CN114149476B

CN114149476B - Polymorphic substance of ribonucleoside analogue, preparation method and application thereof

Info

Publication number: CN114149476B
Application number: CN202111568177.8A
Authority: CN
Inventors: 郑国君; 王亚平; 柏俊; 张伟; 朱礼根; 高亮; 裴冉冉; 周章剑; 刘安友; 陈小峰
Original assignee: Anhui Baker Pharmaceutical Co ltd
Current assignee: Anhui Baker Pharmaceutical Co ltd
Priority date: 2021-04-23
Filing date: 2021-12-21
Publication date: 2023-09-01
Anticipated expiration: 2041-12-21
Also published as: CN113307834A; CN114149476A

Abstract

The application provides a polymorphic form of a ribonucleotide analogue, a preparation method and application thereof. Polymorphs of ribonucleotide analogues (i.e. compound 1): form I, form II, form III;the crystal form provided by the application has high purity, good stability and preservation resistance, and little or no obvious change in impurity content after long-term preservation.

Description

Polymorphic substance of ribonucleoside analogue, preparation method and application thereof

The application claims the priority of the prior application of patent application number 202110440883.8 filed by applicant in 2021, 04 and 23 to China national intellectual property agency, entitled "polymorph of ribonucleoside analogue, preparation method and application thereof". The entire contents of said prior application are incorporated by reference into the present application.

Technical Field

The application belongs to the field of compound crystal forms, and particularly relates to a ribonucleoside analogue polymorph, a preparation method and application thereof.

Background

Monnupiravir (EIDD-2801, CAS number 2349386-89-4) is a drug developed by Emerri university drug Innovation, atlanta, georgia. It is a ribonucleoside analogue that mimics naturally occurring nucleosides, generating errors during replication of single stranded RNA viruses to prevent viral replication, thereby inhibiting viral replication, and has the structure:

the EIDD-2801 has been shown to have therapeutic effects on Murine Hepatitis Virus (MHV), severe acute respiratory syndrome coronavirus (SARS-COV), middle east respiratory syndrome coronavirus (MERS-COV), novel coronavirus (2019-nCoV) and the like.

How to develop a pharmaceutical crystal form of EIDD-2801 which is more suitable for pharmaceutical preparation, in particular to a crystal form with improved stability, hygroscopicity, preservability and/or efficacy, etc., thereby obtaining good effects in pharmaceutical preparation and administration stages, and becoming a technical problem to be solved urgently.

Disclosure of Invention

In order to improve the technical problems described above, the present application provides a polymorph of compound 1 (EIDD-2801):

the present application provides form I of compound 1 having characteristic peaks at 6.45±0.20°, 12.99±0.20°, 16.30±0.20°, 16.95±0.20°, 17.19±0.20°, 21.21±0.20° by X-ray powder diffraction expressed in 2θ using cu—kα radiation.

Preferably, the crystal form I uses Cu-ka radiation, and the X-ray powder diffraction expressed in terms of 2θ has characteristic peaks at 6.45±0.20°, 12.99±0.20°, 16.30±0.20°, 16.95±0.20°, 17.19±0.20°, 21.21±0.20°, 27.96 ±0.20°, 28.39±0.20°.

Also preferably, the crystal form I uses Cu-ka radiation, and the X-ray powder diffraction expressed in terms of 2θ has characteristic peaks at 6.45±0.20°, 12.99±0.20°, 16.30±0.20°, 16.95±0.20°, 17.19±0.20°, 19.60±0.20°, 20.53±0.20°, 21.21±0.20°, 27.96 ±0.20°, 28.39±0.20°, 33.94±0.20°.

More preferably, the crystal form I uses Cu-ka radiation, and the X-ray powder diffraction expressed in terms of 2θ has characteristic peaks at 6.48±0.20°, 12.99±0.20°, 16.30±0.20°, 16.95±0.20°, 17.19±0.20°, 18.03±0.20°, 19.35±0.20°, 19.60±0.20°, 20.31±0.20 °, 20.53±0.20 °, 21.21±0.20 °, 27.96 ±0.20 °, 28.39±0.20°, 28.92 ±0.20°, 31.58±0.20°, 33.94±0.20°.

According to an embodiment of the present application, the form I uses Cu-ka radiation, and X-ray powder diffraction expressed in terms of 2θ angles is shown in table 1, error range ± 0.20 °:

TABLE 1 XRPD resolution data for form I

Peak numbering	2θ[°]	Relative strength%	Peak numbering	2θ[]	Relative strength%
						1	6.451	100.0	25	28.880	12.9
2	9.730	16.0	26	29.659	6.0
						3	12.990	55.8	27	30.081	15.6
4	16.300	57.4	28	30.868	5.4
						5	16.950	54.2	29	31.639	14.7
6	17.189	49.9	30	33.052	6.6
						7	18.010	27.8	31	33.420	12.3
8	18.290	5.7	32	33.941	20.9
						9	19.380	19.6	33	34.373	4.3
10	19.600	22.8	34	36.198	6.6
						11	19.840	12.8	35	36.519	4.1
12	20.260	14.3	36	37.271	2.7
						13	20.530	28.4	37	37.839	4.5
14	21.210	64.0	38	39.118	7.7
						15	22.529	5.7	39	40.168	4.2
16	22.919	5.4	40	40.420	5.2
						17	23.319	16.3	41	40.930	17.8
18	23.910	12.2	42	42.072	6.6
						19	24.199	11.9	43	42.329	4.9
20	25.699	9.6	44	43.502	4.9
						21	26.349	13.6	45	44.110	15.6
22	27.469	4.6	46	45.190	4.6
						23	27.961	40.7	47	47.426	3.4
24	28.389	43.7

。

Preferably, the form I has a powder X-ray diffraction pattern substantially as shown in figure 1.

According to an embodiment of the application, the form I has a DSC profile substantially as shown in figure 2.

According to an embodiment of the application, the form I has a TGA profile substantially as shown in figure 3.

The present application provides form II of compound 1 having characteristic peaks at 3.20±0.20°, 16.94±0.20°, 17.98±0.20°, 20.50±0.20°, 21.24±0.20°, 27.94±0.20° by X-ray powder diffraction expressed in 2θ using cu—kα radiation.

Preferably, the crystal form II uses Cu-K alpha radiation, and X-ray powder diffraction expressed in terms of 2 theta has characteristic peaks at 3.20+/-0.20 degrees, 6.44+/-0.20 degrees, 16.94+/-0.20 degrees, 17.98+/-0.20 degrees, 20.50+/-0.20 degrees, 20.28+/-0.20 degrees, 21.24+/-0.20 degrees and 27.94+/-0.20 degrees.

More preferably, the form II uses Cu-ka radiation and the X-ray powder diffraction expressed in terms of 2θ has characteristic peaks at 3.20±0.20°, 6.44±0.20°, 16.94±0.20°, 17.98±0.20°, 19.36±0.20°, 20.50±0.20°, 20.28±0.20°, 21.24±0.20°, 27.94±0.20°, 28.36±0.20°.

According to an embodiment of the present application, the form II uses Cu-ka radiation, and X-ray powder diffraction expressed in terms of 2θ angles is shown in table 2, error range ± 0.20 °:

TABLE 2 XRPD resolution data for form II

Peak numbering	2θ[°]	Relative strength%	Peak numbering	2θ[°]	Relative strength%
						1	3.198	100.0	27	33.900	8.5
2	6.440	17.0	28	34.362	6.7
						3	9.700	2.7	29	34.956	1.8
4	12.981	9.9	30	35.175	1.6
						5	16.297	8.8	31	36.138	3.1
6	16.940	80.3	32	36.579	3.2
						7	17.980	32.6	33	37.296	2.9
8	18.299	5.8	34	37.818	1.4
						9	19.360	16.2	35	38.684	2.2
10	19.819	9.3	36	39.096	2.2
						11	20.278	18.5	37	40.419	2.2
12	20.501	34.4	38	40.863	3.6
						13	21.241	68.1	39	42.337	2.6
14	22.519	5.0	40	43.539	1.6
						15	23.317	8.1	41	44.177	2.8
16	24.141	7.1	42	45.262	1.0
						17	25.696	3.3	43	48.397	1.3
18	26.359	4.4
						19	27.437	4.2
20	27.942	51.4
						21	28.360	14.9
22	28.839	10.6
						23	30.061	12.1
24	30.822	2.3
						25	31.542	11.1
26	33.662	6.3

。

Preferably, the form II has a powder X-ray diffraction pattern substantially as shown in figure 4.

According to an embodiment of the application, the form II has a DSC profile substantially as shown in figure 5.

According to an embodiment of the application, the form II has a TGA profile substantially as shown in figure 6.

The present application provides form III of compound 1 having characteristic peaks at 16.71±0.20°, 17.57±0.20°, 18.22±0.20°, 19.90±0.20°, 22.31±0.20° by X-ray powder diffraction expressed in 2θ angle using cu—kα radiation.

Preferably, the form III uses Cu-K alpha radiation, and X-ray powder diffraction expressed in terms of 2 theta has characteristic peaks at 14.15+ -0.20 DEG, 16.71+ -0.20 DEG, 17.57+ -0.20 DEG, 18.22+ -0.20 DEG, 19.90+ -0.20 DEG, 20.52+ -0.20 DEG, 22.31+ -0.20 DEG, 24.39+ -0.20 deg.

More preferably, the form III uses Cu-ka radiation, and the X-ray powder diffraction expressed in terms of 2θ has characteristic peaks at 10.10±0.20° 11.10±0.20°, 14.15±0.20°, 16.71±0.20°, 17.57±0.20°, 18.22±0.20°, 19.90±0.20°, 20.52±0.20°, 22.31±0.20°, 24.13±0.20°, 24.39±0.20°, 25.42±0.20°.

According to an embodiment of the present application, the form III uses Cu-ka radiation, and X-ray powder diffraction expressed in terms of 2θ angles is shown in table 3, error range ± 0.20 °:

TABLE 3 XRPD resolution data for form III

Peak numbering	2θ[°]	Relative strength%	Peak numbering	2θ[°]	Relative strength%
						1	10.100	20.3	19	28.890	16.6
2	11.101	19.6	20	30.069	4.4
						3	13.211	13.6	21	30.940	9.3
4	14.150	27.9	22	31.239	4.7
						5	16.710	49.8	23	31.870	5.9
6	17.570	88.7	24	32.390	16.3
						7	18.220	100.0	25	33.870	6.8
8	19.900	41.8	26	35.129	4.2
						9	20.280	17.5	27	36.698	3.5
10	20.520	33.3	28	37.139	5.0
						11	20.870	17.0	29	38.630	4.4
12	22.310	42.5	30	39.060	4.5
						13	22.710	8.0	31	40.970	3.9
14	23.399	5.1	32	41.610	7.6
						15	24.130	20.4	33	42.591	9.0
16	24.390	29.8	34	45.690	2.5
						17	25.421	21.0	35	46.682	2.9
18	27.161	13.8	36	47.479	2.9

Preferably, the form III has a powder X-ray diffraction pattern substantially as shown in figure 7.

According to an embodiment of the application, the form III has a DSC profile substantially as shown in figure 8.

According to an embodiment of the application, the form III has a TGA profile substantially as shown in figure 9.

According to an embodiment of the application, the polymorphs include the non-solvate (anhydrate) of compound 1 and the crystalline forms of the solvates; in some embodiments, the polymorphic form comprises a hydrate, e.g., a hemihydrate, a monohydrate, a dihydrate, a trihydrate, a tetrahydrate.

The application also provides a preparation method of the polymorph of the compound 1 (EIDD-2801), which comprises the steps of mixing the compound 1 with a solvent, and recrystallizing to obtain the polymorph;

the solvent is selected from organic solvents and/or water.

According to an embodiment of the present application, the organic solvent is selected from one, two or more of methanol, ethanol, isopropanol, acetone, acetonitrile, toluene, ethyl acetate, butyl acetate, methyl t-butyl ether, tetrahydrofuran, and the like.

According to an embodiment of the present application, the solvent may be selected from methanol, ethanol, isopropanol, acetone, acetonitrile, toluene, ethyl acetate, butyl acetate, tetrahydrofuran, a mixed solvent of ethyl acetate and methyl t-butyl ether, a mixed solvent of ethanol and water, or a mixed solvent of acetonitrile and water.

According to an embodiment of the application, the mass to volume ratio of the compound 1 to the solvent is 1g (2-10) mL, for example 5g:25mL.

According to an embodiment of the present application, the preparation method includes: and mixing the compound 1 with a solvent, heating and stirring until the system is dissolved, cooling, washing and drying to obtain the polymorph.

According to an embodiment of the present application, the process for the preparation of the polymorph may be selected from any one of the following processes:

the method comprises the following steps: mixing the compound 1 and ethyl acetate according to the mass volume ratio of 1g (2-10) mL, heating, stirring, heating to reflux (preferably reflux time is 1-5 hours), slowly cooling to 10-15 ℃, filtering, leaching with ethyl acetate, and drying (preferably drying temperature is 30-50 ℃), thus obtaining the polymorph;

the second method is as follows: mixing the compound 1 and ethyl acetate according to the mass volume ratio of 1g (2-10) mL, heating, stirring and heating to reflux (preferably reflux for 0.5-2 hours), cooling to 50-55 ℃, and adding (preferably dropwise) methyl tert-butyl ether (preferably after 2 min); continuously cooling to 10-15 ℃, stirring (preferably stirring for 0.5-1 hour), filtering, washing and drying to obtain the polymorphic substance; preferably, the volume ratio of the ethyl acetate to the methyl tertiary butyl ether is 1 (0.8-1.3);

and a third method: the compound 1, absolute ethyl alcohol and water are heated, stirred and warmed to be a system solution according to the mass volume ratio of 1g (2-10) mL (0.3-0.8) mL, slowly cooled to 10-15 ℃, stirred, filtered, washed and dried to obtain the polymorph;

the method four: mixing the compound 1 with absolute ethyl alcohol according to the mass volume ratio of 1g (2-10) mL, heating, stirring and heating to dissolve the system, cooling to 10-15 ℃ in ice water bath, stirring, filtering, washing and drying to obtain the polymorph;

and a fifth method: mixing the compound 1 and acetone according to the mass volume ratio of 1g (2-10) mL, heating, refluxing and pulping, then slowly cooling to 0-5 ℃, stirring, filtering, washing and drying to obtain the polymorph;

the method six: mixing the compound 1 and methanol according to the mass volume ratio of 1g (2-10) mL, heating and stirring until the system is dissolved, then slowly cooling to 0-5 ℃, stirring, filtering, washing and drying to obtain the polymorph;

and a seventh method: mixing the compound 1 and water according to the mass volume ratio of 1g (2-10) mL, heating, stirring and heating to a system solution (for example, heating to 60 ℃), naturally cooling to 10-15 ℃, stirring, filtering, washing and drying to obtain the polymorph;

method eight: mixing the compound 1 and tetrahydrofuran according to the mass volume ratio of 1g (2-10) mL, heating, stirring and heating to a system solution (for example, heating to 60 ℃), naturally cooling to 10-15 ℃, stirring, filtering, washing and drying to obtain the polymorph;

method nine: heating and stirring the compound 1, water and acetonitrile according to the mass volume ratio of (0.3-0.8) mL (0.8-2) mL to dissolve the system, slowly cooling to 0-5 ℃, stirring, filtering, washing and drying to obtain the polymorph;

method ten: heating and stirring the compound 1, water and methanol according to the mass volume of 1g (0.3-0.8) mL (0.8-2) mL until the system is dissolved, slowly cooling to 0-5 ℃, stirring, filtering, washing and drying to obtain the polymorph;

method eleven: mixing the compound 1 and tetrahydrofuran according to the mass volume ratio of (2-10) mL, heating, stirring and heating to dissolve the system, naturally cooling to 10-15 ℃, stirring, filtering, washing and drying to obtain the polymorph;

method twelve: mixing the compound 1 and toluene according to the mass volume ratio of 1g (2-10) mL, heating, stirring, refluxing, naturally cooling to 10-15 ℃, stirring, filtering, washing and drying to obtain the polymorph;

the method is thirteenth: mixing the compound 1 and isopropanol according to the mass volume ratio of 1g (2-10) mL, heating and stirring until the system is dissolved (for example, heating to 70 ℃), naturally cooling to 10-15 ℃, stirring, filtering, washing and drying to obtain the polymorph;

the method is fourteen: mixing the compound 1 and isopropanol according to the mass volume ratio of 1g (2-10) mL, heating and stirring until the system is dissolved, then slowly cooling to 10-15 ℃, stirring, filtering, washing and drying to obtain the polymorph;

fifteen methods: and mixing the compound 1 and n-propanol according to the mass volume ratio of 1g (2-10) mL, heating and stirring until the system is dissolved, then slowly cooling to 10-15 ℃, stirring, filtering, washing and drying to obtain the polymorph.

The application also provides a pharmaceutical composition containing the polymorphic substance.

According to an embodiment of the present application, the pharmaceutical composition may further comprise at least one pharmaceutically acceptable excipient.

According to embodiments of the present application, the pharmaceutical composition may be a formulation known in the art. For example, oral preparations, capsules, lozenges, injections, etc.

The application also provides application of the polymorph or the pharmaceutical composition in preparing antiviral drugs. For example, the antiviral drug may be selected from coronavirus and/or Murine Hepatitis Virus (MHV), e.g., the coronavirus may be selected from severe acute respiratory syndrome coronavirus (SARS-COV), middle east respiratory syndrome coronavirus (MERS-COV) and/or novel coronavirus (2019-nCoV).

Advantageous effects

The crystal form provided by the application has high purity, good stability and preservation resistance, and can be subjected to accelerated experiments, high temperature (such as 50+/-2 ℃, 60+/-2 ℃ and 70+/-2 ℃), high humidity (such as humidity 70+/-5% RH, 80+/-5% RH and 90+/-5% RH) and illumination (such as total illuminance not lower than 1.0X10) ⁶ Lux hr), the crystal transformation is not easy to occur after long-term storage, and the content of impurities (single impurities and/or total impurities) is little or even not obviously changed. Is favorable for further application in improving stability and safety in the preparation process, reducing side effects, increasing bioavailability and the like.

The preparation method of the crystal form has low cost, simple operation and easily controlled reaction conditions, and can stably obtain the target crystal form.

Drawings

FIG. 1 is a powder X-ray diffraction pattern of form I;

FIG. 2 is a DSC profile of form I;

FIG. 3 is a TGA spectrum of form I;

FIG. 4 is a powder X-ray diffraction pattern of form II;

FIG. 5 is a DSC spectrum of form II;

FIG. 6 is a TGA spectrum of form II;

FIG. 7 is a powder X-ray diffraction pattern of form III;

FIG. 8 is a DSC spectrum of form III;

fig. 9 is a TGA profile of form III.

Detailed Description

The technical scheme of the application will be further described in detail below with reference to specific embodiments. It is to be understood that the following examples are illustrative only and are not to be construed as limiting the scope of the application. All techniques implemented based on the above description of the application are intended to be included within the scope of the application.

Unless otherwise indicated, the starting materials and reagents used in the following examples were either commercially available or may be prepared by known methods.

Powder X-ray diffraction was scanned over a range of 4.0-50.0 °,0.01/1s, cu (40 kV,100 mA).

Example 1

Weighing 5g of EIDD-2081 crude product, adding 25mL of ethyl acetate into a reaction bottle, heating, stirring, heating to reflux, refluxing for 3 hours, slowly reducing the temperature to 10-15 ℃, filtering, and leaching with a small amount of ethyl acetate. Scraping, and drying in a blast oven at 45 ℃ for 24 hours to obtain the product. The yield of the product was 95%.

And (5) identifying the product of the embodiment to obtain the crystal form I.

Example 2

Weighing 5g of EIDD-2081 crude product, adding 25mL of absolute ethyl alcohol and 2.5mL of water into a reaction bottle, heating, stirring, heating to 70 ℃, dissolving the solid, slowly cooling to 10-15 ℃, stirring for 0.5h, filtering, and leaching with a small amount of ethanol. Scraping, and drying in a blast oven at 50 ℃ for 24 hours to obtain the product. The yield of the product was 75%.

And (5) identifying the product of the embodiment to obtain the crystal form I.

Example 3

Weighing 5g of EIDD-2081 crude product, adding 25mL of absolute ethyl alcohol into a reaction bottle, heating, stirring and heating to 70 ℃, dissolving the solid, cooling to 10-15 ℃ by ice water bath, stirring for 0.5h, filtering and leaching with a small amount of ice-ethyl alcohol. Scraping, and drying in a blast oven at 50 ℃ for 24 hours to obtain the product. The product yield was 78%.

And (5) identifying the product of the embodiment to obtain the crystal form I.

Example 4

Weighing 5g of EIDD-2081 crude product, adding into a reaction bottle, adding 30ml of acetone, heating, refluxing and pulping for 2h, slowly cooling to 0-5 ℃, stirring for 1h, filtering, washing a filter cake with a small amount of acetone, and drying in a drying oven (T=50 ℃) to obtain 3.52g of white solid. The product yield was 70.4%.

And (5) identifying the product of the embodiment to obtain the crystal form I.

Example 5

Weighing 5g of EIDD-2081 crude product, adding into a reaction bottle, adding 2.5ml of water and 5ml of acetonitrile, heating, dissolving, stirring for 2h, slowly cooling to 0-5 ℃ and stirring for 1h, filtering, washing a filter cake with a small amount of acetonitrile, and drying in a drying oven (T=50 ℃) to obtain 2.23g of white solid. The product yield was 44.6%.

And (5) identifying the product of the embodiment to obtain the crystal form I.

Example 6

Weighing 5g of EIDD-2081 crude product, adding 20mL of tetrahydrofuran into a reaction bottle, heating, stirring, heating to 60 ℃, dissolving the solid, naturally cooling to 10-15 ℃, stirring for 1h, filtering, eluting with a small amount of ice tetrahydrofuran, scraping, and drying in a blast oven at 50 ℃ for 24h to obtain the crystal form II. The yield thereof was found to be 60%.

Example 7

Weighing 5g of EIDD-2081 crude product, adding into a reaction bottle, adding 25ml of ethanol, heating, dissolving, stirring for 2h, slowly cooling under the condition of heat preservation, separating out solid when the temperature is reduced to 60 ℃, preserving the heat for 0.5h under the temperature, and continuously slowly cooling. Cooling to 35 ℃ every half hour, cooling with ice water, continuously cooling to 10-15 ℃ and stirring for 1h, filtering, washing a filter cake with a small amount of ethanol, and drying in a drying oven (T=50 ℃).

Example 8

Polymorphs of EIDD-2081 can also be obtained by any of the following preparation methods.

Weighing 5g of EIDD-2081 crude product, adding 20mL of ethyl acetate into a reaction bottle, heating, stirring, heating to reflux, refluxing for 0.5 hour, cooling to 50-55 ℃, and dropwise adding 20mL of methyl tertiary butyl ether for 2 min. Continuously cooling to 10-15 ℃, stirring for 0.5h, filtering, and leaching with a small amount of ice methyl tertiary butyl ether. Scraping, and drying in a blast oven at 45 ℃ for 24 hours, wherein the product yield is 95%.

Or weighing 5g of EIDD-2081 crude product, adding into a reaction bottle, adding 10ml of methanol, heating, dissolving, stirring for 2 hours, slowly cooling to 0-5 ℃, stirring for 1 hour, filtering, washing a filter cake with a small amount of methanol, and drying in a drying oven (T=50 ℃) to obtain 2.53g of white solid with the yield of=50.6%.

Or weighing 5g of EIDD-2081 crude product, adding 10mL of water into a reaction bottle, heating, stirring, heating to 60 ℃, dissolving the solid, naturally cooling to 10-15 ℃, stirring for 1h, filtering, scraping, and drying in a blast oven at 50 ℃ for 24h. The yield thereof was found to be 76%.

Or weighing 5g of EIDD-2081 crude product, adding into a reaction bottle, adding 2.5ml of water and 5ml of methanol, heating, dissolving, stirring for 2 hours, slowly cooling to 0-5 ℃, stirring for 1 hour, filtering, washing a filter cake with a small amount of methanol, and drying in a drying oven (T=50 ℃) to obtain 2.17g of white solid with the yield of=43.4%.

Or weighing 5g of EIDD-2081 crude product, adding 50mL of butyl acetate into a reaction bottle, heating, stirring, heating to reflux for 1h, naturally cooling to 10-15 ℃, stirring for 1h, filtering, eluting with a small amount of butyl glacial acetate, scraping out, and drying in a blast oven at 50 ℃ for 24h. The yield thereof was found to be 65%.

Or weighing 5g of EIDD-2081 crude product, adding 50mL of toluene into a reaction bottle, heating, stirring, heating to reflux for 1h, naturally cooling to 10-15 ℃, stirring for 1h, filtering, eluting with a small amount of ice toluene, scraping, and drying in a blast oven at 50 ℃ for 24h. The yield thereof was found to be 70%.

Or weighing 5g of EIDD-2081 crude product, adding 50mL of isopropanol into a reaction bottle, heating, stirring and raising the temperature to 70 ℃, naturally cooling to 10-15 ℃ for stirring for 1h, filtering, eluting with a small amount of ice isopropanol, scraping, and drying in a blast oven at 50 ℃ for 24h to obtain the yield of 71%.

Or weighing 5g of EIDD-2081 crude product, adding 50mL of normal propyl alcohol into a reaction bottle, heating, stirring and raising the temperature to 70 ℃, dissolving the solid, naturally cooling to 10-15 ℃, stirring for 1h, filtering, eluting with a small amount of ice normal propyl alcohol, scraping, and drying in a blast oven at 50 ℃ for 24h, wherein the yield is 65%.

Example 9 Crystal form identification test

The XRD spectrum of form I is shown in FIG. 1, and the spectrum analysis data is shown in Table 1. The DSC and TGA spectra of form I are shown in FIGS. 2 and 3, respectively.

The XRD spectrum of form II is shown in FIG. 4, and the spectrum analysis data is shown in Table 2. The DSC and TGA spectra of form II are shown in FIGS. 5 and 6, respectively.

The XRD spectrum of form III is shown in FIG. 7, and the spectrum analysis data is shown in Table 3. The DSC profile and TGA profile of form III are shown in FIGS. 8 and 9, respectively.

TABLE 1 XRPD resolution data for form I

Peak numbering	2θ[°]	Relative strength%	Peak numbering	2θ[°]	Relative strength%
						1	6.451	100.0	25	28.880	12.9
2	9.730	16.0	26	29.659	6.0
						3	12.990	55.8	27	30.081	15.6
4	16.300	57.4	28	30.868	5.4
						5	16.950	54.2	29	31.639	14.7
6	17.189	49.9	30	33.052	6.6
						7	18.010	27.8	31	33.420	12.3
8	18.290	5.7	32	33.941	20.9
						9	19.380	19.6	33	34.373	4.3
10	19.600	22.8	34	36.198	6.6
						11	19.840	12.8	35	36.519	4.1
12	20.260	14.3	36	37.271	2.7
						13	20.530	28.4	37	37.839	4.5
14	21.210	64.0	38	39.118	7.7
						15	22.529	5.7	39	40.168	4.2
16	22.919	5.4	40	40.420	5.2
						17	23.319	16.3	41	40.930	17.8
18	23.910	12.2	42	42.072	6.6
						19	24.199	11.9	43	42.329	4.9
20	25.699	9.6	44	43.502	4.9
						21	26.349	13.6	45	44.110	15.6
22	27.469	4.6	46	45.190	4.6
						23	27.961	40.7	47	47.426	3.4
24	28.389	43.7

TABLE 2 XRPD resolution data for form II

TABLE 3 XRPD resolution data for form III

Example 10 Crystal form Performance test

Solubility (one)

Investigation reagent: purified water, methanol, DMF, ethyl acetate, acetone, dichloromethane, acetonitrile, ethanol;

the experimental steps are as follows:

taking the samples (namely, crystal form I, crystal form II and crystal form III) which are ground into fine powder at room temperature, weighing quantitative samples (see tables 1-3) according to different solubility requirements, shaking in a water bath at 25 ℃, shaking forcefully for 30 seconds every 5 minutes, observing the dissolution condition within 30 minutes, and considering that solute particles are completely dissolved without visual observation.

TABLE 1

TABLE 2

TABLE 3 Table 3

The solubility of forms I-III in different solvents is summarized in Table 4.

TABLE 4 Table 4

(II) hygroscopicity

Experimental instrument: glass weighing bottle with stopper (outer diameter 50mm, height 15 mm).

The experimental steps are as follows:

1. the clean glass weighing bottle with the plug is placed in a proper constant temperature dryer (the lower part of the bottle is placed with ammonium chloride or ammonium sulfate saturated solution) at 25 ℃ for precisely weighing the weight (m 1) the day before the experiment;

2. a proper amount of a test sample is taken and paved in the weighing bottle, the thickness of the test sample is generally about 1mm, and the weight (m 2) is precisely weighed;

3. the weighing bottle is opened and is placed under the constant temperature and humidity condition for 24 hours together with the bottle cap;

4. the lid of the weighing flask was closed, and the weighing flask was precisely weighed (m 3).

The results are shown in Table 5, with forms I-III having slight hygroscopicity.

TABLE 5

(III) stability

Light stability: the crystal forms I-III were subjected to light intensity 4500lx, and samples were taken on days 5 and 12, and the purity was measured by High Performance Liquid Chromatography (HPLC).

High wet stability: forms I-III were taken at room temperature and 92.5% + -5% relative humidity, and samples were taken on days 11 and 30, respectively, and tested for purity by HPLC.

Thermal stability: the crystal forms I-III were sampled at 60.+ -. 2 ℃ on days 11 and 30, respectively, and the purity was checked by HPLC.

Acceleration stability: the crystal forms I-III were sampled at 40+ -2deg.C and 75% + -5% humidity, respectively, and tested for purity by HPLC and for XRPD at month 1 and month 2.

Wherein, the conditions of HPLC detection are as follows:

1.1 chromatographic conditions

Chromatographic column: BDS HypersilC18, 250X 4.6mm,5 μm

Column temperature: 40 DEG C

Flow rate: 1.0mL/min

Detector wavelength: 254nm

Sample injection amount: 10 mu L

Mobile phase a:0.01mol/L potassium dihydrogen phosphate solution

Mobile phase B: methanol

Gradient elution:

1.2 preparation of solutions

Blank solution: 5% methanol

Impurity control stock solution: respectively precisely weighing 10mg of impurities (impurity Imp-G, impurity uridine, impurity Imp-D, impurity Imp-B) into a 10mL volumetric flask, adding 5% methanol, ultrasonically dissolving, diluting to scale, and shaking to obtain stock solution of each impurity (1 mg/mL).

Test solution: precisely weighing 25mg of the sample (i.e. the crystal form I-III), placing in a 25mL volumetric flask, adding 5% methanol, ultrasonically dissolving, diluting to scale, and shaking to obtain (1 mg/mL).

Control solution: precisely weighing 25mg of reference substance, placing in a 25mL volumetric flask, adding 5% methanol, ultrasonically dissolving, diluting to scale, and shaking to obtain reference substance stock solution 1; precisely transferring 1mL of reference substance stock solution 1, placing in a 100mL volumetric flask, adding 5% methanol to dilute to scale, and shaking to obtain reference substance stock solution 2; precisely transferring 1mL of control stock solution 2, placing in a 10mL volumetric flask, adding 5% methanol to dilute to scale, and shaking to obtain control solution (1 μg/mL).

System adaptation solution: and precisely weighing 50mg of the EIDD-2801 reference substance, adding 0.5mL of each impurity stock solution into a 50mL volumetric flask, adding 5% methanol, ultrasonically dissolving and diluting to scale, and shaking uniformly to obtain the (1 mg/mL of the EIDD-2801 reference substance, and 10 mug/mL of each impurity).

The stability test results are shown in tables 6-8.

TABLE 6

TABLE 7

TABLE 8

From the stability test results, the crystal forms I-III have good stability. The crystal form of the test sample was detected (identified by XRPD) after two months of accelerated test, with no change in crystal form.

Therefore, the crystal form of the application has high purity and low impurity content; has good stability and preservation resistance, can be preserved for a long time under the conditions of acceleration experiment, high humidity, high temperature and illumination, is not easy to generate crystal transformation, has small impurity content change and even basically has no obvious change.

The embodiments of the present application have been described above. However, the present application is not limited to the above embodiment. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the protection scope of the present application.

Claims

1. A crystalline form iii of compound I, having a powder X-ray diffraction pattern as shown in figure 7,

2. a pharmaceutical composition comprising form iii of compound I according to claim 1 and at least one pharmaceutically acceptable excipient, said composition being selected from any one of oral, injectable.

3. Use of crystalline form iii of compound I according to claim 1 or of the pharmaceutical composition according to claim 2 for the preparation of a medicament for the treatment of antiviral diseases, said viruses being coronaviruses.

4. The use according to claim 3, wherein the coronavirus is selected from any one of severe acute respiratory syndrome coronavirus (SARS-COV), middle east respiratory syndrome coronavirus (MERS-COV), novel coronavirus (2019-nCoV).