阅读说明：本技术 阿糖胞苷前药mb07133晶型e及其应用 (Cytarabine prodrug MB07133 crystal form E and application thereof ) 是由 郭维博 田丹 张登科 金伟丽 孙江凯 何凯敏 于 2020-05-11 设计创作，主要内容包括：本发明公开了4-氨基-1-[5-O-(2R,4S)-2-氧基-4-(4-吡啶)-1,3,2-二氧磷杂环己烷-2]-β-D-阿拉伯呋喃糖基-2(1H)-嘧啶酮(阿糖胞苷前药,MB07133)的晶型。本发明还提供了包括该晶型的药物组合物以其在制备药物中的应用。(The invention discloses a crystal form of 4-amino-1- [5-O- (2R,4S) -2-oxyl-4- (4-pyridine) -1,3, 2-dioxaphosphorinane-2 ] -beta-D-arabinofuranose-2 (1H) -pyrimidone (cytarabine prodrug, MB 07133). The invention also provides a pharmaceutical composition comprising the crystal form and application thereof in preparing medicaments.)

MB07133 crystal characterized in that it has an X-ray powder diffraction pattern substantially as shown in figure 5.

2. The crystal of claim 1 having a differential thermal analysis curve with a sharp endothermic peak at 247.95 ℃, 251.10 ℃, 246.57 ℃ or 249.26 ℃.

3. The crystal of claim 1, which is a single crystal.

4. A medicament for treating or preventing liver diseases or metabolic diseases, which comprises the crystal of any one of claims 1 to 3 and a pharmaceutically acceptable excipient.

5. The pharmaceutical agent of claim 4, which is an injection.

6. The medicament of claim 4, which is used for treating advanced liver cancer.

7. Use of a crystal according to any one of claims 1 to 3 in the manufacture of a medicament for the treatment or prevention of a liver disease or metabolic disease.

8. The use according to claim 6 in the manufacture of a medicament for the treatment or prevention of hepatitis B.

9. The use according to claim 6 in the manufacture of a medicament for the treatment of advanced liver cancer.

10. A method for detecting a crystal according to any of claims 1 to 3, characterized in that the suspected crystal is subjected to an X-ray powder diffraction detection and the resulting X-ray powder diffraction pattern is compared with the X-ray powder diffraction pattern shown in fig. 5.

Technical Field

The invention belongs to the field of medicinal chemistry, and particularly relates to a crystal form of a cytarabine prodrug MB07133, application thereof and the like.

Background

MB07133, namely 4-amino-1- [5-O- (2R,4S) -2-oxo-4- (4-pyridine) -1,3, 2-dioxaphosphorinane-2 ] -beta-D-arabinofuranosyl-2 (1H) -pyrimidinone, also known as cytarabine prodrug, having the structure shown in formula (I):

the compound is a cytarabine hepdi ect prodrug compound in prodrug compounds disclosed in Chinese patent CN1711278A, and can be used for treating or preventing liver diseases or metabolic diseases, particularly treating late-stage liver cancer. However, this patent does not disclose or suggest studying the crystallography of this compound.

In the case that whether a new (single) crystal form exists in the MB07133, which cannot be expected, the existing solvents and mixed solvents which can be used for crystallization are more astronomical, however, the inventor has no fear and has long-term arduous research, and finds that the MB07133 is crystallized, the crystal distribution is very complex, polycrystal is often crystallized, and single crystal is not easy to obtain, but the inventor finally unexpectedly and effectively crystallizes to obtain a series of crystals, and selects 6 different single crystals from the crystals, so that the stability is advantageous, thereby facilitating the production, storage and transportation and/or improving the safety of liver targeting therapy.

Disclosure of Invention

The technical problem to be solved by the invention is to provide a new crystal form of MB 07133. In addition, the invention also provides a preparation method of the crystal form, a medicament containing the crystal form, application in treatment, a detection method and the like.

In particular, in a first aspect, the present invention provides crystals of MB07133 selected from one of form a, form B, form C, form D, form E or form F.

In this context, unless indicated to the contrary, the terms "crystal" and "crystalline form" are used interchangeably to refer to a solid in which the internal particles are periodically and repeatedly arranged in three dimensions; the terms "(crystal form(s)", "crystal form(s)", and "(crystal form(s)", are used interchangeably to refer to a particular crystal. Preferably the crystal of the first aspect of the invention is a single crystal.

In the crystals of the first aspect of the present invention, the form a crystals have an X-ray powder diffraction pattern substantially as shown in figure 1. In a particular embodiment of the invention, form a has an X-ray powder diffraction pattern at 2 Θ (°, ± 0.2): diffraction peaks exist at 10.6 °, 12.5 °, 13.7 °, 16.0, 16.3 °, 17.2 °, 17.4 °, 18.6 °, 20.3 °, 21.8 °, 22.2 °, 23.1 °, 23.4 °, 24.9 °, 25.6 °, 26.0 °, 28.7 °, 29.4 °, 30.1 °, 31.0 °, 32.9 ° and 37.8 °; in addition, the differential thermal analysis curve of the A-type crystal has a sharp endothermic peak at 239.81 ℃;

the form B crystal has an X-ray powder diffraction pattern substantially as shown in figure 2. In a particular embodiment of the invention, form B has an X-ray powder diffraction pattern at 2 Θ (°, ± 0.2): diffraction peaks exist at 10.4 °, 12.1 °, 15.8 °, 16.1 °, 17.3 °, 18.6 °, 20.2 °, 21.6 °, 22.1 °, 22.9 °, 23.2 °, 24.8 °, 25.4 °, 25.8 °, and 30.8 °; in addition, the differential thermal analysis curve of the B-type crystal has a sharp endothermic peak at 253.21 ℃;

the crystalline form C has an X-ray powder diffraction pattern substantially as shown in figure 3. In a particular embodiment of the invention, form C has an X-ray powder diffraction pattern at 2 Θ (°, ± 0.2): diffraction peaks exist at 10.5 °, 12.4 °, 13.5 °, 15.8 °, 16.3 °, 17.4 °, 18.6 °, 20.2 °, 21.6 °, 22.1 °, 23.0 °, 23.3 °, 24.8 °, 25.5 °, 28.6 °, 29.3 °, and 31.0 °; in addition, the differential thermal analysis curve of the C-type crystal has a sharp endothermic peak at 247.95 ℃;

the crystalline form D has an X-ray powder diffraction pattern substantially as shown in figure 4. In a particular embodiment of the invention, form D has an X-ray powder diffraction pattern at 2 θ (°, ± 0.2): diffraction peaks exist at 12.5 °, 15.5 °, 15.9 °, 16.3 °, 18.2 °, 18.6 °, 19.6 °, 20.3 °, 21.2 °, 23.1 °, 23.4 °, 24.2 °, 24.4 °, 24.8 °, and 27.4 °; in addition, the differential thermal analysis curve of the D-form crystal has a sharp endothermic peak at 251.10 ℃;

the crystalline form E has an X-ray powder diffraction pattern substantially as shown in figure 5. In a particular embodiment of the invention, form E has an X-ray powder diffraction pattern at 2 Θ (°, ± 0.2): diffraction peaks are present at 10.6 °, 12.5 °, 13.7 °, 15.5 °, 16.0 °, 16.5 °, 17.2 °, 17.5 °, 18.2 °, 18.7 °, 20.4 °, 21.7 °, 22.2 °, 23.1 °, 23.4 °, 24.3 °, 24.9 °, 25.6 °, 26.0 °, 27.2 °, 29.4 °, 31.0 °, and 41.5 °; in addition, the differential thermal analysis curve of the E-type crystal has a sharp endothermic peak at 246.59 ℃;

the crystalline form F has an X-ray powder diffraction pattern substantially as shown in figure 5. In a particular embodiment of the invention, form F has an X-ray powder diffraction pattern at 2 θ (°, ± 0.2): diffraction peaks exist at 10.6 °, 12.3 °, 12.5 °, 13.7 °, 16.0 °, 16.5 °, 17.2 °, 17.5 °, 17.7 °, 18.7 °, 20.4 °, 21.8 °, 22.2 °, 23.1 °, 23.4 °, 24.9 °, 25.6 °, 28.7 °, 29.4 °, 30.1 °, and 31.0 °; in addition, the differential thermal analysis curve of the E-form crystal has a sharp endothermic peak at 249.26 ℃.

In the prior art, the variety of solvents used for crystallization is wide, the mixed solvents consisting of solvents of different types and proportions cannot be counted, the crystallization practice is basically still empirical, and the crystallized crystal form cannot be predicted according to the crystallization condition. However, the present inventors have long and arduous studies on MB07133 and, with some luck, finally found a solvent that can be used for crystallizing MB07133 and a method for preparing the same. Thus in a second aspect, the present invention provides a process for the preparation of the crystal of the first aspect of the invention.

For the crystal form A, the preparation method comprises the following steps: dissolving MB07133 in a sulfuric acid solution, stirring at 15-25 ℃, adding sodium dihydrogen phosphate, slowly adding a sodium hydroxide solution, adjusting the pH value to 5.0-8.0, stirring at 10-20 ℃, collecting crystals and drying. MB07133 may be amorphous, or may be of form B, form C, form D, form E, or form F.

For the crystal form B, the preparation method comprises the following steps: MB07133 is heated and stirred in dimethyl sulfoxide until dissolved, and cooled to 0-10 ℃ after dropping toluene, and then stirred, and crystals are collected and dried. Wherein, MB07133 can be amorphous, or can be crystal form A, crystal form C, crystal form D, crystal form E or crystal form F.

For the crystal form C, the preparation method comprises the following steps: MB07133 is heated and stirred in dimethyl sulfoxide until dissolved, and cooled to 0-10 ℃ after dropping acetone, and then stirred, and crystals are collected and dried. Wherein, MB07133 can be amorphous, or can be crystal form A, crystal form B, crystal form D, crystal form E or crystal form F.

For the crystal form D, the preparation method comprises the following steps: MB07133 is heated and stirred in N, N-dimethylformamide until dissolved, and then cooled to 0-10 ℃ after ethyl acetate is dripped, and then the crystal is collected and dried. Wherein, MB07133 can be amorphous, or can be crystal form A, crystal form B, crystal form C, crystal form E or crystal form F.

For the crystal form E, the preparation method comprises the following steps: MB07133 is heated and stirred in N-methyl pyrrolidone and water until dissolved, cooled to 0-10 ℃ after dropping acetone, stirred, collected crystal and dried. Wherein, MB07133 can be amorphous, or can be crystal form A, crystal form B, crystal form C, crystal form D or crystal form F.

For the crystal form F, the preparation method comprises the following steps: MB07133 is heated and stirred in dimethyl sulfoxide and water until dissolved, is cooled to 0-10 ℃ after dropping isopropanol, is stirred, collects crystals and dries. Wherein, MB07133 can be amorphous, or can be crystal form A, crystal form B, crystal form C, crystal form D or crystal form E.

In a third aspect, the present invention provides a medicament for treating or preventing liver diseases or metabolic diseases, in particular a novel targeted drug for treating advanced liver cancer, comprising, preferably consisting of, the crystal of the first aspect of the present invention and a pharmaceutically acceptable excipient. Herein, pharmaceutically acceptable excipients refer to nontoxic fillers, stabilizers, diluents, adjuvants or other formulation excipients. For example, diluents, excipients, such as water, physiological saline, microcrystalline cellulose, and the like; fillers, such as starch, sucrose, and the like; binders, such as starch, cellulose derivatives, alginates, gelatin and/or polyvinylpyrrolidone; humectants, such as glycerol; disintegrating agents, such as agar, calcium carbonate and/or sodium bicarbonate; absorption promoters, such as quaternary ammonium compounds; surfactants such as cetyl alcohol; adsorption carriers such as kaolin and/or bentonite clay; lubricants, such as talc, calcium/magnesium stearate, polyethylene glycol, and the like. In addition, the pharmaceutical composition of the invention can further contain other auxiliary materials, such as flavoring agents, sweetening agents and the like. The medicament of the third aspect of the present invention may further comprise other active ingredients for treating or preventing liver diseases or metabolic diseases, particularly active ingredients for treating advanced liver cancer.

According to the well-known technology in the field, the pharmaceutical composition can be prepared into various dosage forms according to the requirements of treatment purposes and administration routes, preferably the composition is in a unit administration dosage form, such as a freeze-dried preparation, a tablet, a capsule, powder, emulsion, a water injection or a spray, more preferably the pharmaceutical composition is in an injection dosage form or an oral dosage form (such as a tablet or a capsule), and more preferably the pharmaceutical composition is in an injection dosage form (such as a freeze-dried powder injection). The medicaments may be administered by conventional routes, in particular enterally, e.g. orally, e.g. in the form of tablets or capsules; or parenterally, for example in the form of injectable solutions or suspensions; or nasal use.

In a fourth aspect, the invention provides the use of a crystal in the manufacture of a medicament for the treatment or prevention of liver disease or metabolic disease, in particular for the treatment of advanced liver cancer. The medicament of the present invention is administered in an effective dose, wherein the effective dose is usually in the amount of the crystal of the first aspect of the present invention. The effective dose may be the amount in a unit dosage form (e.g., a tablet, a needle, a pill, or a dose) of the drug, or may be a unit dose (e.g., a unit weight dose) of the patient for which treatment/prevention is desired. The pharmaceutical manufacturer can easily convert the unit weight dose of the patient to be treated/prevented into the content of the drug in the unit administration dosage form by the average weight of the patient population to be treated/prevented, for example, the average weight of the adult patient may be 60kg, and thus the content of the drug in the unit administration dosage form for the adult can be obtained by multiplying the average weight by the unit weight dose for the adult.

In this context, the patient may be a mammal, such as a human, rabbit, dog or mouse, preferably a human. The unit weight dose of a human can be derived from the dose of the experimental animal according to the equivalent dose conversion relationship between the experimental animal and the human (generally, refer to guidance suggestions of drug administration such as FDA and SFDA, and refer to 'Huang Tanhua et al equivalent dose conversion between animals and human bodies in pharmacological experiments, Chinese clinical pharmacology and therapeutics, 2004, 9(9): 1069-1072') known by those skilled in the art. For example, for a commonly used experimental animal mouse, the conversion relationship with an adult is about 12: 1; for the commonly used experimental animal rats, the conversion relationship with adults is about 6 according to the above-mentioned literature: 1. the dosage of the product is 300-1200 mg/m²D, intravenous drip for 7 consecutive days, repeat dosing after three weeks of rest. By calculation, the average daily dosage of the patient is about 500-2000 mg. Dose escalation was performed at the following dose levels: 300. 600, 1200, 1800, 2400 and 3000mg/m²D until clear dose-limiting toxicity (DLT) occurs and the Maximum Tolerated Dose (MTD) is determined.

The use of the fourth aspect is in the manufacture of a medicament for the treatment or prophylaxis of hepatitis b. Also preferred is the use of the fourth aspect of the invention in the manufacture of a medicament for the treatment of advanced liver cancer.

In a fifth aspect, the present invention provides a method of detecting a crystal according to the first aspect of the invention, characterised in that the suspected crystal is subjected to X-ray powder diffraction detection, whereby the suspected crystal is detectedThe resulting X-ray powder diffraction pattern was compared to the X-ray powder diffraction pattern shown in figure 1 or 2 or 3 or 4 or 5 or 6. According to spectral line position (usually expressed in degrees of Bragg's 2 theta angle), line height, relative abundance and/or interplanar distance d (usually expressed in degrees of the Bragg's 2 theta angle) of the spectrumExpression), etc., one skilled in the art can compare whether a suspected crystal is the crystal of the first aspect of the invention.

The invention has the beneficial effects that the obtained MB07133 crystal has excellent properties, good stability and high purity, does not contain solvent and moisture, is more convenient for the adaptability of the preparation process and is also convenient for storage.

For ease of understanding, the present invention incorporates patents which are set forth to provide a clear description of the invention and are incorporated herein by reference in their entirety as if fully set forth herein.

The invention will be described in detail below by means of specific embodiments and the accompanying drawings. It is to be expressly understood that the description is illustrative only and is not intended as a definition of the limits of the invention. Many variations and modifications of the present invention will be apparent to those skilled in the art in light of the teachings of this specification.

Drawings

FIG. 1: x-ray powder diffraction pattern of MB07133 form a.

FIG. 2: x-ray powder diffraction pattern of form B of MB 07133.

FIG. 3: x-ray powder diffraction pattern of form C of MB 07133.

FIG. 4: x-ray powder diffraction pattern of form D of MB 07133.

FIG. 5: x-ray powder diffraction pattern of form E of MB 07133.

FIG. 6: x-ray powder diffraction pattern of form F of MB 07133.

FIG. 7: differential thermal analysis curve pattern of MB07133 form A.

FIG. 8: differential thermal analysis curve pattern of MB07133 form B.

FIG. 9: differential thermal analysis curve pattern of MB07133 form C.

FIG. 10: differential thermal analysis curve pattern of MB07133 form D.

FIG. 11: differential thermal analysis curve pattern of MB07133 form E.

FIG. 12: differential thermal analysis curve pattern of MB07133 form F.

Detailed Description

The present invention will be explained in detail with reference to examples, which are only used to illustrate the technical solutions of the present invention, but the scope of the present invention is not limited thereto.

Test instrument for experiments

1. X-ray powder diffraction pattern

The instrument comprises the following steps: PHI-5400X-ray photoelectron analyzer (available from PE corporation, USA)

The test parameters are: voltage: 46kv, current: 40mA, copper k α radiation, λ:

2. thermogram for heat difference analysis (DSC)

The instrument comprises the following steps: SII Nano, EXSTAR, DSC6220

Temperature rise rate: 10 ℃/min

Temperature range: 50 to 250 DEG C

Carrier gas: high purity nitrogen gas

3. Material sources are as follows:

the MB07133 compound is prepared by the preparation method of Chinese patent CN 1711278A. The method comprises the following specific steps:

under the protection of nitrogen, 2, 3-di-O-TBS-cytarabine-N, N-dimethylformamidine (250g, 0.47moL) and tetrahydrofuran (2.5L) were added into the reaction kettle, cooled to 4 ℃, and tert-butyl magnesium chloride solution (617mL, 0.62moL) was added dropwise at a temperature not exceeding 8 ℃, after which the reaction was completed for 1.25 h. Phosphate reagent (262g, 0.78moL) was added in one portion and the reaction stirred at room temperature for 16 h. Ammonium chloride solution (20%, 2.5L) was added dropwise, ethyl acetate (2.5L) was added thereto, the mixture was stirred and separated, the aqueous phase was extracted with ethyl acetate (1.1L), the organic phases were combined, the solution was separated by back-washing with sodium chloride solution (15%, 1.6L), dried over magnesium sulfate (260g), filtered and concentrated to dryness to give 526g of a dark orange slurry.

The dark orange slurry was dissolved in methanol (2.5L) and HCl-dioxane solution (790mL,3.16moL) at 50-55 deg.C, reacted at 50-55 deg.C for 16h, and concentrated under reduced pressure to give viscous orange tar. The tar was partitioned with water (800mL) and ethyl acetate (800mL), sodium bicarbonate solid was added slowly until the pH of the water was 7, the layers were separated, ethyl acetate (800mL) was re-extracted, the aqueous layer was filtered, concentrated under reduced pressure, and taken dry with ethanol to give 445g of oil. Ethanol (700mL) was added, stirred at room temperature for 2h, filtered and dried to give 199g of solid, slurried with water (650mL), and added with hydrochloric acid (20mL), stirred for reaction, and added with sodium bicarbonate solid (36g) to adjust the value to 6-7, filtered and dried to give 160g of MB07133 solid.

Preparation of MB07133 Compound¹H NMR(600MHz,DMSO):δ:8.61(2H,dd)，7.54(1H,d),7.43(2H,dd),7.07-7.14(2H,s),6.12(1H,d),5.75-5.78(1H,m),5.63-5.64(2H,d),5.60(1H,d),4.41-4.59(2H,m),4.25-4.33(2H,m),4.00(1H,m),3.95-3.97(1H,m),3.92(1H,s),2.13-2.28(2H,m)。

Example 1

Preparation and identification of cytarabine prodrug MB07133 crystal form A crystal

30mL of purified water was added to a 100mL three-necked flask, 10g of the white powder obtained in MB07133 was added thereto, and a 3mol/L aqueous solution of sulfuric acid was added dropwise with stirring to adjust the pH to 2.0 to 5.0. Controlling the temperature to be 15-25 ℃, stirring for 1h, adding 0.14g of sodium dihydrogen phosphate, slowly dropwise adding a 14% sodium hydroxide aqueous solution into the reaction system, and adjusting the pH value to be 5.0-8.0. After the pH value is adjusted, the temperature is controlled to be 10-20 ℃, stirring is carried out for 3 hours, centrifugation is carried out, a filter cake is collected, crystals are reserved, and then drying is carried out directly at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal is the crystal form A, and as shown in figures 1 and 7, the stable crystal form A can be obtained by the method.

Example 2

Preparation and identification of cytarabine prodrug MB07133 crystal form A crystal

Adding 24mL of purified water and 8g of MB07133 crystal form B crystals into a 100mL three-necked bottle, dropwise adding 3mol/L sulfuric acid aqueous solution while stirring, and adjusting the pH value to 2.0-5.0. Controlling the temperature to be 15-25 ℃, stirring for 1h, adding 1.2g of sodium dihydrogen phosphate, slowly dropwise adding a 14% sodium hydroxide aqueous solution into the reaction system, and adjusting the pH value to be 5.0-8.0. After the pH value is adjusted, the temperature is controlled to be 10-20 ℃, stirring is carried out for 3 hours, centrifugation is carried out, a filter cake is collected, crystals are reserved, and then drying is carried out directly at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal is a crystal form A.

Example 3

Preparation and identification of cytarabine prodrug MB07133 crystal form A crystal

Adding 24mL of purified water and 8g of MB07133 crystal form C crystals into a 100mL three-necked bottle, dropwise adding 3mol/L sulfuric acid aqueous solution while stirring, and adjusting the pH value to 2.0-5.0. Controlling the temperature to be 15-25 ℃, stirring for 1h, adding 1.2g of sodium dihydrogen phosphate, slowly dropwise adding a 14% sodium hydroxide aqueous solution into the reaction system, and adjusting the pH value to be 5.0-8.0. After the pH value is adjusted, the temperature is controlled to be 10-20 ℃, stirring is carried out for 3 hours, centrifugation is carried out, a filter cake is collected, crystals are reserved, and then drying is carried out directly at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal is a crystal form A.

Example 4

Preparation and identification of cytarabine prodrug MB07133 crystal form A crystal

30mL of purified water and 10g of MB07133 crystal form D crystal are added into a 100mL three-necked bottle, and a 3mol/L sulfuric acid aqueous solution is dropwise added under stirring to adjust the pH value to 2.0-5.0. Controlling the temperature to be 15-25 ℃, stirring for 1h, adding 1.2g of sodium dihydrogen phosphate, slowly dropwise adding a 14% sodium hydroxide aqueous solution into the reaction system, and adjusting the pH value to be 5.0-8.0. After the pH value is adjusted, the temperature is controlled to be 10-20 ℃, stirring is carried out for 3 hours, centrifugation is carried out, a filter cake is collected, crystals are reserved, and then drying is carried out directly at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal is a crystal form A.

Example 5

Preparation and identification of cytarabine prodrug MB07133 crystal form A crystal

Adding 24mL of purified water and 8g of MB07133 crystal form E crystals into a 100mL three-necked bottle, dropwise adding 3mol/L sulfuric acid aqueous solution while stirring, and adjusting the pH value to 2.0-5.0. Controlling the temperature to be 15-25 ℃, stirring for 1h, adding 1.2g of sodium dihydrogen phosphate, slowly dropwise adding a 14% sodium hydroxide aqueous solution into the reaction system, and adjusting the pH value to be 5.0-8.0. After the pH value is adjusted, the temperature is controlled to be 10-20 ℃, stirring is carried out for 3 hours, centrifugation is carried out, a filter cake is collected, crystals are reserved, and then drying is carried out directly at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal is a crystal form A.

Example 6

Preparation and identification of cytarabine prodrug MB07133 crystal form A crystal

30mL of purified water and 10g of MB07133 crystal form F crystal are added into a 100mL three-necked bottle, and a 3mol/L sulfuric acid aqueous solution is dropwise added under stirring to adjust the pH value to 2.0-5.0. Controlling the temperature to be 15-25 ℃, stirring for 1h, adding 1.2g of sodium dihydrogen phosphate, slowly dropwise adding a 14% sodium hydroxide aqueous solution into the reaction system, and adjusting the pH value to be 5.0-8.0. After the pH value is adjusted, the temperature is controlled to be 10-20 ℃, stirring is carried out for 3 hours, centrifugation is carried out, a filter cake is collected, crystals are reserved, and then drying is carried out directly at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal is a crystal form A.

Example 7

Preparation and identification of a cytarabine prodrug MB07133 crystal form B crystal.

Adding 100mL of dimethyl sulfoxide into a 500mL three-necked bottle, adding 10g of the obtained MB07133 white powder, stirring and heating to 130 ℃ to completely dissolve the solid, then dropwise adding 100mL of toluene, cooling to 0-10 ℃ after the addition, keeping the temperature at 0-10 ℃, stirring for 3h, performing suction filtration, retaining crystals, and then directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal is the crystal form B, and as shown in figures 2 and 8, the stable crystal form B can be obtained by the method.

Example 8

Preparation and identification of a cytarabine prodrug MB07133 crystal form B crystal.

Adding 100mL of dimethyl sulfoxide into a 500mL three-necked bottle, adding 10g of MB07133 crystal form A crystals, stirring and heating to 125 ℃ to completely dissolve the solid, then dropwise adding 100mL of toluene, cooling to 0-10 ℃, keeping the temperature at 0-10 ℃, stirring for 3 hours, performing suction filtration, retaining the crystals, and directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal is crystal form B.

Example 9

Preparation and identification of a cytarabine prodrug MB07133 crystal form B crystal.

Adding 100mL of dimethyl sulfoxide into a 500mL three-necked bottle, adding 10g of MB07133 crystal form C crystals, stirring and heating to 130 ℃ to completely dissolve the solid, then dropwise adding 100mL of toluene, cooling to 0-10 ℃, keeping the temperature at 0-10 ℃, stirring for 3 hours, performing suction filtration, retaining the crystals, and then directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal is crystal form B.

Example 10

Preparation and identification of a cytarabine prodrug MB07133 crystal form B crystal.

Adding 100mL of dimethyl sulfoxide into a 500mL three-necked bottle, adding 10g of MB07133 crystal form D crystals, stirring and heating to 128 ℃ to completely dissolve the solid, then dropwise adding 100mL of toluene, cooling to 0-10 ℃, keeping the temperature at 0-10 ℃, stirring for 3 hours, performing suction filtration, retaining the crystals, and directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal is crystal form B.

Example 11

Preparation and identification of a cytarabine prodrug MB07133 crystal form B crystal.

Adding 100mL of dimethyl sulfoxide into a 500mL three-necked bottle, adding 10g of MB07133 crystal form E crystals, stirring and heating to 125 ℃ to completely dissolve the solid, then dropwise adding 100mL of toluene, cooling to 0-10 ℃, keeping the temperature at 0-10 ℃, stirring for 3 hours, performing suction filtration, retaining the crystals, and directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal is crystal form B.

Example 12

Preparation and identification of a cytarabine prodrug MB07133 crystal form B crystal.

Adding 100mL of dimethyl sulfoxide into a 500mL three-necked bottle, adding 10g of MB07133 crystal form F crystals, stirring and heating to 128 ℃ to completely dissolve the solid, then dropwise adding 100mL of toluene, cooling to 0-10 ℃, keeping the temperature at 0-10 ℃, stirring for 3 hours, performing suction filtration, retaining the crystals, and directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal is crystal form B.

Example 13

Preparation and identification of a crystalline form C crystal of a cytarabine prodrug MB 07133.

Adding 100mL of dimethyl sulfoxide into a 500mL three-necked bottle, adding 10g of the obtained MB07133 white powder, stirring and heating to 115 ℃ to completely dissolve the solid, then dropwise adding 150mL of acetone, cooling to 0-10 ℃ after the addition, keeping the temperature at 0-10 ℃, stirring for 3h, performing suction filtration, retaining crystals, and then directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal form B is a crystal form B, and as shown in figures 3 and 9, the stable crystal form B can be obtained by the method.

Example 14

Preparation and identification of a crystalline form C crystal of a cytarabine prodrug MB 07133.

Adding 100mL of dimethyl sulfoxide into a 500mL three-necked bottle, adding 10g of MB07133 crystal form A crystals, stirring and heating to 120 ℃ to completely dissolve the solid, then dropwise adding 150mL of acetone, cooling to 0-10 ℃, keeping the temperature at 0-10 ℃, stirring for 3 hours, performing suction filtration, retaining the crystals, and directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal is crystal form C.

Example 15

Preparation and identification of a crystalline form C crystal of a cytarabine prodrug MB 07133.

Adding 100mL of dimethyl sulfoxide into a 500mL three-necked bottle, adding 10g of MB07133 crystal form B crystals, stirring and heating to 118 ℃ to completely dissolve the solid, then dropwise adding 150mL of acetone, cooling to 0-10 ℃, keeping the temperature at 0-10 ℃, stirring for 3 hours, performing suction filtration, retaining the crystals, and directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal is crystal form C.

Example 16

Preparation and identification of a crystalline form C crystal of a cytarabine prodrug MB 07133.

Adding 100mL of dimethyl sulfoxide into a 500mL three-necked bottle, adding 10g of MB07133 crystal form D crystals, stirring and heating to 118 ℃ to completely dissolve the solid, then dropwise adding 150mL of acetone, cooling to 0-10 ℃, keeping the temperature at 0-10 ℃, stirring for 3 hours, performing suction filtration, retaining the crystals, and directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal is crystal form C.

Example 17

Preparation and identification of a crystalline form C crystal of a cytarabine prodrug MB 07133.

Adding 100mL of dimethyl sulfoxide into a 500mL three-necked bottle, adding 10g of MB07133 crystal form E crystals, stirring and heating to 115 ℃ to completely dissolve the solid, then dropwise adding 150mL of acetone, cooling to 0-10 ℃, keeping the temperature at 0-10 ℃, stirring for 3 hours, performing suction filtration, retaining the crystals, and directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal is crystal form C.

Example 18

Preparation and identification of a crystalline form C crystal of a cytarabine prodrug MB 07133.

Adding 100mL of dimethyl sulfoxide into a 500mL three-necked bottle, adding 10g of MB07133 crystal form F crystals, stirring and heating to 120 ℃ to completely dissolve the solid, then dropwise adding 150mL of acetone, cooling to 0-10 ℃, keeping the temperature at 0-10 ℃, stirring for 3 hours, performing suction filtration, retaining the crystals, and directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal is crystal form C.

Example 19

Preparation and identification of a cytarabine prodrug MB07133 crystal form D crystal.

Adding 100mL LN, N-dimethylformamide into a 500mL three-necked bottle, adding 10g of the obtained MB07133 white powder, stirring and heating to 120 ℃, completely dissolving the solid, then cooling and dropwise adding 200mL ethyl acetate, cooling to 0-10 ℃, keeping the temperature at 0-10 ℃, stirring for 3 hours, performing suction filtration, keeping crystals, and then directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC measurements showed crystalline form D, as shown in figures 4 and 10, indicating that stable crystalline form D can be obtained with this method.

Example 20

Preparation and identification of a cytarabine prodrug MB07133 crystal form D crystal.

Adding 100mL LN, N-dimethylformamide into a 500mL three-neck flask, adding 10g of MB07133 crystal form A crystal, stirring and heating to 125 ℃ to completely dissolve the solid, then cooling and dropwise adding 200mL ethyl acetate, cooling to 0-10 ℃ after adding, keeping the temperature at 0-10 ℃, stirring for 3h, performing suction filtration, retaining the crystal, and then directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal form D is crystal form.

Example 21

Preparation and identification of a cytarabine prodrug MB07133 crystal form D crystal.

Adding 100mL LN, N-dimethylformamide into a 500mL three-neck flask, adding 10g of MB07133 crystal form B crystal, stirring and heating to 120 ℃ to completely dissolve the solid, then cooling and dropwise adding 200mL ethyl acetate, cooling to 0-10 ℃ after adding, keeping the temperature at 0-10 ℃, stirring for 3h, performing suction filtration, retaining the crystal, and then directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal form D is crystal form.

Example 22

Preparation and identification of a cytarabine prodrug MB07133 crystal form D crystal.

Adding 100mL LN, N-dimethylformamide into a 500mL three-necked bottle, adding 10g of MB07133 crystal form C crystal, stirring and heating to 118 ℃ to completely dissolve the solid, then cooling and dropwise adding 200mL ethyl acetate, cooling to 0-10 ℃ after adding, keeping the temperature at 0-10 ℃, stirring for 3h, performing suction filtration, retaining the crystal, and then directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal form D is crystal form.

Example 23

Preparation and identification of a cytarabine prodrug MB07133 crystal form D crystal.

Adding 100mL LN, N-dimethylformamide into a 500mL three-neck flask, adding 10g of MB07133 crystal form E crystal, stirring and heating to 116 ℃ to completely dissolve the solid, then cooling and dropwise adding 200mL ethyl acetate, cooling to 0-10 ℃ after adding, keeping the temperature at 0-10 ℃, stirring for 3h, performing suction filtration, retaining the crystal, and then directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal form D is crystal form.

Example 24

Preparation and identification of a cytarabine prodrug MB07133 crystal form D crystal.

Adding 100mL LN, N-dimethylformamide into a 500mL three-necked bottle, adding 10g of MB07133 crystal form F crystal, stirring and heating to 118 ℃ to completely dissolve the solid, then cooling and dropwise adding 200mL ethyl acetate, cooling to 0-10 ℃ after adding, keeping the temperature at 0-10 ℃, stirring for 3h, performing suction filtration, retaining the crystal, and then directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show that the crystal form D is crystal form.

Example 25

Preparation and identification of crystal form E crystal of cytarabine prodrug MB 07133.

Adding 20mL of N-methylpyrrolidone into a 500mL three-necked bottle, adding 20mL of water, adding 10g of the obtained MB07133 white powder, stirring and heating to 100 ℃ to completely dissolve the solid, then cooling and dropwise adding 100mL of acetone, cooling to 0-10 ℃ after the addition is finished, keeping the temperature at 0-10 ℃, stirring for 3 hours, performing suction filtration, keeping the crystal, and then directly performing forced air drying at 50 ℃. X-ray powder diffraction and DSC measurements showed crystalline form D, as shown in figures 5 and 11, indicating that stable crystalline form E crystals can be obtained with this method.

Example 26

Preparation and identification of crystal form E crystal of cytarabine prodrug MB 07133.

Adding 20mL of N-methylpyrrolidone into a 500mL three-necked bottle, adding 20mL of water, adding 10g of MB07133 crystal form A crystal, stirring and heating to 105 ℃, completely dissolving the solid, then dropwise adding 100mL of acetone, cooling to 0-10 ℃, keeping the temperature at 0-10 ℃, stirring for 3h, performing suction filtration, retaining the crystal, and then directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show crystal form E.

Example 27

Preparation and identification of crystal form E crystal of cytarabine prodrug MB 07133.

Adding 20mL of N-methylpyrrolidone into a 500mL three-necked bottle, adding 20mL of water, adding 10g of MB07133 crystal form B crystal, stirring and heating to 110 ℃ to completely dissolve the solid, then dropwise adding 100mL of acetone, cooling to 0-10 ℃ after the addition, keeping the temperature at 0-10 ℃, stirring for 3h, performing suction filtration, retaining the crystal, and then directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show crystal form E.

Example 28

Preparation and identification of crystal form E crystal of cytarabine prodrug MB 07133.

Adding 20mL of N-methylpyrrolidone into a 500mL three-necked bottle, adding 20mL of water, adding 10g of MB07133 crystal form C crystals, stirring and heating to 110 ℃ to completely dissolve the solid, then dropwise adding 100mL of acetone, cooling to 0-10 ℃ after the addition, keeping the temperature at 0-10 ℃, stirring for 3h, performing suction filtration, retaining the crystals, and then directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show crystal form E.

Example 29

Preparation and identification of crystal form E crystal of cytarabine prodrug MB 07133.

Adding 20mL of N-methylpyrrolidone into a 500mL three-necked bottle, adding 20mL of water, adding 10g of MB07133 crystal form D crystal, stirring and heating to 107 ℃ to completely dissolve the solid, then dropwise adding 100mL of acetone, cooling to 0-10 ℃ after the addition, keeping the temperature at 0-10 ℃, stirring for 3h, performing suction filtration, retaining the crystal, and then directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show crystal form E.

Example 30

Preparation and identification of crystal form E crystal of cytarabine prodrug MB 07133.

Adding 20mL of N-methylpyrrolidone into a 500mL three-necked bottle, adding 20mL of water, adding 10g of MB07133 crystal form F crystal, stirring and heating to 120 ℃ to completely dissolve the solid, then dropwise adding 100mL of acetone, cooling to 0-10 ℃ after the addition, keeping the temperature at 0-10 ℃, stirring for 3h, performing suction filtration, retaining the crystal, and then directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show crystal form E.

Example 31

Preparation and identification of a crystalline form F of a cytarabine prodrug MB 07133.

Respectively adding 10mL of purified water and 10mL of DMSO into a 250mL three-necked bottle, adding 5g of the obtained MB07133 white powder, stirring and heating to 100 ℃ until the solid is completely dissolved, then dropwise adding 150mL of isopropanol, cooling to 0-10 ℃ after the addition is finished, keeping the temperature at 0-10 ℃, stirring for 3h, performing suction filtration, retaining crystals, and then directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC measurements showed crystalline form F, as shown in figures 6 and 12, indicating that stable crystalline form F can be obtained with this method.

Example 32

Preparation and identification of a crystalline form F of a cytarabine prodrug MB 07133.

Respectively adding 20mL of purified water and 20mL of DMSO into a 500mL three-necked bottle, then adding 10g of MB07133 crystal form A crystal, stirring and heating to 110 ℃ until the solid is completely dissolved, then dropwise adding 300mL of isopropanol, cooling to 0-10 ℃ after the addition, keeping the temperature at 0-10 ℃, stirring for 3h, performing suction filtration, retaining the crystal, and then directly performing air blowing drying at 50 ℃. X-ray powder diffraction and DSC detection show crystal form F.

Example 33

Preparation and identification of a crystalline form C crystal of a cytarabine prodrug MB 07133.

Respectively adding 20mL of purified water and 20mL of LDMSO into a 250mL three-necked bottle, then adding 10g of MB07133 crystal form B crystal, stirring and heating to 120 ℃ until the solid is completely dissolved, then dropwise adding 300mL of isopropanol, cooling to 0-10 ℃ after adding, keeping the temperature at 0-10 ℃, stirring for 3h, performing suction filtration, retaining the crystal, and then directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show crystal form F.

Example 34

Preparation and identification of a crystalline form F of a cytarabine prodrug MB 07133.

Respectively adding 20mL of purified water and 20mL of LDMSO into a 250mL three-necked bottle, then adding 10g of MB07133 crystal form C crystals, stirring and heating to 110 ℃ until the solid is completely dissolved, then dropwise adding 300mL of isopropanol, cooling to 0-10 ℃ after adding, keeping the temperature at 0-10 ℃, stirring for 3h, performing suction filtration, retaining the crystals, and then directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show crystal form F.

Example 35

Preparation and identification of a crystalline form F of a cytarabine prodrug MB 07133.

Respectively adding 20mL of purified water and 20mL of LDMSO into a 250mL three-necked bottle, then adding 10g of MB07133 crystal form D crystal, stirring and heating to 115 ℃ until the solid is completely dissolved, then dropwise adding 300mL of isopropanol, cooling to 0-10 ℃ after adding, keeping the temperature at 0-10 ℃, stirring for 3h, performing suction filtration, retaining the crystal, and then directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show crystal form F.

Example 36

Preparation and identification of a crystalline form F of a cytarabine prodrug MB 07133.

Respectively adding 20mL of purified water and 20mL of LDMSO into a 250mL three-necked bottle, then adding 10g of MB07133 crystal form E crystal, stirring and heating to 112 ℃ until the solid is completely dissolved, then dropwise adding 300mL of isopropanol, cooling to 0-10 ℃ after adding, keeping the temperature at 0-10 ℃, stirring for 3h, performing suction filtration, retaining the crystal, and then directly drying by blowing at 50 ℃. X-ray powder diffraction and DSC detection show crystal form F.

Example 37

Stability of crystalline form a compounds

This example describes stability experiments for crystalline form a compounds of the present invention.

The stability test of the crystal compound of the crystal form A under the three conditions of high temperature, high humidity and illumination shows that the results are shown in the following table (table 1), and related substances of a sample are basically unchanged under the conditions of high temperature (60 ℃), high humidity (90% +/-5%) and strong light (4500Lx), which shows that the crystal form A is stable to the conditions of high temperature (60 ℃), high humidity and illumination.

Table 1 table for measuring stability results of crystal form a

The results of the stability test at 40 ℃ over 6 months are shown in the following table (table 2), which indicates that the crystals of the present invention have good stability and are suitable for long-term storage.

Table 2 long-term test measurement result table for crystal form a

Example 38

Stability of crystalline compound of form B

This example describes stability experiments for crystalline form B compounds of the present invention.

The stability test of the crystal compound of the crystal form B under the three conditions of high temperature, high humidity and illumination shows that the results are shown in the following table (3), and the related substances of the sample are basically unchanged under the conditions of high temperature (60 ℃), high humidity (90% +/-5%) and strong light (4500Lx), which shows that the crystal form B is stable to the conditions of high temperature (60 ℃), high humidity and illumination.

Table 3 table for measuring stability results of crystal form B

The results of the stability test at 40 ℃ over 6 months are shown in the following table (table 4), which indicates that the crystals of the present invention have good stability and are suitable for long-term storage.

Table 4 long-term test measurement result table for crystal form B

Example 39

Stability of crystalline form C compounds

This example describes stability experiments for crystalline form C compounds of the present invention.

The stability test of the crystal form C crystal compound under the three conditions of high temperature, high humidity and illumination shows that the results are shown in the following table (5), and the related substances of the sample are basically unchanged under the conditions of high temperature (60 ℃), high humidity (90% +/-5%) and strong light (4500Lx), which indicates that the crystal form B is stable to the conditions of high temperature of 60 ℃, high humidity and illumination.

TABLE 5 table for determining stability results of crystal form C

The results of the stability test at 40 ℃ over 6 months are shown in the following table (table 6), which indicates that the crystals of the present invention have good stability and are suitable for long-term storage.

Table 6 long-term test measurement result table for crystal form C

Example 40

Stability of crystalline form D compounds

This example describes stability experiments for crystalline form D compounds of the present invention.

The stability test of the crystal form D crystal compound under the three conditions of high temperature, high humidity and illumination shows that the results are shown in the following table (7), and the related substances of the sample are basically unchanged under the conditions of high temperature (60 ℃), high humidity (90% +/-5%) and strong light (4500Lx), which indicates that the crystal form D is stable to the conditions of high temperature of 60 ℃, high humidity and illumination.

Table 7 table for measuring stability results of crystal form D

The results of the stability test at 40 ℃ over 6 months are shown in the following table (Table 8), which indicates that the crystals of the present invention are excellent in stability and suitable for long-term storage.

Table 8 table of long-term test measurement results of crystal form D

EXAMPLE 41

Stability of crystalline form E compound

This example describes stability experiments for crystalline form E compounds of the present invention.

The stability test of the crystal form E crystal compound under the three conditions of high temperature, high humidity and illumination shows that the results are shown in the following table (table 9), and the related substances of the sample are basically unchanged under the conditions of high temperature (60 ℃), high humidity (90% +/-5%) and strong light (4500Lx), which indicates that the crystal form E is stable to the conditions of high temperature of 60 ℃, high humidity and illumination.

TABLE 9 table for determining stability results of crystal form E

The results of the stability test for 6 months at 40 ℃ are shown in the following table (Table 10), which indicates that the crystals of the present invention are excellent in stability and suitable for long-term storage.

TABLE 10 Long-term test measurement results of form E

Example 42

Stability of crystalline form F compounds

This example describes stability experiments for crystalline form F compounds of the present invention.

The stability test of the crystal compound of the crystal form F under the three conditions of high temperature, high humidity and illumination shows that the following table (table 11) shows that the related substances of the sample are basically unchanged under the conditions of high temperature (60 ℃), high humidity (90% +/-5%) and strong light (4500Lx), which indicates that the crystal form F is stable to the high temperature of 60 ℃, the high humidity and the illumination.

TABLE 11 table for determining stability results of form F

The results of the stability test for 6 months at 40 ℃ are shown in the following table (table 12), which indicates that the crystals of the present invention are excellent in stability and suitable for long-term storage.

Table 12 table of long-term test measurement results of crystal form F

Example 43

Pharmaceutical composition comprising the crystalline form a of the present invention

Weighing or measuring materials according to the following formula (table 13), adding the crystal form A crystal of MB07133 into injection water at the temperature of 2-8 ℃, stirring, adding phosphoric acid to adjust the pH value to 3.8-4.0, and continuing stirring to clarify the solution. Placing in a low-temperature water bath, keeping the temperature at (2-8 ℃), adding 0.1% (w/v) of activated carbon for injection, stirring for 30min, removing carbon, performing primary filtration through a 0.45 mu m filter membrane, performing fine filtration through a 0.22 mu m filter membrane, filling and half-plugging, placing in a freeze dryer, freeze drying, performing full-pressing plugging, and capping. Obtaining the MB07133 freeze-dried powder injection.

TABLE 13 pharmaceutical formulations of form A crystals

Example 44

Pharmaceutical composition comprising crystalline form B of the present invention

Weighing or measuring materials according to the following formula (table 14), adding the crystal form B crystal of MB07133 into injection water at the temperature of 2-8 ℃, stirring, adding phosphoric acid to adjust the pH value to 3.8-4.0, and continuing stirring to clarify the solution. Placing in a low-temperature water bath, keeping the temperature at (2-8 ℃), adding 0.1% (w/v) of activated carbon for injection, stirring for 30min, removing carbon, performing primary filtration through a 0.45 mu m filter membrane, performing fine filtration through a 0.22 mu m filter membrane, filling and half-plugging, placing in a freeze dryer, freeze drying, performing full-pressing plugging, and capping. Obtaining the MB07133 freeze-dried powder injection.

TABLE 14 pharmaceutical formulations of form B crystals

Example 45

Pharmaceutical composition comprising crystalline form C of the present invention

Weighing or measuring materials according to the following formula (table 15), adding the crystal form C crystal of MB07133 into injection water at the temperature of 2-8 ℃, stirring, adding phosphoric acid to adjust the pH value to 3.8-4.0, and continuing stirring to clarify the solution. Placing in a low-temperature water bath, keeping the temperature at (2-8 ℃), adding 0.1% (w/v) of activated carbon for injection, stirring for 30min, removing carbon, performing primary filtration through a 0.45 mu m filter membrane, performing fine filtration through a 0.22 mu m filter membrane, filling and half-plugging, placing in a freeze dryer, freeze drying, performing full-pressing plugging, and capping. Obtaining the MB07133 freeze-dried powder injection.

TABLE 15 pharmaceutical formulations of form C crystals

Example 46

Pharmaceutical composition comprising crystalline form D of the present invention

Weighing or measuring materials according to the following formula (table 16), adding the crystal form D crystal of MB07133 into injection water at the temperature of 2-8 ℃, stirring, adding phosphoric acid to adjust the pH value to 3.8-4.0, and continuing stirring to clarify the solution. Placing in a low-temperature water bath, keeping the temperature at (2-8 ℃), adding 0.1% (w/v) of activated carbon for injection, stirring for 30min, removing carbon, performing primary filtration through a 0.45 mu m filter membrane, performing fine filtration through a 0.22 mu m filter membrane, filling and half-plugging, placing in a freeze dryer, freeze drying, performing full-pressing plugging, and capping. Obtaining the MB07133 freeze-dried powder injection.

TABLE 16 pharmaceutical formulations of form D crystals

Example 47

Pharmaceutical composition comprising crystalline form E of the invention

Weighing or measuring materials according to the following formula (table 17), adding the crystal form E crystal of MB07133 into injection water at the temperature of 2-8 ℃, stirring, adding phosphoric acid to adjust the pH value to 3.8-4.0, and continuing stirring to clarify the solution. Placing in a low-temperature water bath, keeping the temperature at (2-8 ℃), adding 0.1% (w/v) of activated carbon for injection, stirring for 30min, removing carbon, performing primary filtration through a 0.45 mu m filter membrane, performing fine filtration through a 0.22 mu m filter membrane, filling and half-plugging, placing in a freeze dryer, freeze drying, performing full-pressing plugging, and capping. Obtaining the MB07133 freeze-dried powder injection.

TABLE 17 pharmaceutical formulations of form E crystals

Example 48

Pharmaceutical composition comprising crystalline form F of the present invention

Weighing or measuring materials according to the following formula (table 18), adding the crystal form F crystal of MB07133 into injection water at the temperature of 2-8 ℃, stirring, adding phosphoric acid to adjust the pH value to 3.8-4.0, and continuing stirring to clarify the solution. Placing in a low-temperature water bath, keeping the temperature at (2-8 ℃), adding 0.1% (w/v) of activated carbon for injection, stirring for 30min, removing carbon, performing primary filtration through a 0.45 mu m filter membrane, performing fine filtration through a 0.22 mu m filter membrane, filling and half-plugging, placing in a freeze dryer, freeze drying, performing full-pressing plugging, and capping. Obtaining the MB07133 freeze-dried powder injection.

Table 18 pharmaceutical formulations of form F crystals