CN115819352A - Preparation method of transferase inhibitor and intermediate thereof - Google Patents

Abstract

The invention relates to a preparation method of a transferase inhibitor and an intermediate thereof; the invention provides a preparation method of a histone lysine N-methyltransferase inhibitor compound 1- (4- (4-methoxybenzoyl) oxy) phenethyl) -2- (4- (trifluoromethyl) benzamido) -1H-benzo [ d ] imidazole-5-carboxylic acid and an intermediate compound thereof, namely 2-amino-1- (4- (4-methoxybenzoyl) oxy) phenethyl) -1H-benzo [ d ] imidazole-5-carboxylic acid tert-butyl ester for the first time; except for the starting materials, the intermediate compounds are novel compounds. The preparation method provided by the invention has mild reaction conditions, good yield and low post-treatment difficulty, provides guarantee for subsequent preparation of high-quality medicines, and is suitable for small-amount preparation in a laboratory and industrial large-scale production.

Description

Preparation method of transferase inhibitor and intermediate thereof

Technical Field

The invention belongs to the field of organic synthesis, relates to a preparation method of a transferase inhibitor and an intermediate thereof, and more particularly relates to a preparation method of a histone lysine methyltransferase inhibitor compound 1- (4- (4-methoxybenzoyl) oxy) phenethyl) -2- (4- (trifluoromethyl) benzamido) -1H-benzo [ d ] imidazole-5-carboxylic acid and an intermediate compound 2-amino-1- (4- (4-methoxybenzoyl) oxy) phenethyl) -1H-benzo [ d ] imidazole-5-carboxylic acid tert-butyl ester thereof.

Background

Euchromatin histone lysine N-methyltransferase G9a, also known as EHMT2 (euchromatic histone-lysine N-methyltransferase 2), catalyzes the methylation of lysine 373 (K373) at positions 9 (H3K 9) and p53 of histone H3. Research shows that G9a is closely related to the maintenance of human immunodeficiency virus-1 latency, cocaine addiction, central nervous system disorder, gene expression and transcription, hematopoietic stem cell differentiation and other processes or diseases. In addition, it can also act as a co-activator of p21 transcription leading to apoptosis. G9a is over-expressed in various cancers such as leukemia, prostatic cancer, liver cancer, lung cancer and the like, and the knockout of the G9a gene can inhibit the growth of cells such as prostatic cancer, lung cancer and the like. In a mouse model, a slowing of the progression of acute myelogenous leukemia can be observed when G9a is knocked out. Therefore, the G9a mediated histone methylation is closely related to the occurrence and development of tumors, is considered to be a novel anti-tumor target with a wide prospect, and the development of inhibitors thereof is also receiving more and more attention.

Wherein, the structure of the compound 1- (4- (4-methoxybenzoyl) oxy) phenethyl) -2- (4- (trifluoromethyl) benzamido) -1H-benzo [ d ] imidazole-5-carboxylic acid (product code number BIX-01338) is shown as the following formula, and the compound is a histone lysine N-methyltransferase inhibitor which plays a very important role in the field of medicine, but no synthetic method is reported in the prior art.

Therefore, the design and implementation of a synthetic method which is suitable for industrial production, simple and convenient to operate and high in yield is the key point of research and development of the technicians in the field.

Disclosure of Invention

The invention aims to provide a preparation method of a transferase inhibitor and an intermediate thereof, and particularly aims to provide a preparation method of a histone lysine methyltransferase inhibitor compound 1- (4- (4-methoxybenzoyl) oxy) phenethyl) -2- (4- (trifluoromethyl) benzamido) -1H-benzo [ d ] imidazole-5-carboxylic acid and an intermediate compound thereof, namely 2-amino-1- (4- (4-methoxybenzoyl) oxy) phenethyl) -1H-benzo [ d ] imidazole-5-carboxylic acid tert-butyl ester, so as to solve the problems in the background technology.

To achieve the above object, a first aspect of the present invention provides a process for preparing tert-butyl 2-amino-1- (4- (4-methoxybenzoyl) oxy) phenethyl) -1H-benzo [ d ] imidazole-5-carboxylate, an intermediate compound, comprising the steps of:

step 1: reacting the compound 1 with the compound 2 to obtain a compound 3;

step 2: carrying out reduction reaction on the compound 3 to obtain a compound 4;

and step 3: closing the ring of the compound 4 and cyanogen bromide to obtain a compound 5;

and 4, step 4: reaction of compound 5 with compound 6 affords compound 7.

As a preferred technical scheme of the invention, the preparation method of the step 1 comprises the following steps: dissolving the compound 1 into an organic solvent at room temperature, adding alkali, continuously stirring, adding the compound 2, stirring and reacting for 4-6 h at room temperature, adding water after the TLC monitoring reaction is finished, extracting, combining organic phases, washing, drying and concentrating to obtain the compound; preferably, the organic solvent in step 1 is selected from one or more of acetonitrile, tetrahydrofuran, dichloromethane, dioxane, 2-methyltetrahydrofuran, toluene, N-dimethylformamide, methyl tert-butyl ether, diethyl ether and dimethyl sulfoxide, and more preferably is N, N-dimethylformamide; preferably, the base in step 1 is selected from one or more of sodium hydroxide, sodium carbonate, potassium carbonate, triethylamine and N, N-diisopropylethylamine, and more preferably N, N-diisopropylethylamine; preferably, the molar ratio of the compound 1 to the compound 2 is 1 (1-1.5).

As a preferred technical scheme of the invention, the preparation method of the step 2 comprises the following steps: dissolving the compound 3 in an organic solvent, adding a noble metal catalyst, cooling to-15-10 ℃, reacting for 6-8h under the condition of hydrogen, filtering, concentrating and purifying by column chromatography after TLC monitoring reaction is finished, thus obtaining the compound; or, the preparation method of step 2 comprises the following steps: dissolving the compound 3 into an organic solvent, adding a reducing agent, reacting for 6-8h at the condition of-15-room temperature, after the TLC monitoring reaction is finished, filtering, concentrating, and purifying by column chromatography to obtain the compound; preferably, the organic solvent in step 2 is selected from one or more of dichloromethane, isobutanol, methanol, n-butanol, butyl acetate and ethyl acetate, and more preferably is a combination of methanol and ethyl acetate; when the organic solvent is selected from the combination of methanol and ethyl acetate, the volume ratio of the methanol to the ethyl acetate is preferably 1 (0.8-1.2); preferably, the noble metal catalyst in step 2 is selected from one of palladium carbon, raney nickel and platinum carbon; more preferably palladium carbon, preferably, the mass percent of the noble metal palladium or platinum in the palladium carbon or the platinum carbon is 5-20%, and preferably, the mass percent of the noble metal palladium or platinum in the palladium carbon or the platinum carbon is 20%; the reducing agent in step 2 is selected from one of lithium aluminum hydride, sodium hydrosulfite, iron powder/amine chloride and zinc powder/amine chloride, and more preferably iron powder/amine chloride or zinc powder/amine chloride.

As a preferable technical scheme of the invention, the preparation method of the step 3 comprises the following steps: dissolving a compound 4 into an organic solvent, adding cyanogen bromide to control the temperature in the reaction to be not more than 35 ℃ for reaction, monitoring by TLC (thin layer chromatography) to ensure that the reaction is completely converted, then carrying out reduced pressure concentration to remove the organic solvent, adding a saturated sodium bicarbonate aqueous solution to adjust the pH value to 7-8, stirring for 1-2 h, filtering, and drying to obtain the compound 4; preferably, the organic solvent in step 3 is selected from one or more of dichloromethane, isobutanol, methanol, n-butanol, butyl acetate and ethyl acetate, and more preferably methanol; preferably, the molar ratio of the compound 4 to the cyanogen bromide in the step 3 is 1 (1.2-2).

As a preferred technical solution of the present invention, the preparation method of step 4 comprises the following steps: dissolving a compound 5 into an organic solvent 1, adding alkali, cooling to-3 ℃, slowly dropwise adding a mixed solution of a compound 6 and an organic solvent 2, stirring for 8-15min after dropwise adding, adding water for diluting after TLC monitoring reaction, adjusting the pH to 5-6 by using a dilute hydrochloric acid solution, extracting, combining organic phases, drying, concentrating, and purifying by column chromatography to obtain the compound 5; preferably, the organic solvent 1 in step 4 is selected from one or more of dichloromethane, toluene, tetrahydrofuran, butyl acetate and ethyl acetate, and more preferably dichloromethane; preferably, the base in step 4 is selected from one or more of potassium acetate, pyridine, sodium carbonate, potassium carbonate, triethylamine, N-diisopropylethylamine, potassium phosphate and sodium hydroxide, and more preferably is N, N-diisopropylethylamine; preferably, the molar ratio of the compound 5 to the base in the step 4 is 1 (3-3.5); preferably, the organic solvent 2 in step 4 is an anhydrous solvent selected from one or more of dichloromethane, toluene, tetrahydrofuran, butyl acetate and ethyl acetate, more preferably dichloromethane, and the anhydrous solvent in the present invention is obtained by a conventional method well known to those skilled in the art, for example, by a molecular sieve drying process or directly purchasing a solvent that has been subjected to a drying process; preferably, the molar ratio of the compound 5 to the compound 6 in the step 4 is 1 (0.95-1.05), more preferably, the molar ratio of the compound 5 to the compound 6 in the step 4 is 1.

In a second aspect, the present invention provides a process for the preparation of compound 10, comprising the steps of:

and 5: carrying out condensation reaction on the compound 7 and the compound 8 to obtain a compound 9;

step 6: deprotection of compound 9 under acidic conditions affords compound 10.

As a preferred technical scheme of the invention, the preparation method of the step 5 comprises the following steps: mixing the compound 7, the compound 8, a condensing agent, a catalyst and an organic solvent, stirring for 14-18h, adding water into a reaction system after LCMS monitoring reaction is finished, filtering and drying after all solids are separated out to obtain the compound; preferably, the condensing agent in step 5 is EDCI, DCC, HATU, TBTU, T ₃ P, pyBOp, HOBt, more preferably EDCI; preferably, the catalyst in step 5 is DMAP; preferably, the organic solvent in step 5 is selected from one or more of dioxane, 2-methyltetrahydrofuran, toluene, N-dimethylformamide and methyl tert-butyl ether, and more preferably is N, N-dimethylformamide; preferably, the molar ratio of the compound 7 to the compound 8 in the step 5 is 1 (1-2); preferably, the molar ratio of the compound 7 to the condensing agent in the step 5 is 1 (1-2); preferably, the molar ratio of the condensing agent to the catalyst in step 5 is 1 (0.1 to 0.2).

As a preferred technical solution of the present invention, the preparation method of step 6 comprises the following steps: dissolving the compound 9 in an organic solvent, adding acid, reacting for 4-8h at room temperature, monitoring by LCMS after the reaction is finished, concentrating, adding water and acetonitrile, pulping, filtering and drying to obtain the compound; preferably, the acid in step 6 is selected from hydrochloric acid solution, hydrochloric acid gas, hydrochloric acid methanol solution, hydrochloric acid dioxane solution, and trifluoroacetic acid, and more preferably trifluoroacetic acid; preferably, the organic solvent in step 6 is selected from one or more of dichloromethane, isobutanol, methanol, n-butanol, butyl acetate and ethyl acetate, and more preferably dichloromethane.

A third aspect of the present invention provides an intermediate compound 7 useful in the preparation of compound 10, having the structure:

compared with the prior art, the invention has the beneficial effects that:

1) The invention provides a preparation method of a histone lysine N-methyltransferase inhibitor compound 1- (4- (4-methoxybenzoyl) oxy) phenethyl) -2- (4- (trifluoromethyl) benzamido) -1H-benzo [ d ] imidazole-5-carboxylic acid and an intermediate compound thereof, namely 2-amino-1- (4- (4-methoxybenzoyl) oxy) phenethyl) -1H-benzo [ d ] imidazole-5-carboxylic acid tert-butyl ester for the first time; except for the starting materials, the intermediate compounds are novel compounds.

2) The invention uses easily obtained and cheap initial raw materials to prepare a target product through 6 steps of reaction, wherein the steps 1 and 2 can be carried out in a two-step continuous way, the step 3 has very clean reaction, and a pure compound 5 can be obtained through simple post-treatment; after the reaction in the step 5 is finished, water is added to separate out a pure compound 9, so that the reaction yield of the whole route is high, and the reaction effect is very good; particularly for the step 4, the invention unexpectedly finds that the single product with high yield can be obtained by combining the acyl chloride compound and the addition amount thereof, controlling the type and the proportion of the solvent for the reaction, the screened alkali, the reaction feeding method and the like according to the characteristics of the compound 5, and the invention has the advantages of conventional reagents, environmental friendliness, short reaction time and good atom economy.

3) The preparation method provided by the invention has mild reaction conditions, good yield and low post-treatment difficulty, provides guarantee for subsequent preparation of high-quality medicines, and is suitable for small-amount preparation in a laboratory and industrial large-scale production.

Detailed Description

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental methods in the following examples, which are not specified under specific conditions, are generally carried out under conventional conditions.

The starting materials or reagents used in the examples are, unless otherwise specified, commercially available.

The room temperature mentioned in the examples is 10 to 35 ℃. Unless otherwise indicated, the reagents were used without purification. All solvents were purchased from commercial suppliers, such as Aldrich (Aldrich), and used without treatment.

The reaction was analyzed by TLC and/or by LCMS, and termination of the reaction was judged by consumption of starting material. Thin Layer Chromatography (TLC) for analysis was performed on glass plates (EMD Chemicals) precoated with silica gel 60F254 0.25 mm plates, visualized with UV light (254 nm) and/or iodine on silica gel, and/or heated with TLC stains such as alcoholic phosphomolybdic acid, ninhydrin solution, potassium permanganate solution, or ceric sulfate solution.

The abbreviations used in the present invention have the usual meaning in the art, such as: DMF means N, N-dimethylformamide; EA represents ethyl acetate; DIPEA represents N, N-diisopropylethylamine; PE represents petroleum ether; DCC denotes N, N' -dicyclohexylcarbodiimide; EDCI represents 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride; HATU represents 2- (7-azabenzotriazole) -N, N' -tetramethyluronium hexafluorophosphate; TBTU represents 2- (1H-benzotriazol L-1-yl) -1, 3-tetramethyluronium tetrafluoroborate; t is ₃ P represents 1-propylphosphoric anhydride; pyBOp represents 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate; HOBt represents 1-hydroxybenzotriazole; DMAP represents 4-dimethylaminopyridine; meOH denotes methanol.

Example 1

Dissolving compound 1 (24.9g, 103.2mmol) in DMF (250 mL) at room temperature, adding DIPEA (26.7g, 206.4mmol), stirring for 5min, adding compound 2 (15.6g, 113.5mmol), stirring at room temperature for 5h, monitoring by TLC, after the reaction is finished, adding water (1250 mL), EA (500 mL. Times.3) extracting, combining organic phases, washing with saturated saline (1.5L), washing aqueous phase with EA (600 mL), combining organic phases, and using anhydrous Na ₂ SO ₄ Drying, concentration and suction drying gave crude compound 3 (37 g).

Example 2

The crude compound 3 (37 g) obtained in example 1 was dissolved in a mixed solution of MeOH (400 mL) and EA (400 mL), palladium on carbon (7 g, palladium content 20%) was added, the temperature was reduced to-15 ℃, hydrogen balloon was added, gas was replaced three times, and then the reaction was stirred for 7h, after completion of the tlc monitoring reaction, filtration, concentration, and column chromatography purification (PE: EA =10, 1) were performed to obtain a pure solid compound 4 (31.9 g, two-step yield 94%, LCMS: ES ⁺ m/z＝329[M+H] ⁺ ； ¹ H-NMR(400MHz,DMSO-d6)δ9.18(s,1H),7.15(dd,J＝8.0,1.6Hz,1H),7.10(d,J＝2.0Hz,1H),7.07(d,J＝8.4Hz,2H),6.69(d,J＝8.4Hz,2H),6.45(d,J＝8.4Hz,1H),5.18(t,J＝6.4Hz,1H),4.68(s,2H),3.25(dd,J＝13.2,7.2Hz,2H),2.76(t,J＝8.0Hz,2H),1.49(s,9H))。

Example 3

Compound 4 (31.9 g, 97.1mmol) was dissolved in MeOH (200 mL), cyanogen bromide (12.3 g,116.5 mmol) was added and the reaction was carried out at 30 deg.C, TLC monitored that the reaction was completely converted, methanol was removed by concentration under reduced pressure, and saturated NaHCO was used ₃ Adjusting the pH of the water phase to 7-8, stirring for 1h, filtering, and vacuum drying to obtain pure solid compound 5 (31.6g, 92%, LCMS: ES) ⁺ m/z＝354[M+H] ⁺ ； ¹ H-NMR(400MHz,DMSO-d6)δ9.30(s,1H),9.02(s,2H),7.86(d,J＝1.2Hz,1H),7.75(dd,J＝8.4,1.6Hz,1H),7.44(d,J＝8.4Hz,1H),6.99(d,J＝8.4Hz,2H),6.62(d,J＝8.4Hz,2H),4.37(t,J＝7.2Hz,2H),2.88(t,J＝7.2Hz,2H),1.55(s,9H))。

Example 4

Dissolving compound 5 (20g, 56.6mmol) in DCM (800 mL), adding DIPEA (25.6g, 198.1mmol), slowly adding a solution of compound 6 (9.7g, 56.6mmol) in ultra-dry DCM (200 mL) when the temperature is reduced to 0 ℃ under ice bath conditions, and after dropwise adding, continuing to addStirring was continued for 10min, TLC was carried out after completion of the reaction, water (400 mL) was added, the pH of the aqueous phase was adjusted to 5-6 with dilute hydrochloric acid solution (0.6 mol/L), extraction was carried out with DCM (600 mL. Times.3), the organic phases were combined, and anhydrous Na was used ₂ SO ₄ Drying, filtration, concentration under reduced pressure, and column chromatography purification (DCM: meOH = 40) ⁺ m/z＝488[M+H] ⁺ ； ¹ H-NMR(400MHz,DMSO-d6)δ8.26(s,2H),8.06(d,J＝8.8Hz,2H),7.80(d,J＝1.2Hz,1H),7.68(dd,J＝8.4,1.6Hz,1H),7.41(d,J＝8.4Hz,1H),7.37(d,J＝8.4Hz,2H),7.16(d,J＝8.4Hz,2H),7.11(d,J＝9.2Hz,2H),4.39(t,J＝7.2Hz,2H),3.88(s,3H),3.01(t,J＝7.2Hz,2H),1.55(s,9H))。

Example 5

Compound 7 (18.5g, 37.9mmol), compound 8 (10.8g, 56.9mmol), EDCI (10.9g, 56.9mmol), DMAP (1.39g, 11.4mmol) were dissolved in DMF (370 mL), the reaction was stirred for 1693 h, after completion of the reaction, water (2L) was added to the reaction mixture to precipitate a large amount of solid, which was filtered and dried to obtain pure compound 9 (23.5g, 94%, LCMS: ES) as a solid ⁺ m/z＝660[M+H] ⁺ ； ¹ H-NMR(400MHz,DMSO-d6)δ12.94(s,1H),8.41(d,J＝8.0Hz,2H),8.10(d,J＝1.2Hz,1H),8.03(d,J＝8.8Hz,2H),7.87(d,J＝8.4Hz,2H),7.79(dd,J＝8.4,1.2Hz,1H),7.55(d,J＝8.4Hz,1H),7.35(d,J＝8.4Hz,2H),7.13(d,J＝8.4Hz,2H),7.09(d,J＝8.8Hz,2H),4.56(t,J＝7.2Hz,2H),3.86(s,3H),3.18(t,J＝7.2Hz,2H),1.57(s,9H))。

Example 6

Dissolving compound 9 (20g, 30.3mmol) in DCM (500 mL), adding trifluoroacetic acid (100 mL), reacting at room temperature for 6h, monitoring reaction by LCMS, removing DCM by concentration under reduced pressure, adding water (200 mL), pulping acetonitrile (200 mL) for 16h, filtering, and dryingTo obtain pure compound 10 (17.6 g,96%, LCMS: ES) ⁺ m/z＝604[M+H] ⁺ ， ¹ H-NMR(400MHz,DMSO-d6)δ12.97(m,2H),8.42(d,J＝8.4Hz,2H),8.12(d,J＝1.2Hz,1H),8.04(d,J＝8.8Hz,2H),7.88(d,J＝8.4Hz,2H),7.85(dd,J＝8.8,1.6Hz,1H),7.58(d,J＝8.8Hz,1H),7.38(d,J＝8.4Hz,2H),7.15(d,J＝8.4Hz,2H),7.10(d,J＝8.8Hz,2H),4.56(t,J＝6.8Hz,2H),3.87(s,3H),3.19(t,J＝6.8Hz,2H))。

All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes and modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the present invention as defined by the appended claims.

Claims (10)

1. A method for preparing compound 7, comprising the steps of:

step 1: reacting the compound 1 with the compound 2 to obtain a compound 3;

step 2: carrying out reduction reaction on the compound 3 to obtain a compound 4;

and step 3: closing the ring of the compound 4 and cyanogen bromide to obtain a compound 5;

and 4, step 4: reaction of compound 5 with compound 6 affords compound 7.

2. The method of claim 1, wherein the method of step 1 comprises the steps of: dissolving the compound 1 into an organic solvent at room temperature, adding alkali, continuously stirring, adding the compound 2, stirring and reacting for 4-6 h at room temperature, after TLC monitoring reaction is finished, adding water for dilution, extracting, combining organic phases, washing, drying and concentrating to obtain the compound.

3. The method of claim 1, wherein the step 2 method comprises the steps of: dissolving the compound 3 into an organic solvent, adding a noble metal catalyst, cooling to-15-10 ℃, reacting for 6-8h under the condition of hydrogen, filtering, concentrating, and purifying by column chromatography after TLC monitoring reaction is finished, thus obtaining the compound.

4. The method of claim 1, wherein the step 3 comprises the steps of: dissolving compound 4 in organic solvent, adding cyanogen bromide to control reaction temperature not to exceed 35 deg.C for reaction, monitoring by TLC for complete reaction conversion, concentrating under reduced pressure to remove organic solvent, adding saturated NaHCO ₃ Adjusting the pH value of the aqueous solution to 7-8, stirring for 1-2 h, filtering and drying to obtain the water-soluble organic fertilizer.

5. The preparation method according to claim 4, wherein the molar ratio of the compound 4 to the cyanogen bromide is 1 (1.2-2).

6. The method of claim 1, wherein the step 4 comprises the steps of: dissolving a compound 5 into an organic solvent 1, adding alkali, cooling to-3 ℃, slowly dropwise adding a mixed solution of a compound 6 and an organic solvent 2, stirring for 8-15min after dropwise adding, adding water for diluting after TLC monitoring reaction, adjusting the pH to 5-6 by using a dilute hydrochloric acid solution, extracting, combining organic phases, drying, concentrating, and purifying by column chromatography to obtain the compound.

7. The preparation method according to claim 6, wherein the base is selected from one or more of potassium acetate, pyridine, sodium carbonate, potassium carbonate, triethylamine, N-diisopropylethylamine, potassium phosphate, and sodium hydroxide; the molar ratio of the compound 5 to the alkali is 1 (3-3.5).

8. The method according to claim 6, wherein the molar ratio of the compound 5 to the compound 6 is 1 (0.95 to 1.05).

9. A method of preparing compound 10, comprising the steps of:

and 5: carrying out condensation reaction on the compound 7 and the compound 8 to obtain a compound 9;

and 6: deprotection of compound 9 under acidic conditions affords compound 10.

10. An intermediate compound 7 useful in the preparation of compound 10, having the formula: