CN113816965B

CN113816965B - Compound with IDO (indomethacin diphosphate) inhibitory activity and preparation method thereof

Info

Publication number: CN113816965B
Application number: CN202111389705.3A
Authority: CN
Inventors: 苏小庭; 孙学涛; 刘兰祥; 戴信敏
Original assignee: Beijing Xinkaiyuan Pharmaceuticals Co Ltd
Current assignee: Beijing Xinkaiyuan Pharmaceuticals Co Ltd
Priority date: 2021-11-23
Filing date: 2021-11-23
Publication date: 2022-03-08
Anticipated expiration: 2041-11-23
Also published as: CN113816965A

Abstract

The invention belongs to the field of medicines, and provides a compound with IDO (indomethacin diphosphate) inhibitory activity and a preparation method thereof. The compound with IDO inhibitory activity is a compound with a structure shown in a formula I, and pharmaceutically acceptable salt, hydrate, solvate, stereoisomer or isotope compound thereof. The structure of the formula I is as follows:

Description

Compound with IDO (indomethacin diphosphate) inhibitory activity and preparation method thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a compound with IDO (indomethacin diphosphate) inhibitory activity and a preparation method thereof.

Background

Indoleamine-2, 3-dioxygenase (IDO) is an inflammatory cytokine-induced rate-limiting enzyme of tryptophan (Trp) catabolism, which is involved in the inhibition of intracellular pathogen replication and in immune regulation. To date, three types of tryptophan metabolizing enzymes have been identified: IDO1, IDO2, and tryptophan-2, 3-dioxygenase 2. Abnormal increase in IDO expression or activity is closely related to the pathogenesis of various diseases, and is an important factor leading to various diseases. The IDO inhibitor is used as a medicine with a new medicine target and a new mechanism, can be used for treating a plurality of major diseases such as tumors, Alzheimer diseases, depression, cataract and the like, and has wide social and economic benefit prospects. Meanwhile, IDO is over-expressed in various cancer cells such as lung cancer, prostatic cancer, pancreatic cancer, colorectal cancer and the like, and can help cancer cells escape from an immune system by reducing the level of L-tryptophan in a cell microenvironment, so that the over-expression of tumor cells is inhibited, and the aim of arousing the autoimmune system of a human body can be achieved by inhibiting the expression of IDO.

Disclosure of Invention

In order to search for a novel IDO inhibitor, the invention designs and synthesizes a series of (5H-imidazo [5,1-a ] isoindol-5-yl) methylamine derivatives which have novel structures, high safety and high IDO inhibition activity through extensive and intensive research, and researches the antitumor activity of the compounds.

An object of the present invention is to provide a (5H-imidazo [5,1-a ] isoindol-5-yl) methylamine derivative.

Another object of the present invention is to provide a process for the preparation of the above compounds.

It is another object of the present invention to provide a pharmaceutical composition comprising the above compound.

Another object of the present invention is to provide the use of the above-mentioned compounds and the above-mentioned pharmaceutical compositions as IDO inhibitors for the treatment of tumors.

In order to achieve the purpose, the invention adopts the technical scheme that:

in a first aspect, a compound having the structure of formula i, a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, or isotopic compound thereof:

Ⅰ

wherein R represents H, F, Cl, Br, CN; x represents CH and N; n represents 0 to 2.

Further, the compound is selected from compounds having the following structures:

。

in a second aspect, the present invention provides a process for the preparation of a compound having the structure of formula i, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer or isotopic compound thereof, when X represents N.

When X represents N, the structure of the compound is as follows:

the preparation method comprises the following steps:

wherein halo represents F, Cl, Br, I;

and (3) synthesizing an intermediate IV:

reacting a compound II with a structure shown in a formula II and a compound III with a structure shown in a formula III in a first solvent at a first reaction temperature to generate an intermediate IV with a structure shown in a formula IV;

and (3) synthesis of an intermediate V:

the intermediate IV and 4-piperidone react in a second solvent under the action of an acid-binding agent at a second reaction temperature to generate an intermediate V with a structure of a formula V;

synthesis of compounds having the structure of formula la:

and reacting the intermediate V and (5H-imidazo [5,1-a ] isoindol-5-yl) methylamine in a third solvent under the action of a reducing agent at a third reaction temperature to generate the compound with the structure of the formula Ia.

As a preferable scheme of the first preparation method provided by the invention, the first reaction temperature is-20-80 ℃;

and/or the second reaction temperature is-20 to 100 ℃;

and/or the third reaction temperature is 0-25 ℃.

Further preferably, the first reaction temperature is 50 to 60 ℃.

Further preferably, the second reaction temperature is 20-30 ℃.

As another preferable embodiment of the first preparation method provided by the present invention, the first solvent is at least one selected from dioxane, tetrahydrofuran, acetonitrile, and toluene;

and/or the second solvent is at least one selected from dioxane, tetrahydrofuran, acetonitrile and dichloromethane;

and/or the acid-binding agent is at least one selected from potassium carbonate, sodium bicarbonate, triethylamine and N, N-diisopropylethylamine;

and/or the third solvent is at least one selected from dichloromethane, methanol, ethanol and acetonitrile;

and/or the reducing agent is selected from at least one of sodium borohydride, sodium cyanoborohydride and sodium triacetoxyborohydride.

Further preferably, the first solvent is tetrahydrofuran.

Further preferably, the second solvent is acetonitrile.

Further preferably, the acid-binding agent is potassium carbonate.

Further preferably, the third solvent is methanol.

Further preferably, the reducing agent is sodium borohydride.

The first preparation method provided by the invention takes a compound II and a compound III as initial raw materials, and finally prepares the compound with the structure of formula Ia by synthesizing an intermediate IV and an intermediate V. The whole preparation process is simple to operate, easy to control, low in requirements on production equipment and suitable for industrial large-scale production.

In a third aspect, the present invention provides a process for the preparation of a compound having the structure of formula i, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer or isotopic compound thereof, when X represents CH.

When X represents CH, the structure of the compound is as follows:

the preparation method comprises the following steps:

and (3) synthesizing an intermediate VII:

reacting a compound II with a structure shown in a formula II and a compound VI with a structure shown in a formula VI in a fourth solvent in the presence of alkali under the action of a condensing agent at a fourth reaction temperature to generate an intermediate VII with a structure shown in a formula VII;

synthesis of intermediate viii:

the intermediate VII and an oxidant react in a fifth solvent at a fifth reaction temperature to generate an intermediate VIII with a structure of a formula VIII;

synthesis of compounds having the structure of formula ib:

and reacting the intermediate VIII and (5H-imidazo [5,1-a ] isoindol-5-yl) methylamine in a third solvent under the action of a reducing agent at a third reaction temperature to generate the compound with the structure of the formula Ib.

As a preferable scheme of the second preparation method provided by the invention, the fourth reaction temperature is 20-120 ℃;

and/or the fifth reaction temperature is-80-25 ℃;

and/or the third reaction temperature is 0-25 ℃.

Further preferably, the fourth reaction temperature is 20-30 ℃.

Further preferably, the fifth reaction temperature is 15-25 ℃.

As another preferable embodiment of the second preparation method provided by the present invention, the fourth solvent is at least one selected from dichloromethane, N-dimethylformamide, and N, N-dimethylaniline;

and/or the base is at least one of triethylamine and diisopropylethylamine;

and/or the condensing agent is at least one selected from the group consisting of a mixed solution of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and 1-hydroxybenzotriazole, 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate and O-benzotriazol-tetramethylurea hexafluorophosphate;

and/or the fifth solvent is at least one selected from dichloromethane, trichloromethane and 1, 2-dichloroethane;

and/or, the oxidant is selected from at least one of 2, 3-dichloro-5, 6-dicyan p-benzoquinone (DDQ), pyridinium chlorochromate (PCC), pyridinium chlorodichromate (PDC), Dess-Martin oxidant (Dess-Martin oxidation), Stevens oxidant (Swern oxidation);

Further preferably, the fourth solvent is N, N-dimethylformamide.

More preferably, the condensing agent is a mixed solution of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and 1-hydroxybenzotriazole.

Further preferably, the fifth solvent is dichloromethane.

Further preferably, the oxidant is dess-martin oxidant.

Further preferably, the third solvent is methanol.

Further preferably, the reducing agent is sodium borohydride.

The second preparation method provided by the invention takes a compound II and a compound VI as initial raw materials, and finally prepares the compound with the structure of formula Ib by synthesizing an intermediate VII and an intermediate VIII. The whole preparation process is simple to operate, easy to control, low in requirements on production equipment and suitable for industrial large-scale production.

The conditions in the two production methods provided by the present invention, such as reactants, solvents, bases, acid-binding agents, oxidizing agents, reducing agents, condensing agents, the amounts of the compounds used, the reaction temperature, and the like, are not limited to the above explanations. The compounds of formula i provided herein may also be conveniently prepared by optionally combining the various synthetic procedures described in this specification or known in the art, such combinations being readily performed by those skilled in the art to which the invention pertains.

In a fourth aspect, the present invention provides a pharmaceutical composition comprising a compound having the structure of formula i, a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, or isotopic compound thereof.

In the present invention, a compound having a structure of formula i, a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer or isotopic compound thereof is used as an active ingredient in a pharmaceutical composition.

The present invention provides pharmaceutical compositions comprising, in addition to a therapeutically effective amount of at least one compound having the structure of formula i, a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, or isotopic compound thereof:

one or more pharmaceutical excipients;

and/or one or more pharmaceutically active substances having an IDO inhibitory activity other than the compound having the structure of formula i, a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer or isotopic compound thereof.

Methods of preparing the pharmaceutical compositions provided herein will be apparent to those skilled in the art and include conventional mixing, dissolving, lyophilizing and like techniques.

The pharmaceutical composition provided by the invention can be prepared into various common dosage forms, such as tablets, pills, capsules, granules, oral solutions, oral suspensions, oral emulsions, injections and the like, according to conventional preparation methods in the pharmaceutical field. It is convenient to provide the patient with clinical use by administering to the patient by various common modes of administration, such as oral or parenteral administration (by intravenous, intramuscular, topical or subcutaneous routes).

In a fifth aspect, the present invention provides a compound having a structure of formula i, a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, or isotopic compound thereof, and a pharmaceutical composition comprising a compound having a structure of formula i, a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, or isotopic compound thereof, for use as an IDO inhibitor in the preparation of a medicament for treating a tumor.

For the application of the compound and the pharmaceutical composition provided by the invention in preparing the medicine for treating the tumor, a mode of applying a therapeutically effective amount of the compound or the pharmaceutical composition provided by the invention to a patient needing to treat the tumor is adopted to realize the treatment effect. The compound or the pharmaceutical composition provided by the invention can be used alone as a treatment means for treating tumors, and can also be used in combination with other conventional treatment means for treating tumors, such as surgery, radiotherapy, chemotherapy and the like.

The therapeutically effective amount of the compounds or pharmaceutical compositions provided herein for treating tumors depends on a number of factors. May vary depending on the particular type of tumor that is to be treated, as can be determined by one skilled in the art without undue experimentation. The actual treatment will also take into account factors such as the age and weight of the patient, the severity of the condition, the particular mode of administration, etc., and will ultimately depend on the discretion of the attendant physician or clinician.

Further, the tumor is selected from:

lung cancer, liver cancer, colon cancer, pancreatic cancer, breast cancer, prostate cancer, brain cancer, ovarian cancer, cervical cancer, testicular cancer, kidney cancer, head and neck cancer, lymphoma, melanoma, or leukemia.

Some of the terms involved in the expression of the present invention are defined as follows:

the term "pharmaceutically acceptable salts" refers 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; the inorganic acid comprises hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, phosphoric acid, sulfuric acid, perchloric acid and the like; the organic acid includes acetic acid, oxalic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, tartaric acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, fumaric acid, gluconic acid, glutamic acid, isethionic acid, lactic acid, mandelic acid, mucic acid, pamoic acid, pantothenic acid, succinic acid, malonic acid, or the like.

Or, a salt formed when an acid proton present in the parent compound is replaced with a metal ion or coordinated with an organic base; the metal ions include alkali metal ions, alkaline earth ions, aluminum ions and the like; the organic base includes ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, etc.

The term "hydrate" refers to a substance in which water molecules are coordinately or covalently bound to cations or anions in a compound, or to a substance in which water ions are not directly bound to cations or anions but exist at certain positions in a solid crystal lattice in a certain proportion.

The term "solvate" refers to a substance formed by association of a compound with molecules of a pharmaceutically acceptable solvent, which typically includes ethanol, acetic acid, and the like.

The term "stereoisomer" refers to compounds having the same molecular formula, but differing in the arrangement of the atoms in the molecule in space. Since the compounds provided by the present invention may have one or more asymmetric centers, the compounds may be prepared as (R) -stereoisomers alone or (S) -stereoisomers alone or as mixtures thereof. Unless otherwise indicated, the description or designation of a particular compound in the present invention is intended to include the individual enantiomers as well as racemic or other mixtures thereof. Methods for determining stereochemical configuration and separating stereoisomers are routine in the art (see the discussion in chapter 4 of Advanced Organic Chemistry, 4 th edition, j. march, John Wiley and Sons, New York, 1992). Thus, the present invention also encompasses any stereoisomeric form, its corresponding enantiomers (D-and L-isomers or (+) and (-) isomers) and diastereomers thereof and mixtures thereof, having the ability to modulate IDO kinase activity, and is not limited to any one stereoisomeric form.

The term "pharmaceutical composition" refers to a mixture of one or more of the compounds provided herein with other chemical ingredients (e.g., pharmaceutical excipients). The purpose of the pharmaceutical composition is to facilitate the administration of the compound to the organism.

The term "pharmaceutical excipient" refers to a substance that has been reasonably evaluated in terms of safety, does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the compound administered, and is included in a pharmaceutical preparation, in addition to an active ingredient. The pharmaceutic adjuvant has important functions of solubilization, dissolution assistance, sustained and controlled release and the like besides excipient, carrier and stability improvement, and is an important component which may influence the quality, safety and effectiveness of the pharmaceutical composition. The pharmaceutical excipients include, but are not limited to, carriers, diluents, excipients, solubilizers, binders, disintegrants, penetration enhancers, pH adjusters, buffers, release retardants, flavoring agents, preservatives, antioxidants, and the like.

The beneficial effects of the invention include:

(1) the invention provides (5H-imidazo [5,1-a ] isoindol-5-yl) methylamine derivatives which can be used as effective IDO inhibitors and have strong inhibitory activity and small side effect.

(2) The invention also provides a pharmaceutical composition containing the (5H-imidazo [5,1-a ] isoindol-5-yl) methylamine derivative, which has good anti-tumor pharmacological activity.

Detailed Description

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

In the preparation method of the compound with the structure shown in the formula I, the pharmaceutically acceptable salt, the hydrate, the solvate, the stereoisomer or the isotope compound thereof, when X represents N, the structure of the compound is as follows:

the preparation method comprises the following steps:

wherein halo represents F, Cl, Br, I;

in the first step, intermediate IV is synthesized: and dissolving the compound II and the compound III in a first solvent, and reacting at-20-80 ℃ to generate an intermediate IV. Wherein the first solvent can be at least one selected from dioxane, tetrahydrofuran, acetonitrile and toluene.

For example, 1mol of compound II and 1mol of compound III are dissolved in 1L of dioxane and reacted with stirring at 30 ℃ to give intermediate IV. After the reaction is finished, the intermediate IV can be obtained by conventional treatment processes such as extraction, filtration, concentration, purification and the like.

The second step is the synthesis of intermediate v: and dissolving the intermediate IV, 4-piperidone and an acid-binding agent in a second solvent, and stirring at-20-100 ℃ to react to generate an intermediate V. Wherein the acid-binding agent can be at least one of potassium carbonate, sodium bicarbonate, triethylamine and N, N-diisopropylethylamine; the second solvent may be at least one selected from dioxane, tetrahydrofuran, acetonitrile, and dichloromethane.

For example, 1mol of intermediate IV, 1mol of 4-piperidone and 1mol of sodium bicarbonate are dissolved in 2L of dioxane and the reaction is stirred at 50 ℃ to give intermediate V. After the reaction is finished, the intermediate V can be obtained by conventional treatment processes such as extraction, filtration, concentration, purification and the like.

The third step is the synthesis of a compound having the structure of formula Ia: dissolving the intermediate V, (5H-imidazo [5,1-a ] isoindol-5-yl) methylamine and a reducing agent in a third solvent, and reacting at 0-25 ℃ to generate the compound with the structure of the formula Ia. Wherein, the reducing agent can be at least one of sodium borohydride, sodium cyanoborohydride and sodium triacetoxyborohydride. The third solvent may be at least one selected from dichloromethane, methanol, ethanol, and acetonitrile.

For example, 1mol of intermediate V, 1mol of ((5H-imidazo [5,1-a ] isoindol-5-yl) methylamine and 1.5mol of sodium triacetoxyborohydride are dissolved in 1L of dichloromethane, and the mixture is stirred and reacted at 10 ℃ to generate the compound with the structure of formula Ia.

In the preparation method of the compound with the structure shown in the formula I, the pharmaceutically acceptable salt, the hydrate, the solvate, the stereoisomer or the isotope compound thereof, when X represents CH, the structure of the compound is as follows:

the preparation method comprises the following steps:

in the first step, intermediate VII is synthesized: and dissolving the compound II, the compound VI, a condensing agent and alkali in a fourth solvent, and reacting at 20-120 ℃ to generate an intermediate VII. Wherein the fourth solvent can be at least one selected from dichloromethane, N-dimethylformamide and N, N-dimethylaniline. The base can be at least one of triethylamine and diisopropylethylamine. The condensing agent can be at least one selected from a mixed solution of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and 1-hydroxybenzotriazole, 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate and O-benzotriazol-tetramethylurea hexafluorophosphate.

For example, 1mol of the compound II, 1mol of the compound VI, 1mol of 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethyluronium hexafluorophosphate and 1mol of triethylamine are dissolved in 0.3L of dichloromethane and stirred at 50 ℃ to react to produce an intermediate VII. After the reaction is finished, the intermediate VII can be obtained by conventional treatment processes such as extraction, filtration, concentration, purification and the like.

The second step is the synthesis of intermediate VIII: and dissolving the intermediate VII and an oxidant in a fifth solvent, and stirring at-80-25 ℃ to react to generate the intermediate VIII. Wherein, the fifth solvent can be at least one of dichloromethane, trichloromethane and 1, 2-dichloroethane. The oxidant may be at least one selected from 2, 3-dichloro-5, 6-dicyan-p-benzoquinone (DDQ), pyridinium chlorochromate (PCC), pyridinium chlorodichromate (PDC), Dess-Martin oxidant (Dess-Martin oxidation), Stevens oxidant (Swern oxidation).

For example, 1mol of intermediate VII and 1mol of chlorodichromate are dissolved in 1.5L of chloroform, and the reaction is stirred at 0 ℃ to give intermediate VIII. After the reaction is finished, the intermediate VIII can be obtained by conventional treatment processes such as extraction, filtration, concentration, purification and the like.

The third step is the synthesis of a compound having the structure of formula lb: dissolving the intermediate VIII, (5H-imidazo [5,1-a ] isoindol-5-yl) methylamine and a reducing agent in a third solvent, and reacting at 0-25 ℃ to generate the compound with the structure shown in the formula Ib. Wherein, the reducing agent can be at least one of sodium borohydride, sodium cyanoborohydride and sodium triacetoxyborohydride. The third solvent may be at least one selected from dichloromethane, methanol, ethanol, and acetonitrile.

For example, 1mol of intermediate VIII, 1mol of (5H-imidazo [5,1-a ] isoindol-5-yl) methylamine and 1.5mol of sodium cyanoborohydride are dissolved in 1L of acetonitrile and reacted with stirring at 15 ℃ to give the compound having the structure of formula Ib. After the reaction is finished, the compound with the structure of the formula Ib can be obtained by the conventional treatment processes of extraction, filtration, concentration, purification and the like.

The invention is described in more detail by referring to a part of the tests, which are carried out in sequence, and the following detailed description is given by combining specific examples:

unless defined otherwise, technical terms used in the following examples have the same meanings as commonly understood by one of ordinary skill in the art to which the present invention belongs. The reagents used in the following examples, unless otherwise specified, are all conventional biochemical reagents; the raw materials, instruments, equipment and the like used in the following examples are either commercially available or available by existing methods; the dosage of the reagent is the dosage of the reagent in the conventional experiment operation if no special description exists; the experimental methods are conventional methods unless otherwise specified.

Example 1

4- (((5H-imidazo [5,1-a ] isoindol-5-yl) methyl) amino) -N- (4-chlorophenyl) piperidine-1-carboxamide (I-01)

The first step is as follows: synthesis of intermediate V-1

4-chloroaniline (12.76 g, 0.1 mol) and 4-piperidone (9.90 g, 0.1 mol) were added to 100mL of Tetrahydrofuran (THF), potassium carbonate (6.95 g, 0.05 mol) was added, stirring was continued, the temperature was reduced to 0-5 ℃ and COCl was passed through₂(9.81 g, 0.1 mol), stirred for 30 minutes, then warmed to 20-25 ℃, reacted for 2 hours, the reaction was completed, a saturated sodium carbonate solution was added to adjust pH =7-8, ethyl acetate (100 mL × 2) was extracted, liquid separation was performed, the organic phase was dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: ethyl acetate/petroleum ether = 1/2) to obtain 15.40g of intermediate v-1 with a yield of 60.94%. Intermediate V-1 was a brown oil.

The second step is that: synthesis of Compound I-01

Intermediate v-1 (10.11 g, 0.04 mol) and (5H-imidazo [5,1-a ] isoindol-5-yl) methylamine (7.41 g, 0.04 mol) were added to 80mL of methanol, sodium borohydride (2.27 g, 0.06 mol) was added in portions at room temperature, the reaction was stirred at room temperature for 2 hours, after the completion of the reaction, a saturated ammonium chloride solution (20 mL) was added to quench the reaction, methanol was removed by concentration under reduced pressure, 100mL of water and ethyl acetate (200 mL × 2) were added to the residue, extraction, liquid separation, the organic phase was dried over anhydrous sodium sulfate, filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol = 10/1) to obtain 3.26g of an off-white solid (compound i-01) with a yield of 19.32% and ESI (+) m/z ═ 422.2.

Example 2

4- (((5H-imidazo [5,1-a ] isoindol-5-yl) methyl) amino) -N- (4-fluorophenyl) piperidine-1-carboxamide (I-02)

The first step is as follows: synthesis of intermediate V-2

4-fluoroaniline (11.1 g, 0.1 mol) and 4-piperidone (9.90 g, 0.1 mol) were added to 100mL of Tetrahydrofuran (THF), potassium carbonate (6.95 g, 0.05 mol) was added thereto, the mixture was stirred, the temperature was then reduced to 0-5 ℃ and COCl was introduced thereinto₂(9.81 g, 0.1 mol), stirred for 30 minutes, then warmed to 20-25 ℃, reacted for 2 hours, the reaction was completed, a saturated sodium carbonate solution was added to adjust pH =7-8, ethyl acetate (100 mL × 2) was extracted, liquid separation was performed, the organic phase was dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: ethyl acetate/petroleum ether = 1/2) to obtain 16.50g of intermediate v-2 with a yield of 69.84%. Intermediate V-2 was a brown oil.

The second step is that: synthesis of Compound I-02

Intermediate v-2 (9.45 g, 0.04 mol) and (5H-imidazo [5,1-a ] isoindol-5-yl) methylamine (7.41 g, 0.04 mol) were added to 80mL of methanol, sodium borohydride (2.27 g, 0.06 mol) was added in portions at room temperature, the reaction was stirred at room temperature for 2 hours, after the completion of the reaction, a saturated ammonium chloride solution (20 mL) was added to quench the reaction, methanol was removed by concentration under reduced pressure, 100mL of water and ethyl acetate (200 mL × 2) were added to the residue, extraction, liquid separation, the organic phase was dried over anhydrous sodium sulfate, filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol = 10/1) to obtain 4.25g of an off-white solid (compound i-02) with a yield of 26.20% and ESI (+) m/z of 406.2.

Example 3

3- (4- (((5H-imidazo [5,1-a ] isoindol-5-yl) methyl) amino) piperidin-1-yl) -N- (3-fluorophenyl) propanamide (I-03)

The first step is as follows: synthesis of intermediate IV-3

Adding 3-fluoroaniline (11.11 g, 0.1 mol) into 100mL Tetrahydrofuran (THF), cooling to 0-5 ℃, dropwise adding 3-bromopropionyl chloride (17.14 g, 0.1 mol), stirring for 30 minutes, then heating to 50-60 ℃, reacting for 2 hours, finishing the reaction, adding saturated sodium carbonate solution to adjust the pH to be 7-8, extracting with ethyl acetate (100 mL multiplied by 2), separating liquid, drying an organic phase with anhydrous sodium sulfate, filtering, concentrating the filtrate under reduced pressure, and purifying the residue by column chromatography (eluent: ethyl acetate/petroleum ether = 1/2) to obtain 15.30g of an intermediate IV-3 with the yield of 62.17%. The intermediate IV-3 is a white-like solid.

The second step is that: synthesis of intermediate V-3

Intermediate iv-3 (12.30 g, 0.05 mol) and 4-piperidone (4.96 g, 0.05 mol) were added to 100mL of acetonitrile, potassium carbonate (6.95 g, 0.05 mol) was added at room temperature, the reaction was stirred at room temperature overnight, after completion of the reaction, insoluble matter was removed by filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol = 50/1) to give 11.89g of intermediate v-3 with a yield of 89.97%. Intermediate V-3 was a brown oily solid.

The third step: synthesis of Compound I-03

Intermediate v-3 (10.57 g, 0.04 mol) and (5H-imidazo [5,1-a ] isoindol-5-yl) methylamine (7.41 g, 0.04 mol) were added to 80mL of methanol, sodium borohydride (2.27 g, 0.06 mol) was added in portions at room temperature, the reaction was stirred at room temperature for 2 hours, after the completion of the reaction, a saturated ammonium chloride solution (20 mL) was added to quench the reaction, methanol was removed by concentration under reduced pressure, 100mL of water and ethyl acetate (200 mL × 2) were added to the residue, extraction, liquid separation, the organic phase was dried over anhydrous sodium sulfate, filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol = 10/1) to obtain 8.32g of an off-white solid (compound i-03) with a yield of 47.98% and ESI (+) m/z of 434.2.

Example 4

2- (4- (((5H-imidazo [5,1-a ] isoindol-5-yl) methyl) amino) piperidin-1-yl) -N- (4-bromophenyl) acetamide (I-04)

The first step is as follows: synthesis of intermediate IV-4

Adding 4-bromoaniline (17.20 g, 0.1 mol) into 100mL Tetrahydrofuran (THF), cooling to 0-5 ℃, dropwise adding bromoacetyl chloride (15.74 g, 0.1 mol), stirring for 30 minutes, then heating to 50-60 ℃, reacting for 2 hours, finishing the reaction, adding saturated sodium carbonate solution to adjust pH =7-8, extracting with ethyl acetate (100 mL × 2), separating, drying an organic phase with anhydrous sodium sulfate, filtering, concentrating a filtrate under reduced pressure, and purifying a residue by column chromatography (eluent: ethyl acetate/petroleum ether = 1/2) to obtain 25.38g of intermediate IV-4, wherein the yield is 86.63%. The intermediate IV-4 is a white-like solid.

The second step is that: synthesis of intermediate V-4

Intermediate iv-4 (14.65 g, 0.05 mol) and 4-piperidone (4.96 g, 0.05 mol) were added to 100mL of acetonitrile, potassium carbonate (6.95 g, 0.05 mol) was added at room temperature, the reaction was stirred at room temperature overnight, after completion of the reaction, insoluble matter was removed by filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol = 50/1) to give 12.50g of intermediate v-4 with a yield of 80.34%. Intermediate V-4 was a brown oily solid.

The third step: synthesis of Compound I-04

Intermediate v-4 (12.45 g, 0.04 mol) and (5H-imidazo [5,1-a ] isoindol-5-yl) methylamine (7.41 g, 0.04 mol) were added to 80mL of methanol, sodium borohydride (2.27 g, 0.06 mol) was added portionwise at room temperature, the reaction was stirred at room temperature for 2 hours, after the completion of the reaction, a saturated ammonium chloride solution (20 mL) was added to quench the reaction, methanol was removed by concentration under reduced pressure, 100mL of water and ethyl acetate (200 mL × 2) were added to the residue, extraction, liquid separation, the organic phase was dried over anhydrous sodium sulfate, filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol = 10/1) to give 4.48g of an off-white solid (compound i-04) with a yield of 23.31% and ESI (+) m/z ═ 480.1.

Example 5

2- (4- (((5H imidazo [5,1-a ] isoindol-5-yl) methyl) amino) piperidin-1-yl) -N- (4-cyano) acetamide (I-05)

The first step is as follows: synthesis of intermediate IV-5

Adding 4-cyanoaniline (11.80 g, 0.1 mol) into 100mL Tetrahydrofuran (THF), cooling to 0-5 ℃, dropwise adding bromoacetyl chloride (15.74 g, 0.1 mol), stirring for 30 minutes, then heating to 50-60 ℃, reacting for 2 hours, finishing the reaction, adding saturated sodium carbonate solution to adjust pH =7-8, extracting with ethyl acetate (100 mL × 2), separating, drying the organic phase with anhydrous sodium sulfate, filtering, concentrating the filtrate under reduced pressure, and purifying the residue by column chromatography (eluent: ethyl acetate/petroleum ether = 1/2) to obtain 20.71g of intermediate IV-5, wherein the yield is 86.63%. The intermediate IV-5 is a white-like solid.

The second step is that: synthesis of intermediate V-5

Intermediate iv-5 (11.95 g, 0.05 mol) and 4-piperidone (4.96 g, 0.05 mol) were added to 100mL of acetonitrile, potassium carbonate (6.95 g, 0.05 mol) was added at room temperature, the reaction was stirred at room temperature overnight, after the completion of the reaction, insoluble matter was removed by filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol = 50/1) to give 10.55g of intermediate v-5 with a yield of 82.00%. Intermediate V-5 was a brown oily solid.

The third step: synthesis of Compound I-05

Intermediate v-5 (10.29 g, 0.04 mol) and (5H-imidazo [5,1-a ] isoindol-5-yl) methylamine (7.41 g, 0.04 mol) were added to 80mL of methanol, sodium borohydride (2.27 g, 0.06 mol) was added in portions at room temperature, the reaction was stirred at room temperature for 2 hours, after the completion of the reaction, a saturated ammonium chloride solution (20 mL) was added to quench the reaction, methanol was removed by concentration under reduced pressure, 100mL of water and ethyl acetate (200 mL × 2) were added to the residue, extraction, liquid separation, the organic phase was dried over anhydrous sodium sulfate, filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol = 10/1) to obtain 7.68g of an off-white solid (compound i-05), yield 45.02%, ESI (+) m/z ═ 427.2.

Example 6

3- (4- (((5H-imidazo [5,1-a ] isoindol-5-yl) methyl) amino) piperidin-1-yl) -N- (4-fluorophenyl) propanamide (I-06)

The first step is as follows: synthesis of intermediate IV-6

Adding 4-fluoroaniline (11.11 g, 0.1 mol) into 100mL Tetrahydrofuran (THF), cooling to 0-5 ℃, dropwise adding 3-bromopropionyl chloride (17.14 g, 0.1 mol), stirring for 30 minutes, then heating to 50-60 ℃, reacting for 2 hours, finishing the reaction, adding saturated sodium carbonate solution to adjust the pH to be 7-8, extracting with ethyl acetate (100 mL multiplied by 2), separating liquid, drying an organic phase with anhydrous sodium sulfate, filtering, concentrating the filtrate under reduced pressure, and purifying the residue by column chromatography (eluent: ethyl acetate/petroleum ether = 1/2) to obtain 15.45g of an intermediate IV-6 with the yield of 62.78%. The intermediate IV-6 is a white-like solid.

The second step is that: synthesis of intermediate V-6

Intermediate iv-6 (12.30 g, 0.05 mol) and 4-piperidone (4.96 g, 0.05 mol) were added to 100mL of acetonitrile, potassium carbonate (6.95 g, 0.05 mol) was added at room temperature, the reaction was stirred at room temperature overnight, after completion of the reaction, insoluble matter was removed by filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol = 50/1) to give 12.48g of intermediate v-6 with a yield of 94.44%. Intermediate V-6 was a brown oily solid.

The third step: synthesis of Compound I-06

Intermediate v-6 (10.57 g, 0.04 mol) and (5H-imidazo [5,1-a ] isoindol-5-yl) methylamine (7.41 g, 0.04 mol) were added to 80mL of methanol, sodium borohydride (2.27 g, 0.06 mol) was added in portions at room temperature, the reaction was stirred at room temperature for 2 hours, after the completion of the reaction, a saturated ammonium chloride solution (20 mL) was added to quench the reaction, methanol was removed by concentration under reduced pressure, 100mL of water and ethyl acetate (200 mL × 2) were added to the residue, extraction, liquid separation, the organic phase was dried over anhydrous sodium sulfate, filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol = 10/1) to obtain 8.35g of an off-white solid (compound i-06), yield 48.15%, ESI (+) m/z ═ 434.2.

Example 7

2- (4- (((5H-imidazo [5,1-a ] isoindol-5-yl) methyl) amino) cyclohexyl) -N- (2-fluorophenyl) acetamide (I-07)

The first step is as follows: synthesis of intermediate VII-7

2-fluoroaniline (11.11 g, 0.1 mol) and 4-hydroxycyclohexylacetic acid (15.82 g, 0.1 mol) were added to N, N-Dimethylformamide (DMF) (30 mL), a solution of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI) (21.09 g, 0.11 mol) and 1-hydroxybenzotriazole (HOBt) (14.86 g, 0.11 mol) in DMF (30 mL) and triethylamine (20.24 g, 0.2 mol) were added thereto at room temperature, the reaction was stirred at room temperature overnight, and after completion of the reaction, 200mL of a saturated sodium carbonate solution was added, followed by extraction with ethyl acetate (100 mL. times.2), liquid separation, drying of the organic phase over anhydrous sodium sulfate, filtration, concentration of the filtrate under reduced pressure, and column chromatography purification of the residue (eluent: ethyl acetate/petroleum ether = 1/1) gave 24.50g of intermediate VII-7 with a yield of 97.49%. The intermediate VII-7 is a white-like solid.

The second step is that: synthesis of intermediate VIII-7

Intermediate vii-7 (12.57 g, 0.05 mol) and dess-martin oxidant (25.45 g, 0.06 mol) were added to 80mL Dichloromethane (DCM) and stirred at room temperature overnight, after completion of the reaction, insoluble material was removed by filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol = 100/1) to give 10.04g of intermediate viii-7 in 80.55% yield. Intermediate VIII-7 was a yellow oily solid.

The third step: synthesis of Compound I-07

Intermediate viii-7 (9.97 g, 0.04 mol) and (5H-imidazo [5,1-a ] isoindol-5-yl) methylamine (7.41 g, 0.04 mol) were added to 80mL of methanol, sodium borohydride (2.27 g, 0.06 mol) was added in portions at room temperature, the reaction was stirred at room temperature for 2 hours, after the completion of the reaction, a saturated ammonium chloride solution (20 mL) was added to quench the reaction, the methanol was removed by concentration under reduced pressure, 100mL of water and ethyl acetate (200 mL × 2) were added to the residue, extraction, liquid separation, the organic phase was dried over anhydrous sodium sulfate, filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol = 20/1) to obtain 6.28g of an off-white solid (compound i-07), yield 37.51%, ESI (+) m/z ═ 419.2.

Example 8

2- (4- (((5H-imidazo [5,1-a ] isoindol-5-yl) methyl) amino) cyclohexyl) -N- (2-bromophenyl) acetamide (I-08)

The first step is as follows: synthesis of intermediate VII-8

2-bromoaniline (17.20 g, 0.1 mol) and 4-hydroxycyclohexylacetic acid (15.82 g, 0.1 mol) were added to N, N-Dimethylformamide (DMF) (30 mL), a solution of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI) (21.09 g, 0.11 mol) and 1-hydroxybenzotriazole (HOBt) (14.86 g, 0.11 mol) in DMF (30 mL) and triethylamine (20.24 g, 0.2 mol) were added thereto at room temperature, the reaction was stirred at room temperature overnight, and after completion of the reaction, 200mL of a saturated sodium carbonate solution was added, followed by extraction with ethyl acetate (100 mL. times.2), liquid separation, drying of the organic phase over anhydrous sodium sulfate, filtration, concentration of the filtrate under reduced pressure, and column chromatography purification of the residue (eluent: ethyl acetate/petroleum ether = 1/1) gave 28.55g of intermediate VII-8 in 91.44% yield. The intermediate VII-8 is a white-like solid.

The second step is that: synthesis of intermediate VIII-8

Intermediate vii-8 (15.61 g, 0.05 mol) and dess-martin oxidant (25.45 g, 0.06 mol) were added to 80mL Dichloromethane (DCM) and stirred at room temperature overnight, after completion of the reaction, insoluble material was removed by filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol = 100/1) to give 11.25g of intermediate viii-8 in 72.54% yield. Intermediate VIII-8 was a yellow oily solid.

The third step: synthesis of Compound I-08

Intermediate viii-8 (12.41 g, 0.04 mol) and (5H-imidazo [5,1-a ] isoindol-5-yl) methylamine (7.41 g, 0.04 mol) were added to 60mL of methanol, sodium borohydride (2.27 g, 0.06 mol) was added in portions at room temperature, the reaction was stirred at room temperature for 2 hours, after the completion of the reaction, a saturated ammonium chloride solution (20 mL) was added to quench the reaction, methanol was removed by concentration under reduced pressure, 100mL of water and ethyl acetate (200 mL × 2) were added to the residue, extraction, liquid separation, the organic phase was dried over anhydrous sodium sulfate, filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol = 20/1) to obtain 7.95g of an off-white solid (compound i-08), yield 41.46%, and ESI (+) m/z ═ 479.1.

Example 9

2- (4- (((5H-imidazo [5,1-a ] isoindol-5-yl) methyl) amino) cyclohexyl) -N- (4-cyanophenyl) acetamide (I-09)

The first step is as follows: synthesis of intermediate VII-9

4-cyanoaniline (11.81 g, 0.1 mol) and 4-hydroxycyclohexylacetic acid (15.82 g, 0.1 mol) were added to N, N-Dimethylformamide (DMF) (30 mL), a solution of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI) (21.09 g, 0.11 mol) and 1-hydroxybenzotriazole (HOBt) (14.86 g, 0.11 mol) in DMF (30 mL) and triethylamine (20.24 g, 0.2 mol) were added thereto at room temperature, the reaction was stirred at room temperature overnight, 200mL of a saturated sodium carbonate solution was added thereto, ethyl acetate (100 mL. times.2) was extracted, the organic phase was separated, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: ethyl acetate/petroleum ether = 1/1) to obtain 24.52g of intermediate VII-9 with a yield of 94.92%. The intermediate VII-9 is a white-like solid.

The second step is that: synthesis of intermediate VIII-9

Intermediate vii-9 (12.92 g, 0.05 mol) and dess-martin oxidant (25.45 g, 0.06 mol) were added to 80mL of Dichloromethane (DCM) and the reaction was stirred overnight at room temperature, after completion of the reaction, insoluble matter was removed by filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol = 100/1) to give 8.95g of intermediate viii-9 in 69.84% yield. Intermediate VIII-9 was a yellow oily solid.

The third step: synthesis of Compound I-09

Intermediate viii-9 (10.25 g, 0.04 mol) and (5H-imidazo [5,1-a ] isoindol-5-yl) methylamine (7.41 g, 0.04 mol) were added to 60mL of methanol, sodium borohydride (2.27 g, 0.06 mol) was added in portions at room temperature, the reaction was stirred at room temperature for 2 hours, after the completion of the reaction, a saturated ammonium chloride solution (20 mL) was added to quench the reaction, the methanol was removed by concentration under reduced pressure, 100mL of water and ethyl acetate (200 mL × 2) were added to the residue, extraction, liquid separation, the organic phase was dried over anhydrous sodium sulfate, filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol = 20/1) to obtain 6.85g of an off-white solid (compound i-09) with a yield of 40.24% and ESI (+) m/z ═ 426.2.

Example 10

4- (((5H-imidazo [5,1-a ] isoindol-5-yl) methyl) amino) -N- (4-chlorophenyl) cyclohexane-1-carboxamide (I-10)

The first step is as follows: synthesis of intermediate VII-10

4-chloroaniline (12.76 g, 0.1 mol) and 4-hydroxycyclohexanecarboxylic acid (14.42 g, 0.1 mol) were added to N, N-Dimethylformamide (DMF) (30 mL), a solution of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI) (21.09 g, 0.11 mol) and 1-hydroxybenzotriazole (HOBt) (14.86 g, 0.11 mol) in DMF (30 mL) and triethylamine (20.24 g, 0.2 mol) were added thereto at room temperature, the reaction was stirred at room temperature overnight, and at the end of the reaction, 200mL of a saturated sodium carbonate solution was added, followed by extraction with ethyl acetate (100 mL. times.2), liquid separation, drying of the organic phase over anhydrous sodium sulfate, filtration, concentration of the filtrate under reduced pressure, and column chromatography purification of the residue (eluent: ethyl acetate/petroleum ether = 1/1) gave 19.50g of intermediate VII-10 with a yield of 76.85%. The intermediate VII-10 is a white-like solid.

The second step is that: synthesis of intermediate VIII-10

Intermediate VII-10 (12.69 g, 0.05 mol) and dess-martin oxidant (25.45 g, 0.06 mol) were added to 80mL of Dichloromethane (DCM) and the reaction was stirred overnight at room temperature, after completion of the reaction, insoluble matter was removed by filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol = 100/1) to give 9.88g of intermediate VIII-10 with a yield of 78.50%. Intermediate VIII-10 was a yellow oily solid.

The third step: synthesis of Compound I-10

Intermediate viii-10 (10.07 g, 0.04 mol) and (5H-imidazo [5,1-a ] isoindol-5-yl) methylamine (7.41 g, 0.04 mol) were added to 60mL of methanol, sodium borohydride (2.27 g, 0.06 mol) was added in portions at room temperature, the reaction was stirred at room temperature for 2 hours, after the completion of the reaction, a saturated ammonium chloride solution (20 mL) was added to quench the reaction, the methanol was removed by concentration under reduced pressure, 100mL of water and ethyl acetate (200 mL × 2) were added to the residue, extraction, liquid separation, the organic phase was dried over anhydrous sodium sulfate, filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol = 10/1) to obtain 8.46g of an off-white solid (compound i-10) with a yield of 50.24% and ESI (+) m/z ═ 421.2.

Example 11

4- (((5H-imidazo [5,1-a ] isoindol-5-yl) methyl) amino) -N-phenylcyclohexane-1-carboxamide (I-11)

The first step is as follows: synthesis of intermediate VII-11

Aniline (9.31 g, 0.1 mol) and 4-hydroxycyclohexyl carboxylic acid (14.42 g, 0.1 mol) were added to N, N-Dimethylformamide (DMF) (30 mL), a DMF (30 mL) solution of 1-ethyl- (3-dimethylaminopropyl) carbonyldiimine hydrochloride (EDCI) (21.09 g, 0.11 mol) and 1-hydroxybenzotriazole (HOBt) (14.86 g, 0.11 mol) and triethylamine (20.24 g, 0.2 mol) were added at room temperature, the reaction was stirred at room temperature overnight, and at the end of the reaction, 200mL of a saturated sodium carbonate solution was added, ethyl acetate (100 mL × 2) was extracted, the organic phase was separated, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: ethyl acetate/petroleum ether = 1/1) to give 16.52g of intermediate vii-11 with a yield of 75.33%. The intermediate VII-11 is a white-like solid.

The second step is that: synthesis of intermediate VIII-11

Intermediate VII-11 (10.96 g, 0.05 mol) and dess-martin oxidizer (25.45 g, 0.06 mol) were added to 80mL of Dichloromethane (DCM) and the reaction was stirred overnight at room temperature, after completion of the reaction, insoluble matter was removed by filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol = 100/1) to give 8.96g of intermediate VIII-11 with a yield of 82.48%. Intermediate VIII-11 was a yellow oily solid.

The third step: synthesis of Compound I-11

Intermediate viii-11 (8.69 g, 0.04 mol) and (5H-imidazo [5,1-a ] isoindol-5-yl) methylamine (7.41 g, 0.04 mol) were added to 60mL of methanol, sodium borohydride (2.27 g, 0.06 mol) was added in portions at room temperature, the reaction was stirred at room temperature for 2 hours, after the completion of the reaction, a saturated ammonium chloride solution (20 mL) was added to quench the reaction, methanol was removed by concentration under reduced pressure, 100mL of water and ethyl acetate (200 mL × 2) were added to the residue, extraction, liquid separation, the organic phase was dried over anhydrous sodium sulfate, filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol = 50/1) to obtain 6.59g of an off-white solid (compound i-11) with a yield of 42.63% and ESI (+) m/z ═ 387.2.

Example 12

4- (((5H-imidazo [5,1-a ] isoindol-5-yl) methyl) amino) -N- (4-fluorophenyl) cyclohexane-1-carboxamide (I-12)

The first step is as follows: synthesis of intermediate VII-12

4-fluoroaniline (11.11 g, 0.1 mol) and 4-hydroxycyclohexylcarboxylic acid (14.42 g, 0.1 mol) were added to N, N-Dimethylformamide (DMF) (30 mL), a solution of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI) (21.09 g, 0.11 mol) and 1-hydroxybenzotriazole (HOBt) (14.86 g, 0.11 mol) in DMF (30 mL) and triethylamine (20.24 g, 0.2 mol) were added thereto at room temperature, the reaction was stirred at room temperature overnight, and at the end of the reaction, 200mL of a saturated sodium carbonate solution was added, followed by extraction with ethyl acetate (100 mL. times.2), liquid separation, drying of the organic phase over anhydrous sodium sulfate, filtration, concentration of the filtrate under reduced pressure, and column chromatography purification of the residue (eluent: ethyl acetate/petroleum ether = 1/1) gave 18.50g of intermediate VII-12 with a yield of 77.97%. The intermediate VII-12 is a white-like solid.

The second step is that: synthesis of intermediate VIII-12

Intermediate vii-12 (11.86 g, 0.05 mol) and dess-martin oxidant (25.45 g, 0.06 mol) were added to 80mL Dichloromethane (DCM) and stirred at room temperature overnight, after completion of the reaction, insoluble material was removed by filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol = 100/1) to give 10.05g of intermediate viii-12 in 85.44% yield. Intermediate VIII-12 was a yellow oily solid.

The third step: synthesis of Compound I-12

Intermediate viii-12 (9.41 g, 0.04 mol) and (5H-imidazo [5,1-a ] isoindol-5-yl) methylamine (7.41 g, 0.04 mol) were added to 60mL of methanol, sodium borohydride (2.27 g, 0.06 mol) was added in portions at room temperature, the reaction was stirred at room temperature for 2 hours, after the completion of the reaction, a saturated ammonium chloride solution (20 mL) was added to quench the reaction, the methanol was removed by concentration under reduced pressure, 100mL of water and ethyl acetate (200 mL × 2) were added to the residue, extraction, liquid separation, the organic phase was dried over anhydrous sodium sulfate, filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol = 50/1) to obtain 7.23g of an off-white solid (compound i-12) with a yield of 44.69% and ESI (+) m/z of 405.2.

IDO inhibitor IC₅₀Biological evaluation

The compounds I-01 to I-12 and comparative example Navoximod were tested for their IDO inhibitory effect.

Among them, Navoximod is a known IDO inhibitor having a CAS No. of 1402837-78-8 and a structural formula shown below:

human IDO1/HEK293 cells were used for IDO kynurenine analysis as follows:

human IDO1/HEK293 cells were seeded in 384-well black-wall clear-bottom tissue culture plates using RPM 1/phenol red-free medium containing 10% FBS at 10.000 cells/50 μ L/well, and then 125nL of a specific concentration of compound was added to each well using the ECHO liquid handling system. Incubate with 5% CO at 37 deg.C₂Cells were incubated for 20 hours and compound treatment was stopped by adding trichloroacetic acid to a final concentration of 0.2%. The cells were further incubated at 50 ℃ for 30 minutes. Equal volumes of supernatant (20 μ L) and 0.2% Ehrlich reagent (4-dimethylaminobenzaldehyde) in glacial acetic acid were mixed in a new bottoming 384-well plate. The plate was then incubated at room temperature for 30 minutes. Absorbance at 490nm was measured on an Envision microplate reader.

Compound IC was confronted with 100% inhibition using a 500nm count reference standard treatment, counting no compound and 0% inhibition using a dimethyl sulfoxide (DMSO) treatment₅₀The value is calculated.

The results of the experiment are shown in the following table:

。

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 compound having the structure of formula I:

Ⅰ

2. The compound of claim 1 having the structure of formula i or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of compounds having the structure:

。

3. a process for the preparation of a compound having the structure of formula i, or a pharmaceutically acceptable salt thereof, as claimed in claim 1 or claim 2, wherein when X represents N, the compound has the structure:

the preparation method comprises the following steps:

wherein halo represents F, Cl, Br, I;

and (3) synthesizing an intermediate IV:

and (3) synthesis of an intermediate V:

synthesis of compounds having the structure of formula la:

4. The method according to claim 3, wherein the first reaction temperature is-20 to 80 ℃;

and/or the second reaction temperature is-20 to 100 ℃;

and/or the third reaction temperature is 0-25 ℃.

5. The production method according to claim 3, wherein the first solvent is at least one selected from the group consisting of dioxane, tetrahydrofuran, acetonitrile, and toluene;

6. A process for the preparation of a compound having the structure of formula i, or a pharmaceutically acceptable salt thereof, as claimed in claim 1 or claim 2, wherein when X represents CH, the compound has the structure:

the preparation method comprises the following steps:

and (3) synthesizing an intermediate VII:

synthesis of intermediate viii:

synthesis of compounds having the structure of formula ib:

7. The preparation method according to claim 6, wherein the fourth reaction temperature is 20 to 120 ℃;

and/or the fifth reaction temperature is-80-25 ℃;

and/or the third reaction temperature is 0-25 ℃.

8. The method according to claim 6, wherein the fourth solvent is at least one selected from the group consisting of dichloromethane, N-dimethylformamide, and N, N-dimethylaniline;

and/or the base is at least one of triethylamine and diisopropylethylamine;

and/or the oxidant is at least one selected from 2, 3-dichloro-5, 6-dicyan p-benzoquinone, pyridinium chlorochromate, pyridinium chlorodichromate, dess-martin oxidant and Stevens oxidant;

9. A pharmaceutical composition comprising a compound of formula i as claimed in claim 1 or claim 2 or a pharmaceutically acceptable salt thereof.

10. Use of a compound having the structure of formula i or a pharmaceutically acceptable salt thereof according to claim 1 or 2 or a pharmaceutical composition according to claim 9 as an IDO inhibitor in the manufacture of a medicament for the treatment of a tumour.