject, the severity of the disease condition and the mode of administration, etc., can be readily determined by one of ordinary skill in the art. The specific dosage will vary depending on the following factors: the particular compound selected, the dosing regimen according to, whether to administer in combination with other compounds, the timing of administration, the tissue to which it is administered, and the physical delivery system carried.

Those skilled in the art will understand that: pharmaceutically acceptable carriers refer to pharmaceutical carriers conventional in the pharmaceutical art, such as: diluents such as water and the like; fillers, such as starch, sucrose, and the like; binders such as cellulose derivatives, alginates, gelatin, polyvinylpyrrolidone; humectants, such as glycerol; disintegrating agents such as agar, calcium carbonate and sodium bicarbonate; absorption promoters, such as quaternary ammonium compounds; surfactants such as cetyl alcohol; adsorption carriers such as kaolin and bentonite clay; lubricants, such as talc, calcium stearate and magnesium stearate, and polyethylene glycol, and the like. In addition, other adjuvants such as flavoring agent and sweetener can also be added into the above medicinal composition.

In a fourth aspect, the present example provides a PDE4 inhibitor comprising a therapeutically effective amount of a compound selected from those described in the first aspect, or a pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable carrier.

In a fifth aspect, the use of a compound according to the first aspect or a pharmaceutically acceptable salt thereof as a PDE4 inhibitor for the manufacture of a medicament for the prevention and/or treatment of an allergic disease, an autoimmune disease or an inflammatory disease.

Further, the autoimmune disease is selected from diseases such as rheumatoid arthritis, osteoarthritis, gouty arthritis, spondylitis, and the like.

The invention is described in further detail with reference to a part of the test results, which are described in detail below with reference to specific examples.

Example 1

Preparation of N- (2- ((4-methoxy-3- (2-methoxyethoxy) phenyl) sulfonyl) isoindolin-4-yl) acetamide

Synthesis of intermediate 1 b:

dissolving compound 1a (40.4 g, 200.0 mmol), 1-bromo-2-methoxyethane (27.6 g, 200.0 mmol) and potassium carbonate (33.1 g, 240.0 mmol) in DMF (500 mL), heating to 60 deg.C, stirring for 6 hours, detecting by TLC, adding water (500 mL) after the reaction is finished, quenching the reaction, extracting with ethyl acetate (500 mL x 2), drying the organic layer, filtering, concentrating, and separating by column chromatography to obtain off-white solid 38.5g with yield of 74.0%.

Synthesis of intermediate 1 c:

dissolving compound 1b (38.0 g, 146.2 mmol) in THF (400 mL), -adding n-butyllithium (175 mmol) at 78 deg.C, stirring for 2 hr, adding sulfur dioxide tetrahydrofuran solution (292.4 mmol) while maintaining temperature, then heating to room temperature for reaction overnight, adding SO under ice bath₂Cl₂(23.5 g, 175.4 mmol) and then stirred at room temperature for 1 hour, extracted with saturated ammonium chloride solution, extracted with methyl tert-butyl ether (300 mL x 2), the organic layer concentrated and isolated by column chromatography to give 30.3g of an off-white solid in 74.1% yield.

Synthesis of Compound 1:

intermediate 1c (1.4 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 1d (880 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.3g of off-white solid with yield 61.9%, ESI (+) m/z = 421.1.

Example 2

Preparation of N- (2- ((2- ((4-methoxy-3- (2-methoxyethoxy) phenyl) sulfonyl) isoindolin-4-yl) pivaloamide

The synthesis of intermediate 1b and intermediate 1c was performed as in example 1.

Synthesis of Compound 2:

intermediate 1c (1.4 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 2a (1090 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.0g of off-white solid in 43.3% yield ESI (+) m/z = 463.2.

Example 3

Preparation of 2- ((4-methoxy-3- (2-methoxyethoxy) phenyl) sulfonyl) isoindoline-4-carbonitrile

Synthesis of Compound 3:

intermediate 1c (1.4 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 3a (720 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.4g of off-white solid in 72.2% yield ESI (+) m/z = 389.1.

Example 4

Preparation of 2- ((4-methoxy-3- (2-methoxyethoxy) phenyl) sulfonyl) isoindoline-4-carboxamide

Synthesis of Compound 4:

intermediate 1c (1.4 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 4a (810 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.2g of off-white solid in 59.1% yield, ESI (+) m/z = 407.1.

Example 5

Preparation of 2- ((4-methoxy-3- (2-methoxyethoxy) phenyl) sulfonyl) isoindoline-4-sulfonamide

Synthesis of Compound 5:

intermediate 1c (1.4 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 5a (990 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.5g of off-white solid in 67.9% yield ESI (+) m/z = 443.1.

Example 6

Preparation of N- (2- ((3- (2- (dimethylamino) ethoxy) -4-methoxyphenyl) sulfonyl) isoindolin-4-yl) acetamide

Synthesis of intermediate 6 a:

compound 1a (40.4 g, 200.0 mmol), 2-bromo-N, N-dimethylethane-1-amine (30.2 g, 200.0 mmol) and potassium carbonate (33.1 g, 240.0 mmol) were dissolved in DMF (500 mL), heated to 60 ℃ and stirred to react for 6 hours, the reaction was detected by TLC, after completion of the reaction, water (500 mL) was added to quench the reaction, ethyl acetate (500 mL 2) was extracted, the organic layer was dried, filtered, concentrated, and separated by column chromatography to give 40.2g of an off-white solid with a yield of 73.6%.

Synthesis of intermediate 6 b:

dissolving compound 6a (38.2 g, 140.0 mmol) in THF (400 mL), -adding n-butyllithium (168.0 mmol) at 78 deg.C, stirring for 2 hr, adding sulfur dioxide tetrahydrofuran solution (280.0 mmol) while maintaining temperature, then heating to room temperature for reaction overnight, adding SO under ice bath₂Cl₂(22.5 g, 168.0 mmol) and stirring at room temperature for 1 hour, extracting with saturated ammonium chloride solution, extracting with methyl tert-butyl ether (300 mL x 2), concentrating the organic layer, and separating by column chromatography31.4g of an off-white solid was obtained in a yield of 76.6%.

Synthesis of Compound 6:

intermediate 6b (1.5 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 1d (880 mg, 5.0 mmol) were placed in dichloromethane (30 mL), stirred at room temperature for 8 hours, monitored by TLC, concentrated after completion of the reaction, and separated by column chromatography to give off-white solid 1.1g, yield 50.8%. ESI (+) m/z = 434.2.

Example 7

Preparation of N- (2- ((3- (2- (dimethylamino) ethoxy) -4-methoxyphenyl) sulfonyl) isoindolin-4-yl) pivaloamide

The synthesis of intermediate 6a and intermediate 6b was performed as in example 6.

Synthesis of compound 7:

intermediate 6b (1.5 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 2a (1090 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.3g of off-white solid in 54.7% yield and ESI (+) m/z = 476.2.

Example 8

Preparation of 2- ((3- (2- (dimethylamino) ethoxy) -4-methoxyphenyl) sulfonyl) isoindoline-4-carbonitrile

Synthesis of compound 8:

intermediate 6b (1.5 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 3a (720 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.6g of off-white solid in 79.8% yield and ESI (+) m/z = 402.1.

Example 9

Preparation of 2- ((3- (2- (dimethylamino) ethoxy) -4-methoxyphenyl) sulfonyl) isoindoline-4-carboxylic acid amide

Synthesis of compound 9:

intermediate 6b (1.5 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 4a (810 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.2g of off-white solid in 57.1% yield ESI (+) m/z = 420.2.

Example 10

Preparation of 2- ((3- (2- (dimethylamino) ethoxy) -4-methoxyphenyl) sulfonyl) isoindoline-4-sulfonamide

Synthesis of compound 10:

intermediate 6b (1.5 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 5a (990 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.5g of off-white solid with a yield of 65.9%, ESI (+) m/z = 456.1.

Example 11

Preparation of 3- (5- ((4-acetamidoindolin-2-yl) sulfonyl) -2-methoxyphenoxy) propionamide

Synthesis of intermediate 11 a:

compound 1a (40.4 g, 200.0 mmol), 3-bromopropionamide (30.2 g, 200.0 mmol) and potassium carbonate (33.1 g, 240.0 mmol) were dissolved in DMF (500 mL), heated to 60 ℃ and stirred for reaction for 6 hours, the reaction was detected by TLC, after the reaction was completed, the reaction was quenched by addition of water (500 mL), extracted with ethyl acetate (500 mL x 2), the organic layer was dried, filtered, concentrated, and column chromatographed to give 42.6g of an off-white solid in 78.0% yield.

Synthesis of intermediate 11 b:

dissolving compound 11a (38.2 g, 140.0 mmol) in THF (400 mL), -adding n-butyllithium (168.0 mmol) at 78 deg.C, stirring for 2 hr, adding sulfur dioxide tetrahydrofuran solution (280.0 mmol) while maintaining temperature, then heating to room temperature for overnight reaction, adding SO under ice bath₂Cl₂(22.5 g, 168.0 mmol) and then stirred at room temperature for 1 hour, extracted with saturated ammonium chloride solution, extracted with methyl tert-butyl ether (300 mL x 2), the organic layer concentrated and isolated by column chromatography to give 29.5g of an off-white solid in 72.0% yield.

Synthesis of compound 11:

intermediate 11b (1.5 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 1d (880 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.7g of off-white solid in 78.5% yield ESI (+) m/z = 434.1.

Example 12

Preparation of N- (2- ((3- (3-amino-3-oxopropoxy) -4-methoxyphenyl) sulfonyl) isoindolin-4-yl) pivaloyl amide

The synthesis of intermediate 11a and intermediate 11b was performed as in example 11.

Synthesis of compound 12:

intermediate 11b (1.5 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 2a (1090 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.4g of off-white solid in 58.9% yield, ESI (+) m/z = 476.2.

Example 13

Preparation of 3- (5- ((4-cyanoisoindolin-2-yl) sulfonyl) -2-methoxyphenoxy) propionamide

Synthesis of compound 13:

intermediate 11b (1.5 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 3a (720 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.3g of off-white solid with a yield of 64.8% and ESI (+) m/z = 402.1.

Example 14

Preparation of 2- ((3- (3-amino-3-oxopropoxy) -4-methoxyphenyl) sulfonyl) isoindoline-4-carboxamide

The synthesis of intermediate 1c and intermediate 1d was performed as in example 1.

Synthesis of compound 14:

intermediate 11b (1.5 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 4a (810 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.5g of off-white solid with yield 71.6% ESI (+) m/z = 420.1.

Example 15

Preparation of 3- (2-methoxy-5- ((4-aminosulfonylisoindolin-2-yl) sulfonyl) phenoxy) propanamide

Synthesis of compound 15:

intermediate 11b (1.5 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 5a (990 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.6g of off-white solid with a yield of 70.3% and ESI (+) m/z = 456.1.

Example 16

Preparation of N- (2- ((4-methoxy-3- (3-methoxypropoxy) phenyl) sulfonyl) isoindolin-4-yl) acetamide

Synthesis of intermediate 16 a:

dissolving compound 1a (40.4 g, 200.0 mmol), 1-bromo-3-methoxypropane (30.2 g, 200.0 mmol) and potassium carbonate (33.1 g, 240.0 mmol) in DMF (500 mL), heating to 60 deg.C, stirring for reaction for 6 hours, detecting the reaction by TLC, adding water (500 mL) after the reaction is finished, quenching the reaction, extracting with ethyl acetate (500 mL x 2), drying the organic layer, filtering, concentrating, and separating by column chromatography to obtain off-white solid 39.8g, with a yield of 72.6%.

Synthesis of intermediate 16 b:

dissolving compound 16a (38.4 g, 140.0 mmol) in THF (400 mL), -adding n-butyllithium (168.0 mmol) at 78 deg.C, stirring for 2 hr, adding sulfur dioxide tetrahydrofuran solution (280.0 mmol) while maintaining temperature, then heating to room temperature for overnight reaction, adding SO under ice bath₂Cl₂(22.5 g, 168.0 mmol) and then stirred at room temperature for 1 hour, extracted with saturated ammonium chloride solution, extracted with methyl tert-butyl ether (300 mL x 2), the organic layer concentrated and isolated by column chromatography to give an off-white solid 25.6g, 62.2% yield.

Synthesis of compound 16:

intermediate 16b (1.5 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 1d (880 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.4g of off-white solid with a yield of 64.5% ESI (+) m/z = 435.2.

Example 17

Preparation of N- (2- ((4-methoxy-3- (3-methoxypropoxy) phenyl) sulfonyl) isoindolin-4-yl) pivaloamide

The synthesis of intermediate 16a and intermediate 16b was performed as in example 16.

Synthesis of compound 17:

intermediate 16b (1.5 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 2a (1090 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.3g of off-white solid in 51.3% yield, ESI (+) m/z = 477.3.

Example 18

Preparation of 2- ((4-methoxy-3- (3-methoxypropoxy) phenyl) sulfonyl) isoindoline-4-carbonitrile

Synthesis of compound 18:

intermediate 16b (1.5 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 3a (720 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.6g of off-white solid in 79.6% yield and ESI (+) m/z = 403.1.

Example 19

Preparation of 2- ((4-methoxy-3- (3-methoxypropoxy) phenyl) sulfonyl) isoindoline-4-carboxamide

Synthesis of compound 19:

intermediate 16b (1.5 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 4a (810 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.4g of off-white solid in 66.7% yield ESI (+) m/z = 421.1.

Example 20

Preparation of 2- ((4-methoxy-3- (3-methoxypropoxy) phenyl) sulfonyl) isoindoline-4-sulfonamide

Synthesis of compound 20:

intermediate 16b (1.5 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 5a (990 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.5g of off-white solid with a yield of 65.8%, ESI (+) m/z = 457.1.

Example 21

Preparation of N- (2- ((3- (difluoromethoxy) -4-methoxyphenyl) sulfonyl) isoindolin-4-yl) acetamide

Synthesis of intermediate 21 a:

compound 1a (40.4 g, 200.0 mmol), difluoroiodomethane (35.6 g, 200.0 mmol) and potassium carbonate (33.1 g, 240.0 mmol) were dissolved in DMF (500 mL), stirred at 25 ℃ for 6 hours, checked by TLC, quenched after completion of the reaction with water (500 mL), extracted with ethyl acetate (500 mL x 2), the organic layer was dried, filtered, concentrated, and isolated by column chromatography to give 30.5g of an off-white solid with a yield of 60.5%.

Synthesis of intermediate 21 b:

dissolving compound 21a (25.2 g, 100.0 mmol) in THF (300 mL), -adding n-butyllithium (120.0 mmol) at 78 deg.C, stirring for 2 hr, adding sulfur dioxide tetrahydrofuran solution (240.0 mmol) while maintaining temperature, then heating to room temperature for overnight reaction, adding SO under ice bath₂Cl₂(16.1 g, 120.0 mmol) and then stirred at room temperature for 1 hour, extracted with saturated ammonium chloride solution, extracted with methyl tert-butyl ether (300 mL x 2), the organic layer concentrated and isolated by column chromatography to give an off-white solid 19.5g, 71.7% yield.

Synthesis of compound 21:

intermediate 21b (1.4 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 1d (880 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.5g of off-white solid in 72.8% yield ESI (+) m/z = 413.1.

Example 22

Preparation of N- (2- ((3- (difluoromethoxy) -4-methoxyphenyl) sulfonyl) isoindolin-4-yl) pivaloyl amide

The synthesis of intermediate 21a and the synthesis of intermediate 21b were performed according to example 21.

Synthesis of compound 22:

intermediate 21b (1.4 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 2a (1090 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.7g of off-white solid in 74.9% yield, ESI (+) m/z = 455.1.

Example 23

Preparation of 2- ((3- (difluoromethoxy) -4-methoxyphenyl) sulfonyl) isoindoline-4-carbonitrile

Synthesis of compound 23:

intermediate 21b (1.4 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 3a (720 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.3g of off-white solid with a yield of 68.4%, ESI (+) m/z = 381.1.

Example 24

Preparation of 2- ((3- (difluoromethoxy) -4-methoxyphenyl) sulfonyl) isoindoline-4-carboxamide

Intermediate 21b (1.4 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 4a (810 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.6g of off-white solid with yield 80.4%, ESI (+) m/z = 399.1.

Example 25

Preparation of 2- ((3- (difluoromethoxy) -4-methoxyphenyl) sulfonyl) isoindoline-4-sulfonamide

Intermediate 21b (1.4 g, 5.0 mmol), triethylamine (758 mg, 7.5 mmol) and compound 5a (990 mg, 5.0 mmol) were placed in dichloromethane (30 mL), the reaction was stirred at room temperature for 8 hours, monitored by TLC, after completion of the reaction, concentrated and isolated by column chromatography to give 1.3g of off-white solid in 59.8% yield, ESI (+) m/z = 435.0.

Biological evaluation

PED4 in vitro Activity assay

The experimental method comprises the following steps:

the compounds were stored in DMSO as 1mmol standards, which were then diluted in DMSO to different gradient concentrations.

The diluted compound solution was added to a 96-well plate, followed by addition of 1. mu.L of 1 Xkinase buffer (1 Xkinase buffer: 50mM HEPES, pH 7.5, 3mM MgCl)₂1mM EGTA, 100mM NaCl, 0.03% CHAPS, 2mM DTT), incubated for 10 minutes, then incubated with ATP reaction buffer in 384 well plates at room temperature for 2 hours, the reaction was terminated, and IC was calculated₅₀The specific data are as follows:

A＜50nM，50 nM≤B≤500nM，500nM＜C。

the above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.

Claims

1. A method of preparing a compound having the structure of formula 12, comprising:

synthesis of intermediate 11 a: dissolving 40.4g and 200.0mmol of compound 1a, 30.2g and 200.0mmol of 3-bromopropionamide and 33.1g and 240.0mmol of potassium carbonate in 500mL of DMF, heating to 60 ℃, stirring for reaction for 6 hours, detecting by TLC, quenching the reaction by adding 500mL of water after the reaction is finished, extracting for 2 times by using 500mL of ethyl acetate, drying an organic layer, filtering, concentrating, and carrying out column chromatography separation to obtain 42.6g of white-like solid with the yield of 78.0%;

synthesis of intermediate 11 b: dissolving 38.2g 140.0mmol of compound 11a in 400mL of THF, adding 168.0mmol of n-butyllithium at-78 deg.C, stirring for 2 hr, adding 280.0mmol of sulfur dioxide-tetrahydrofuran solution while maintaining the temperature, heating to room temperature for overnight reaction, adding 22.5g 168.0mmol of SO under ice bath₂Cl₂Then stirring for 1 hour at room temperature, extracting and killing the reaction by using a saturated ammonium chloride solution, extracting for 2 times by using 300mL of methyl tert-butyl ether, concentrating an organic layer, and carrying out column chromatography separation to obtain 29.5g of off-white solid with the yield of 72.0%;

synthesis of compound 12: 1.5g, 5.0mmol of intermediate 11b, 758mg, 7.5mmol of triethylamine, 1090mg, 5.0mmol of compound 2a were placed in 30mL of dichloromethane, stirred at room temperature for 8 hours, and subjected to TLC monitoring, after completion of the reaction, concentration and column chromatography to obtain 1.4g of off-white solid with yield of 58.9%, ESI (+) m/z =476.2, wherein the compound 12 has IC of 12₅₀＜50nM。

2. A method of preparing a compound having the structure of formula 13, comprising:

synthesis of compound 13: 1.5g of 5.0mmol of intermediate 11b, 758mg of 7.5mmol of triethylamine and 720mg of 5.0mmol of compound 3a are placed in 30mL of dichloromethane, stirred at room temperature for 8 hours, monitored by TLC, concentrated after the reaction is finished, and separated by column chromatography to obtain 1.3g of off-white solid with yield of 64.8%, ESI (+) m/z =402.1, wherein the compound 13 has IC (+) m/z =402.1₅₀＜50nM。