New crystalline forms of spinosad and methods for their preparation

Abstract

This invention discloses novel crystal forms of spinosad and their preparation methods, belonging to the field of biomedicine. The novel crystal forms SN-II and SN-III of this invention overcome the instability of commercially available amorphous spinosad, and are also easier for industrial production, offering better economic benefits.

Description

Invention Field

[0001] This invention belongs to the field of biomedicine, specifically relating to a new crystal form of spinosad and its preparation method. Background of the Invention

[0002] Spinosad, whose main components are spinosyn A and spinosyn D, is a secondary metabolite produced by the aerobic fermentation of the soil actinomycete *Spinosaemonas spinosa*. It belongs to the macrolide class of insecticidal antibiotics and was developed by Dow AgroSciences in the early 1990s. Spinosad exhibits high insecticidal selectivity and environmental compatibility, primarily targeting the binding site on the nicotinic acetylcholine receptor (nAChR) in the insect nervous system. It has a broad insecticidal spectrum, showing high insecticidal activity against pests including Lepidoptera, Thysanoptera, Coleoptera, Diptera, Hymenoptera, and Isoptera, but exhibits extremely low toxicity to mammals, fish, and amphibians. Compared to other chemical and biological pesticides, it possesses high selective toxicity and environmental safety advantages. Therefore, spinosad has broad market application prospects and is currently widely used for pest control in crops such as cotton, tea, and vegetables.

[0003] Spinosad is composed of two components, spinosyn A and spinosyn D, with spinosyn A comprising approximately 85% and spinosyn D approximately 15%. Its chemical name is:

[0004] (2R,3aS,5aR,5bS,9S,13S,14R,16aS,16bR)-13-(((2R,5S,6R)-5-(dimethylamino)-6-methyltetrahydro-2H-pyran-2-yl)oxy)-9-ethyl-14-methyl-2-(((1R,2S,3R,4S,5R) -2,3,4-trimethoxy-5-methylcyclohexyl)oxy)-2,3,3a,5a,5b,6,9,10,11,12,13,14,16a,16b-tetradecahydro-1H-as-indaceno[3,2-d][1]oxacyclododecine-7,15-dione

[0005] The chemical structure is:

[0006]

[0007] Spinosad A has R=H and its molecular formula is C. 41 H 65 NO10 Spinosad D: R=CH3, molecular formula is C 42 H 67 NO 10 .

[0008] Polymorphism of drugs refers to the phenomenon where a drug composed of the same chemical substance has two or more spatial arrangements and cell parameters, forming multiple crystal forms. Research has shown that the compound spinosad can exist in different crystal forms and is therefore classified as a polymorphic compound.

[0009] Patent EP1062345 discloses a process for preparing spinosad, in which the prepared spinosad exists in an amorphous state. It has been found that amorphous spinosad exhibits high aggregation during long-term storage, making it unsuitable for economical formulations.

[0010] Patent CN117586328A discloses a variant I crystal form of spinosad. The variant I crystal form is obtained by dissolving amorphous spinosad in a saturated carbonate solvent and then precipitating it out upon cooling. However, this method consumes a large amount of energy, is costly, and can easily cause harm to humans and the environment.

[0011] There is still a need in this field to develop new crystal forms of spinosad and to find new crystal forms of spinosad that are suitable for industrial production and have better drug performance. Summary of the Invention

[0012] The purpose of this invention is to provide novel crystal forms of spinosad compounds and methods for their preparation.

[0013] This invention provides two novel crystal forms of spinosad compound, named SN-II and SN-III, along with methods for preparing these two crystal forms. The methods for preparing the spinosad compound crystal forms provided by this invention are applicable to drug research and industrial production.

[0014] The SN-II crystal form provided by this invention has an X-ray powder diffraction pattern at a 2θ value of 14.0. ° ±0.2 ° 15.4 ° ±0.2 ° 15.8 ° ±0.2 ° 16.8 ° ±0.2 ° 17.4 ° ±0.2 ° 21.0 ° ±0.2 ° 21.4 ° ±0.2 ° 24.1° ±0.2 ° It has a characteristic peak.

[0015] Furthermore, the SN-II crystal form provided by this invention has an X-ray powder diffraction pattern with a 2θ value of 7.3. ° ±0.2 ° 9.5 ° ±0.2 ° 10.2 ° ±0.2 ° 12.3 ° ±0.2 ° 14.0 ° ±0.2 ° 14.6 ° ±0.2 ° 15.4 ° ±0.2 ° 15.8 ° ±0.2 ° 16.8 ° ±0.2 ° 17.4 ° ±0.2 ° 21.0 ° ±0.2 ° 21.4 ° ±0.2 ° 22.9 ° ±0.2 ° 24.1 ° ±0.2 ° It has a characteristic peak.

[0016] Furthermore, the SN-II crystal form provided by this invention has an X-ray powder diffraction pattern with a 2θ value of 7.3. ° ±0.2 ° 9.5 ° ±0.2 ° 10.2 ° ±0.2 ° 12.3 ° ±0.2 ° 14.0 ° ±0.2 ° 14.6 ° ±0.2 ° 15.4 ° ±0.2 ° 15.8 ° ±0.2 ° 16.8 ° ±0.2 ° 17.4 ° ±0.2 ° 18.4° ±0.2 ° 19.9 ° ±0.2 ° 20.4 ° ±0.2 ° 21.0 ° ±0.2 ° 21.4 ° ±0.2 ° 22.9 ° ±0.2 ° 23.6 ° ±0.2 ° 24.1 ° ±0.2 ° It has a characteristic peak.

[0017] Furthermore, the SN-II crystal form provided by this invention has an X-ray powder diffraction pattern that is essentially as follows: Figure 1 As shown.

[0018] This invention provides a method for preparing the above-mentioned SN-II crystal form, the preparation method comprising: adding spinosad to solvent A, heating and stirring to dissolve, then adding solvent B and continuing to keep warm and stirring to react, after the reaction is completed, cooling to crystallize, after crystallization is complete, filtering, drying, and obtaining spinosad SN-II crystal form.

[0019] Furthermore, solvent A is one or a combination of glacial acetic acid, N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, ketone solvents, ether solvents, alcohol solvents, ester solvents, halogenated hydrocarbons, and nitrated hydrocarbons.

[0020] Furthermore, the heating temperature is 30~85℃.

[0021] Furthermore, the solvent B is an aliphatic hydrocarbon or water, wherein the aliphatic hydrocarbon is preferably n-heptane.

[0022] Furthermore, the volume ratio of solvent A to solvent B is 1:0.2~4.

[0023] Furthermore, the reaction time is 0.5 to 2 hours.

[0024] Furthermore, the crystallization temperature is -5~15℃, and the crystallization time is 2~8h.

[0025] The SN-III crystal form provided by this invention has an X-ray powder diffraction pattern at a 2θ value of 11.5. ° ±0.2 ° 14.0 ° ±0.2 ° 15.4 ° ±0.2° 16.8 ° ±0.2 ° 17.4 ° ±0.2 ° 21.0 ° ±0.2 ° 22.9 ° ±0.2 ° 24.1 ° ±0.2 ° It has a characteristic peak.

[0026] Furthermore, the SN-III crystal form provided by this invention has an X-ray powder diffraction pattern with a 2θ value of 7.3. ° ±0.2 ° 9.5 ° ±0.2 ° 10.2 ° ±0.2 ° 11.5 ° ±0.2 ° 12.3 ° ±0.2 ° 14.0 ° ±0.2 ° 14.6 ° ±0.2 ° 15.4 ° ±0.2 ° 16.8 ° ±0.2 ° 17.4 ° ±0.2 ° 21.0 ° ±0.2 ° 22.9 ° ±0.2 ° 24.1 ° ±0.2 ° 25.0 ° ±0.2 ° It has a characteristic peak.

[0027] Furthermore, the SN-III crystal form provided by this invention has an X-ray powder diffraction pattern with a 2θ value of 7.3. ° ±0.2 ° 9.5 ° ±0.2 ° 10.2 ° ±0.2 ° 11.5 ° ±0.2 ° 12.3 ° ±0.2 ° 14.0 °±0.2 ° 14.6 ° ±0.2 ° 15.4 ° ±0.2 ° 16.8 ° ±0.2 ° 17.4 ° ±0.2 ° 18.4 ° ±0.2 ° 19.9 ° ±0.2 ° 20.4 ° ±0.2 ° 21.0 ° ±0.2 ° 22.9 ° ±0.2 ° 23.6 ° ±0.2 ° 24.1 ° ±0.2 ° 25.0 ° ±0.2 ° 29.9 ° ±0.2 ° It has a characteristic peak.

[0028] Furthermore, the SN-III crystal form provided by this invention has an X-ray powder diffraction pattern that is essentially as follows: Figure 2 As shown.

[0029] The present invention provides a method for preparing the above-mentioned SN-III crystal form, the preparation method comprising: adding spinosad to solvent C, heating and stirring to dissolve, heating and concentrating under vacuum, cooling to crystallize after the reaction is completed, filtering and drying to obtain spinosad SN-III crystal form.

[0030] Furthermore, the solvent C is one or a combination of alcohol solvents, ether solvents, ketone solvents, and ester solvents. Ethanol or ethyl acetate is preferred.

[0031] Furthermore, the heating temperature is 50~85℃, and the heating time is 1~4h.

[0032] Furthermore, the vacuum level is less than -0.9 MPa, and the concentration temperature is 40~70℃.

[0033] Furthermore, the volume ratio of the concentrated solution to the original solution is 0.2~0.8:1, preferably 0.4~0.6:1.

[0034] Furthermore, the crystallization temperature is -5~15℃, and the crystallization time is 0.5~2h.

[0035] Furthermore, in the above methods for preparing SN-II and SN-III crystal forms, the spinosad used is amorphous spinosad.

[0036] Beneficial effects

[0037] Through in-depth research, the inventors unexpectedly discovered two crystal forms of spinosad, SN-II and SN-III. The new crystal forms of the present invention overcome the instability of commercially available amorphous spinosad and are also easier to industrialize, thus having better economic benefits.

[0038] This invention develops a new preparation route for spinosad's new crystal form. The SN-II and SN-III crystal forms can be prepared using this route. The preparation process is simple, easy to operate, suitable for production, and has stable and controllable quality. The product has high yield, high purity, and good chemical stability, making it well-suited for the drug performance research of spinosad.

[0039] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Attached Figure Description

[0040] Figure 1 XRPD pattern of spinosad SN-II crystal form;

[0041] Figure 2 XRPD pattern of spinosad SN-III crystal form;

[0042] Figure 3 The XRPD pattern of spinosad amorphous form. Detailed Implementation

[0043] The present invention will be further illustrated below through embodiments. It should be understood that the embodiments of the present invention are merely illustrative and not intended to limit the invention. Therefore, any simple modifications to the present invention based on the method described herein fall within the scope of protection claimed by the present invention.

[0044] Materials used in the experiment: Spinosad can be purchased or prepared according to the method disclosed in patent EP1062345; other physical materials used in the experiment whose source and specifications are not specified are commercially available analytical grade or chemically pure.

[0045] This invention uses HPLC to determine the purity of spinosad, and the chromatographic conditions are as follows:

[0046] Column temperature: 50℃;

[0047] Detection wavelength: 220nm;

[0048] Flow rate: 1.3 ml / min;

[0049] Injection volume: 10 μL;

[0050] Isocratic elution;

[0051] Example 1

[0052] Approximately 10g of spinosad was added to 100ml of N,N-dimethylacetamide. The mixture was heated to 70℃ and stirred until dissolved. Then, approximately 200ml of water was added and the mixture was kept at 70℃ and stirred for 1 hour. After the reaction was completed, the reaction solution was cooled to -5~15℃ and stirred for 2~8 hours to allow crystals to precipitate. After crystallization, the mixture was filtered and dried to obtain the SN-II crystal form of spinosad, with a yield of 95.6% and a purity of 97.3%.

[0053] Spinosad crystal form characterization

[0054] The X-ray powder diffraction testing instrument and testing conditions involved in this invention are as follows: X-ray powder diffractometer: PANalytical EMPYERA; Sample stage: flat plate; Incident light path: BBHD; Diffraction light path: PLXCEL; Voltage: 45kV; Current: 40mA; Divergence slit: 1; Solar slit: 0.04rad; Step size: 0.5s; Scanning range: 3~50 ° .

[0055] The obtained sample is spinosad SN-II crystal form, and the characteristic peaks in its corresponding X-ray powder diffraction pattern (Cu-Ka) are detailed in the appendix. Figure 1 And Table 1.

[0056]

[0057] Example 2

[0058] Add approximately 10g of spinosad to 100ml of ethyl acetate, heat to 50℃ and stir until dissolved, then add approximately 180ml of n-heptane and maintain the temperature at 50℃ with stirring for 1 hour. After the reaction is complete, cool the reaction solution to -5~15℃ and stir to crystallize for 2~8 hours. After crystallization, filter and dry to obtain spinosad SN-II crystal form, with a yield of 94.7% and a purity of 98.1%.

[0059] Example 2 has the same X-ray powder diffraction pattern as Example 1.

[0060] Example 3

[0061] Add approximately 10g of spinosad to 100ml of diethyl ether, heat to 50℃ and stir until dissolved, then add approximately 160ml of n-heptane and maintain the temperature at 50℃ with stirring for 1 hour. After the reaction is complete, cool the reaction solution to -5~15℃ and stir to crystallize for 2~8 hours. After crystallization, filter and dry to obtain the SN-II crystal form of spinosad, with a yield of 95.2% and a purity of 97.9%.

[0062] Example 3 has the same X-ray powder diffraction pattern as Example 1.

[0063] Example 4

[0064] Approximately 10g of spinosad was added to 100ml of ethyl acetate. The mixture was heated to 70℃ and stirred until dissolved. The solution was then concentrated under reduced pressure at 70℃ for 1 hour (vacuum degree less than -0.9 MPa) until the volume of the solution was 0.5 times that of the original solution. After concentration, the reaction solution was cooled to -5~15℃ and stirred to induce crystallization. After crystallization for 2.5 hours, the solution was filtered and dried to obtain spinosad SN-III crystals, with a yield of 91.4% and a purity of 98.3%.

[0065] The obtained sample is spinosad SN-III crystal form, and the characteristic peaks in its corresponding X-ray powder diffraction pattern (Cu-Ka) are detailed in the appendix. Figure 2 And Table 2.

[0066]

[0067] Example 5

[0068] Approximately 10g of spinosad was added to 100ml of isopropanol. The mixture was heated to 70℃ and stirred until dissolved. The solution was then concentrated under reduced pressure at 70℃ for 1.5 hours (vacuum degree less than -0.9 MPa). Concentration was stopped when the volume of the solution reached 0.4 times that of the original solution. After concentration, the reaction solution was cooled to -5~15℃ and stirred to induce crystallization. Crystallization was completed after 2 hours. The solution was filtered, dried, and the SN-III crystal form of spinosad was obtained, with a yield of 90.3% and a purity of 98.6%.

[0069] Example 5 and Example 4 have the same X-ray powder diffraction pattern.

[0070] Comparative Example 1

[0071] Approximately 10g of spinosad was added to 100ml of pure water. The pH was adjusted to 3.0 with 2% hydrochloric acid solution, stirred to dissolve, and filtered to obtain the solution. The pH was then adjusted to 9.5 with 4% sodium hydroxide solution, stirred to allow the product to precipitate, and filtered to obtain a filter cake. The filter cake was rinsed with 100ml of pure water and dried to obtain an amorphous spinosad sample with a yield of 85.5% and a purity of 93.6%. The characteristic peaks in its corresponding X-ray powder diffraction pattern (Cu-Ka) are detailed in the appendix. Figure 3 .

[0072] The above description is merely an embodiment of the present invention and does not limit the patent scope of the present invention. Any equivalent modifications made based on the content of the present invention specification and drawings, or direct or indirect applications in related technical fields, are similarly included within the patent protection scope of the present invention.

Claims

1. The SN-II crystal form of spinosad, characterized in that: Its X-ray powder diffraction pattern is at a 2θ value of 14.

0. ° ±0.2 ° 15.4 ° ±0.2 ° 15.8 ° ±0.2 ° 16.8 ° ±0.2 ° 17.4 ° ±0.2 ° 21.0 ° ±0.2 ° 21.4 ° ±0.2 ° 24.1 ° ±0.2 ° It has a characteristic peak.

2. The SN-II crystal form of spinosad as described in claim 1, characterized in that: Its X-ray powder diffraction pattern has a 2θ value of 7.

3. ° ±0.2 ° 9.5 ° ±0.2 ° 10.2 ° ±0.2 ° 12.3 ° ±0.2 ° 14.0 ° ±0.2 ° 14.6 ° ±0.2 ° 15.4 ° ±0.2 ° 15.8 ° ±0.2 ° 16.8 ° ±0.2 ° 17.4 ° ±0.2 ° 21.0 ° ±0.2 ° 21.4 ° ±0.2 ° 22.9 ° ±0.2 ° 24.1 ° ±0.2 ° It has a characteristic peak.

3. The SN-II crystal form of spinosad as described in claim 1, characterized in that: Its X-ray powder diffraction pattern has a 2θ value of 7.

3. ° ±0.2 ° 9.5 ° ±0.2 ° 10.2 ° ±0.2 ° 12.3 ° ±0.2 ° 14.0 ° ±0.2 ° 14.6 ° ±0.2 ° 15.4 ° ±0.2 ° 15.8 ° ±0.2 ° 16.8 ° ±0.2 ° 17.4 ° ±0.2 ° 18.4 ° ±0.2 ° 19.9 ° ±0.2 ° 20.4 ° ±0.2 ° 21.0 ° ±0.2 ° 21.4 ° ±0.2 ° 22.9 ° ±0.2 ° 23.6 ° ±0.2 ° 24.1 ° ±0.2 ° It has a characteristic peak.

4. The SN-II crystal form of spinosad as described in any one of claims 1-3, characterized in that: It has a basic X-ray powder diffraction pattern as shown in Figure 1.

5. The SN-III crystal form of spinosad, characterized in that: Its X-ray powder diffraction pattern is at a 2θ value of 11.

5. ° ±0.2 ° 14.0 ° ±0.2 ° 15.4 ° ±0.2 ° 16.8 ° ±0.2 ° 17.4 ° ±0.2 ° 21.0 ° ±0.2 ° 22.9 ° ±0.2 ° 24.1 ° ±0.2 ° It has a characteristic peak.

6. The SN-III crystal form of spinosad as described in claim 5, characterized in that: Its X-ray powder diffraction pattern has a 2θ value of 7.

3. ° ±0.2 ° 9.5 ° ±0.2 ° 10.2 ° ±0.2 ° 11.5 ° ±0.2 ° 12.3 ° ±0.2 ° 14.0 ° ±0.2 ° 14.6 ° ±0.2 ° 15.4 ° ±0.2 ° 16.8 ° ±0.2 ° 17.4 ° ±0.2 ° 21.0 ° ±0.2 ° 22.9 ° ±0.2 ° 24.1 ° ±0.2 ° 25.0 ° ±0.2 ° It has a characteristic peak.

7. The SN-III crystal form of spinosad as described in claim 5, characterized in that: Its X-ray powder diffraction pattern has a 2θ value of 7.

3. ° ±0.2 ° 9.5 ° ±0.2 ° 10.2 ° ±0.2 ° 11.5 ° ±0.2 ° 12.3 ° ±0.2 ° 14.0 ° ±0.2 ° 14.6 ° ±0.2 ° 15.4 ° ±0.2 ° 16.8 ° ±0.2 ° 17.4 ° ±0.2 ° 18.4 ° ±0.2 ° 19.9 ° ±0.2 ° 20.4 ° ±0.2 ° 21.0 ° ±0.2 ° 22.9 ° ±0.2 ° 23.6 ° ±0.2 ° 24.1 ° ±0.2 ° 25.0 ° ±0.2 ° 29.9 ° ±0.2 ° It has a characteristic peak.

8. The SN-III crystal form of spinosad as described in any one of claims 5-7, characterized in that: It has a basic X-ray powder diffraction pattern as shown in Figure 2.

9. A method for preparing spinosad SN-II crystal form, characterized in that: The method includes the following steps: Add spinosad to solvent A, heat and stir until dissolved, then add solvent B and continue to keep warm and stir the reaction. After the reaction is complete, cool down to allow crystals to precipitate. After crystallization is complete, filter and dry to obtain spinosad SN-II crystal form.

10. The method as described in claim 9, characterized in that: Solvent A is one or a combination of glacial acetic acid, N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, ketone solvents, ether solvents, alcohol solvents, ester solvents, halogenated hydrocarbons, and nitrated hydrocarbons.

11. The method as described in claim 9, characterized in that: The heating temperature is 30~85℃.

12. The method as described in claim 9, characterized in that: Solvent B is an aliphatic hydrocarbon or water.

13. The method as described in claim 9, characterized in that: The volume ratio of solvent A to solvent B is 1:0.2~4.

14. The method as described in claim 9, characterized in that: The reaction time is 0.5 to 2 hours.

15. The method as described in claim 9, characterized in that: The crystallization temperature is -5~15℃, and the crystallization time is 2~8h.

16. A method for preparing spinosad SN-III crystal form, characterized in that: The method includes the following steps: Add spinosad to solvent C, heat and stir to dissolve, then concentrate under vacuum. Once the reaction is complete, cool to allow crystals to precipitate. After crystallization, filter and dry to obtain spinosad SN-III crystal form.

17. The method as described in claim 16, characterized in that: The solvent C is one or a combination of alcohol solvents, ether solvents, ketone solvents, and ester solvents.

18. The method as described in claim 16, characterized in that: The heating temperature is 50~85℃, and the heating time is 1~4h.

19. The method as described in claim 16, characterized in that: The vacuum degree is less than -0.9 MPa, and the concentration temperature is 40~70℃.

20. The method as described in claim 16, characterized in that: The volume ratio of the concentrated solution to the original solution is 0.2~0.8:

1.

21. The method as described in claim 16, characterized in that: The crystallization temperature is -5~15℃, and the crystallization time is 0.5~2h.

22. The spinosad SN-II crystal form according to any one of claims 1-3, characterized in that: It can be obtained by the method described in any one of claims 9-15.

23. The spinosad SN-III crystal form according to any one of claims 5-7, characterized in that: It can be obtained by the method of any one of claims 16-21.

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