Preparation method of Beclin1-ATG14L interaction inhibitor

Abstract

The invention belongs to the field of organic synthesis, and particularly relates to a preparation method of a Beclin1-ATG14L interaction inhibitor, the reaction formula is as follows: # imgabs0 #, and the preparation method comprises the following steps: step (1), dissolving a compound A in an organic solvent, adding acid for catalysis, and carrying out ring closing reaction to obtain a compound B; (2) reacting the compound B with a compound C in the presence of an organic solvent and alkali to obtain a compound D; and step (3): performing deprotection on the compound D in the presence of acid and an organic solvent to obtain a compound E. The synthesis process route is original, the blank that no compound E preparation method exists at home and abroad is filled, the synthesis route is simple, the requirement for reaction equipment is low, the reaction condition is mild, operation is easy and convenient, the yield is high, multiple brand-new intermediates are obtained in the synthesis process, the intermediates are stable in property and good in reproducibility, and industrial large-scale production is easy to achieve.

Description

Technical Field

[0001] The invention relates to a method for preparing a Beclin1-ATG14L interaction inhibitor, and belongs to the technical field of medicine and chemistry. Background Art

[0002] VPS34 participates in the formation of two different complexes: autophagy initiation complex I (VPS15-VPS34-ATG14L-Beclin 1) and endosomal transport complex II (VPS15-VPS34-UVRAG-Beclin 1). ATG14L interacts with Beclin 1 through a coiled-coil domain (CCD). The structural formula of the Beclin1-ATG14L interaction inhibitor is as follows:

[0003]

[0004] This compound can disrupt the formation of autophagy initiation complex I, thereby inhibiting autophagy to a certain extent, and will not disrupt the Beclin 1-UVRAG interaction, which has a certain selectivity. Based on the autophagic and non-autophagic functions of Beclin-1, the targeted Beclin-1 regulation strategy has good development prospects in cancer treatment.

[0005] There is no report on the preparation method of the compound in the prior art, so there is an urgent need for a synthesis method of the compound that is economical, fast, easy to operate, simple to post-process, high in yield and high in purity. Summary of the invention

[0006] In order to solve the technical problems in the above reports, the present invention provides a method for synthesizing a Beclin1-ATG14L interaction inhibitor.

[0007] In order to overcome the defects of the prior art and achieve the requirements of commercial scale-up production, the present invention adopts the following preferred technical solutions:

[0008] The first aspect of the present invention provides a method for preparing a compound of formula E, and the reaction formula is as follows:

[0009]

[0010] The steps include:

[0011] Step (1): dissolving compound A in an organic solvent, adding an acid catalyst to carry out a ring-closure reaction to obtain compound B;

[0012] Step (2): reacting compound B with compound C in the presence of an organic solvent and a base to obtain compound D;

[0013] Step (3): Compound D is deprotected in the presence of an acid and an organic solvent to obtain Compound E.

[0014] As a further improvement of the present invention, including but not limited to, the acid used in the ring-closing reaction in step (1) is selected from one or more of acetic acid, formic acid, hydrochloric acid, etc., preferably acetic acid;

[0015] As a further improvement of the present invention, including but not limited to, the molar ratio of compound A to acid in step (1) is 1:(0.1-1), preferably 1:(0.3-0.5);

[0016] As a further improvement of the present invention, including but not limited to, the organic solvent for the ring-closing reaction in step (1) is selected from one or more of ethyl acetate, isopropyl acetate, dichloromethane, tetrahydrofuran, dioxane, toluene, acetonitrile, N,N-dimethylformamide, and N,N-dimethylacetamide, preferably N,N-dimethylformamide;

[0017] As a further improvement of the present invention, including but not limited to, the volume of the organic solvent used in the ring-closure reaction in step (1) is 2 to 15 times, preferably 6 to 13 times, the mass of compound A in g;

[0018] As a further improvement of the present invention, including but not limited to, the reaction temperature of the ring-closing reaction in step (1) is 100 to 145° C., preferably 125 to 140° C.;

[0019] As a further improvement of the present invention, including but not limited to, the reaction time of the ring-closure reaction in step (1) is 9 to 32 hours, preferably 15 to 20 hours;

[0020] As a further improvement of the present invention, the ring-closing reaction in step (1) comprises the following steps: Compound A is selectively reacted in an organic solvent for a period of time, and then an acid catalyst is added to carry out a ring-closing reaction.

[0021] Preferably, the selective reaction period is selected from not less than 3 hours, further not less than 6 hours, not less than 9 hours, not less than 12 hours, not less than 15 hours, for example 6 to 20 hours, 8 to 18 hours, such as 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours or 18 hours.

[0022] As a further improvement of the present invention, in the step (1) the ring-closing reaction, preferably, the reaction time of adding an acid catalyst for the ring-closing reaction is not less than 1 hour, further not less than 3 hours, further not less than 6 hours, not less than 9 hours, for example 3 to 12 hours, such as 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours or 12 hours.

[0023] As a further improvement of the present invention, the inventors used a single organic solvent or acid reflux respectively. No target product was generated when a single organic solvent was refluxed, proving that the ring closure could not be completed. The target compound could also be synthesized when a single acid was used as solvent reflux, but the reaction yield was unstable. Therefore, the conditions of mixing an organic solvent with an acid were selected. Under the conditions of the present invention, the yield was relatively stable and no post-treatment was required, and the product could be directly purified on a column.

[0024] As a further improvement of the present invention, including but not limited to, the base in step (2) is selected from one or more of organic bases such as triethylamine, diisopropylethylamine, pyridine, etc.; sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, cesium carbonate, potassium phosphate, lithium hydroxide, sodium hydroxide, potassium hydroxide, one or more of inorganic bases, preferably potassium carbonate;

[0025] As a further improvement of the present invention, including but not limited to, the molar ratio of compound B to base in step (2) is 1:(1-5), preferably 1:(1.5-2.5);

[0026] As a further improvement of the present invention, including but not limited to, the molar ratio of compound B to compound C in step (2) is 1:(1-2), preferably 1:(1.1-1.3);

[0027] As a further improvement of the present invention, including but not limited to, the organic solvent in step (2) is selected from one or more of ethyl acetate, isopropyl acetate, dichloromethane, tetrahydrofuran, dioxane, toluene, acetonitrile, N,N-dimethylformamide, N,N-dimethylacetamide, preferably N,N-dimethylformamide;

[0028] As a further improvement of the present invention, including but not limited to, the volume amount of the organic solvent in step (2) mL is 2 to 15 times the mass amount of compound C g, preferably 6 to 12 times;

[0029] As a further improvement of the present invention, including but not limited to, the reaction temperature of step (2) is 30 to 90° C., preferably 55 to 65° C.;

[0030] As a further improvement of the present invention, including but not limited to, the reaction time of step (2) is 10 to 24 hours, preferably 15 to 20 hours.

[0031] As a further improvement of the present invention, including but not limited to, the deprotection reaction in step (3) comprises: dissolving compound D in an organic solvent, adding an acid and a catalyst to carry out a deprotection reaction to obtain compound E;

[0032] As a further improvement of the present invention, including but not limited to, in the deprotection reaction of step (3), the acid is selected from one or more of trifluoroformic acid, trifluoroacetic acid, p-toluenesulfonic acid, hydrochloric acid, and sulfuric acid, preferably trifluoroacetic acid;

[0033] As a further improvement of the present invention, including but not limited to, the molar ratio of compound D to acid in step (3) is 1:(5-12), preferably 1:(8-11);

[0034] As a further improvement of the present invention, including but not limited to, in the deprotection reaction of step (3), the catalyst is selected from anisole;

[0035] As a further improvement of the present invention, including but not limited to, the molar ratio of compound D to catalyst in step (3) is 1:(0.1-0.6), preferably 1:(0.2-0.4);

[0036] As a further improvement of the present invention, including but not limited to, the organic solvent in step (3) is selected from one or more of toluene, methanol, ethanol, tetrahydrofuran, ethyl acetate, dichloromethane, dioxane, and acetonitrile, preferably dichloromethane;

[0037] As a further improvement of the present invention, including but not limited to, the volume amount of the organic solvent in step (3) mL is 2 to 15 times the mass amount of compound D g, preferably 6 to 10 times;

[0038] As a further improvement of the present invention, including but not limited to, the reaction temperature of step (3) is 20 to 60°C, preferably 35 to 45°C

[0039] As a further improvement of the present invention, including but not limited to, the reaction time of step (3) is 5 to 15 hours, preferably 6 to 10 hours.

[0040] As a further improvement of the present invention, the first aspect of the present invention uses the PMB amino protecting group, which has a more stable yield and a higher yield than not adding PMB protection or replacing the benzyl protecting group. Therefore, the use of the PMB amino protecting substrate under the reaction conditions of the present invention has a higher reaction activity and a higher product yield, which is conducive to industrial scale-up production.

[0041] The second aspect of the present invention provides a method for preparing a compound of formula C, and the reaction formula is as follows:

[0042]

[0043] The steps include:

[0044] Step (4): Compound M and Compound N are subjected to a substitution reaction in the presence of an organic solvent and a base to obtain Compound C;

[0045] As a further improvement of the present invention, including but not limited to, the base in step (4) is selected from one or more of organic bases such as triethylamine, diisopropylethylamine, pyridine, etc.; sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, cesium carbonate, lithium hydroxide, sodium hydroxide, potassium hydroxide, one or more of inorganic bases, preferably pyridine;

[0046] As a further improvement of the present invention, including but not limited to, the molar ratio of compound N to base in step (4) is 1:(1-5), preferably 1:(2-3.5);

[0047] As a further improvement of the present invention, including but not limited to, the molar ratio of compound N to compound M in step (4) is 1:(1-2), preferably 1:(1.1-1.3);

[0048] As a further improvement of the present invention, including but not limited to, the organic solvent in step (4) is selected from one or more of dichloromethane, tetrahydrofuran, dioxane, toluene, acetonitrile, N,N-dimethylformamide, N,N-dimethylacetamide, preferably dichloromethane;

[0049] As a further improvement of the present invention, including but not limited to, the volume amount mL of the organic solvent in step (4) is 8 to 25 times the mass amount g of the compound M;

[0050] As a further improvement of the present invention, including but not limited to, the reaction temperature of step (4) is 10 to 30° C., preferably 15 to 25° C.;

[0051] As a further improvement of the present invention, including but not limited to, the reaction time of step (4) is 8 to 20 hours, preferably 10 to 15 hours;

[0052] The third aspect of the present invention provides a compound C and a compound D, the structures of which are as follows:

[0053]

[0054] The present invention has the following beneficial effects:

[0055] (1) The synthetic process route of the present invention is original, which fills the gap of the lack of a preparation method for compound E at home and abroad. In addition, the synthetic route of the present invention is simple, has low requirements for reaction equipment, mild reaction conditions, simple operation, high yield, and is suitable for industrial scale-up production.

[0056] (2) After many experiments, the inventors surprisingly found that in step (2), when compound C is not protected with PMB, compound B will react with the exposed amino group first, resulting in a very low yield of only about 16%; when compound C is replaced with benzyl protection, the yield is only about 34%, and when the benzyl protection is removed, the by-product of the connected compound B will be removed together. Therefore, the use of compound C protected with PMB amino group not only has high reaction activity and high yield, but also has the advantages of stable product, less impurities and high purity, which is conducive to industrial scale-up production.

[0057] (3) The inventors used a single organic solvent or acid reflux in step (1). No target product was generated when a single organic solvent was refluxed, indicating that the ring closure could not be completed. The target compound could be synthesized when a single acid was used as solvent reflux, but the reaction yield was unstable. Therefore, the condition of mixing an organic solvent with an acid was selected. The yield under this condition was relatively stable and no post-treatment was required, and the product could be directly purified by column purification.

[0058] (4) The preparation method of the present invention obtains multiple new intermediates during the synthesis process, and the intermediates have stable properties and good reproducibility. The intermediate compounds of the present invention are judged according to the method for producing subsequent API products or the resulting products:

[0059] The novel intermediate compound of the present invention contributes to a creative method for producing subsequent API products therefrom, and the preparation method of the novel intermediate compound of the present invention is simple to operate, has a high yield, good product quality, and is easy to realize large-scale industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0060] Figure 1 is the compound D of Example 6 of the present invention 1 H-NMR spectrum;

[0061] Figure 2 is the compound E of Example 7 of the present invention 1 H-NMR spectrum; DETAILED DESCRIPTION

[0062] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0063] Unless otherwise specified, the experimental methods used in the following examples are all conventional methods; the reagents, materials, etc. used in the following examples, unless otherwise specified, can be obtained from commercial channels.

[0064] Example 1

[0065] Synthesis of compound A:

[0066]

[0067] Compound G (780 mg, 7.19 mmol) was dissolved in N,N-dimethylformamide (10 mL), and cyclobutanecarboxylic acid (863 mg, 8.62 mmol) and DIPEA (1.39 g, 10.78 mmol) were added dropwise in sequence, and HCTU (4.45 g, 10.78 mmol) was added and heated to 80°C for 4 h to allow the raw materials to react completely. The solvent was then evaporated and purified by column to obtain compound A (790 mg, yield 57.7%).

[0068] Example 2

[0069] Synthesis of compound B:

[0071]

[0072] Compound A (790 mg, 4.14 mmol) was dissolved in N,N-dimethylformamide (10 mL), reacted in an oil bath at 135°C for 12 h, then acetic acid (99.6 mg, 1.66 mmol) was added dropwise, and the reaction was continued for 6 h. After the reaction was completed, the product was purified by column chromatography to obtain compound B (550 mg, yield 77%).

[0073] Example 3-4 The preparation method of compound B is as shown in Example 2, and the conditions and results are shown in Table 1 below;

[0074]

[0075]

[0076] It can be seen from Table 1 that when only N,N-dimethylformamide is used as a single solvent, no product is obtained; when only acetic acid is used as a single solvent, the yield is only 51%; when N,N-dimethylformamide + acetic acid are used as a mixture, the yield can reach 77%.

[0077] Example 5

[0078] Synthesis of compound C:

[0079]

[0080] Compound M (912 mg, 3.78 mmol) was dissolved in dichloromethane (20 mL), 3-oxo-3,4-dihydro-2H-1,4-benzoxazine-6-sulfonyl chloride (780 mg, 3.15 mmol) was added, and pyridine (747 mg, 9.45 mmol) was added dropwise. The mixture was reacted at room temperature for 12 h, and a new spot of Rf=0.4 appeared on TLC (PE:DCM=3:1). The reaction solution was quenched with water (20 mL), extracted twice with dichloromethane, and then washed with water. The solution was evaporated to dryness and purified by column to obtain compound C (910 mg, yield 63.85%).

[0081] Example 6

[0082] Synthesis of compound D:

[0083]

[0084] Compound C (912 mg, 2.02 mmol) was dissolved in N,N-dimethylformamide (10 mL), and compound B (290 mg, 1.68 mmol) and potassium carbonate (464 mg, 3.36 mmol) were added in sequence. The mixture was heated to 60 °C for 18 h. After filtration, part of the solvent was evaporated and the mixture was directly loaded on the sample for column purification to obtain compound D (920 mg, yield 93.02%). 1 H-NMR spectrum Figure 1 shown.

[0085] Example 7

[0086] Synthesis of compound E:

[0087]

[0088] Compound D (1.01 g, 1.72 mmol) was dissolved in dichloromethane (10 mL), and then TFA (1.96 g, 17.16 mmol) and anisole (55.7 mg, 0.51 mmol) were added in sequence. The mixture was heated to 40°C for 8 h. After the reaction, the solvent was evaporated and purified by column to obtain compound E (780 mg, yield 97.03%). 1 H-NMR spectrum Figure 2 shown.

[0089] Comparative Example 1

[0090] Synthesis of compound C1:

[0091]

[0092] Compound M1 (499 mg, 4.12 mmol) was dissolved in dichloromethane (18 mL), 3-oxo-3,4-dihydro-2H-1,4-benzoxazine-6-sulfonyl chloride (850 mg, 3.43 mmol) was added, and pyridine (815 mg, 10.3 mmol) was added dropwise. The reaction was carried out at room temperature for 12 h, and the resulting reaction solution was quenched with water (18 mL), extracted twice with dichloromethane (20 mL), and then washed with water. The resulting solution was evaporated to dryness, and purified by column chromatography with dichloromethane to obtain compound C1 (180 mg, yield 15.8%).

[0093] Comparative Example 2

[0094] Synthesis of compound C2:

[0095]

[0096] Compound M2 (768 mg, 3.63 mmol) was dissolved in dichloromethane (20 mL), 3-oxo-3,4-dihydro-2H-1,4-benzoxazine-6-sulfonyl chloride (0.75 g, 3.03 mmol) was added, and pyridine (719 mg, 9.1 mmol) was added dropwise. The reaction was carried out at room temperature for 12 h, and the resulting reaction solution was quenched with water (18 mL), extracted twice with dichloromethane (20 mL), and then washed with water. The resulting solution was evaporated to dryness, and purified by column chromatography with dichloromethane to obtain compound C2 (440 mg, yield 34%).

Claims

1. A method for preparing compound E, characterized in that: The reaction formula is as follows: The steps include: Step (1): dissolving compound A in an organic solvent, adding an acid catalyst to carry out a ring-closure reaction to obtain compound B; Step (2): reacting compound B with compound C in the presence of an organic solvent and a base to obtain compound D; Step (3): Compound D is deprotected in the presence of an acid and an organic solvent to obtain Compound E.

2. The preparation method according to claim 1, characterized in that: The acid used in the ring-closing reaction in step (1) is selected from one or more of acetic acid, formic acid, hydrochloric acid, etc., preferably acetic acid.

3. The preparation method according to claim 1, characterized in that: The organic solvent for the ring-closing reaction in step (1) is selected from one or more of ethyl acetate, isopropyl acetate, dichloromethane, tetrahydrofuran, dioxane, toluene, acetonitrile, N,N-dimethylformamide, and N,N-dimethylacetamide, preferably N,N-dimethylformamide; And / or, preferably, the molar ratio of compound A to acid in step (1) is 1:(0.1-1), preferably 1:(0.3-0.5); and / or, preferably, the volume amount mL of the organic solvent used in the ring-closure reaction in step (1) is 2-15 times the mass amount g of compound A, preferably 6-13 times; And / or, preferably, the reaction temperature of the ring-closure reaction in step (1) is 100 to 145° C., preferably 125 to 140° C.; And / or, preferably, the reaction time of the ring-closure reaction in step (1) is 9 to 32 hours, preferably 15 to 20 hours.

4. The preparation method according to any one of claims 1 to 3, characterized in that: The step (1) of ring-closing reaction comprises the following steps: compound A is reacted in an organic solvent for a period of time, and then an acid catalyst is added to carry out a ring-closing reaction; and / or, preferably, the reaction may be selectively carried out for a period of time selected from not less than 3 hours, preferably 6 to 20 hours; And / or, preferably, the reaction time of adding acid catalyst to carry out the ring-closure reaction is not less than 1 hour, preferably 3 to 12 hours.

5. The preparation method according to claim 1, characterized in that: The base in step (2) is selected from one or more organic bases such as triethylamine, diisopropylethylamine, pyridine, etc.; one or more inorganic bases such as sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, cesium carbonate, potassium phosphate, lithium hydroxide, sodium hydroxide, potassium hydroxide, preferably potassium carbonate; Step (2) satisfies one or more of the following conditions: And / or, preferably, the molar ratio of compound B to base in step (2) is 1:(1-5), preferably 1:(1.5-2.5); And / or, preferably, the molar ratio of compound B to compound C in step (2) is 1:(1-2), preferably 1:(1.1-1.3); and / or, preferably, the organic solvent in step (2) is selected from one or more of ethyl acetate, isopropyl acetate, dichloromethane, tetrahydrofuran, dioxane, toluene, acetonitrile, N,N-dimethylformamide, N,N-dimethylacetamide, preferably N,N-dimethylformamide; And / or, preferably, the volume amount mL of the organic solvent in step (2) is 2 to 15 times the mass amount g of compound C, preferably 6 to 12 times; And / or, preferably, the reaction temperature of step (2) is 30 to 90°C, preferably 55 to 65°C; And / or, preferably, the reaction time of step (2) is 10 to 24 hours, preferably 15 to 20 hours.

6. The preparation method according to claim 1, characterized in that: The deprotection reaction in step (3) comprises: dissolving compound D in an organic solvent, adding an acid and a catalyst to carry out a deprotection reaction, and obtaining compound E; Step (3) satisfies one or more of the following conditions: And / or, preferably, in the deprotection reaction of step (3), the acid is selected from one or more of trifluoroformic acid, trifluoroacetic acid, p-toluenesulfonic acid, hydrochloric acid, and sulfuric acid, preferably trifluoroacetic acid; And / or, preferably, the molar ratio of compound D to acid in step (3) is 1:(5-12), preferably 1:(8-11); And / or, preferably, in the deprotection reaction of step (3), the catalyst is selected from anisole; And / or, preferably, the molar ratio of compound D to catalyst in step (3) is 1:(0.1-0.6), preferably 1:(0.2-0.4); and / or, preferably, the organic solvent in step (3) is selected from one or more of toluene, methanol, ethanol, tetrahydrofuran, ethyl acetate, dichloromethane, dioxane, acetonitrile, preferably dichloromethane; And / or, preferably, the volume amount mL of the organic solvent in step (3) is 2 to 15 times the mass amount g of compound D, preferably 6 to 10 times; And / or, preferably, the reaction temperature of step (3) is 20-60°C, preferably 35-45°C And / or, preferably, the reaction time of step (3) is 5 to 15 hours, preferably 6 to 10 hours.

7. The preparation method according to claim 1, characterized in that: The preparation method of the compound C is as follows: The steps include: Step (4): Compound M and Compound N are subjected to a substitution reaction in the presence of an organic solvent and a base to obtain Compound C.

8. The preparation method according to claim 6, characterized in that: The base in step (4) is selected from one or more organic bases such as triethylamine, diisopropylethylamine, and pyridine; one or more inorganic bases such as sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, cesium carbonate, lithium hydroxide, sodium hydroxide, and potassium hydroxide, preferably pyridine; Step (4) satisfies one or more of the following conditions: And / or, preferably, the molar ratio of compound N to base in step (4) is 1:(1-5), preferably 1:(2-3.5); And / or, preferably, the molar ratio of compound N to compound M in step (4) is 1:(1-2), preferably 1:(1.1-1.3); and / or, preferably, the organic solvent in step (4) is selected from one or more of dichloromethane, tetrahydrofuran, dioxane, toluene, acetonitrile, N,N-dimethylformamide, N,N-dimethylacetamide, preferably dichloromethane; And / or, preferably, the volume amount mL of the organic solvent in step (4) is 8 to 25 times the mass amount g of compound M; And / or, preferably, the reaction temperature of step (4) is 10 to 30°C, preferably 15 to 25°C; And / or, preferably, the reaction time of step (4) is 8 to 20 hours, preferably 10 to 15 hours.

9. A compound C and a compound D, the structures of which are as follows:

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