A method for preparing a beclin1-atg14l interaction inhibitor

Abstract

The application belongs to the field of organic synthesis and particularly relates to a preparation method of a Beclin1-ATG14L interaction inhibitor, a reaction formula of which is as follows: comprising the following steps: step (1): compound A is dissolved in an organic solvent, an acid catalyst is added to perform a ring closing reaction to obtain compound B; step (2): compound B is reacted with compound C in the presence of an organic solvent and a base to obtain compound D; and step (3): compound D is deprotected in the presence of an acid and an organic solvent to obtain compound E. The synthetic process route of the application is original, fills the blank of no preparation method of compound E at home and abroad, is simple in synthetic route, low in reaction equipment requirement, mild in reaction condition, easy to operate, high in yield, obtains multiple novel intermediates in the synthesis process, and the intermediates are stable in property, good in reusability and easy to realize industrialized scale production.

Description

TECHNICAL FIELD

[0001] The application relates to a preparation method of a Beclin1-ATG14L interaction inhibitor and belongs to the technical fields of medicines and chemistry. BACKGROUND

[0002] VPS34 is involved in the formation of two different complexes: autophagy initiation complex I (VPS15-VPS34-ATG14L-Beclin 1) and endosome 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] The compound can destroy the formation of autophagy initiation complex I to inhibit autophagy to a certain extent and does not destroy the interaction of Beclin 1-UVRAG, and has a certain selectivity. Based on the autophagy and non-autophagy functions of Beclin-1, the Beclin-1 regulation strategy has a good development prospect in cancer treatment.

[0005] There is no report on the preparation method of the compound in the prior art, and therefore there is an urgent need for an economic, fast, simple, high-yield and high-purity synthesis method of the compound. SUMMARY

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

[0007] In order to overcome the defects of the prior art and realize the requirements of commercial scale production, the following preferred technical scheme is adopted in the application.

[0008] The application provides a preparation method of a compound of formula E, and the reaction formula is as follows:

[0009]

[0010] The method comprises the following steps:

[0011] Step (1): compound A is dissolved in an organic solvent, an acid catalyst is added to perform a ring-closing reaction, and compound B is obtained;

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

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

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

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

[0016] As a further improvement of the present application, 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 application, including but not limited to, the volume of the organic solvent for the ring-closing reaction in step (1) is 2-15 times, preferably 6-13 times, the mass of compound A;

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

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

[0020] As a further improvement of the present application, the ring-closing reaction in step (1) comprises the following steps: reacting compound A in an organic solvent for a certain period of time, and then adding an acid to catalyze the ring-closing reaction.

[0021] Preferably, the certain period of time is selected from no less than 3 hours, further no less than 6 hours, no less than 9 hours, no less than 12 hours, no less than 15 hours, for example, 6-20 hours, 8-18 hours, and for example, 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 application, the reaction time of the step (1) cyclization reaction, preferably, the cyclization reaction catalyzed by the addition of acid, 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, 3-12 hours, further for example, 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 application, the present inventors respectively use single organic solvent or acid reflux, wherein the single organic solvent reflux has no target product generated, which proves that the cyclization cannot be completed, and when refluxing in a single acid solvent, the target compound can also be synthesized, but the reaction yield is unstable, so the conditions of mixing organic solvent and acid are selected, and the yield under the conditions of the present application is more stable and can be directly purified on the column without post-treatment.

[0024] As a further improvement of the present application, including but not limited to, the base in the step (2) is selected from one or more of the following: triethylamine, diisopropyl ethylamine, pyridine and other organic bases; one or more of the following: sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, cesium carbonate, potassium phosphate, lithium hydroxide, sodium hydroxide, potassium hydroxide inorganic bases, preferably potassium carbonate;

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

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

[0027] As a further improvement of the present application, including but not limited to, the organic solvent in the step (2) is selected from one or more of the following: 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 application, including but not limited to, the volume of the organic solvent used in the step (2) is 2-15 times, preferably 6-12 times, the amount of compound C in grams;

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

[0030] As a further improvement of the present application, including but not limited to, the reaction time in the step (2) is 10-24 h, preferably 15-20 h;

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

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

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

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

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

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

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

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

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

[0040] As a further improvement of the present application, the first aspect of the present application is selected from PMB amino protecting group, which is less stable than PMB protection or replacement of benzyl protecting group, and has high yield. Therefore, the substrate selected from PMB amino protecting group has higher reaction activity under the reaction conditions of the present application, and the product yield is higher, which is beneficial to industrialization and scale-up production.

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

[0042]

[0043] comprising the following steps:

[0044] Step (4): substituting compound M and compound N in the presence of an organic solvent and a base to obtain compound C;

[0045] As a further improvement of the present application, including but not limited to, the base of the step (4) is selected from one or more of the following: triethylamine, diisopropylethylamine, pyridine and other organic bases; one or more of the following inorganic bases: sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, cesium carbonate, lithium hydroxide, sodium hydroxide, potassium hydroxide, preferably pyridine;

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

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

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

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

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

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

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

[0053]

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

[0055] (1) The synthetic process route of the present application has originality, fills the blank of no preparation method of compound E at home and abroad, and the synthetic route of the present application is simple, the reaction equipment requirement is low, the reaction condition is mild, the operation is simple, the yield is high, and is suitable for industrialized scale production.

[0056] (2) The present inventors surprise to find that, in step (2), when compound C is not protected by PMB, compound B will react with the bare amino group first, resulting in a very low yield of only about 16%; when compound C is replaced by benzyl protection, the yield is also only about 34%, and during the debenzyl protection, the compound B connected will also be removed as a byproduct, therefore, the compound C selected with PMB amino protection not only has high reactivity and yield, but also has the advantages of stable product, less impurities and high purity, which is beneficial to industrialized scale production.

[0057] (3) The present inventors use single organic solvent or acid reflux in step (1) respectively, wherein no target product is generated in single organic solvent reflux, which proves that the ring closure cannot be completed, and when single acid is used as a solvent for reflux, the target compound can also be synthesized, but the reaction yield is unstable, therefore, the condition of mixing organic solvent and acid is selected, and under this condition, the yield is stable and direct column purification after treatment is not needed.

[0058] (4) The preparation method of the present application obtains a plurality of novel intermediates in the synthesis process, and the intermediates have stable properties and good reusability, the intermediate compounds in the present application are judged according to the method of producing subsequent API products or the obtained products:

[0059] The novel intermediate compounds of the present application contribute to the creative method of producing subsequent API products, and the preparation method of the novel intermediate compounds of the present application is simple in operation, high in yield, good in product quality, and easy to realize industrialized mass production. BRIEF DESCRIPTION OF DRAWINGS

[0060] Figure 1 The H-NMR spectrum of compound D in Example 6 of the present application is shown in Figure 1. 1

[0061] Figure 2 The H-NMR spectrum of compound E in Example 7 of the present application is shown in Figure 2. 1 DETAILED DESCRIPTION

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

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

[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), then cyclobutyl formic acid (863 mg, 8.62 mmol) and DIPEA (1.39 g, 10.78 mmol) were added dropwise, followed by HCTU (4.45 g, 10.78 mmol) and heated to 80 °C for 4 h until the raw material was completely reacted, then the solvent was evaporated, and column purification was performed to obtain compound A (790 mg, yield 57.7%).

[0068] Example 2

[0069] Synthesis of compound B:

[0070]

[0071] Compound A (790 mg, 4.14 mmol) was dissolved in N,N-dimethylformamide (10 mL), and after 12 h of reaction in an oil bath at 135 °C, acetic acid (99.6 mg, 1.66 mmol) was added dropwise, and the reaction was continued for 6 h. After the reaction was completed, column purification was performed to obtain compound B (550 mg, yield 77%).

[0072] 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.

[0073]

[0074] As can be seen from Table 1 above, when only a single N,N-dimethylformamide solvent is used, no product is obtained; when only a single acetic acid solvent is used, the yield is only 51%; and when N,N-dimethylformamide + acetic acid is used, the yield can reach 77%.

[0075] Example 5

[0076] Synthesis of compound C:

[0077]

[0078] 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 reaction was allowed to proceed at room temperature for 12 h, and TLC (PE:DCM = 3:1) monitoring showed a new spot with Rf = 0.4. The reaction was quenched with water (20 mL), extracted with dichloromethane twice, washed with water, and the resulting solution was evaporated to dryness and purified by column chromatography to give compound C (910 mg, yield 63.85%).

[0079] Example 6

[0080] Synthesis of compound D:

[0081]

[0082] 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 sequentially. The reaction was allowed to proceed at 60°C for 18 h, and the resulting solution was filtered and evaporated to dryness. The residue was directly loaded onto a column and purified by column chromatography to give compound D (920 mg, yield 93.02%). 1 The H-NMR spectrum is shown in Figure 1 .

[0083] Example 7

[0084] Synthesis of compound E:

[0085]

[0086] Compound D (1.01 g, 1.72 mmol) was dissolved in dichloromethane (10 mL), and TFA (1.96 g, 17.16 mmol) and anisole (55.7 mg, 0.51 mmol) were added sequentially. The reaction was allowed to proceed at 40°C for 8 h, and the resulting solution was evaporated to dryness and purified by column chromatography to give compound E (780 mg, yield 97.03%). 1 The H-NMR spectrum is shown in Figure 2 .

[0087] Comparative Example 1

[0088] Synthesis of compound C1:

[0089]

[0090] Compound Ml (499 mg, 4.12 mmol) was dissolved in dichloromethane (18 mL), 3-oxo-3,4-dihydro-2H-l,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 allowed to proceed at room temperature for 12 h. The resulting reaction solution was quenched with water (18 mL) and extracted twice with dichloromethane (20 mL) and washed with water. The resulting solution was evaporated and purified by column chromatography using dichloromethane to give compound CI (180 mg, 15.8% yield).

[0091] Comparative Example 2

[0092] Synthesis of compound C2:

[0093]

[0094] Compound M2 (768 mg, 3.63 mmol) was dissolved in dichloromethane (20 mL), 3-oxo-3,4-dihydro-2H-l,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 allowed to proceed at room temperature for 12 h. The resulting reaction solution was quenched with water (18 mL) and extracted twice with dichloromethane (20 mL) and washed with water. The resulting solution was evaporated and purified by column chromatography using dichloromethane to give compound C2 (440 mg, 34% yield).

Claims

1. A method for preparing compound E, characterized in that, The reaction formula is shown below: Includes the following steps: Step (1): Dissolve compound A in an organic solvent, add acid to catalyze the cyclization reaction, and obtain compound B; Step (2): Compound B and compound C are reacted in the presence of an organic solvent and a base to obtain compound D; Step (3): Compound D is deprotected in the presence of acid and organic solvent to obtain compound E; The acid used in the cyclization reaction of step (1) is selected from one or more of acetic acid, formic acid, and hydrochloric acid; The base in step (2) is selected from one or more of triethylamine, diisopropylethylamine, and pyridine organic bases, or one or more of sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, cesium carbonate, potassium phosphate, lithium hydroxide, sodium hydroxide, and potassium hydroxide inorganic bases. The acid in step (3) is selected from one or more of trifluoroformic acid, trifluoroacetic acid, p-toluenesulfonic acid, hydrochloric acid, and sulfuric acid.

2. The preparation method according to claim 1, characterized in that, The acid used in the cyclization reaction of step (1) is acetic acid.

3. The preparation method according to claim 1, characterized in that, The organic solvent for the cyclization 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. And / or, in step (1), the molar ratio of compound A to acid is 1:(0.1-1); And / or, the volume of the organic solvent used in the cyclization reaction of step (1) is 2 to 15 times the mass of compound A in g; And / or, the reaction temperature for the ring-closing reaction in step (1) is 100–145 °C; And / or, the reaction time for the ring-closing reaction in step (1) is 9 to 32 hours.

4. The preparation method according to claim 3, characterized in that, The organic solvent for the cyclization reaction in step (1) is N,N-dimethylacetamide; And / or, in step (1), the molar ratio of compound A to acid is 1:(0.3 to 0.5); And / or, the volume of the organic solvent used in the cyclization reaction of step (1) is 6 to 13 times the mass of compound A in g; And / or, the reaction temperature of the ring-closing reaction in step (1) is 125–140 °C; And / or, the reaction time for the ring-closing reaction in step (1) is 15 to 20 hours.

5. The preparation method according to any one of claims 1 to 4, characterized in that, The cyclization reaction in step (1) includes the following steps: after reacting compound A in an organic solvent for a period of time, acid is added to catalyze the cyclization reaction.

6. The preparation method according to claim 5, characterized in that, The reaction time is no less than 3 hours.

7. The preparation method according to claim 6, characterized in that, The reaction takes 6 to 20 hours.

8. The preparation method according to claim 5, characterized in that, The reaction time for the ring-closing reaction catalyzed by the addition of acid shall not be less than 1 hour.

9. The preparation method according to claim 8, characterized in that, The reaction time is 3 to 12 hours.

10. The preparation method according to claim 1, characterized in that, The alkali used in step (2) is potassium carbonate.

11. The preparation method according to claim 1, characterized in that, Step (2) satisfies one or more of the following conditions: In step (2), the molar ratio of compound B to the base is 1:(1-5); And / or, in step (2), the molar ratio of compound B to compound C is 1:(1-2); And / or, the organic solvent in step (2) is selected from ethyl acetate, isopropyl acetate, dichloromethane, tetrahydrofuran, dioxane, etc. One or more of toluene, acetonitrile, N,N-dimethylformamide, and N,N-dimethylacetamide; And / or, the volume of the organic solvent used in step (2) is 2 to 15 times the mass of compound C in g; And / or, the reaction temperature in step (2) is 30–90 °C; And / or, the reaction time for step (2) is 10 to 24 hours.

12. The preparation method according to claim 11, characterized in that, Step (2) satisfies one or more of the following conditions; In step (2), the molar ratio of compound B to the base is 1:(1.5-2.5); And / or, in step (2), the molar ratio of compound B to compound C is 1:(1.1 to 1.3); And / or, the organic solvent in step (2) is N,N-dimethylformamide; And / or, the volume of the organic solvent used in step (2) is 6 to 12 times the mass of compound C in g; And / or, the reaction temperature of step (2) is 55-65°C; And / or, the reaction time of step (2) is 15 to 20 hours.

13. The preparation method according to claim 1, characterized in that, The deprotection reaction in step (3) includes: dissolving compound D in an organic solvent, adding acid and a catalyst to carry out the deprotection reaction to obtain compound E; step (3) satisfies one or more of the following conditions: The acid used in the deprotection reaction in step (3) is trifluoroacetic acid; And / or, in step (3), the molar ratio of compound D to acid is 1:(5-12); And / or, in step (3) the deprotection reaction, the catalyst is anisole; And / or, in step (3), the molar ratio of compound D to catalyst is 1:(0.1 to 0.6); And / or, the organic solvent in step (3) is selected from one or more of toluene, methanol, ethanol, tetrahydrofuran, ethyl acetate, dichloromethane, dioxane, and acetonitrile; And / or, the volume of the organic solvent used in step (3) is 2 to 15 times the mass of compound D in g; And / or, the reaction temperature in step (3) is 20–60 °C; And / or, the reaction time for step (3) is 5 to 15 hours.

14. The preparation method according to claim 13, characterized in that, Step (3) satisfies one or more of the following conditions: In step (3), the molar ratio of compound D to acid is 1:(8-11); And / or, in step (3), the molar ratio of compound D to catalyst is 1:(0.2-0.4); And / or, the organic solvent in step (3) is dichloromethane; And / or, the volume of the organic solvent used in step (3) is 6 to 10 times the mass of compound D (g); And / or, the reaction temperature of step (3) is 35–45°C. And / or, the reaction time of step (3) is 6 to 10 hours.

15. The preparation method according to claim 1, characterized in that, The preparation method of compound C is as follows: Includes the following steps: 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.

16. The preparation method according to claim 15, characterized in that, The base in step (4) is selected from one or more of triethylamine, diisopropylethylamine, and pyridine organic bases, or one or more of sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, cesium carbonate, lithium hydroxide, sodium hydroxide, and potassium hydroxide inorganic bases.

17. The preparation method according to claim 15, characterized in that, The step (4) satisfies one or more of the following conditions: the base in the step (4) is pyridine; And / or, in step (4), the molar ratio of compound N to base is 1:(1-5); And / or, in step (4), the molar ratio of compound N to compound M is 1:(1-2); And / or, the organic solvent in step (4) is selected from one or more of dichloromethane, tetrahydrofuran, dioxane, toluene, acetonitrile, N,N-dimethylformamide, and N,N-dimethylacetamide; And / or, the volume of the organic solvent used in step (4) is 8 to 25 times the mass of compound M in g; And / or, the reaction temperature in step (4) is 10–30 °C; And / or, the reaction time for step (4) is 8 to 20 hours.

18. The preparation method according to claim 17, characterized in that, Step (4) satisfies one or more of the following conditions: In step (4), the molar ratio of compound N to the base is 1:(2-3.5); And / or, in step (4), the molar ratio of compound N to compound M is 1:(1.1 to 1.3); And / or, the organic solvent in step (4) is dichloromethane; And / or, the reaction temperature of step (4) is 15-25°C; And / or, the reaction time of step (4) is 10 to 15 hours.

19. Compound C and compound D have the following structures:

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