CN106995441B - Crystal form, preparation method, pharmaceutical composition and the purposes of imidazolone compounds - Google Patents

Abstract

The present invention relates to the crystal form of imidazolone compounds, preparation method, pharmaceutical composition and purposes, belong to medical compounds field of crystals.Crystal form provided by the invention is with good stability, is included in good stability under three kinds of extreme conditions such as high temperature, high humidity and intense light irradiation, and good stability is also kept in tableting processes.Crystal form provided by the invention has good body absorption metabolisming property, including blood concentration, Drug-time curve AUC, half-life period etc..Moreover, crystal form solution rate of the invention is improved, be conducive to the application on preparation.

Description

Crystal form of imidazolone compound, preparation method, pharmaceutical composition and application thereof

Technical Field

The invention relates to a crystal form of imidazolone compounds with PI3K/mTOR dual-inhibition activity, a preparation method, a pharmaceutical composition and application thereof, and belongs to the field of pharmaceutical compound crystals.

Background art:

mammalian target of rapamycin (mTOR), a classical serine/threonine protein kinase, belongs to the phosphatidylinositol-3 kinase (PI 3K) related kinase family members, and is a major signaling molecule for cellular functions such as intracellular synthesis and catabolism. mTOR signaling pathways are closely related to nutrition, energy status and growth factors. It regulates a number of cellular processes including autophagy, protein, lipid, lysosomal synthesis and energy metabolism, cytoskeletal organization, cell survival, and the like. mTOR regulates the switch of anabolism and catabolism under changing mammalian cell peripheral nutritional conditions, thereby enabling cells to grow and survive under different nutritional conditions. Due to the important role of mTOR in cells, aberrant or deregulated mTOR signaling can lead to the development of human diseases (e.g., diseases such as cancer). Therefore, the mTOR signaling pathway is becoming an important target for designing anticancer drugs.

The PI3K/Akt/mTOR signal pathway activation is closely related to the occurrence of various tumors, and mTOR can accelerate the cell cycle, reduce apoptosis and promote the migration of tumor cells in brain glioma, breast cancer and ovarian cancer. Activation of mTOR initiates with growth factor receptors on the surface of some cells that are activated by ligands, such as epidermal growth factor receptor and insulin-like growth factor-1 and-2 (IGF-1 and-2). Activation of the receptor results in activation of PI3K kinase, which in turn results in activation of downstream effector Akt proteins. Akt is a regulatory factor that regulates cell survival at multiple levels. Phosphorylation of Akt inhibits the downstream TSC1/2 complex, resulting in activation of mTOR by Rheb. Downstream of the signaling pathways for PI3K/Akt and PEN/Akt and Ras/Erk1/2, the TSC1/2 complex plays a critical role in regulating mTOR activation.

It has now been found that two different mTOR protein complexes, mTORC1 and mTORC2, exist within the cell. These two protein complexes comprise unique proteins that interact with mTOR and are each regulated by different mechanisms. Significant progress has been made in the development of mTOR inhibitor drugs. Rapamycin was the first mTOR inhibitor discovered and showed better cancer inhibitory effects in various cancer models. Although rapamycin analogues with better pharmacological properties have been developed, clinically useful rapamycin analogues are limited to a few cancers. Akt is an important kinase for cancer cell survival, and mTORC2 can directly phosphorylate Akt, which provides a new idea for the research of mTORC2 on anticancer aspect, and promotes the research and development of second generation anticancer drugs simultaneously acting on two targets of mTORC1 and mTORC 2. Simultaneous inhibition of the activity of both mTOR complexes (mTORC1 and mTORC2) in cancer cells has a broader and more potent anticancer effect.

Compound 1, chemically 1- ((1s,4s) -4-hydroxycyclohexyl) -3-methyl-8- (6- (1-methyl-1H-pyrazol-4-yl) pyridin-3-yl) -1H-imidazo [4,5-c ] quinolin-2 (3H) -one, is a dual inhibitor of the protein kinase PI3K/mTOR and has the structure shown in the following formula:

compound 1

Compound 1 and its pharmaceutically acceptable salts have been disclosed in WO2015074516a1, which reportedly show good pharmaceutical activity in cellular and animal models. Therefore, the development of a more stable and more suitable preparation and a crystal form of the compound 1 with better absorption and metabolism has important significance for clinical application.

Disclosure of Invention

The present invention provides a crystal of a pharmaceutically acceptable salt (e.g., hydrochloride) of compound 1 represented by the following formula:

compound 1.

The name of compound 1 is 1- ((1s,4s) -4-hydroxycyclohexyl) -3-methyl-8- (6- (1-methyl-1H-pyrazol-4-yl) pyridin-3-yl) -1H-imidazo [4,5-c ] quinolin-2 (3H) -one. The preparation of said compound 1 and its hydrochloride is described for example in WO2015074516a1, example 18. The WO2015074516a1 publication is incorporated herein by reference in its entirety.

The invention provides a compound 1 hydrochloride monohydrate of form I characterized by X-ray powder diffraction characteristic peaks expressed in 2 theta angles (°) using Cu-K α radiation that may include 9.028 ± 0.2, 11.196 ± 0.2, 17.393 ± 0.2, 22.504 ± 0.2.

According to the invention, the X-ray powder diffraction characteristic peaks of the form I, which are radiated by Cu-K α and expressed by 2 theta angle (°), can comprise 9.028 +/-0.2, 11.196 +/-0.2, 15.406 +/-0.2, 16.380 +/-0.2, 17.393 +/-0.2, 18.066 +/-0.2, 18.739 +/-0.2, 20.894 +/-0.2, 22.504 +/-0.2 and 22.955 +/-0.2.

Preferably, the X-ray powder diffraction characteristic peaks expressed by 2 theta angle (°) of the crystal form I irradiated by Cu-K α comprise 9.028 +/-0.2, 11.196 +/-0.2, 15.406 +/-0.2, 16.380 +/-0.2, 17.393 +/-0.2, 18.066 +/-0.2, 18.739 +/-0.2, 20.894 +/-0.2, 22.504 +/-0.2, 22.955 +/-0.2, 26.312 +/-0.2, 26.918 +/-0.2, 27.556 +/-0.2 and 35.168 +/-0.2.

More preferably, the characteristic peaks of X-ray powder diffraction of the form I radiated by Cu-K α and expressed by 2 theta angle (DEG) comprise 9.028 + -0.2, 11.196 + -0.2, 12.200 + -0.2, 15.406 + -0.2, 16.380 + -0.2, 16.828 + -0.2, 17.393 + -0.2, 18.066 + -0.2, 18.739 + -0.2, 20.036 + -0.2, 20.894 + -0.2, 22.504 + -0.2, 22.955 + -0.2, 24.973 + -0.2, 25.505 + -0.2, 26.312 + -0.2, 26.918 + -0.2, 27.556 + -0.2, 28.403 + -0.2, 29.176 + -0.2, 31.586 + -0.2 and 35.168 + -0.2.

Preferably, form I has a Cu-K α radiation X-ray powder diffraction pattern substantially as shown in figure 1.

The present invention also provides a process a for preparing form I of compound 1 hydrochloride monohydrate, comprising:

1) dissolving compound 1 hydrochloride in water;

2) adding sodium chloride into the solution of the step 1);

3) and cooling, crystallizing, filtering and drying to obtain the crystal form I.

According to the present invention, it is preferable that,

in step 1), water may be heated before or after addition of compound 1 hydrochloride to dissolve compound 1 hydrochloride; wherein, the amount of water may be 2 to 80 times, for example, 4 to 70 times, 6 to 60 times, 8 to 50 times or 10 to 25 times of the weight of the hydrochloride of the compound 1; the water can be heated, for example, to 70-100 deg.C, such as 75 deg.C, 80 deg.C, 85 deg.C, 90 deg.C, or 95 deg.C;

in the step 2), sodium chloride can be added while keeping the temperature of the solution in the step 1); as an example, the amount of sodium chloride may be controlled so that it is 0.1 to 26%, such as 0.5 to 20%, 0.8 to 15%, or 1 to 10%, such as 3 to 5%, of the total weight of the solution;

the sodium chloride may be in its suitable form, for example a sodium chloride solution or a sodium chloride solid may be used. Preferably, sodium chloride is added and stirred to dissolve; the sodium chloride solution is preferably an aqueous sodium chloride solution, wherein the weight percentage of sodium chloride may be from 10% to a saturation concentration, e.g., 12%, 15%, 16%, 17%, 18%, 20%, 22%, 24%, 25%, 26%;

in the step 3), the temperature can be slowly reduced to below 60 ℃ (such as 20-50 ℃) for crystallization under stirring, and the crystal form I of the hydrochloride monohydrate of the compound 1 can be obtained by suction filtration, leaching and vacuum drying at 15-35 ℃ (such as 20 ℃, 25 ℃ and 30 ℃).

The present invention also provides a process B for preparing form I of compound 1 hydrochloride monohydrate, comprising:

1) dissolving compound 1 hydrochloride in an aqueous ethanol solution;

2) and cooling, crystallizing, filtering and drying to obtain the crystal form I.

According to the present invention, it is preferable that,

in step 1), the aqueous ethanol solution may be heated before or after the addition of the compound 1 hydrochloride to dissolve the compound 1 hydrochloride; wherein, the amount of the ethanol aqueous solution can be 5 to 80 times of the weight of the hydrochloride of the compound 1, such as 10 to 70 times, 20 to 60 times or 30 to 50 times; the aqueous ethanol solution can be heated, for example, to 50-100 deg.C, such as 55 deg.C, 60 deg.C, 65 deg.C, 70 deg.C, 75 deg.C, 80 deg.C, 85 deg.C, 90 deg.C or 95 deg.C;

the mass percentage of ethanol in the ethanol water solution can be, for example, 30-99%, such as 40-98%, 45-95%, 46-85%, 48-80%, 50-75% or 60-70%;

in the step 2), the temperature can be slowly reduced to below 40 ℃ (such as 20-30 ℃) for crystallization under stirring, suction filtration is carried out, and vacuum drying is carried out at 15-35 ℃ (such as room temperature 25 ℃) to obtain the crystal form I of the hydrochloride monohydrate of the compound 1.

The present invention also provides crystalline form II of the hydrochloride monohydrate of compound 1 characterized by X-ray powder diffraction characteristic peaks expressed in 2 Θ angles (°) using radiation of Cu-K α may include 8.934 ± 0.2, 11.126 ± 0.2, 15.367 ± 0.2, 22.437 ± 0.2.

According to the invention, the characteristic peaks of the crystal form II in X-ray powder diffraction expressed by 2 theta angle (°) by using Cu-K α radiation comprise 8.934 +/-0.2, 11.126 +/-0.2, 12.161 +/-0.2, 15.367 +/-0.2, 16.289 +/-0.2, 17.369 +/-0.2, 18.037 +/-0.2, 18.667 +/-0.2, 20.896 +/-0.2, 22.437 +/-0.2, 22.928 +/-0.2, 24.995 +/-0.2, 26.269 +/-0.2, 26.890 +/-0.2 and 27.574 +/-0.2.

Preferably, the characteristic peaks of X-ray powder diffraction of the crystal form II irradiated by Cu-K α and expressed by 2 theta angle (DEG) comprise 8.486 + -0.2, 8.934 + -0.2, 11.126 + -0.2, 12.161 + -0.2, 13.317 + -0.2, 15.367 + -0.2, 16.289 + -0.2, 16.742 + -0.2, 17.369 + -0.2, 18.037 + -0.2, 18.667 + -0.2, 19.966 + -0.2, 20.896 + -0.2, 22.437 + -0.2, 22.928 + -0.2, 24.995 + -0.2, 25.467 + -0.2, 26.269 + -0.2, 26.890 + -0.2, 27.213 + -0.2, 27.574 + -0.2, 28.366 + -0.2, 29.075 + -0.2 and 35.001 + -0.2.

Preferably, form II has a Cu-K α radiation X-ray powder diffraction pattern substantially as shown in figure 8.

The present invention also provides a process for preparing crystalline form II of hydrochloride monohydrate of compound 1 comprising:

1) mixing saturated solutions of compound 1 hydrochloride in two different solvents;

2) volatilizing the solvent from the mixture of step 1) to obtain form II.

According to the production method of the present invention, preferably,

in the step 1), the two saturated solutions can be mixed at the temperature of 10-35 ℃, preferably 20-25 ℃; the solvent is selected from organic solvents, for example from one or more of the following: ester solvents (e.g., ethyl acetate, methyl acetate, ethyl formate, methyl formate), ketone solvents (e.g., acetone, 2-butanone), ether solvents (e.g., tetrahydrofuran, 1, 4-dioxane, methyl tert-butyl ether, methyl isopropyl ether, methyl ethyl ether, diethyl ether), nitrile solvents (e.g., acetonitrile, propionitrile); preferably, the ratio of the total volume of each of the two saturated solutions is 2:1 to 1:2, such as 1: 1; for example, the two saturated solutions can be mixed in a 96-well plate;

in the step 2), the mixture obtained in the step 1) can be placed in an atmospheric environment to slowly volatilize the solvent; as an example, form II can be obtained by covering a 96-well plate with a perforated sealing film, placing in a fume hood, and naturally evaporating to dryness in the atmosphere.

The present invention also provides form III of the hydrochloride salt of compound 1, characterized in that X-ray powder diffraction peaks expressed in 2 Θ angles (°) using Cu-K α radiation may comprise 6.396 ± 0.2, 7.115 ± 0.2, 8.972 ± 0.2, 10.803 ± 0.2, 11.870 ± 0.2, 18.542 ± 0.2, 23.071 ± 0.2.

According to the present invention, X-ray powder diffraction characteristic peaks of said crystalline form III in 2 θ degrees (°) may include: 6.396 + -0.2, 7.115 + -0.2, 8.972 + -0.2, 10.803 + -0.2, 11.147 + -0.2, 11.870 + -0.2, 12.139 + -0.2, 15.417 + -0.2, 16.297 + -0.2, 16.559 + -0.2, 17.374 + -0.2, 18.074 + -0.2, 18.542 + -0.2, 19.310 + -0.2, 22.464 + -0.2, 23.071 + -0.2, 24.550 + -0.2, 25.843 + -0.2, 26.903 + -0.2, 28.737 + -0.2, 29.664 + -0.2 and 35.016 + -0.2.

Preferably, form III has a Cu-K α radiation X-ray powder diffraction pattern substantially as shown in figure 10.

The invention also provides a preparation method of the compound 1 hydrochloride crystal form III, which comprises the following steps:

1) mixing saturated solutions of compound 1 hydrochloride in two different solvents;

2) volatilizing the solvent from the mixture of step 1) to obtain form III.

According to the production method of the present invention, preferably,

in the step 1), the two saturated solutions can be mixed at a temperature below 35 ℃, preferably 20-25 ℃; the solvent is selected from organic solvents, for example from one or more of the following: alcohol solvents (such as methanol, ethanol, n-propanol, isopropanol, n-butanol), ether solvents (such as tetrahydrofuran, 1, 4-dioxane, methyl tert-butyl ether, methyl isopropyl ether, methyl ethyl ether, diethyl ether); preferably, the ratio of the total volume of each of the two saturated solutions is 2:1 to 1:2, such as 1: 1; for example, the two saturated solutions can be mixed in a 96-well plate;

in the step 2), the mixture obtained in the step 1) can be placed in an atmospheric environment to slowly volatilize the solvent; as an example, form III can be obtained by covering a 96-well plate with a perforated sealing film, placing in a fume hood, and naturally evaporating to dryness in the atmosphere.

The invention also provides a crystal form IV of the hydrochloride of the compound 1, which is characterized in that X-ray powder diffraction characteristic peaks expressed by 2 theta angle (°) by using Cu-K α radiation can comprise 6.178 +/-0.2, 8.996 +/-0.2, 11.170 +/-0.2, 15.393 +/-0.2, 16.343 +/-0.2, 17.349 +/-0.2, 18.064 +/-0.2, 18.708 +/-0.2, 19.479 +/-0.2, 19.994 +/-0.2, 20.901 +/-0.2, 22.470 +/-0.2, 22.935 +/-0.2, 24.964 +/-0.2, 25.504 +/-0.2, 26.287 +/-0.2, 26.920 +/-0.2 and 27.545 +/-0.2.

According to the invention, the X-ray powder diffraction characteristic peaks of the crystal form IV radiated by Cu-K α and expressed by 2 theta angle (DEG) can comprise 6.178 +/-0.2, 6.614 +/-0.2, 7.181 +/-0.72, 7.181 +/-0.2, 7.181 +/-0.72, 7.181, 3672.363 and 7.181 +/-0.72, 7.181 +/-0.2.

Preferably, form IV has a Cu-K α radiation X-ray powder diffraction pattern substantially as shown in figure 11.

The present invention also provides a process for preparing form IV of the hydrochloride salt of compound 1 comprising:

1) mixing saturated solutions of compound 1 hydrochloride in two different solvents;

2) volatilizing the solvent from the mixture of step 1) to obtain form IV.

According to the production method of the present invention, preferably,

in the step 1), the two saturated solutions can be mixed at a temperature below 35 ℃, preferably 20-25 ℃; the solvent is selected from organic solvents, for example from one or more of the following: aromatic hydrocarbon solvents (such as benzene, toluene, xylene, chlorobenzene), ester solvents (such as ethyl acetate, methyl acetate, ethyl formate, methyl formate); preferably, the ratio of the total volume of each of the two saturated solutions is 2:1 to 1:2, such as 1: 1; for example, the two saturated solutions can be mixed in a 96-well plate;

in the step 2), the mixture obtained in the step 1) can be placed in an atmospheric environment to slowly volatilize the solvent; as an example, form IV can be obtained by covering a 96-well plate with a perforated sealing film, placing in a fume hood, and naturally evaporating to dryness in the atmosphere.

The present invention also provides form V of compound 1 hydrochloride dihydrate characterized by X-ray powder diffraction characteristic peaks expressed in 2 Θ angles (°) using radiation of Cu-K α may include 6.181 ± 0.2, 8.318 ± 0.2, 18.223 ± 0.2, 31.778 ± 0.2.

According to the present invention, the X-ray powder diffraction characteristic peaks expressed in 2 theta angle (°) of the form V irradiated using Cu-K α may include 6.181 + -0.2, 7.226 + -0.2, 8.318 + -0.2, 9.524 + -0.2, 10.496 + -0.2, 12.037 + -0.2, 18.223 + -0.2, 27.421 + -0.2, 31.778 + -0.2.

Preferably, form V has a Cu-K α radiation X-ray powder diffraction pattern substantially as shown in figure 13.

The present invention also provides a process for preparing form V of compound 1 hydrochloride dihydrate comprising:

1) preparing a saturated solution of the hydrochloride salt of compound 1 in brine;

2) standing the solution obtained in the step 1), crystallizing, and performing suction filtration to obtain a crystal form V.

According to the production method of the present invention, preferably,

in the step 1), the saturated saline solution of the hydrochloride of the compound 1 is a saturated sodium chloride solution of the hydrochloride of the compound 1; the weight percent of sodium chloride in the saturated solution can be 1% to a saturated concentration, e.g., 1%, 5%, 10%, 12%, 15%, 16%, 17%, 18%, 20%, 22%, 24%, 25%, 26%;

preferably, the saturated solution of step 1) is obtained from the mother liquor filtered in step 3) of the preparation method of the crystal form I;

in step 2), the solution may be left at 40 ℃ or lower, for example 30 ℃ or lower, such as 20-25 ℃ for 8 hours or more, for example overnight, or for 24 hours or more, for example 36 hours or more, 48 hours or more, or seven days or more.

The invention also provides a compound 1 hydrochloride dihydrate form VI characterized in that X-ray powder diffraction peaks expressed in 2 theta angle (°) using Cu-K α radiation can include 7.489 + -0.2, 8.897 + -0.2, 11.140 + -0.2, 11.638 + -0.2, 13.348 + -0.2, 13.755 + -0.2, 16.110 + -0.2, 17.152 + -0.2, 18.782 + -0.2, 19.865 + -0.2, 20.891 + -0.2, 21.477 + -0.2, 25.245 + -0.2, 26.184 + -0.2, 26.431 + -0.2, 27.242 + -0.2, 28.489 + -0.2.

According to the invention, the X-ray powder diffraction characteristic peaks of the crystal form VI radiated by Cu-K α and expressed by 2 theta angle (°) can comprise 7.489 + -0.2, 8.153 + -0.2, 8.897 + -0.2, 11.140 + -0.2, 11.638 + -0.2, 13.348 + -0.2, 13.755 + -0.2, 14.985 + -0.2, 15.467 + -0.2, 16.110 + -0.2, 17.152 + -0.2, 18.240 + -0.2, 18.782 + -0.2, 19.865 + -0.2, 20.891 + -0.2, 21.477 + -0.2, 22.333 + -0.2, 22.888 + -0.2, 25.245 + -0.2, 26.184 + -0.2, 26.431 + -0.2, 27.242 + -0.2, 28.489 + -0.2 and 29.710 + -0.2.

Preferably, form VI has a Cu-K α radiation X-ray powder diffraction pattern substantially as shown in figure 15.

The present invention also provides a process for preparing crystalline form VI of hydrochloride dihydrate of compound 1 comprising:

1) dissolving compound 1 hydrochloride in a mixture of water and acetonitrile;

2) stirring, separating out crystals, filtering and drying to obtain the crystal form VI.

According to the present invention, it is preferable that,

in the step 1), the hydrochloride of the compound 1 can be dissolved in a mixed solution of water and acetonitrile at a temperature of below 40 ℃, for example below 30 ℃, such as 20-25 ℃; the volume percentage of acetonitrile in the mixed solution of water and acetonitrile can be, for example, 5-99%, such as 10-95%, 15-75%, 20-60% or 25-50%;

in the step 2), stirring for 20 hours at a temperature below 40 ℃, for example below 30 ℃, such as 20-25 ℃, to precipitate a large amount of white crystals, performing suction filtration, and vacuum-drying the obtained solid at 25 ℃ to obtain the crystal form VI.

The present invention also provides crystalline form VII of the hydrochloride dihydrate of compound 1 characterized by X-ray powder diffraction characteristic peaks expressed in 2 Θ angles (°) using radiation of Cu-K α may include 6.264 ± 0.2, 6.760 ± 0.2, 7.556 ± 0.2, 14.455 ± 0.2, 20.123 ± 0.2, 26.373 ± 0.2.

According to the invention, the X-ray powder diffraction characteristic peaks of the crystal form VII irradiated by Cu-K α and expressed by 2 theta angle (°) can comprise 6.264 +/-0.2, 6.760 +/-0.2, 7.556 +/-0.2, 11.414 +/-0.2, 11.743 +/-0.2, 12.488 +/-0.2, 13.419 +/-0.2, 14.455 +/-0.2, 17.246 +/-0.2, 18.099 +/-0.2, 20.123 +/-0.2, 21.082 +/-0.2, 25.370 +/-0.2, 26.373 +/-0.2 and 27.294 +/-0.2.

Preferably, form VII has a Cu-K α radiation X-ray powder diffraction pattern substantially as shown in figure 17.

The present invention also provides a process a for preparing crystalline form VII of the hydrochloride dihydrate of compound 1, comprising:

1) dissolving compound 1 hydrochloride in water;

2a) cooling and crystallizing the aqueous solution obtained in the step 1); or 2b) adding sodium chloride into the aqueous solution obtained in the step 1), and stirring for crystallization;

3) filtering and drying to obtain a crystal form VII.

According to the method of the present invention, it is preferred,

in step 1), the amount of water is 10 to 500 times, for example 20 to 200 times, such as 60 or 180 times, the weight of the hydrochloride of the compound 1; preferably, the water is heated before or after the addition of the hydrochloride salt of compound 1; preferably, the method further comprises the step of removing insoluble substances by hot filtration; the water can be heated, for example, to 70-100 deg.C, such as 75 deg.C, 80 deg.C, 85 deg.C, 90 deg.C, or 95 deg.C;

in step 2a), the aqueous solution of step 1) is cooled to below 40 ℃, for example below 30 ℃, such as 20-25 ℃ for crystallization;

in step 2b), stirring at a temperature below 40 ℃, for example below 30 ℃, for example 20-25 ℃ for crystallization; the sodium chloride may be in its suitable form, for example a sodium chloride solution or a sodium chloride solid may be used. Preferably, sodium chloride is added and stirred to dissolve; for example, the amount of sodium chloride may be 1 to 15 times, for example 4 to 8 times, the amount of the hydrochloride of compound 1;

wherein steps 2a) and 2b) are alternatively performed;

in the step 3), the filtration may be suction filtration, and the drying may be vacuum drying.

The present invention also provides a process B for preparing crystalline form VII of the hydrochloride dihydrate of compound 1, comprising:

and carrying out mixed spinning pulping on one or more of the crystal forms I-VI by using water for 3 days to obtain a crystal form VII.

Preferably, the method is one in which the kneading and beating are carried out at a temperature of less than 40 ℃, for example less than 30 ℃, such as 20-25 ℃; the dosage of the water in the mixed spinning pulping is 20-200 times, for example 100 times, of the total weight of the crystal forms I-VI.

The invention also provides a pharmaceutical composition comprising one or more of the above crystals or crystal forms.

According to the present invention, the pharmaceutical composition may further comprise a pharmaceutically acceptable carrier.

A pharmaceutically acceptable carrier is preferably one that is relatively non-toxic and non-injurious to a patient at concentrations consistent with effective activity of the active ingredient, such that any side effects caused by the carrier do not destroy the beneficial effects of the active ingredient. A pharmaceutically effective amount of a compound or a pharmaceutically acceptable salt thereof is preferably an amount that results in, or affects, the particular condition being treated. The compounds of the present invention may be administered together with pharmaceutically acceptable carriers well known in the art in any effective conventional dosage unit form including immediate release, sustained release and timed release formulations in the following manner: oral, parenteral, topical, nasal, ocular, sublingual, rectal, vaginal, and the like.

For oral administration, the compound or a pharmaceutically acceptable salt thereof may be formulated into solid or liquid preparations such as capsules, pills, tablets, troches (troche), dragees (lozenes), melt gels (melt), powders, solutions, suspensions or emulsions, and may be prepared according to methods known in the art for preparing pharmaceutical compositions. The solid unit dosage form may be a capsule, which may be of the ordinary hard or soft capsule type, containing, for example, surfactants, lubricants, and inert fillers (e.g., lactose, sucrose, calcium phosphate, and corn starch).

In another embodiment, a compound of the invention or a pharmaceutically acceptable salt thereof may be compressed into a tablet with a conventional tablet base (e.g., lactose, sucrose and corn starch) and in combination with: binders (e.g., acacia, corn starch or gelatin), disintegrating agents to aid in the disintegration and dissolution of the tablet after administration (e.g., potato starch, alginic acid, corn starch and guar gum, gum tragacanth, acacia), lubricants to improve the flowability of the tablet granulation and to prevent adhesion of the tablet materials to the surfaces of the tablet die and punch (e.g., talc, stearic acid or magnesium stearate, calcium stearate or zinc stearate), dyes, colorants, and flavoring agents (e.g., peppermint, oil of wintergreen or cherry flavoring) to improve the organoleptic properties of the tablets and make them more acceptable to the patient. Suitable excipients for oral liquid dosage forms include dicalcium phosphate and diluents such as water and alcohols (e.g., ethanol, benzyl alcohol, and polyvinyl alcohol), with or without the addition of pharmaceutically acceptable surfactants, suspending or emulsifying agents. Various other materials may be present as coatings or to modify the physical form of the dosage unit. For example, tablets, pills, or capsules may be coated with shellac, sugar or both.

The compounds of the invention may also be administered parenterally, i.e., subcutaneously, intravenously, intraocularly, intrasynovially, intramuscularly or intraperitoneally, as an injection of the compound, preferably in a physiologically acceptable diluent with a pharmaceutical carrier, which may be a sterile liquid or a mixture of liquids, such as water, saline, aqueous dextrose and related sugar solutions, alcohols such as ethanol, isopropanol or cetyl alcohol, glycols such as propylene glycol or polyethylene glycol, glycerol ketals such as 2, 2-dimethyl-1, 1-dioxolane-4-methanol, ethers such as polyethylene glycol 400(PEG400), oils, fatty acids, fatty acid esters or glycerides or acetylated glycerides, with or without the addition of pharmaceutically acceptable surfactants such as soaps or detergents, suspending agents such as pectin, carbomer, methylcellulose, hypromellose or carboxymethylcellulose, or emulsifying agents and other pharmaceutically acceptable adjuvants.

Exemplary surfactants for parenteral formulations are polyethylene sorbitan fatty acid esters, such as sorbitan monooleate, and the high molecular weight adducts of ethylene oxide with a hydrophobic base formed by the condensation of propylene oxide and propylene glycol.

The compositions of the present invention may also be administered in the form of suppositories for rectal administration of the drug. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and therefore will melt in the rectum to release the drug. Such materials are, for example, cocoa butter and polyethylene glycols.

Controlled release formulations for parenteral administration include liposomal microspheres, polymeric microspheres, and polymeric gel formulations known in the art.

It may be desirable or necessary to deliver the pharmaceutical composition to a patient by a mechanical delivery device. The construction and use of mechanical delivery devices for delivering pharmaceutical agents is well known in the art. Direct techniques such as administering drugs directly to the brain typically involve placing a drug delivery catheter into the ventricular system of the patient to bypass the blood brain barrier.

The compounds of the present invention may be administered as a single agent or in combination with one or more other agents, wherein the combination does not cause unacceptable adverse effects. The invention also relates to such combinations. For example, the compounds of the present invention can be combined with known chemotherapeutic or anti-cancer agents (e.g., agents that combat hyperproliferative diseases or other indications, etc.), as well as with mixtures and combinations thereof. Other indications include, but are not limited to, anti-angiogenic agents, mitotic inhibitors, alkylating agents, anti-metabolites, DNA-intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzyme inhibitors, topoisomerase inhibitors, biological response modifiers, or anti-hormones.

The invention also provides one or more of the crystalline forms for use in the treatment or prevention of a disease associated with protein kinase activity.

The invention also provides methods of modulating (e.g., down-regulating) the activity of a protein kinase comprising contacting the protein kinase with an effective amount of one or more of the crystalline forms described above. The method can be used in vivo or in vitro. Preferably, the protein kinase is selected from at least one of mTOR and PI 3K.

According to another aspect of the present application, there is provided a method of treating a disease associated with protein kinase activity, the method comprising administering to a subject in need thereof an effective amount of one or more of the crystalline forms described above. The subject may be a mammal, such as a human.

Diseases associated with protein kinase activity described herein (e.g., diseases treated or prevented by inhibiting one or both of mTOR and PI3K) can be tumors, such as leukemia, malignant lymphoma, multiple myeloma, gastrointestinal stromal tumor, colon cancer, rectal cancer, breast cancer, liver cancer, stomach cancer, ovarian cancer, uterine cancer, cervical cancer, vaginal cancer, choriocarcinoma, lung cancer, kidney cancer, prostate cancer, bladder cancer, pancreatic cancer, glioblastoma, mast cell tumors, brain tumors, germ cell tumors, melanoma, sarcomas, including dermatofibrosarcoma protruberans, osteosarcoma. The diseases associated with protein kinase activity described herein may also be metabolic diseases (e.g. diabetes, obesity) and cardiovascular diseases (e.g. atherosclerosis).

The invention also provides the use of one or more of the crystalline forms for the manufacture of a medicament for the treatment or prevention of a disease or condition which may be one associated with protein kinase activity, including for example diseases which may be treated or prevented by inhibition of one or both of mTOR and PI 3K.

The invention also provides application of one or more of the crystal forms in preparing a medicament for inhibiting one or two of mTOR and PI3K kinase.

Including diseases caused by uncontrolled cell growth, proliferation and/or survival, an inappropriate cellular immune response, or an inappropriate cellular inflammatory response, or diseases with uncontrolled cell growth, proliferation and/or survival, inappropriate cellular immune responses or inappropriate cellular inflammatory responses, in particular, the disease is, for example, hematological tumors, solid tumors and/or their metastases, such as leukemia and myelodysplastic syndrome, malignant lymphomas, head and neck tumors including brain tumors and brain metastases, breast tumors including non-small cell lung tumors and small cell lung tumors, gastrointestinal tumors, endocrine tumors, breast tumors and other gynecological tumors, urological tumors including kidney tumors, bladder tumors and prostate tumors, skin tumors and sarcomas, and/or their metastases.

The invention also provides the use of the compounds of the invention and compositions thereof in the manufacture of a medicament for the treatment of hyperproliferative disorders in a mammal. The compounds may be used to inhibit, block, reduce, etc., cell proliferation and/or cell division and/or induce apoptosis. Hyperproliferative disorders include, but are not limited to, psoriasis, keloids and other hyperplasia affecting the skin, benign prostatic hyperplasia (BpH), solid tumors such as breast cancer, respiratory tract cancer, lung cancer, brain cancer, reproductive organ cancer, digestive tract cancer, urinary tract cancer, eye cancer, liver cancer, skin cancer, head and neck cancer, thyroid cancer, parathyroid cancer and their distant metastases. Such conditions also include lymphomas, sarcomas and leukemias.

As used herein, "metabolic disease" refers to a disease caused by metabolic problems, including metabolic disorders and metabolic hyperactivity, and includes mainly the following diseases: diabetes, diabetic ketoacidosis, hyperglycemic hyperosmolar syndrome, hypoglycemia, gout, protein-energy malnutrition, vitamin A deficiency, scurvy, vitamin D deficiency, osteoporosis, etc.

As used herein, "cardiovascular disease" is also known as circulatory disease and is a series of diseases involving the circulatory system, which refers to the organs and tissues that transport blood in the human body, mainly including the heart, blood vessels (arteries, veins, microvessels), and can be subdivided into acute and chronic, and is generally associated with arteriosclerosis. Cardiovascular diseases include: heart disease, hypotension, hypertension, hyperglycemia, stroke, myocardial infarction, thrombosis, arteriosclerosis, etc.

These conditions have been well characterized in humans, but also exist in other mammals with similar etiologies, and can be treated by administering the pharmaceutical compositions of the present invention.

In some embodiments, the pharmaceutical composition may be tablets, capsules, pills, powders, sustained release formulations, solutions and suspensions for oral administration, sterile solutions, suspensions or emulsions for parenteral injection, ointments or creams for topical use, or suppositories for rectal administration. In other embodiments, the pharmaceutical composition is in unit dosage form suitable for single administration of a precise dose. In other embodiments, the amount of the compound ranges from about 0.001mg/kg body weight/day to about 1000mg/kg body weight/day. In other embodiments, the amount of the compound ranges from about 0.5mg/kg body weight/day to about 50mg/kg body weight/day. In some embodiments, the amount of the compound is from about 0.001 g/day to about 7 g/day. In other embodiments, the amount of the compound is from about 0.002 g/day to about 6 g/day. In other embodiments, the amount of the compound is from about 0.005 g/day to about 5 g/day. In other embodiments, the amount of the compound is from about 0.01 g/day to about 5 g/day. In other embodiments, the amount of the compound is from about 0.02 g/day to about 5 g/day. In other embodiments, the amount of the compound is from about 0.05 g/day to about 2.5 g/day. In other embodiments, the amount of the compound is from about 0.1 g/day to about 1 g/day. In other embodiments, dosage levels below the lower limit of the aforesaid range may be more than adequate. In other embodiments, dosage levels above the upper limit of the range recited above may be desired. In some embodiments, the compound is administered in a single dose, once per day. In other embodiments, the compound is administered in multiple doses, more than once per day. In some embodiments, the compound is administered twice daily. In other embodiments, the compound is administered three times per day. In other embodiments, the compound is administered four times per day. In other embodiments, the compound is administered four or more times per day. In some embodiments, the subject to which the pharmaceutical composition is administered is a mammal. In other embodiments, the mammal is a human. In other embodiments, the pharmaceutical composition further comprises at least one therapeutic agent (i.e., formulated as a dosage form). In some embodiments, the pharmaceutical composition and the at least one therapeutic agent are combined in separate dosage forms into a combination product, such as a kit of parts (kit of parts).

The crystal form provided by the invention has good stability under three extreme conditions of high temperature, high humidity, strong illumination and the like, and also keeps good stability in the tabletting process. The crystal form provided by the invention has good in vivo absorption metabolic properties, including blood concentration, drug time curve AUC, half-life period and the like. Moreover, the dissolution speed of the crystal form is improved, and the crystal form is beneficial to application in preparations.

Pharmaceutical terms

Certain pharmaceutical terms as used herein with respect to the terms "subject", "patient" or "individual" refer to an individual suffering from a disease, disorder or condition, and the like, including mammals and non-mammals. Examples of mammals include, but are not limited to, any member of the class mammalia: humans, non-human primates (e.g., chimpanzees and other apes and monkeys); livestock, such as cattle, horses, sheep, goats, pigs; domestic animals such as rabbits, dogs, and cats; laboratory animals, including rodents, such as rats, mice, and guinea pigs, and the like. Examples of non-human mammals include, but are not limited to, birds, fish, and the like. In one embodiment related to the methods and compositions provided herein, the mammal is a human.

As used herein, the term "treating" and other similar synonyms include alleviating, or ameliorating a symptom of a disease or disorder, preventing other symptoms, ameliorating, or preventing an underlying metabolic cause of a symptom, inhibiting a disease or disorder, e.g., arresting the development of a disease or disorder, alleviating a disease or disorder, ameliorating a disease or disorder, alleviating a symptom of a disease or disorder, or discontinuing a symptom of a disease or disorder, and further, the term encompasses prophylactic purposes. The term also includes obtaining a therapeutic effect and/or a prophylactic effect. The therapeutic effect refers to curing or ameliorating the underlying disease being treated. In addition, a cure or amelioration of one or more physiological symptoms associated with the underlying disease is also a therapeutic effect, e.g., an improvement in the condition of the patient is observed, although the patient may still be affected by the underlying disease. For prophylactic effect, the composition can be administered to a patient at risk of developing a particular disease, or to a patient presenting with one or more physiological symptoms of the disease, even if a diagnosis of the disease has not yet been made.

The terms "effective amount," "therapeutically effective amount," or "pharmaceutically effective amount" as used herein, refer to an amount of at least one agent or compound that is sufficient to alleviate one or more symptoms of the disease or disorder being treated to some extent after administration. The result may be a reduction and/or alleviation of signs, symptoms, or causes, or any other desired change in a biological system. For example, an "effective amount" for treatment is the amount of a composition comprising a compound disclosed herein that is clinically necessary to provide a significant remission effect of the condition. An effective amount suitable in any individual case can be determined using techniques such as a dose escalation assay.

The terms "administering," "administration," "administering," and the like as used herein refer to a method capable of delivering a compound or composition to a desired site for biological action. These methods include, but are not limited to, oral routes, via the duodenal route, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intraarterial injection or infusion), topical and rectal administration. Administration techniques useful for the compounds and methods described herein are well known to those skilled in the art, for example, in Goodman and Gilman, the pharmacological Basis of Therapeutics, current ed.; pergamon and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, Pa. In preferred embodiments, the compounds and compositions discussed herein are administered orally.

The term "acceptable" as used herein with respect to a formulation, composition or ingredient means that there is no long-term deleterious effect on the general health of the subject being treated.

Drawings

FIG. 1 is an X-ray powder diffraction (XRPD) pattern of form I from example I-1 using Cu-K α radiation.

FIG. 2 is a thermogravimetric analysis (TGA) spectrum of form I of example I-1.

FIG. 3 is a Differential Scanning Calorimetry (DSC) spectrum of form I of example I-1.

FIG. 4 is an XRPD pattern of a solid of form I in example I-4 at 60 ℃.

FIG. 5 is an XRPD pattern of a solid of form I at 25 ℃/90% RH as in example I-4.

FIG. 6 is an XRPD spectrum of a solid of form I under light conditions as in example I-4.

Figure 7 is a comparison of XRPD before and after tableting form I of example I-5.

FIG. 8 is an XRPD spectrum of form II from example II-1.

FIG. 9 is a TGA spectrum of form II of example II-1.

FIG. 10 is an XRPD spectrum of form III of example III-1.

FIG. 11 is an XRPD pattern of form IV from example IV-1.

Figure 12 is a TGA profile of form IV from example IV-1.

FIG. 13 is an XRPD spectrum of form V from example V-1.

FIG. 14 is a TGA spectrum of form V of example V-1.

FIG. 15 is an XRPD spectrum of form VI in example VI-1.

FIG. 16 is a TGA profile of form VI of example VI-1.

Figure 17 is an XRPD spectrum of form VII in example VII-1.

Figure 18 is a TGA profile of crystalline form VII from example VII-1.

FIG. 19 is a Differential Scanning Calorimetry (DSC) profile of form VII in example VII-1.

Detailed Description

The technical solution of the present invention will be described in detail below with reference to examples in order to better understand the technical solution and essence of the present invention. The embodiments are merely illustrative and should not be construed as limiting the scope of the invention. Variations or changes that may be made by those skilled in the art in light of this summary and the following examples are intended to be within the scope of the invention as claimed.

In the following examples the X-ray powder diffraction was determined by means of an X-ray powder diffractometer, model Bruker D8advance, equipped with a LynxEye detector. The 2 theta scan angle of the sample was from 3 deg. to 40 deg., the scan step was 0.02 deg., and the tube voltage and tube current were 40KV and 40mA, respectively. The sample pan used for sample measurement was a zero background sample pan.

Differential Scanning Calorimetry (DSC) analysis in the following examples was performed using a TA DSC Q200, the standard sample used for calibration being indium. 2-3mg of sample was accurately weighed and placed in a TA DSC sample pan and the exact mass of the sample was recorded. The sample was heated to 200 ℃ and 250 ℃ in a nitrogen flow of 50mL/min at a heating rate of 10 ℃/min. Thermogravimetric analysis in the following examples was performed using TA TGA Q500. 2-3mg of sample was placed in an equilibrated aluminum sample pan and the sample mass was automatically weighed in a TGA oven. The sample was heated to 200-300 ℃ at a rate of 10 ℃/min. During the test, the nitrogen flow rates to the balance chamber and sample chamber were 40mL/min and 60mL/min, respectively.

Unless otherwise indicated, the starting materials, substrates, or reagents in the following examples are commercially available (e.g., the absolute ethanol used is commercially available analytically pure absolute ethanol), and can also be prepared by methods known in the art.

EXAMPLE I-1 preparation of form I

Dissolving 96g of compound 1 hydrochloride in 860ml of water at the bath temperature of 100 ℃, adding 43g of sodium chloride solid while the solution is hot, dissolving the sodium chloride solid under stirring, slowly cooling to 30 ℃ for crystallization, performing suction filtration, leaching with 100ml of water, and performing vacuum drying on the obtained solid at 25 ℃ to obtain 86g of yellow-green crystals, wherein the crystal form I of the compound 1 hydrochloride monohydrate is tested.

Example I-2 preparation of form I

Dissolving 110g of hydrochloride of the compound 1 in 2L of water at the bath temperature of 100 ℃, adding a solution of 100g of sodium chloride in 500ml of water while the solution is hot, slowly cooling to 20 ℃ under stirring for crystallization, performing suction filtration, leaching with 100ml of water, and performing vacuum drying on the obtained solid at the temperature of 30 ℃ to obtain 97g of yellow-green crystals, wherein the crystals are tested to be the crystal form I of the hydrochloride monohydrate of the compound 1.

Example I-3 preparation of form I

Dissolving 0.5g of hydrochloride of the compound 1 in a mixed solution of 10ml of ethanol and 5.5ml of water at the bath temperature of 85 ℃, naturally cooling to 25 ℃ under stirring for crystallization, performing suction filtration, and drying the solid in vacuum at 25 ℃ to obtain 0.36g of yellow-green crystals, wherein the crystal form I of the hydrochloride monohydrate of the compound 1 is tested.

Example I-4 stability test of form I

A small amount of the crystalline form I of the hydrochloride monohydrate of the compound 1 of example I-1 was taken and placed in a drug stability laboratory box, stability tests were conducted under the conditions listed in Table 1, the results of purity and content are shown in Table I-1, and the results of the crystalline form tests are shown in FIGS. 4 to 6.

As shown in the table I-1, under three extreme conditions of high temperature, high humidity, strong illumination and the like, the purity and the content of the compound of the crystal form I have no obvious change (the purity floats within 0.2 percent, and the content floats within 1 percent), and the stability of the crystal form I is proved to be better.

As shown in fig. 4 to 6, under three extreme conditions of high temperature, high humidity, strong light and the like, the crystal form I remains unchanged, which proves that the stability of the crystal form I is better.

TABLE I-1 solid stability test results for crystalline form I

Examples I-5 form I tableting stability experiments

A small amount of form I of compound 1 hydrochloride monohydrate of example 1 was manually compressed into tablets having a diameter of 8mm, the tablets were lightly crushed, the powder was analyzed for XRPD and the results compared to the XRPD results before compression, if shown in FIG. 7, to see if compression had an effect on the form. The XRPD patterns of the powder before and after tabletting of the crystal form I are compared, and the result shows that the XRPD patterns of the powder before and after tabletting are the same without changing the crystal form I in the tabletting process.

Examples I-6 oral administration of form I in SD rats for metabolism

11.392mg of form I Compound 1 hydrochloride salt suspended in 5.274mL of 0.5% sodium carboxymethylcellulose, vortexed to form a homogeneous suspension, and sonicated for 2min to finally form a homogeneous suspension solution at a concentration of 2mg/mL (ready to use on the day of administration, storage not longer than 4 hours). Three SD rats were gavaged at a dose of 10mg/kg body weight, blood was taken at the set time points and analyzed, and the results are shown in the following Table I-2:

table I-2: experimental data of gavage drug metabolism of rats with crystal form I

From table I-2, it can be seen that the maximum plasma concentration (Cmax) of 14600ng/mL and AUC last of 190696h ng/mL after the administration of the form I drug are very high values in drug metabolism, and the half-life of 5.5h is also an ideal value in drug metabolism, which all prove that the absorption and metabolism properties of the form I drug in animals are good.

Example II-1 preparation of form II

At room temperature, 100 microliters of each of the ethyl acetate saturated solution of the compound 1 hydrochloride and the 2-butanone saturated solution were mixed in a 96-well plate, covered with a perforated sealing film, placed in a fume hood, and naturally volatilized to obtain crystals, which were tested to be the compound 1 hydrochloride monohydrate in crystal form II.

Example II-2 preparation of form II

At room temperature, 100 microliters of each of the tetrahydrofuran saturated solution and the acetonitrile saturated solution of the compound 1 hydrochloride was mixed in a 96-well plate, covered with a perforated sealing film, placed in a fume hood, and naturally volatilized under atmospheric conditions to obtain crystals, which were tested to be the compound 1 hydrochloride monohydrate in crystal form II.

Example II-3 preparation of form II

At room temperature, 100 microliters of each of the tetrahydrofuran saturated solution and the acetone saturated solution of the compound 1 hydrochloride was mixed in a 96-well plate, covered with a perforated sealing film, placed in a fume hood, and naturally volatilized under atmospheric conditions to obtain crystals, which were tested to be the compound 1 hydrochloride monohydrate in crystal form II.

EXAMPLE III-1 preparation of form III

Mixing 100 microliters of ethanol saturated solution and isopropanol saturated solution of compound 1 hydrochloride in a 96-well plate at room temperature, covering with a perforated sealing film, placing in a fume hood, and naturally volatilizing in an atmospheric environment to obtain a crystal, wherein the crystal is tested to be the crystal form III of the compound 1 hydrochloride.

EXAMPLE III-2 preparation of form III

Mixing 100 microliters of the isopropanol saturated solution and the methyl tert-butyl ether saturated solution of the compound 1 hydrochloride in a 96-well plate at room temperature, covering the mixture with a perforated sealing film, placing the covered plate in a fume hood, and naturally volatilizing the covered plate in an atmospheric environment to obtain crystals, wherein the crystals are tested to be the crystal form III of the compound 1 hydrochloride.

EXAMPLE IV-1 preparation of form IV

At room temperature, 100 microliters of each of the toluene saturated solution of the compound 1 hydrochloride and the isobutyl acetate saturated solution was mixed in a 96-well plate, covered with a perforated sealing film, placed in a fume hood, and naturally volatilized under atmospheric conditions to obtain crystals, which were tested to be the crystalline form IV of the compound 1 hydrochloride.

EXAMPLE V-1 preparation of form V

The mother liquor of the example I-1 is placed for more than 48 hours at 20-25 ℃, white crystals are separated out, and the crystals are obtained by suction filtration and tested to be the crystal form V of the compound 1 hydrochloride dihydrate.

EXAMPLE V-2 preparation of form V

The mother liquor of the example I-2 is placed at 20-25 ℃ for a long time, white crystals are separated out, and the crystals are obtained by suction filtration and tested to be the crystal form V of the hydrochloride dihydrate of the compound 1.

EXAMPLE VI-1 preparation of form VI

Dissolving 0.5g of hydrochloride of the compound 1 in a mixed solution of 50ml of water and 20ml of acetonitrile at 20-25 ℃, stirring for 20 hours at room temperature, separating out a large amount of white crystals, performing suction filtration, and performing vacuum drying on the obtained solid at 25 ℃ to obtain 0.27g of white crystals, wherein the crystal form VI of the hydrochloride dihydrate of the compound 1 is tested.

Example VII-1 preparation of form VII

Dissolving 0.5g of hydrochloride of the compound 1 in 90ml of water heated to 90 ℃, cooling to room temperature (20-25 ℃), adding 3g of sodium chloride while stirring, stirring for 20 hours at room temperature, separating out a large amount of yellow-white solid, performing suction filtration, and performing vacuum drying on the obtained solid at 25 ℃ to obtain 0.46g of white crystals, which are tested to be the crystal form VII of the hydrochloride dihydrate of the compound 1.

Example VII-2 preparation of form VII

Dissolving 0.5g of hydrochloride of the compound 1 in 12ml of water heated to 100 ℃, carrying out hot filtration to remove a small amount of insoluble substances, naturally cooling the mother liquor to room temperature (20-25 ℃) while stirring, carrying out suction filtration, and carrying out vacuum drying on the obtained solid at 25 ℃ to obtain 0.21g of white crystals, wherein the crystal form VII of the hydrochloride dihydrate of the compound 1 is tested.

Examples VII-3 preparation of form VII

Taking 10mg of the crystal form I, adding 1ml of water, and carrying out mixed spinning and pulping at the temperature of 20-25 ℃ for 3 days to obtain white crystals, wherein the white crystals are tested to be the crystal form VII of the compound 1 hydrochloride dihydrate.

Examples VII-4 preparation of form VII

Form VII was obtained according to example VII-3, but replacing form I with one of the forms II, III, IV, V, VI described above.

Examples VII-5 dissolution Rate testing of form VII

Weighing 3mg of the chemical crystal form VII, the chemical crystal form I and the chemical crystal form VI respectively, putting the chemical crystal form VII, the chemical crystal form I and the chemical crystal form VI into 3 plastic centrifuge tubes of 1ml respectively, adding 1ml of distilled water into each tube, placing the tubes after shaking for 5 seconds at 20-25 ℃, and observing the dissolution phenomenon:

form VII was completely clear within 10 seconds;

the crystal form I is basically dissolved and cleared within 2 hours and is completely dissolved and cleared within 5 hours;

form VI did not dissolve clear after 5 hours.

Therefore, the crystal form VII is an instant crystal form and has good application value in the aspect of manufacturing an instant solvent type.

Claims (9)

1. A hydrochloride monohydrate crystal form I of a compound 1 shown as a formula is characterized in that Cu-K α is used for radiation, and X-ray powder diffraction characteristic peaks expressed by a 2 theta angle (DEG) comprise 9.028 +/-0.2, 11.196 +/-0.2, 12.200 +/-0.2, 15.406 +/-0.2, 16.380 +/-0.2, 16.828 +/-0.2, 17.393 +/-0.2, 18.066 +/-0.2, 18.739 +/-0.2, 20.036 +/-0.2, 20.894 +/-0.2, 22.504 +/-0.2, 22.955 +/-0.2, 24.973 +/-0.2, 25.505 +/-0.2, 26.312 +/-0.2, 26.918 +/-0.2, 27.556 +/-0.2, 28.403 +/-0.2, 29.176 +/-0.2, 31.586 +/-0.2 and 35.168 +/-0.2;

2. form I according to claim 1, characterized in that form I has a Cu-K α radiation X-ray powder diffraction pattern as shown in figure 1.

3. A process for the preparation of form I according to claim 1 or 2, selected from one of the following preparation process a or preparation process B:

preparation method A, comprising:

1) dissolving compound 1 hydrochloride in water, heating the water to 85-100 ℃ before or after adding compound 1 hydrochloride to dissolve compound 1 hydrochloride; wherein the amount of water is 10-25 times of the weight of the hydrochloride of the compound 1;

2) adding sodium chloride into the solution obtained in the step 1) at the temperature of the solution obtained in the step 1), and stirring to dissolve the sodium chloride; the sodium chloride is a sodium chloride aqueous solution or a sodium chloride solid, the amount of the sodium chloride is controlled to be 3-5% of the total weight of the solution, and when the sodium chloride aqueous solution is the sodium chloride aqueous solution, the weight percentage of the sodium chloride is 10% to the saturated concentration;

3) slowly cooling to 20-50 ℃ under stirring for crystallization, carrying out suction filtration, leaching, and vacuum drying at 15-35 ℃ to obtain a crystal form I of the hydrochloride monohydrate of the compound 1;

or,

preparation method B, comprising:

1) dissolving compound 1 hydrochloride in an aqueous ethanol solution; wherein the aqueous ethanol solution is heated to 50-100 ℃ before or after the addition of the hydrochloride of compound 1 to dissolve the hydrochloride of compound 1; the dosage of the ethanol water solution is 30-50 times of the weight of the hydrochloride of the compound 1; the mass percentage of the ethanol in the ethanol water solution is 60-75%;

2) slowly cooling to 20-30 ℃ under stirring for crystallization, performing suction filtration, and performing vacuum drying at 15-35 ℃ to obtain the crystal form I of the hydrochloride monohydrate of the compound 1.

4. A method for preparing form V of hydrochloride dihydrate of compound 1, wherein said compound 1 has the chemical structure of claim 1, said form V having a Cu-K α radiation X-ray powder diffraction pattern substantially as shown in figure 13, said method comprising:

1) preparing a saturated solution of the hydrochloride salt of compound 1, wherein the saturated solution is from the filtered mother liquor of step 3) of preparation a according to claim 3;

2) standing the solution obtained in the step 1) at a temperature of below 20-25 ℃ for more than 48h, crystallizing, and performing suction filtration to obtain a crystal form V.

5. A method of preparing form VII of hydrochloride dihydrate of compound 1, wherein said compound 1 has the chemical structure of claim 1, form VII having a Cu-K α radiation X-ray powder diffraction pattern substantially as shown in figure 17, said method of preparation comprising:

carrying out mixed-spinning pulping on the crystal form I of claim 1 or 2 at 20-25 ℃ for 3 days by using water to obtain a crystal form VII; 10mg of form I and 1mL of water.

6. A pharmaceutical composition comprising the crystalline form I of claim 1 or 2.

7. Use of the crystalline form I of claim 1 or 2 for the preparation of a medicament for the treatment or prevention of disease, which medicament is an agent that inhibits one or both of mTOR and PI3K kinase.

8. Use of the crystalline form I of claim 1 or 2 for the preparation of a medicament for the treatment or prevention of a disease or disorder associated with protein kinase activity.

9. The use of claim 8, wherein the medicament treats or prevents the disease by inhibiting one or both of mTOR and PI3K kinases.