HK1260821B - Crystals of aniline pyrimidine compound serving as egfr inhibitor - Google Patents

Description

Crystallization of anilinopyrimidine compounds as EGFR inhibitors

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to and the benefit of Chinese Patent Application No. 201610470835.2, filed with the State Intellectual Property Office of China on June 24, 2016, the disclosure of which is incorporated herein by reference in its entirety.

Technical Field

The present application belongs to the field of medicinal chemistry. Specifically, the present application relates to the crystallization, crystal composition, pharmaceutical composition, preparation method and use of the anilinopyrimidine compound N-(2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxy-5-(4-(3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)pyrimidin-2-ylamino)phenyl)acrylamide hydrochloride as an EGFR inhibitor.

Background Art

EGFR (Epidermal Growth Factor Receptor) is a receptor for epidermal growth factor (EGF) cell proliferation and signaling, also known as HER1 and ErbB1. EGFR belongs to the ErbB receptor family, which includes EGFR (ErbB-1), HER2/c-neu (ErbB-2), HER3 (ErbB-3), and HER4 (ErbB-4). EGFR is a glycoprotein, a tyrosine kinase receptor that penetrates the cell membrane and has a molecular weight of 170 kDa.

EGFR is located on the cell membrane surface and is activated by binding to ligands, including EGF and TGFα. Upon activation, EGFR converts from a monomer to a dimer. The dimerization involves both the binding of two receptor molecules of the same species (homodimerization) and the binding of different members of the human EGF-related receptor (HER) tyrosine kinase family (heterodimerization). EGFR dimerization activates its intracellular kinase pathways, including phosphorylation of key tyrosine residues in the intracellular domain, and stimulates numerous intracellular signaling pathways involved in cell proliferation and survival.

EGFR is overexpressed or aberrantly expressed in many solid tumors. EGFR is associated with tumor cell proliferation, angiogenesis, tumor invasion, metastasis, and inhibition of apoptosis. Possible mechanisms include: elevated EGFR expression leading to enhanced downstream signaling; increased expression of mutant EGFR receptors or ligands leading to sustained EGFR activation; enhanced autocrine loops; disruption of receptor downregulation mechanisms; and activation of aberrant signaling pathways. EGFR overexpression plays a crucial role in the progression of malignant tumors, and has been found in tissues such as glial cells, renal cancer, lung cancer, prostate cancer, pancreatic cancer, and breast cancer.

Abnormal expression of EGFR and Erb-B2 plays a key role in tumor transformation and growth. Taking lung cancer as an example, EGFR is expressed in 50% of non-small cell lung cancer (NSCLC) cases, and its expression is correlated with poor prognosis. These two factors make EGFR and its family members prime candidates for targeted therapies. Two small molecule inhibitors targeting EGFR, gefitinib and erlotinib, received rapid approval from the US FDA for the treatment of patients with advanced NSCLC who have lost their response to conventional chemotherapy.

Early clinical data suggest that 10% of NSCLC patients respond to gefitinib and erlotinib. Molecular biological analysis has shown that, in most cases, patients who respond to the drugs harbor specific mutations in the gene encoding EGFR: deletions of amino acids 747–750 in exon 19 account for 45% of mutations, and another 10% occur in exons 18 and 20. The most common activating EGFR mutations (L858R and delE746_A750) result in increased affinity for small-molecule tyrosine kinase inhibitors (TKIs) and decreased affinity for adenosine triphosphate (ATP) relative to wild-type (WT) EGFR. The T790M mutation is a point mutation in EGFR exon 20 that confers acquired resistance to treatment with gefitinib or erlotinib. The latest research shows that the affinity of L858R combined with T790M mutation for ATP is stronger than that of L858R alone. TKI is an ATP-competitive kinase inhibitor, which leads to a lower binding rate of TKI to the kinase region.

Because the above mutations play an important role in the resistance mechanism of targeted EGFR therapy, it is necessary to provide EGFR-L858R/T790M double mutant inhibitors for clinical treatment. At the same time, because inhibition of EGFR-WT can lead to multiple clinical toxic side effects, it is also necessary to provide inhibitors that are selective for activating mutant forms of EGFR (such as EGFR-L858R mutant, delE746_A750 mutant or Exon19 deletion EGFR mutant) and/or resistant mutant forms of EGFR (such as EGFR-T790M mutant) relative to EGFR-WT for clinical treatment.

Currently, there have been reports on a variety of selective EGFR inhibitors. Among them, Chinese patent application No. 201510419018.X, filed on July 16, 2015, discloses a number of EGFR inhibitors (incorporated herein by reference in its entirety), including the hydrochloride salt of N-(2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxy-5-(4-(3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)pyrimidin-2-ylamino)phenyl)acrylamide as shown in the following formula I:

In addition to therapeutic efficacy, drug developers strive to provide active molecules with suitable forms that possess pharmaceutical properties. Chemical stability, solid-state stability, and shelf life of the active ingredient are crucial factors for achieving a commercially viable production process or producing a pharmaceutical composition containing the active compound. Therefore, providing a form with the desired properties is crucial to drug development.

SUMMARY OF THE INVENTION

On the one hand, the present application provides a hydrochloride crystal A of a compound of formula I:

The X-ray diffraction (XRD) pattern of the hydrochloride crystal A of the compound of formula I has diffraction peaks at 2θ of 8.96°±0.2°, 14.11°±0.2°, 14.87°±0.2°, 16.52°±0.2°, 18.67°±0.2°, 21.93°±0.2° and 27.09°±0.2°.

On the other hand, the present application provides a method for preparing a hydrochloride crystal A of a compound of formula I, the method comprising the following steps:

1) contacting the compound of formula I with hydrochloric acid; and

2) crystallizing from a crystallization solvent, wherein the crystallization solvent is selected from acetonitrile, methanol, isopropanol or an ethanol/water mixture.

On the other hand, the present application provides a crystalline composition, wherein the hydrochloride crystals A of the compound of formula I account for more than 50% by weight of the crystalline composition, preferably more than 80%, more preferably more than 90%, and most preferably more than 95%.

Another aspect of the present application provides a pharmaceutical composition comprising a therapeutically effective amount of hydrochloride crystal A of the compound of formula I or the above-mentioned crystalline composition.

Another aspect of the present application provides use of the hydrochloride crystal A of the compound of formula I or the above crystal composition or the above pharmaceutical composition in the preparation of a drug for treating EGFR-mediated diseases.

In another aspect, the present application provides a hydrochloride crystal B of a compound of formula I:

The X-ray diffraction (XRD) pattern of the hydrochloride crystal B of the compound of formula I has diffraction peaks at 2θ of 9.17°±0.2°, 9.93°±0.2°, 14.07°±0.2°, 20.31°±0.2°, 21.44°±0.2° and 26.10°±0.2°.

On the other hand, the present application provides a method for preparing the hydrochloride crystal B of the compound of formula I, which comprises the following steps:

1) contacting the compound of formula I with hydrochloric acid; and

2) crystallizing from a crystallization solvent, wherein the crystallization solvent is selected from ethanol.

On the other hand, the present application provides a crystalline composition, wherein the hydrochloride crystals B of the compound of formula I account for more than 50% by weight of the crystalline composition, preferably more than 80%, more preferably more than 90%, and most preferably more than 95%.

Another aspect of the present application provides a pharmaceutical composition comprising a therapeutically effective amount of hydrochloride crystal B of the compound of formula I or the above-mentioned crystalline composition.

Another aspect of the present application provides use of the hydrochloride crystal B of the compound of formula I or the above-mentioned crystalline composition or the above-mentioned pharmaceutical composition in the preparation of a drug for treating EGFR-mediated diseases.

In another aspect, the present application provides a hydrochloride crystal C of a compound of formula I:

The X-ray diffraction (XRD) pattern of the hydrochloride crystal C of the compound of formula I has diffraction peaks at 2θ of 7.68°±0.2°, 8.21°±0.2°, 10.89°±0.2°, 15.95°±0.2°, 19.10°±0.2°, 20.52°±0.2° and 21.54°±0.2°.

On the other hand, the present application provides a method for preparing Crystal C, which comprises the following steps:

1) contacting the compound of formula I with hydrochloric acid; and

2) crystallizing from a crystallization solvent, wherein the crystallization solvent is selected from tetrahydrofuran, acetone or dioxane.

On the other hand, the present application provides a crystalline composition, wherein the hydrochloride crystals C of the compound of formula I account for more than 50% by weight of the crystalline composition, preferably more than 80%, more preferably more than 90%, and most preferably more than 95%.

Another aspect of the present application provides a pharmaceutical composition comprising a therapeutically effective amount of the hydrochloride crystal C of the compound of formula I or the above-mentioned crystalline composition.

Another aspect of the present application provides use of the hydrochloride crystal C of the compound of formula I or the above-mentioned crystalline composition or the above-mentioned pharmaceutical composition in the preparation of a drug for treating EGFR-mediated diseases.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 shows the XRD pattern of the hydrochloride crystal A of the compound of formula I (method 1 in Example 3).

Figure 2 shows the DSC spectrum of the hydrochloride crystal A of the compound of formula I (Example 3 Method 1).

Figure 3 shows the XRD pattern of the hydrochloride crystal B of the compound of formula I (method 5 of Example 4).

Figure 4 shows the DSC spectrum of the hydrochloride crystal B of the compound of formula I (Example 4, Method 5).

Figure 5 shows the XRD pattern of the hydrochloride crystal C of the compound of formula I (Example 5, Method 6).

FIG6 DSC spectrum of the hydrochloride crystal C of the compound of formula I (Example 5, Method 6).

Detailed Description of the Invention

On the one hand, the present application provides a hydrochloride crystal A of a compound of formula I:

The X-ray diffraction (XRD) pattern of the hydrochloride crystal A of the compound of formula I has diffraction peaks at 2θ of 8.96°, 14.11°, 14.87°, 16.52°, 18.67°, 21.93° and 27.09°±0.2°; typically has diffraction peaks at 2θ=8.33°, 8.96°, 12.16°, 14.11°, 14.87°, 16.52°, 17.66°, 18.67°, 21.93° and 27.09°±0.2°; more typically has diffraction peaks at 2θ=8.33°, 8.96°, 12.16°, 14.11°, 14.87°, 16.52°, 17.66°, 18.67°, 21.93° and 27.09°±0.2°. The diffraction peaks of the present invention are as follows: 14.11°, 14.87°, 16.52°, 17.66°, 18.23°, 18.67°, 21.93°, 22.65° and 27.09°±0.2°; and further typically have diffraction peaks of 2θ=8.33°, 8.96°, 11.74°, 12.16°, 14.11°, 14.87°, 16.52°, 17.66°, 18.23°, 18.67°, 19.48°, 19.92°, 21.93°, 22.65°, 24.95°, 27.09° and 27.55°±0.2°.

In some embodiments of the present application, the X-ray diffraction peaks of the hydrochloride crystal A of the compound of formula I described in the present application have the following characteristics:

Serial number 2θ±0.2(°) Relative strength (%) Serial number 2θ±0.2(°) Relative strength (%) 1 8.33 14.3 12 19.92 23.0 2 8.96 59.7 13 21.93 100.0 3 11.74 18.5 14 22.65 31.1 4 12.16 23.1 15 23.05 17.2 5 14.11 70.6 16 24.04 17.3 6 14.87 61.7 17 24.95 25.4 7 16.52 90.1 18 26.18 17.3 8 17.66 35.8 19 27.09 65.3 9 18.23 31.5 20 27.55 24.0 10 18.67 54.6 twenty one 28.74 16.4 11 19.48 27.2 twenty two 29.11 13.2

In some embodiments of the present application, the X-ray diffraction pattern of the hydrochloride crystal A of the compound of formula I described in the present application is shown in FIG1 .

In some embodiments of the present application, the DSC spectrum of the hydrochloride crystal A of the compound of formula I described herein has a peak at about 271°C.

In some embodiments of the present application, the DSC spectrum of the hydrochloride crystal A of the compound of formula I described in the present application is shown in FIG2 .

On the other hand, the present application provides a method for preparing a hydrochloride crystal A of a compound of formula I, the method comprising the following steps:

1) contacting the compound of formula I with hydrochloric acid; and

2) crystallizing from the crystallization solvent and optionally filtering;

The crystallization solvent is selected from acetonitrile, methanol, isopropanol or ethanol/water mixture.

In some embodiments of the present application, when the crystallization solvent for preparing the hydrochloride crystals A of the compound of formula I is selected from an ethanol/water mixture, the ratio of ethanol to water (calculated by volume) is 9:1 to 1:9, preferably 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8 or 1:9, and more preferably 3:1.

In some embodiments of the present application, the molar ratio of hydrochloric acid to the compound of formula I in the preparation method A of the hydrochloride crystal of the compound of formula I is selected from 1:0.5 to 1.5, preferably from 1:0.8 to 1.2, and more preferably from 1:1.

In some embodiments of the present application, the compound of formula I is contacted with hydrochloric acid in a crystallization solvent.

Another aspect of the present application provides a crystalline composition of the hydrochloride crystals A of the compound of formula I. In some embodiments of the present application, the hydrochloride crystals A of the compound of formula I account for more than 50% by weight of the crystalline composition, preferably more than 80%, more preferably more than 90%, and most preferably more than 95%.

Another aspect of the present application provides a pharmaceutical composition of the hydrochloride crystals A of the compound of Formula I, comprising a therapeutically effective amount of the hydrochloride crystals A of the compound of Formula I, or a crystalline composition of the hydrochloride crystals A of the compound of Formula I. In addition, the pharmaceutical composition may contain or not contain a pharmaceutically acceptable carrier, excipient, and/or medium.

Another aspect of the present application provides the use of the hydrochloride crystal A of the compound of formula I or the above-mentioned crystalline composition or the above-mentioned pharmaceutical composition in the preparation of a drug for treating EGFR-mediated diseases.

Another aspect of the present application provides a method for treating EGFR-mediated diseases, comprising administering a therapeutically effective amount of the hydrochloride crystalline A of the compound of formula I, or the crystalline composition, or the pharmaceutical composition to a mammal in need thereof.

Another aspect of the present application provides the hydrochloride crystal A of the compound of formula I, or the crystal composition, or the pharmaceutical composition for treating EGFR-mediated diseases.

On the one hand, the present application provides a hydrochloride crystal B of a compound of formula I:

The X-ray diffraction (XRD) pattern of the hydrochloride crystal B of the compound of formula I has diffraction peaks at 2θ of 9.17°, 9.93°, 14.07°, 20.31°, 21.44° and 26.10°±0.2°; typically has 2θ=9.17°, 9.93°, 10.65°, 13.46°, 14.07°, 20.31°, 21.44°, 22.33°, 2 The invention discloses a diffraction peak having 2θ=4.93° and 26.10°±0.2°; more typically, the diffraction peak has 2θ=6.71°, 9.17°, 9.93°, 10.65°, 11.44°, 13.46°, 14.07°, 18.94°, 20.31°, 21.44°, 21.66°, 22.33°, 24.93°, 25.73°, and 26.10°±0.2°.

In some embodiments of the present application, the X-ray diffraction peaks of the hydrochloride crystal B of the compound of formula I described in the present application have the following characteristics:

Serial number 2θ±0.2(°) Relative strength (%) Serial number 2θ±0.2(°) Relative strength (%) 1 6.71 12.2 10 20.31 48.5 2 9.17 35.9 11 20.72 14.4 3 9.93 100.0 12 21.44 58.3 4 10.65 23.9 13 21.66 52.9 5 11.44 13.0 14 22.33 26.6 6 13.46 23.6 15 23.77 10.7 7 14.07 79.8 16 24.93 38.7 8 18.38 10.1 17 25.73 39.5 9 18.94 18.5 18 26.10 57.2

In some embodiments of the present application, the X-ray diffraction pattern of the hydrochloride crystal B of the compound of formula I described in the present application is shown in FIG3 .

In some embodiments of the present application, the DSC spectrum of the hydrochloride crystal B of the compound of formula I described herein has a peak at about 259°C.

In some embodiments of the present application, the DSC spectrum of the hydrochloride crystal B of the compound of formula I described in the present application is shown in FIG4 .

On the other hand, the present application provides a method for preparing hydrochloride crystals B of the compound of formula I, which comprises the following steps:

1) contacting the compound of formula I with hydrochloric acid; and

2) crystallizing from the crystallization solvent and optionally filtering;

Wherein the crystallization solvent is selected from ethanol.

In some embodiments of the present application, the molar ratio of hydrochloric acid to the compound of formula I in the preparation method B of the hydrochloride crystal of the compound of formula I is selected from 1:0.5 to 1.5, preferably from 1:0.8 to 1.2, and more preferably from 1:1.

In some embodiments of the present application, the compound of formula I is contacted with hydrochloric acid in a crystallization solvent.

Another aspect of the present application provides a crystalline composition of the hydrochloride crystals B of the compound of formula I. In some embodiments of the present application, the hydrochloride crystals B of the compound of formula I account for more than 50% by weight of the crystalline composition, preferably more than 80%, more preferably more than 90%, and most preferably more than 95%.

Another aspect of the present application provides a pharmaceutical composition of the hydrochloride crystals B of the compound of Formula I, comprising a therapeutically effective amount of the hydrochloride crystals B of the compound of Formula I, or a crystalline composition of the hydrochloride crystals B of the compound of Formula I. In addition, the pharmaceutical composition may contain or not contain a pharmaceutically acceptable carrier, excipient, and/or medium.

Another aspect of the present application provides the use of the hydrochloride crystal B of the compound of formula I or the above-mentioned crystalline composition or the above-mentioned pharmaceutical composition in the preparation of a drug for treating EGFR-mediated diseases.

Another aspect of the present application provides a method for treating EGFR-mediated diseases, comprising administering a therapeutically effective amount of the hydrochloride crystalline B of the compound of formula I, or the crystalline composition, or the pharmaceutical composition to a mammal in need thereof.

Another aspect of the present application provides the hydrochloride crystal B of the compound of formula I, or the crystal composition, or the pharmaceutical composition for treating EGFR-mediated diseases.

On the one hand, the present application provides a hydrochloride crystal C of a compound of formula I:

The X-ray diffraction (XRD) pattern of the hydrochloride crystal C of the compound of formula I has diffraction peaks at 2θ of 7.68°, 8.21°, 9.55°, 10.89°, 15.95°, 19.10°, 20.52° and 21.54°±0.2°; The diffraction peaks of the present invention are as follows: 1.08°, 21.54° and 28.22°±0.2°; more typically, the diffraction peaks of 2θ=7.68°, 8.21°, 9.55°, 10.89°, 14.22°, 14.95°, 15.95°, 19.10°, 20.52°, 21.08°, 21.54°, 23.05°, 26.23° and 28.22°±0.2°.

In some embodiments of the present application, the X-ray diffraction peaks of the hydrochloride crystal C of the compound of formula I described in the present application have the following characteristics:

Serial number 2θ±0.2(°) Relative strength (%) Serial number 2θ±0.2(°) Relative strength (%) 1 7.68 85.7 10 20.52 43.6 2 8.21 80.2 11 21.08 39.2 3 9.55 35.6 12 21.54 59.8 4 10.89 63.5 13 22.23 15.9 5 14.22 24.9 14 23.05 26.1 6 14.95 18.2 15 23.74 12.6 7 15.95 100.0 16 26.23 29.8 8 2019.10 53.7 17 26.99 12.5 9 19.77 11.0 18 28.22 30.4

In some embodiments of the present application, the X-ray diffraction pattern of the hydrochloride crystal C of the compound of formula I described in the present application is shown in FIG5 .

In some embodiments of the present application, the DSC spectrum of the hydrochloride crystal C of the compound of formula I described herein has peaks at about 175°C and 262°C.

In some embodiments of the present application, the DSC spectrum of the hydrochloride crystal C of the compound of formula I described in the present application is shown in Figure 6.

On the other hand, the present application provides a method for preparing a hydrochloride crystal C of a compound of formula I, the method comprising the following steps:

1) contacting the compound of formula I with hydrochloric acid; and

2) crystallizing from the crystallization solvent and optionally filtering;

The crystallization solvent is selected from tetrahydrofuran, acetone or dioxane.

In some embodiments of the present application, the molar ratio of HCl to the compound of formula I in the preparation method C of the hydrochloride crystal of the compound of formula I is selected from 1:0.5 to 1.5, preferably from 1:0.8 to 1.2, and more preferably from 1:1.

In some embodiments of the present application, the compound of formula I is contacted with hydrochloric acid in a crystallization solvent.

Another aspect of the present application provides a crystalline composition of the hydrochloride crystals C of the compound of formula I. In some embodiments of the present application, the hydrochloride crystals C of the compound of formula I account for more than 50% by weight of the crystalline composition, preferably more than 80%, more preferably more than 90%, and most preferably more than 95%.

Another aspect of the present application provides a pharmaceutical composition of the hydrochloride crystals C of the compound of Formula I, comprising a therapeutically effective amount of the hydrochloride crystals C of the compound of Formula I, or a crystalline composition of the hydrochloride crystals C of the compound of Formula I. In addition, the pharmaceutical composition may contain or not contain a pharmaceutically acceptable carrier, excipient, and/or medium.

Another aspect of the present application provides the use of the hydrochloride crystal C of the compound of formula I or the above-mentioned crystalline composition or the above-mentioned pharmaceutical composition in the preparation of a drug for treating EGFR-mediated diseases.

Another aspect of the present application provides a method for treating EGFR-mediated diseases, comprising administering a therapeutically effective amount of the hydrochloride crystalline C of the compound of formula I, or the crystalline composition, or the pharmaceutical composition to a mammal in need thereof.

Another aspect of the present application provides the hydrochloride crystal C of the compound of formula I, or the crystal composition, or the pharmaceutical composition for treating EGFR-mediated diseases.

In some embodiments of the present application, the EGFR-mediated disease is selected from a disease mediated by an EGFR-L858R activating mutation. In some embodiments of the present application, the EGFR-mediated disease is selected from a disease mediated by an EGFR-T790M activating mutation. In some embodiments of the present application, the EGFR-mediated disease is selected from a disease mediated by a combined EGFR-L858R and EGFR-T790M double mutation. In some embodiments of the present application, the EGFR-mediated disease is cancer; the cancer is selected from ovarian cancer, cervical cancer, colorectal cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer, leukemia, lymphoma, non-Hodgkin's lymphoma, gastric cancer, lung cancer, hepatocellular carcinoma, gastric cancer, gastrointestinal stromal tumor, thyroid cancer, bile duct cancer, endometrial cancer, renal cancer, anaplastic large cell lymphoma, acute myeloid leukemia, multiple myeloma, melanoma, mesothelioma; the lung cancer can be selected from non-small cell lung cancer, small cell lung cancer, lung adenocarcinoma, and lung squamous cell carcinoma.

When testing the stability of the crystals described herein, the crystals can be placed under high temperature, high humidity, or light conditions. The high temperature condition can be 40-60°C, the high humidity condition can be 75%-92.5% relative humidity, and the light condition can be 5000 Lux. When evaluating the stability of the crystals, multiple parameters such as the sample content, total impurities, and water content can be examined, and these parameters can be comprehensively evaluated based on the product properties.

In this application, the X-ray diffraction patterns were measured using the following method: instrument: Bruker D2 X-ray diffractometer; method: target: Cu; tube voltage: 30 kV; tube current: 10 mA; scanning range: 4-40°; scanning speed: 0.1 s per step, 0.02° per step.

In this application, differential scanning calorimetry (DSC) analysis was performed using the following method: instrument: Mettler DSC-1 differential scanning calorimeter; method: a sample (~5 mg) was placed in a DSC aluminum pan for testing, the method being: 30°C-300°C, with a heating rate of 10°C/min.

It should be noted that in X-ray diffraction spectroscopy, the diffraction pattern obtained from a crystalline compound is often characteristic of a specific crystal form. The relative intensities of the spectral bands (especially at low angles) may vary due to preferential orientation effects caused by differences in crystallization conditions, particle size, and other measurement conditions. Therefore, the relative intensities of the diffraction peaks are not characteristic of the intended crystal form. When determining whether a crystal form is identical to a known crystal form, the relative positions of the peaks, rather than their relative intensities, should be considered. Furthermore, for any given crystal form, the peak positions may have slight errors, which is well known in the art of crystallography. For example, peak positions can shift due to temperature fluctuations, sample movement, or instrument calibration during sample analysis, and the measurement error of 2θ values can sometimes be as much as ±0.2°. Therefore, this error should be taken into account when determining the structure of each crystal form. In XRD patterns, peak positions are typically expressed in 2θ angles or interplanar distances d. The two have a simple conversion relationship: d = λ/2sinθ, where d represents the interplanar distance, λ represents the wavelength of the incident X-ray, and θ is the diffraction angle. For the same compound and the same crystal form, the peak positions of their XRD spectra are generally similar, but the relative intensity errors may be large. It should also be noted that in the identification of mixtures, some diffraction lines may be missing due to factors such as a decrease in content. In this case, it is not necessary to rely on all the bands observed in a high-purity sample; even a single band may be characteristic for a given crystal.

It should be noted that DSC measures the transition temperature when a crystal absorbs or releases heat due to structural changes or melting. For the same crystalline form of the same compound, the error in thermal transition temperatures and melting points in consecutive analyses is typically within approximately 5°C. When a compound is said to have a given DSC peak or melting point, this refers to the DSC peak or melting point ±5°C. DSC provides an auxiliary method for distinguishing different crystalline forms. Different crystal forms can be identified based on their distinct transition temperature characteristics.

As used herein, the term "pharmaceutical composition" refers to a formulation of one or more compounds of the present invention with carriers, excipients, and/or vehicles generally accepted in the art for delivering biologically active compounds to an organism, such as a human. The purpose of a pharmaceutical composition is to facilitate administration of the compounds of the present invention to an organism.

The term "carrier" is defined as a chemical compound that facilitates the introduction of a compound into cells or tissues.

"Pharmaceutically acceptable carrier" includes, but is not limited to, any adjuvant, excipient, glidant, sweetener, diluent, preservative, dye/colorant, flavor enhancer, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier approved by the national drug administration agency as acceptable for use in humans or livestock.

A "therapeutically effective amount" refers to an amount of a compound of the present invention that, when administered to a mammal, preferably a human, is sufficient to achieve treatment of a viral infection in the mammal, preferably a human, as defined below. The amount of a compound of the present invention that constitutes a "therapeutically effective amount" varies depending on the compound, the disease state and its severity, the mode of administration, and the age of the mammal to be treated, but can be routinely determined by one of ordinary skill in the art based on their knowledge and this disclosure.

As used herein, "treatment" encompasses the treatment of a viral infection in a mammal, preferably a human, suffering from the viral infection, and includes:

(i) inhibit viral infection, i.e., prevent its development;

(ii) alleviate the viral infection, i.e. cause recovery of the viral infection; or

(iii) Relieve symptoms caused by viral infection.

All solvents used in this application are commercially available and can be used without further purification. The reaction is generally carried out under inert nitrogen and in anhydrous solvents.

The compounds of this application were named manually or by software, and commercially available compounds were named using supplier catalog names.

In this application, proton nuclear magnetic resonance data were recorded on a BRUKER AVANCE III HD 500M spectrometer, and chemical shifts are expressed in ppm downfield from tetramethylsilane; mass spectra were measured on a Waters ACQUITY UPLC+XEVO G2 QTof. The mass spectrometer was equipped with an electrospray ionization source (ESI) operating in positive or negative mode.

The hydrochloride crystals A, B, and C of the compound represented by Formula I provided herein have the advantages of high purity, high crystallinity, and good stability. At the same time, the preparation method of the hydrochloride crystals A, B, and C of the compound represented by Formula I provided herein is simple, the solvent is cheap and readily available, and the crystallization conditions are mild, which is suitable for industrial production.

The following examples provide further non-limiting details of the technical solutions of this application. They should not be considered as limiting the scope of this application, but are merely illustrative and representative of this application. The solvents, reagents, and raw materials used in this application were all commercially available chemically pure or analytically pure products.

Example 1 Hydrochloride of N-(2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxy-5-(4-(3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)pyrimidin-2-ylamino)phenyl)acrylamide (I)

Step 1: N ¹ -(2-chloropyrimidin-4-yl)benzene-1,2-diamine

Disperse o-phenylenediamine (3.24 g, 30 mmol) and 2,4-dichloropyrimidine (4.47 g, 30 mmol) in anhydrous ethanol (60 mL). Add diisopropylethylamine (7.74 g, 60 mmol) and heat under reflux for 3 hours. Concentrate under vacuum to remove the solvent. The residue is dissolved in dichloromethane (100 mL), washed with water and saturated brine, and concentrated under vacuum to remove the solvent. The residue is separated by column chromatography (EA:PE = 1:2) to obtain the title compound (5.32 g, 80%).

¹ H NMR (CDCl ₃ ): δ 8.08 (1H, d, J = 5.6Hz), 7.20-7.12 (2H, m), 6.85-6.78 (2H, m), 6.74 (1H, s), 6.24 (1H, d, J = 5.6Hz), 3.82 (2H, br).

Step 2: 1-(2-chloropyrimidin-4-yl)-1H-benzo[d]imidazol-2(3H)-one

Dissolve N ¹ -(2-chloropyrimidin-4-yl)phenyl-1,2-diamine (2.21 g, 10 mmol) in DMF (15 mL), add carbonyldiimidazole (2.43 g, 15 mmol), stir at room temperature for 1 hour, pour into water (50 mL), continue stirring for 10 minutes, filter, wash with water (30 mL x 3), and dry to obtain the title compound (2.23 g, 90%).

¹ H NMR (DMSO-d ₆ ): δ 11.64 (1H, br), 8.78 (1H, d, J = 5.6Hz), 8.43 (1H, d, J = 5.6Hz), 8.26 (1H, d, J = 7.6Hz), 7.22-7.10 (3H, m).

Step 3: 1-(2-chloropyrimidin-4-yl)-3-methyl-1H-benzo[d]imidazol-2(3H)-one

Disperse 1-(2-chloropyrimidin-4-yl)-1H-benzo[d]imidazol-2(3H)-one (600 mg, 2.43 mmol) in anhydrous DMF (10 mL) and cool in an ice-water bath. Add sodium hydride (116 mg, 60%, 2.90 mmol) and stir for 1 hour. Add iodomethane (345 mg, 2.43 mmol) dropwise and continue stirring for 1 hour. Pour the reaction mixture into water (50 mL), stir for 30 minutes, filter, wash with water (30 mL x 3), and dry to obtain the title compound (459 mg, 72%).

¹ H NMR (DMSO-d ₆ ): δ 8.79 (1H, d, J = 5.6Hz), 8.44 (1H, d, J = 6.0Hz), 8.29 (1H, d, J = 8.0Hz), 7.30-7.28 (2H, m), 7.24-7.19 (1H, m), 3.39 (3H, s).

Step 4: 1-(2-(4-Fluoro-2-methoxy-5-nitrophenylamino)pyrimidin-4-yl)-3-methyl-1H-benzo[d]imidazol-2(3H)-one p-toluenesulfonate

1-(2-Chloropyrimidin-4-yl)-3-methyl-1H-benzo[d]imidazol-2(3H)-one (459 mg, 1.76 mmol), 4-fluoro-2-methoxy-5-nitroaniline (360 mg, 1.93 mmol), and p-toluenesulfonic acid monohydrate (551 mg, 2.89 mmol) were dispersed in 2-pentanol (10 mL) and stirred at 105°C overnight. After cooling, the mixture was filtered and the filter cake was washed three times with a small amount of 2-pentanol and dried to give the title compound (440 mg, 43%).

¹ H NMR (CDCl ₃ ): δ10.95 (1H, br), 8.49 (1H, d, J = 7.6Hz), 8.39 (1H, d, J = 7.2Hz), 8.21 (1H, d, J = 7.2Hz), 7.87 (2H, d, J = 8.4Hz), 7.68 ( 1H, d, J=8.4Hz), 7.28-7.23 (2H, m), 7.04 (2H, d, J=7.6Hz), 6.91-6.85 (2H, m), 3.92 (3H, s), 3.46 (3H, s), 2.38 (3H, s).

Step 5: 1-(2-(4-((2-(dimethylamino)ethyl)(methyl)amino)-2-methoxy-5-nitrophenylamino)pyrimidin-4-yl)-3-methyl-1H-benzo[d]imidazol-2(3H)-one

1-(2-(4-Fluoro-2-methoxy-5-nitrophenylamino)pyrimidin-4-yl)-3-methyl-1H-benzo[d]imidazol-2(3H)-one p-toluenesulfonate (440 mg, 0.76 mmol) was dissolved in NMP (5 mL). Diisopropylethylamine (206 mg, 1.59 mmol) and ^N₁ , ^N₁ , ^N₂ -trimethylethane-1,2-diamine (116 mg, 1.14 mmol) were added. The reaction mixture was stirred at 85°C overnight. After cooling, the reaction solution was poured into water (50 mL), filtered, rinsed with a small amount of methanol, and dried to obtain the title compound (326 mg, 88%).

¹ H NMR (CDCl ₃ ): δ8.92 (1H, s), 8.51 (1H, d, J = 5.6Hz), 8.27 (1H, d, J = 7.6Hz), 7.82 (1H, d, J = 5.6Hz), 7.47 (1H, s), 7.29-7.19 (1H, m), 7.17-7.13 (1H, m) , 7.04 (1H, d, J = 7.6Hz), 6.69 (1H, s), 3.98 (3H, s), 3.47 (3H, s), 3.27 (2H, t, J = 7.2Hz), 2.89 (3H, s), 2.88 (2H, t, J = 7.2Hz), 2.26 (6H, s).

Step 6: 1-(2-(5-amino-4-((2-(dimethylamino)ethyl)(methyl)amino)-2-methoxyphenylamino)pyrimidin-4-yl)-3-methyl-1H-benzo[d]imidazol-2(3H)-one

1-(2-(4-((2-(dimethylamino)ethyl)(methyl)amino)-2-methoxy-5-nitrophenylamino)pyrimidin-4-yl)-3-methyl-1H-benzo[d]imidazol-2(3H)-one (326 mg, 0.66 mmol) was dissolved in methanol (10 mL), Pd/C (10%, 30 mg) was added, and the hydrogen atmosphere was replaced three times. The system was stirred under a hydrogen atmosphere overnight and filtered. The product was easily oxidized. The filtrate was quickly concentrated in vacuo and directly used for the next reaction.

Step 7: N-(2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxy-5-(4-(3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)pyrimidin-2-ylamino)phenyl)acrylamide hydrochloride

Dissolve 1-(2-(5-amino-4-((2-(dimethylamino)ethyl)(methyl)amino)-2-methoxyphenylamino)pyrimidin-4-yl)-3-methyl-1H-benzo[d]imidazol-2(3H)-one, obtained in the previous step, in anhydrous dichloromethane (10 mL). Add diisopropylethylamine (129 mg, 1.00 mmol) and cool in an ice-water bath. Slowly add a solution of acryloyl chloride (60 mg, 0.66 mmol) in anhydrous dichloromethane (2 mL) dropwise to the system over 15 minutes. After stirring for 15 minutes, the reaction mixture was poured into petroleum ether (50 mL) and stirred for 10 minutes. Filter with suction, and rinse the filter cake with petroleum ether. The resulting crude product was isolated by column chromatography (DCM:MeOH = 20:1) to afford the title compound (164 mg, 45% yield for both steps).

¹ H NMR (DMSO-d ₆ ): δ10.15 (1H, br), 9.72 (1H, br), 8.70 (1H, s), 8.41 (1H, d, J = 5.6Hz), 8.16-8.12 (2H, m), 7.67 (1H, d, J = 5.6Hz), 7.22-712 (2H, m), 6.99-6.92 (3H, m), 6.19 (1H, dd, J = 2.0Hz, 17.2Hz), 5.68 (1H, dd, J = 2.0Hz, 10.4Hz), 3.77 (3H, s), 3.34 (3H, s), 3.28 (4H, br), 2.72 (6H, s), 2.60 (3H, s).

Example 2: N-(2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxy-5-(4-(3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)pyrimidin-2-ylamino)phenyl)acrylamide (I)

1-(2-(5-amino-4-((2-(dimethylamino)ethyl)(methyl)amino)-2-methoxyphenylamino)pyrimidin-4-yl)-3-methyl-1H-benzo[d]imidazol-2(3H)-one (82 g) obtained in Step 6 of Example 1 was added to THF (800 mL) and water (80 mL), stirred to dissolve, and 3-chloropropionyl chloride (24.8 g) was added dropwise. After TLC showed the disappearance of the starting material, triethylamine (358.2 g) was added and the temperature was raised to 65°C for reaction. After completion of the reaction, the reaction solution was concentrated to dryness, dissolved in 1 L of dichloromethane, and separated twice with water (500 mL). The organic phase was collected and concentrated to obtain 88 g of a crude product. The crude product was separated by column chromatography (DCM:MeOH=20:1) to obtain 62.5 g of the title compound.

ESI-MS [M+H] ⁺ : 517.2677.

¹ H NMR (DMSO-d ₆ ): δ10.05 (1H, s), 8.67 (1H, s), 8.5 (1H, s), 8.44 (1H, d, J = 5.6Hz), 8.12 (1H, d, J = 7.6Hz) , 7.13 (2H, m), 6.9 (1H, t, J = 6.4Hz), 7.7 (1H, d, J = 5.6Hz), 7.05 (1H, s), 6.4 (1H, dd, J = 10 .15Hz, 16.9Hz), 6.21 (1H, dd, J=1.6Hz, 16.9Hz), 5.72 (1H, brd, J=11.50Hz), 3.77 (3H, s ), 3.35 (3H, s), 2.91 (2H, t, J = 5.65Hz), 2.75 (3H, s), 2.34 (2H, t, J = 5.7Hz), 2.21 (6H, s).

Example 3: Crystallization of the hydrochloride salt of the compound of formula I A

Method 1

10 g of the compound obtained in Example 2 was added to a 500 mL reaction kettle. 150 mL of ethanol was added and stirred. 10 mL of 2N hydrochloric acid was slowly added. After dissolving, the mixture was stirred for 2 h and filtered. The filter cake was dried under vacuum at 45-50°C. The collected solid (8.3 g) was dissolved in 49.8 mL of ethanol/water (ethanol:water = 3:1) solution. After stirring at 80°C and dissolving, the mixture was cooled to 25-30°C and filtered. The filter cake was dried under vacuum at 45-50°C to obtain the corresponding crystals.

Method 2

1 g of the compound obtained in Example 2 was added to a 25 mL reactor, followed by 10 mL of acetonitrile. Stir thoroughly, then slowly add 1 mL of 2N hydrochloric acid, causing the solid to gradually dissolve. Stirring continued for 10 minutes until solids precipitated. After thorough stirring for 12 hours, the mixture was filtered, and the filter cake was rinsed with 2 mL of acetonitrile and dried under vacuum at 45°C to obtain the corresponding crystals.

Method 3

1 g of the compound obtained in Example 2 was added to a 25 mL reactor. 5 mL of methanol was added and stirred thoroughly. 1 mL of 2N hydrochloric acid was slowly added, and the solid gradually dissolved. Stirring continued for 10 minutes until solid precipitated. After thorough stirring for 12 hours, the mixture was filtered, and the filter cake was rinsed with 2 mL of methanol and dried under vacuum at 45°C to obtain the corresponding crystals.

Method 4

1 g of the compound obtained in Example 2 was added to a 25 mL reaction kettle. 5 mL of isopropanol was added and stirred thoroughly. 1 mL of 2N hydrochloric acid was slowly added. The solid gradually dissolved and solid precipitated quickly. After thorough stirring for 12 h, the mixture was filtered. The filter cake was rinsed with 2 mL of isopropanol and dried under vacuum at 45°C to obtain the corresponding crystals.

Example 4: Crystallization of the hydrochloride salt of the compound of formula I B

Method 5

Add 10 g of the compound obtained in Example 2 to a 500 mL reactor, add 150 mL of ethanol, and stir at 25°C. If the reaction system remains unclear, slowly add 2N hydrochloric acid, stir for 2 h, and filter. Dry the filter cake in a vacuum at 45-50°C to obtain the desired crystalline form.

Example 5 Crystalline Hydrochloride of Compound I C

Method 6

1 g of the compound obtained in Example 2 was added to a 25 mL reaction kettle. 5 mL of tetrahydrofuran was added and stirred thoroughly. 1 mL of 2N hydrochloric acid was slowly added. The solid gradually dissolved, but soon became turbid. After thorough stirring for 12 h, the mixture was filtered. The filter cake was rinsed with 2 mL of tetrahydrofuran and dried under vacuum at 45°C to obtain the corresponding crystals.

Method 7

1 g of the compound obtained in Example 2 was added to a 25 mL reaction kettle. 5 mL of acetone was added and stirred thoroughly. 1 mL of 2N hydrochloric acid was slowly added, and the solid gradually dissolved. Stirring continued for 10 minutes until solid precipitated. After thorough stirring for 12 hours, the mixture was filtered, and the filter cake was rinsed with 2 mL of acetone and dried under vacuum at 45°C to obtain the corresponding crystals.

Method 8

1 g of the compound obtained in Example 2 was added to a 25 mL reaction kettle. 5 mL of 1,4-dioxane was added and stirred thoroughly. 1 mL of 2N hydrochloric acid was slowly added, and the solid gradually dissolved. Stirring continued for 10 minutes until solid precipitated. After thorough stirring for 12 hours, the mixture was filtered, and the filter cake was rinsed with 2 mL of 1,4-dioxane and dried in vacuo at 45°C to obtain the corresponding crystals.

Example 6: Stability test

Crystal A obtained by method 1 of Example 3 was placed at room temperature and protected from light. Samples were taken for testing at 1.5 months, 2 months, 5 months, and 6 months respectively. The test results were compared with the initial test results at day 0. The test results are shown in the following table:

Crystal B obtained by method 5 of Example 4 was placed in a high temperature environment of 60°C, a high humidity environment of 75% RH and a high humidity environment of 92.5% RH, and an illumination environment of 5000 Lux. Samples were taken for testing on the 5th, 10th, and 30th days, and the test results were compared with the initial test results on day 0. The test results are shown in the following table:

Example 7: In vitro activity test

1. In vitro enzymatic assay

EGFR and EGFR (T790M, L858R) kinases are expressed and purified through an insect expression system or purchased from commercial products.

A homogeneous time-resolved fluorescence (HTRF) assay platform for EGFR and EGFR (T790M, L858R) kinase activity was established using Cisbio's homogeneous time-resolved fluorescence (HTRF) method to determine compound activity. Compounds were serially diluted 10-fold starting at 1 μM in 100% DMSO. 4 μl of each dilution was added to 96 μl of reaction buffer (50 mM HEPES (pH 7.0), 0.02% _NaN₃ , 0.01% BSA, 0.1 mM Orthovanadate, 5 _mM MgCl₂, 50 nM SEB, 1 mM DTT). 2.5 μl of the diluted solution was then added to a 384-well plate (OptiPlate-384, PerkinElmer). 2.5 μl of kinase was then added, the mixture was centrifuged, and 5 μl of ATP and TK substrate-biotin were added to initiate the reaction. After incubating the 384-well plate in a 23°C incubator for a specified period of time, 5 μl of Eu ³⁺ -Cryptate-labeled TK-Antibody and 5 μl of streptavidin-XL665 were added to terminate the reaction. After incubation for 1 hour, fluorescence was read on an Envision (PerkinElmer) microscope. IC ₅₀ values for the compounds were calculated using GraphPad Prism 5.0 software.

2. Cell Proliferation Assay

Human non-small cell lung cancer cells NCI-H1975 were cultured in RPMI-1640 medium (supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin) in a cell culture incubator (37°C, 5% _CO₂ ). 2000 cells (195 μl) were seeded per well of a 96-well plate and cultured overnight. The following day, compounds were added in a three-fold serial dilution starting at 10 mM. 4 μl of each dilution was added to 96 μl of culture medium, and then 5 μl was added to the cell culture medium (final DMSO concentration of 0.1%, v/v). After 72 hours of treatment, the medium was aspirated and 30 μl of reagent (Promega) was added. Fluorescence signals were read on an Envison (Perkin Elmer) instrument, and _IC₅₀ values for cell proliferation inhibition were calculated using GraphPad Prism 5.0.

Human skin squamous cell carcinoma cell line A431 was cultured in a cell culture incubator (37°C, 5% _CO₂ ) using DMEM supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin. For compound testing, the substrate concentration was 0.6%. After reselection with 0.3% low-melting-point agar, 2,000 cells were plated per well (100 μl) in a 96-well plate. Compounds were serially diluted three-fold from 10 mM, with 2 μl of each concentration added to 98 μl of culture medium. Subsequently, 5.3 μl was added to the cell culture medium (final DMSO concentration of 0.1%, v/v). After one week (7 days), 20 μl of the reagent (Promega) was added, and the cells were incubated at 37°C for 4 hours. Fluorescence signals were read on an Envison (Perkin Elmer) and _IC₅₀ values for cell proliferation inhibition were calculated using GraphPad Prism 5.0.

Biological activity list

AZD9291 was prepared according to Example 28 of WO2013014448.

Example 8: Pharmacokinetic Evaluation

The test compound was administered orally to healthy adult male rats using a single dose of 10 mg/kg adjuvanted with 20% sulfobutyl ether-β-cyclodextrin. Animals receiving oral administration were fasted overnight and from 10 hours before to 4 hours after administration. Blood was collected at 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after oral administration. Approximately 0.3 mL of whole blood was collected from the orbital venous plexus into a heparinized tube. The sample was centrifuged at 4000 rpm for 5 minutes at 4°C. Plasma was transferred to a centrifuge tube and stored at -80°C until analysis. Plasma concentrations in the test compound samples were analyzed using a non-validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. Plasma concentration-time data for individual animals were analyzed using WinNonlin (Professional Edition, Version 6.3; Pharsight). A non-compartmental model was used for concentration analysis. Pharmacokinetic parameters of the test compound were calculated.

Claims (15)

1. Crystallization of the hydrochloride salt of compound I, A: The X-ray diffraction pattern of the hydrochloride crystal A of the compound of formula I has diffraction peaks at 2θ of 8.96°±0.2°, 14.11°±0.2°, 14.87°±0.2°, 16.52°±0.2°, 18.67°±0.2°, 21.93°±0.2° and 27.09°±0.2°. 2. The hydrochloride crystal A of the compound of formula I as described in claim 1, wherein the X-ray diffraction pattern has diffraction peaks with 2θ values of 8.33°±0.2°, 8.96°±0.2°, 12.16°±0.2°, 14.11°±0.2°, 14.87°±0.2°, 16.52°±0.2°, 17.66°±0.2°, 18.67°±0.2°, 21.93°±0.2°, and 27.09°±0.2°. 3. The hydrochloride crystal A of the compound of formula I as described in claim 1, wherein the X-ray diffraction pattern has diffraction peaks with 2θ values of 8.33°±0.2°, 8.96°±0.2°, 11.74°±0.2°, 12.16°±0.2°, 14.11°±0.2°, 14.87°±0.2°, 16.52°±0.2°, 17.66°±0.2°, 18.23°±0.2°, 18.67°±0.2°, 21.93°±0.2°, 22.65°±0.2°, and 27.09°±0.2°. 4. The hydrochloride crystal A of the compound of formula I as described in claim 1, having an X-ray diffraction pattern with 2θ values of 8.33°±0.2°, 8.96°±0.2°, 11.74°±0.2°, 12.16°±0.2°, 14.11°±0.2°, 14.87°±0.2°, 16.52°±0.2°, 17.66°±0.2°, 18.23°±0.2°, 18.67°±0.2°, 19.48°±0.2°, 19.92°±0.2°, 21.93°±0.2°, 22.65°±0.2°, 24.95°±0.2°, and 27.09°±0.2°. Diffraction peaks at 0.2° and 27.55°±0.2°. 5. The hydrochloride crystal A of the compound of formula I as described in claim 1, wherein the DSC spectrum shows a peak at 271°C. 6. A method for preparing the hydrochloride crystal A of the compound of formula I as described in claim 1, comprising the following steps: 1) Contacting the compound of formula I with hydrochloric acid; and 2) Crystallization from the crystallization solvent and optional filtration; The crystallization solvent is selected from acetonitrile, methanol, isopropanol, or an ethanol/water mixture. 7. A crystalline composition wherein the hydrochloride crystals A of the compound of formula I according to claim 1 or 5 constitute more than 50% by weight of the crystalline composition. 8. The crystalline composition of claim 7, wherein the hydrochloride crystal A of the compound of formula I of claim 1 or 5 accounts for more than 80% by weight of the crystalline composition. 9. The crystalline composition of claim 7, wherein the hydrochloride crystal A of the compound of formula I of claim 1 or 5 accounts for more than 90% by weight of the crystalline composition. 10. The crystalline composition of claim 7, wherein the hydrochloride crystal A of the compound of formula I of claim 1 or 5 accounts for more than 95% by weight of the crystalline composition. 11. A pharmaceutical composition comprising a therapeutically effective amount of a hydrochloride crystal of the compound of formula I as claimed in claim 1 or 5, or a crystalline composition as claimed in claim 7. 12. Use of the hydrochloride crystals A of the compound of formula I as claimed in claim 1 or 5, or the crystalline composition as claimed in claim 7, or the pharmaceutical composition as claimed in claim 11, in the preparation of a medicament for treating EGFR-mediated diseases. 13. The use as described in claim 12, wherein the EGFR-mediated disease is selected from ovarian cancer, cervical cancer, colorectal cancer, breast cancer, pancreatic cancer, glioma, melanoma, prostate cancer, leukemia, lymphoma, gastric cancer, lung cancer, hepatocellular carcinoma, gastrointestinal stromal tumor, thyroid cancer, bile duct cancer, endometrial cancer, kidney cancer, multiple myeloma, or mesothelioma. 14. The use as described in claim 13, wherein the EGFR-mediated disease is selected from lung cancer. 15. The use as described in claim 14, wherein the lung cancer is selected from non-small cell lung cancer.