HK40037665A

HK40037665A - Egfr inhibitors

Info

Publication number: HK40037665A
Application number: HK62021027699.9A
Authority: HK
Inventors: 代星; 江岳恒
Original assignee: 益方生物科技(上海)股份有限公司
Priority date: 2018-05-15
Filing date: 2019-05-14
Publication date: 2021-06-11

Description

EGFR inhibitors

Background

Technical Field

In various embodiments, the present invention relates generally to EGFR inhibitors, pharmaceutical compositions comprising the inhibitors, and methods of making and using the same.

Background

Epidermal Growth Factor Receptor (EGFR) is a receptor tyrosine protein kinase, a transmembrane protein in the ErbB receptor family.

EGFR regulates the proliferation, survival, adhesion, migration and differentiation of cells, which are over-activated or maintained in a variety of tumor cells such as lung cancer cells, breast cancer cells, prostate cancer cells, and the like. Abnormal activation of EGFR plays a key role in tumor transformation and growth. Blockade of activation of EGFR has been clinically proven to be one of the effective targeted therapies for the treatment of cancer. EGFR has been found to be expressed in 50% of NSCLC (non-small cell lung cancer) patients, making EGFR and its family members candidate targets for targeted therapy. Gefitinib and erlotinib are first-generation EGFR small molecule inhibitors, primarily used as drugs for the treatment of advanced NSCLC. Clinical results indicate that gefitinib or erlotinib is effective in about 10% of caucasian NSCLC patients and about 35% of asian NSCLC patients. The analysis also showed that in most NSCLC patients with EGFR-activating mutants, the response rate to EGFR-Tyrosine Kinase Inhibitors (TKI) was significantly higher than that of NSCLC patients with wild-type EGFR alone.

Clinical studies further indicate that many patients develop resistance to these EGFR small molecule inhibitors quickly (12-14 months), i.e., acquired resistance. The Gatekeeper residue mutation (T790M mutation) occurs in EGFR exon 20, which is one of the major mechanisms responsible for drug resistance. New generations of inhibitors of these EGFR mutants have recently succeeded. Afatinib is a potent and irreversible dual inhibitor of EGFR and human epidermal growth factor receptor 2(HER2) tyrosine kinases. Other similar multi-target, highly active and irreversible inhibitors, such as canertinib and dacomitinib, are also in clinical trials. These novel second generation irreversible inhibitors have potent EGFR inhibition on L858R and T790M mutants and have significant impact on gefitinib or erlotinib resistant cancer patients. However, these second generation EGFR mutant inhibitors also have strong inhibitory effects on wild type EGFR (WT-EGFR). Clinical studies have shown that inhibition of wild-type EGFR can lead to drug toxicity and side effects in most patients, such as human rash or diarrhea.

To overcome the toxicity and side effects of second generation EGFR inhibitors, it is desirable to reduce the inhibitory effect on wild type EGFR (WT-EGFR). The third-generation EGFR inhibitor can keep strong inhibition effect on an EGFR L858R mutant, an Exon19 deletion mutant and/or a T790M mutant, and has relatively low inhibition effect on WT-EGFR and other tyrosine protein kinase receptors. Such compounds may be used to treat not only cancer patients resistant to EGFR L858R mutant and Exon19 deletion mutant, but also cancer patients having EGFR-T790M mutant. One of the third generation EGFR inhibitors AZD9291 has beneficial clinical effects. However, its major metabolite AZ5104 has strong inhibitory effect on wild-type EGFR (WT-EGFR), which may induce most common side effects, such as clinically common skin rash, diarrhea, etc.

Brief description of the invention

U.S. publication No. 2017/0355696a1 describes various pyrimidine compounds that are effective in selectively inhibiting EGFR variants or mutants and are useful in the treatment of diseases or disorders mediated by such EGFR variants or mutants, such as cancer. In various embodiments, the present invention relates to EGFR inhibitors and pharmaceutically acceptable salts thereof, e.g., in crystalline form and/or as substantially pure compounds, pharmaceutical compositions comprising the same, methods of making the same, and methods of use thereof.

Certain embodiments of the present invention relate to the compound 4(N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (1- (2,2, 2-trifluoroethyl) -1H-indol-3-yl) pyrimidin-2-yl) amino) phenyl) acrylamide), or a pharmaceutically acceptable salt thereof.

Some embodiments of the present invention relate to a method of preparing compound 4 or a pharmaceutically acceptable salt thereof. In some embodiments, the method comprises:

(1) converting compound 1 or a salt thereof to a compound of formula III or a salt thereof under amide forming conditions, wherein Lg in formula III is a leaving group; and

(2) converting a compound of formula III or a salt thereof to compound 4 under elimination reaction conditions; in some embodiments, the method further comprises reacting compound 4 with a suitable acid to form a pharmaceutically acceptable salt of compound 4. In some embodiments, Lg in formula III is a halogen or an oxygen-containing leaving group, e.g., Lg in formula III can be chloro. In some embodiments, converting compound 1 to a compound of formula III, or a salt thereof, comprises contacting compound 1 with a compound having formula IV,wherein Lg is a leaving group, e.g., chloro. In some embodiments, converting the compound of formula III or a salt thereof to compound 4 comprises contacting the compound of formula III with a base (e.g., a base, such as potassium hydroxide). Compound 1 can be prepared by various methods, for example, as described herein.

Compound 4 can be prepared in high purity according to the methods disclosed herein. Typically, compound 4 prepared according to the methods herein has less than 2% (e.g., less than 1.5%, less than 1%, less than 0.8%, less than 0.5%, less than 0.2%) total impurities as measured by HPLC. In some embodiments, compound 4 does not contain more than 1% (e.g., no more than 0.8%, no more than 0.5%, no more than 0.2%) of a single impurity as measured by HPLC.

In some embodiments, the present disclosure provides compound 4 or a pharmaceutically acceptable salt produced by any of the synthetic methods herein. In some embodiments, the present disclosure relates to substantially pure compound 4 or a pharmaceutically acceptable salt thereof. In some embodiments, the HPLC purity of compound 4 is at least 95% (e.g., about 96%, about 97%, about 98%, about 98.5%, about 99%, or greater than 99%). In some embodiments, compound 4 may be substantially free of impurities characterized by a relative retention time of about 1.02 (impurity a) for HPLC. In some embodiments, compound 4 may be substantially free of impurities characterized by a relative retention time of about 0.88 for HPLC (impurity B).

In some embodiments, the present disclosure relates to a substantially pure pharmaceutically acceptable salt of compound 4. Generally, a pharmaceutically acceptable salt of substantially pure compound 4 can be prepared by mixing substantially pure compound 4 with a suitable acid (e.g., hydrochloric acid, methanesulfonic acid, and the like). In some embodiments, the substantially pure pharmaceutically acceptable salt of compound 4 can be a substantially pure mesylate salt of compound 4, e.g., a monomethanesulfonate salt of compound 4, referred to herein as compound 5.

Compound 5 described herein can be substantially pure. For example, compound 5 is characterized by a purity (by weight and/or HPLC area) of at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or at least 99%). In some embodiments, substantially pure compound 5 is characterized by an amount of its methanesulfonic acid that is close to the theoretical methanesulfonic acid content calculated based on the molecular formula of compound 5. In any of the embodiments described herein, substantially pure compound 5 can comprise, consist essentially of, or consist of compound 5 in crystalline form I described herein. In some embodiments, form I of compound 5 can be characterized by its particle size distribution, e.g., as described herein.

In some embodiments, the present invention provides a pharmaceutical composition comprising compound 4 (e.g., substantially pure compound 4 herein) or a pharmaceutically acceptable salt thereof, e.g., a mesylate salt. In some embodiments, the present invention provides a pharmaceutical composition comprising, consisting essentially of, or consisting essentially of substantially pure compound 4 or a pharmaceutically acceptable salt thereof, e.g., compound 5, and optionally a pharmaceutically acceptable excipient or carrier.

The pharmaceutical compositions described herein may be formulated for any suitable route of administration. In some embodiments, the pharmaceutical composition may be formulated for oral administration. For example, in any of the embodiments described herein, the pharmaceutical composition can be formulated in the form of a tablet or capsule. In some embodiments, the pharmaceutical composition may be enterically coated. However, in some embodiments, the pharmaceutical composition may be a non-enteric coating.

Certain embodiments of the present invention relate to methods of treating various diseases or disorders in a subject in need thereof using the compounds, salts, solid forms and/or pharmaceutical compositions herein. In some embodiments, the disease or disorder is mediated by EGFR variants or mutants, such as L858R mutants, Exon19 deletion mutants, and/or T790M mutants. In some embodiments, the invention provides a method of treating cancer. In some embodiments, the cancer is 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, liver cancer, gastrointestinal stromal tumor, thyroid cancer, bile duct cancer, endometrial cancer, kidney cancer, anaplastic large cell lymphoma, acute myelogenous leukemia, multiple myeloma, and mesothelioma. In some embodiments, the cancer is lung cancer. In some embodiments, the cancer is non-small cell lung cancer. In some embodiments, the subject is characterized as resistant to one or more EGFR inhibitors, e.g., compound 4 or a salt thereof, e.g., gefitinib, erlotinib and/or icotinib.

The methods described herein generally include administering to a subject a therapeutically effective amount of substantially pure compound 4 or a pharmaceutically acceptable salt thereof, e.g., compound 5. The methods described herein are not limited to any particular route of administration. For example, in any of the embodiments described herein, the administration can be oral administration.

In any of the embodiments described herein, compound 4, e.g., substantially pure compound 4, or a pharmaceutically acceptable salt thereof, e.g., compound 5, can be used in a monotherapy wherein the active agent consists of or consists essentially of compound 4 or a pharmaceutically acceptable salt thereof, e.g., compound 5, said compound 4 can be in crystalline form I, amorphous form, or a combination thereof. However, in some embodiments, the methods described herein may also be used in combination with other therapies, including additional active agents. For example, the methods herein may be used in combination with one or more selected from the group consisting of surgical procedures (e.g., conventional anti-cancer surgical therapies), radiation therapy, chemotherapy, and anti-tumor immunotherapy.

Drawings

Figure 1 shows a representative 1H NMR spectrum of compound 5 in d6 DMSO.

Figure 2A shows a representative X-ray powder diffraction (XRPD) spectrum of form I of compound 5. Figure 2B shows representative thermogravimetric analysis (TGA) and Differential Scanning Calorimetry (DSC) analysis of form I of compound 5.

Figure 3 shows the XRPD spectrum obtained from a heat treatment experiment using form I. The XRPD overlay of the sample (after treatment) and the initial drug showed that form I converted to amorphous form after heat treatment.

Figure 4A shows a representative HPLC line of compound 4 produced by the methods herein without a step of recrystallization in Isopropanol (IPA) and water. Figure 4B shows a representative HPLC line for compound 4 after recrystallization from IPA and water.

Figure 5A shows a representative HPLC line of compound 5 obtained from the salt form of compound 4, which was further purified without recrystallization in IPA and water. Figure 5B shows a representative HPLC line of compound 5 obtained from the salt form of compound 4, which was further purified by recrystallization from IPA and water.

Detailed Description

In various embodiments, compounds and pharmaceutically acceptable salts, e.g., crystalline forms or substantially pure compounds, are provided. Pharmaceutical compositions, methods of making and methods of using the same are also provided.

Compounds and salts

In various embodiments, the invention relates to compounds and/or salts of compounds (e.g., monomethanesulfonates) that are potent inhibitors of EGFR variants or mutants and that are useful for treating various diseases and disorders, such as those mediated by EGFR variants or mutants. Examples of such compounds are previously described in U.S. patent publication nos. 2017/0355696a1 and 15/524,228, which are incorporated herein by reference in their entirety.

In some particular embodiments, the present invention provides substantially pure compound 4 or a pharmaceutically acceptable salt thereof. In some embodiments, the present invention provides a methanesulfonic acid addition salt of compound 4. The chemical name of compound 4 is N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (1- (2,2, 2-trifluoroethyl) -1H-indol-3-yl) pyrimidin-2-yl) amino) phenyl) acrylamide. Compound 4 is a potent and selective inhibitor of EGFR variants or mutants (e.g., Exon19 deletion mutants and/or T790M mutants) and has the structure shown below:

compound 4 has one or more basic nitrogen atoms and can form acid addition salts. In some particular embodiments, the present invention relates to the monomethanesulfonate salt of compound 4. In some embodiments, the monomethanesulfonate salt of compound 4 is referred to herein as compound 5, and is represented by the following structure:

for the avoidance of doubt, as used herein, the monomethanesulfonate salt of compound 4 is understood to be formed from a base (compound 4) and methanesulfonic acid in a ratio of 1: 1 molar ratio. Those skilled in the art will appreciate that the exact protonation sites of methane sulfonic acid for compound 4 may vary and are in equilibrium with each other. Compound 5 as shown does not show which nitrogen is protonated, but reflects that it is 1: 1.

In some embodiments, the present disclosure relates to substantially pure compound 4 or a pharmaceutically acceptable salt thereof. In some embodiments, the HPLC purity of compound 4 is at least 95% (e.g., about 96%, about 97%, about 98%, about 98.5%, about 99%, or greater than 99%). In some embodiments, compound 4 may be substantially free of impurities characterized by a relative retention time of about 1.02 (impurity a) for HPLC. For example, in some embodiments, impurity a may be present in an amount of less than 1%, less than 0.8%, less than 0.5%, less than 0.2%, less than 0.1%, as measured by HPLC. In some embodiments, compound 4 can be characterized by an HPLC line that includes a peak representing compound 4, and a peak representing impurity a, wherein the area percent of the peak representing impurity a is less than 1%, less than 0.2%, or less than 0.1%. In some embodiments, compound 4 can be characterized by an HPLC line that includes a peak representing compound 4 at an area percentage of at least 95% (e.g., about 96%, about 97%, about 98%, about 98.5%, about 99%, or greater than 99%), wherein the HPLC line does not include an identifiable peak representing impurity a. In some embodiments, compound 4 may be substantially free of impurities characterized by a relative retention time of about 0.88 for HPLC (impurity B). For example, in some embodiments, impurity B may be present in an amount of less than 0.5%, less than 0.2%, or less than 0.1%, as measured by HPLC. In some embodiments, compound 4 can be characterized by an HPLC line that includes a peak representing compound 4, and a peak representing impurity B, wherein the area percent of the peak representing impurity B is less than 0.5%, less than 0.2%, or less than 0.1%. In some embodiments, compound 4 can be characterized by an HPLC line that includes a peak representing compound 4 at an area percentage of at least 95% (e.g., about 96%, about 97%, about 98%, about 98.5%, about 99%, or greater than 99%), wherein the HPLC line does not include an identifiable peak representing impurity B. In some embodiments, compound 4 may be substantially free of impurity a and impurity B, e.g., less than 0.5%, less than 0.2%, or less than 0.1% of the total content of impurities a and B as measured by HPLC. In some embodiments, compound 4 can be characterized by an HPLC line that includes a peak representing compound 4, a peak representing impurity a, and a peak representing impurity B, wherein the total area percentage of the peaks representing impurity a and the peaks representing impurity B is less than 0.5%, less than 0.2%, or less than 0.1%. In some embodiments, compound 4 can be characterized by an HPLC line that includes a peak representing compound 4 at least 95% by area (e.g., about 96%, about 97%, about 98%, about 98.5%, about 99% or greater than 99%), wherein the HPLC line does not include an identifiable peak representing impurity B, does not include an identifiable peak representing impurity a, or does not include an identifiable peak representing either impurity a or B. In some embodiments, the term "measured by HPLC" refers to measurement using the HPLC method shown in example 1, e.g., using 220nm as the detection wavelength. In some embodiments, the term "characterized by HPLC lines" refers to HPLC lines obtained by the HPLC method as set forth in example 1, e.g., using 220nm as the detection wavelength. The examples section details an exemplary method for preparing compound 4. The figure shows a representative HPLC line for purity analysis of compound 4.

In some embodiments, the present disclosure relates to a substantially pure pharmaceutically acceptable salt of compound 4. Generally, a pharmaceutically acceptable salt of substantially pure compound 4 can be prepared by mixing substantially pure compound 4 with a suitable acid (e.g., hydrochloric acid, methanesulfonic acid, and the like). The pharmaceutically acceptable salt of compound 4 may generally be a single salt, in a molar ratio to the corresponding acid of about 1: 1, or double salt, in a molar ratio to the corresponding acid of about 1: 2 (i.e. about 2 moles of acid per mole of compound 4). Non-limiting examples of suitable acids include any of those that can form pharmaceutically acceptable acid addition salts with compound 4. Examples of suitable acids include 1-hydroxy-2-naphthoic acid, 2-dichloroacetic acid, 2-hydroxyethanesulfonic acid, 2-oxoglutaric acid, 4-acetamidobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, ascorbic acid (L), aspartic acid (L), benzenesulfonic acid, benzoic acid, camphoric acid (+), camphor-10-sulfonic acid (+), capric acid (n-capric acid), caproic acid (n-caproic acid), caprylic acid (caprylic acid), carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane 1, 2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptoic acid (D), gluconic acid (D), glucuronic acid (D), glutamic acid, glutaric acid, glycerophosphoric acid, glyceric acid, Glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isobutyric acid, lactic acid (DL), lactobionic acid, lauric acid, maleic acid, malic acid (L), malonic acid, mandelic acid (DL), methanesulfonic acid, naphthalene 1, 5-disulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, nitric acid, oleic acid, oxalic acid, palmitic acid, pamoic acid (pamoic acid), phosphoric acid, propionic acid, pyroglutamic acid (-L), salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, tartaric acid (+ L), thiocyanic acid, toluenesulfonic acid (p), and undecylenic acid. In some embodiments, the substantially pure pharmaceutically acceptable salt of compound 4 can be a substantially pure mesylate salt of compound 4.

The monomethanesulfonate salt compound 5 described herein may be substantially pure. For example, compound 5 is characterized by a purity (by weight and/or HPLC area) of at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or at least 99%). In some embodiments, compound 5 is characterized by a purity (by weight and/or HPLC area) of about 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99%, or any range between the specified values. Substantially pure compound 5 can be prepared from substantially pure compound 4. Unless otherwise apparent from the context, anything other than a compound or salt, or a solvate or hydrate form thereof, is considered an impurity, including, for example, residual solvent, moisture content, etc., for the purpose of calculating the weight percent of the compound/salt in a substantially pure compound or salt. For the avoidance of doubt, a composition comprising a substantially pure compound or salt herein and one or more other ingredients is to be understood as a composition obtained by mixing a substantially pure compound or salt herein and one or more other ingredients, such as water, pharmaceutically acceptable excipients and the like, either directly or indirectly.

Substantially pure compound 5 described herein may comprise an amount of methanesulfonic acid that approximates the theoretical methanesulfonic acid content calculated based on the formula for compound 5. In some embodiments, substantially pure compound 5 is characterized by a methanesulfonic acid content of about 13 to 15 weight percent. For example, methods for determining the methanesulfonic acid content by titration using a Mettler Toledo T50 titrator are known.

Substantially pure compound 5 herein may comprise no or substantially no compound 4, and/or may comprise no or substantially no other salts of compound 4. In some embodiments, substantially pure compound 5 is substantially free of compound 4, e.g., in an amount less than 5% by weight (e.g., less than 3%, less than 1%, less than 0.2%, less than 0.1%, or less than 0.05%). In some embodiments, substantially pure compound 5 does not comprise compound 4, except in an amount that may be present by equilibrium. In some embodiments, substantially pure compound 5 is free of detectable amounts of compound 4. In some embodiments, substantially pure compound 5 is substantially free of other salts of compound 4, such as the bis-mesylate salt, in an amount less than 5% by weight (e.g., less than 3%, less than 1%, less than 0.2%, less than 0.1%, or less than 0.05%). In some embodiments, substantially pure compound 5 does not contain detectable amounts of other salts of compound 4. In some embodiments, substantially pure compound 5 does not comprise the bis-mesylate salt of compound 4, except in an amount that can be present by equilibrium.

Compound 5 can exist in various solid states. For example, in some embodiments, compound 5 can be in a crystalline form. In some embodiments, compound 5 may be an amorphous solid. In some embodiments, compound 5 can be a mixture of crystalline and amorphous forms.

In some particular embodiments, the present invention provides form I of compound 5, which is the only identifiable stable crystalline form.

As used herein, form I of compound 5 refers to a crystalline form of compound 5, which can be characterized by an XRPD pattern substantially the same as figure 2A; an XRPD pattern having the major peaks of fig. 2A (e.g., peaks with relative intensities of 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more); an XRPD pattern having one or more (e.g., 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, and 23) of the following peaks: 2 θ angles of 7.8, 10.2, 10.8, 11.2, 13.6, 13.9, 15.0, 16.4, 17.7, 18.4, 18.7, 18.9, 19.2, 19.8, 20.6, 20.8, 21.1, 22.6, 23.7, 23.9, 24.9, 25.9 and 27.8 degrees ± 0.2 °; a DSC plot with an endothermic peak and a peak temperature of about 210.1 ℃; a DSC curve substantially the same as shown in figure 2B; a TGA profile substantially the same as shown in figure 2B; or a combination thereof. As used herein, the main peak of an XRPD spectrum refers to a peak having a diffraction angle of 4 to 30 degrees (2. theta.) and a relative intensity of 10% or more. In some embodiments, the main peaks of the XRPD spectrum may refer to peaks having a relative intensity of 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more. Some details of form I of compound 5 are also described in U.S. patent application provisional application No. 62/678,634, filed on 31/5/2018, which application claims priority thereto, and the contents of which are incorporated herein by reference in their entirety.

For example, in some embodiments, form I of compound 5 is characterized by an XRPD pattern having four or more of the following peaks: the 2 θ angles are 7.8, 10.2, 10.8, 11.2, 13.6, 13.9, 15.0, 16.4, 17.7, 18.4, 18.7, 18.9, 19.2, 19.8, 20.6, 20.8, 21.1, 22.6, 23.7, 23.9, 24.9, 25.9, and 27.8 degrees ± 0.2 °. In some embodiments, form I of compound 5 is characterized by an XRPD pattern having eight or more of the following peaks: the 2 θ angles are 7.8, 10.2, 10.8, 11.2, 13.6, 13.9, 15.0, 16.4, 17.7, 18.4, 18.7, 18.9, 19.2, 19.8, 20.6, 20.8, 21.1, 22.6, 23.7, 23.9, 24.9, 25.9, and 27.8 degrees ± 0.2 °. In some embodiments, form I of compound 5 is characterized by an XRPD pattern having twelve or more of the following peaks: the 2 θ angles are 7.8, 10.2, 10.8, 11.2, 13.6, 13.9, 15.0, 16.4, 17.7, 18.4, 18.7, 18.9, 19.2, 19.8, 20.6, 20.8, 21.1, 22.6, 23.7, 23.9, 24.9, 25.9, and 27.8 degrees ± 0.2 °. In some embodiments, form I of compound 5 is characterized by an XRPD pattern having sixteen or more of the following peaks: the 2 θ angles are 7.8, 10.2, 10.8, 11.2, 13.6, 13.9, 15.0, 16.4, 17.7, 18.4, 18.7, 18.9, 19.2, 19.8, 20.6, 20.8, 21.1, 22.6, 23.7, 23.9, 24.9, 25.9, and 27.8 degrees ± 0.2 °. In some embodiments, form I of compound 5 is characterized by an XRPD pattern having all of the following peaks: the 2 θ angles are 7.8, 10.2, 10.8, 11.2, 13.6, 13.9, 15.0, 16.4, 17.7, 18.4, 18.7, 18.9, 19.2, 19.8, 20.6, 20.8, 21.1, 22.6, 23.7, 23.9, 24.9, 25.9, and 27.8 degrees ± 0.2 °. In some embodiments, form I of compound 5 is characterized by an XRPD pattern substantially the same as shown in figure 2A. In some embodiments, form I of compound 5 may also be characterized by a DSC profile having an endothermic peak with a peak temperature of about 210.1 ℃, and/or substantially the same DSC curve as shown in figure 2B. In some embodiments, form I of compound 5 can also be characterized by a TGA profile substantially the same as that shown in figure 2B.

The size of the crystals used to prepare the tablets or capsules containing the compounds of the invention may be important. If the crystals are too small, they may stick to the plunger of the tablet press or have undesirable flow properties. On the other hand, they cannot be too large. As the crystal size increases, the rate of dissolution in the intestine decreases. Thus, if the crystals are too large, the bioavailability of the compound may be compromised. The particle size distribution can be described using quantiles, such as D10%, D50%, D90%, and the like. For the sake of brevity, "%" may be omitted in this application. Thus, D10 was identical to D10%. As used herein, "particle size distribution" refers to the cumulative volume particle size distribution of equivalent spherical diameters as determined by a Malvern Mastersizer 3000 particle size analyzer.

In some embodiments, the crystals of form I of compound 5 are characterized by a particle size distribution of 1) D90: about 150 μm to about 250 μm; d50: about 90 μm to about 140 μm; and D10 is about 40 μm to about 75 μm; 2) d90: about 100 μm to about 150 μm; d50: about 20 μm to about 60 μm; and D10 is about 5 μm to about 15 μm; 3) d90: about 120 μm to about 200 μm; d50: about 50 μm to about 100 μm; and D10 is from about 10 μm to about 20 μm; 4) d90: about 140 μm to about 180 μm; d50: about 90 μm to about 130 μm; and D10 is about 55 μm to about 85 μm; 5) d90: about 50 μm to about 100 μm; d50: about 20 μm to about 30 μm; and D10 is about 3.5 μm to about 9 μm; 6) d90: about 70 μm to about 95 μm; d50: about 25 μm to about 40 μm; and D10 is about 5 μm to about 15 μm; 7) d90: about 120 μm to about 160 μm; d50: about 55 μm to about 85 μm; and D10 is about 5 μm to about 15 μm; or 8) D90: about 90 μm to about 130 μm; d50: about 35 μm to about 60 μm; and D10 is about 10 μm to about 15 μm. In some embodiments, the crystals of compound 5 in form I are characterized by a particle size distribution of D90: about 80 μm to about 120 μm; and D50: about 30 μm to about 60 μm. The unit "μm" herein refers to microns.

The particle size distribution herein can be adjusted by crystallization conditions. For example, as detailed in the examples section, the inventors have found that gradual slow cooling in the presence of seeds can help achieve the desired particle size distribution. Generally, following this gradual cooling procedure, the crystallization process described herein may produce form I with a narrower particle size distribution, which facilitates further formulation processing. The seeds can be prepared by the methods described herein without the need for a seeding step. In some embodiments, the seeds used in the gradual cooling procedure may be sieved to obtain the desired particle size distribution of the seeds themselves. For example, in some embodiments, the seeds may be prepared by passing the bulk crystals through a 200 mesh screen and collecting the crystals that remain as seeds, i.e., those that do not pass through the screen. In some embodiments, the seed crystals are used without a sieving procedure. In some embodiments, other sized screens may be used to obtain different seed particle size distributions. In some embodiments, the particle size distribution may be adjusted by milling, such as wet milling as described herein.

In any of the embodiments described herein, substantially pure compound 5 can consist essentially of form I of compound 5. In any of the embodiments described herein, compound 5 may be present as crystalline form I. In any of the embodiments described herein, compound 5 may also be present in amorphous form. In any of the embodiments described herein, substantially pure compound 5 or a pharmaceutical composition comprising compound 5 may comprise compound 5 only in the form of crystalline form I, i.e., no other solid forms of compound 5 can be identified by XRPD. In any of the embodiments described herein, substantially pure compound 5 or a pharmaceutical composition comprising compound 5 can further comprise compound 5 in a mixture of crystalline form I and amorphous form. In some embodiments, substantially pure compound 5 or a pharmaceutical composition comprising compound 5 may comprise compound 5 in amorphous form.

Exemplary methods of preparing crystalline compound 5 are described herein. Typically, substantially pure compound 4 is dissolved in a solvent (e.g., a mixture of ethanol and ethyl acetate) to form a solution; less than about 1 equivalent (e.g., 0.95 equivalent) of methanesulfonic acid can be added to the solution to form compound 5. The addition of methane sulfonic acid may be carried out, for example, at elevated temperatures, e.g., about 50-80 c, about 55-65 c. In some embodiments, compound 5 may precipitate out after addition of methanesulfonic acid, e.g., after addition of a majority of methanesulfonic acid. In some embodiments, compound 5 can be precipitated, for example, by cooling the solution or by reducing the amount of solvent by evaporation or adding an anti-solvent. In some embodiments, seeds of form I may be added to facilitate/control the crystallization process. Examples of the preparation of form I of compound 5 are provided in the examples section.

In some embodiments, compound 5 may be recrystallized under suitable conditions. Suitable solvents for recrystallization include, but are not limited to, tetrahydrofuran, toluene, methanol, ethanol, n-propanol, isopropanol, isobutanol, methyl tert-butyl ether, diethyl ether, isoamyl alcohol (isoamyl alcohol), 3-methyl-1-butanol (isopentanol), butyl acetate, ethyl formate, 1, 4-dioxane, n-butanol, tert-butanol, n-heptane, cyclohexane, dichloromethane, methyl isobutyl ketone, dimethyl benzene, isobutyl acetate, 2-butanone, acetonitrile, acetone, ethyl acetate, isopropyl acetate, and water. The solvents may be used alone or in combination of plural kinds. Recrystallization techniques are well known in the art.

Preparation method

U.S. publication No. 2017/0355696a1 describes a process for preparing compound 4 and its various pharmaceutically acceptable salts. An exemplary synthetic method in U.S. publication No. 2017/0355696a1 includes a two-step conversion from an aniline compound (corresponding to compound 1 of the present disclosure) to the disulfonate ester of compound 4 with low yield.

As shown herein, representative methods of preparing compound 4 or a pharmaceutically acceptable salt (or alternatively referred to as synthetic methods) can provide the desired compound 4 or a pharmaceutically acceptable salt in improved yield and high purity, and are suitable for large-scale production.

In various embodiments, the present invention provides a novel process for preparing compound 4 or a pharmaceutically acceptable salt thereof. The process generally comprises, typically under elimination reaction conditions, converting a compound of formula III, or a salt thereof, to compound 4:

wherein Lg in formula III is a leaving group, such as halogen (e.g., chloro).

In some embodiments, the compound of formula III may be prepared from compound 1 or a salt thereof. Accordingly, in some embodiments, the method comprises 1) converting compound 1 or a salt thereof to a compound of formula III or a salt thereof, e.g., under amide forming conditions, wherein Lg in formula III is a leaving group;

and 2) converting the compound of formula III or a salt thereof to compound 4, for example under elimination reaction conditions.

Although the prior art method of synthesizing compound 4 from compound 1 can be achieved by a one-step reaction with acryloyl chloride or acrylic anhydride, it has been found that a two-step conversion by the intermediate compound of formula III provides significant advantages. For example, by using this two-step conversion process, the final yield of compound 4 or a pharmaceutically acceptable salt can be increased. Purification of compound 4 or a pharmaceutically acceptable salt can also be simplified, which makes the synthetic route of the present invention more suitable for large scale production. In general, compound 1 or a salt thereof may be reacted with an acid of formula IVAcid reaction of, or activated forms thereof, e.g. of formula IVWherein Lg is a leaving group, under amide forming conditions to form a compound of formula III. The formation of amides is well known in the art, and various conditions may be applicable to the synthetic methods herein. For example, in some embodiments, compound 1 can be combined with an acid chloride of formula IV in a suitable solvent (e.g., an ethereal solvent, such as tetrahydrofuran, which can be combined with water in a ratio of about 10: 1). Generally, the reaction may be carried out at a temperature below room temperature, for example, about 0-10 ℃; an external base is generally not required. Exemplary conditions are shown in the examples section. The Lg in formula IV-acid or formula IV is generally the same as Lg in formula III. For example, typically, the formula III, formula IV-acid or Lg in formula IV can each be a leaving group selected from halogen and oxygen-containing leaving groups, e.g., -O-SO₂R, wherein R is alkyl (e.g., methyl, CF)₃) Or an aryl group (e.g., phenyl, p-tolyl, etc.). In some embodiments, the formula III, formula IV-acid or Lg in formula IV can be chloro. However, in some embodiments, the formula IV-acid or Lg in formula IV may also be different from and may be converted to Lg in formula III.

In some embodiments, the compound of formula III is not isolated prior to proceeding to the next step. In some embodiments, the compound of formula III may also be isolated and then used in the next step.

The compounds of formula III are also novel compositions of the invention. In some embodiments, the present invention also provides a compound of formula III or a salt thereof (e.g., a pharmaceutically acceptable salt thereof),

wherein Lg is a leaving group (e.g., as defined herein). In some embodiments, Lg is halogen, e.g., chlorine. In some embodiments, Lg is an oxygen-containing leaving group as described herein. In some embodiments, Lg can also be hydroxyl or protected hydroxyl. In some embodiments, Lg can also be an alkoxy group, e.g., C_1-6Alkoxy (e.g., methoxy, ethoxy, isopropoxy, etc.). In some embodiments, the disclosure also provides compositions comprising compound 4, or a pharmaceutically acceptable salt thereof, and a compound of formula III (e.g., as described herein, e.g., Lg is chloro or hydroxy, etc.), or a salt thereof. In some embodiments, the composition is substantially free of a compound of formula III or salt thereof, e.g., less than 1%, less than 0.5%, less than 0.2%, less than 0.1% as determined by HPLC (e.g., the HPLC method described in example 1, e.g., using 220nm as the detection wavelength). In some embodiments, the composition is a pharmaceutical composition, which in some embodiments comprises a therapeutically effective amount of compound 4 or a pharmaceutically acceptable salt thereof, e.g., mesylate compound 5.

The compound of formula III can generally be converted to compound 4 by treatment with a base, e.g., a base, such as an alkaline hydroxide, e.g., sodium hydroxide, potassium hydroxide, and the like. In some embodiments, the conversion of the compound of formula III to compound 4 may comprise contacting the compound of formula III with potassium hydroxide. Typically, the elimination reaction can be carried out in the same solvent as described above for the amide formation, e.g., using a mixture of tetrahydrofuran and water (e.g., in a 10: 1 ratio). The amount of base can be adjusted and is usually in excess, for example 2 to 5 equivalents. The elimination reaction may be carried out under heating. In some embodiments, other elimination conditions may also be used to effect the conversion. Reactions that eliminate the leaving group from the β -position of the amide are well known and may be suitable for use in the synthetic methods herein.

Exemplary conditions for converting compound 1 to compound 4 are shown in the examples section. As described above, the overall yield of compound 4 from compound 1 is greatly improved and exceeds 70%. Thus, the synthetic methods herein, as well as other advantages, at least greatly facilitate the large scale production of compound 4.

In some embodiments, the method further comprises purifying compound 4 by recrystallization, e.g., in a mixture of isopropanol and water. In some embodiments, recrystallization can further increase the purity of compound 4. For example, as described herein, in some embodiments, recrystallization in a mixture of isopropanol and water may reduce the content of impurities a and B (as defined herein). Exemplary conditions are shown in the examples.

In some embodiments, a pharmaceutically acceptable salt of compound 4 is desired. In such embodiments, the method further comprises the step of reacting compound 4 with a suitable acid to form a pharmaceutically acceptable salt of compound 4. In some embodiments, compound 4 can be prepared in high purity by the methods described herein, and when used as a starting material, it can also provide pharmaceutically acceptable salts with high purity. Thus, in some embodiments, the present disclosure also provides methods of preparing pharmaceutically acceptable salts by reacting substantially pure compound 4 (e.g., as described herein) with a suitable acid (e.g., any of those described herein, e.g., hydrochloric acid, methanesulfonic acid, and the like). In some embodiments, substantially pure compound 4 has an HPLC purity of at least 95% (e.g., about 96%, about 97%, about 98%, about 98.5%, about 99%, or greater than 99%). In some embodiments, substantially pure compound 4 may be substantially free of impurities characterized by a relative retention time of about 1.02 (impurity a) for HPLC. For example, in some embodiments, impurity a may be present in an amount of less than 1%, less than 0.8%, less than 0.5%, less than 0.2%, less than 0.1%, as measured by HPLC. In some embodiments, impurity a is not detected by HPLC (or is below the detection limit). In some embodiments, substantially pure compound 4 may be substantially free of impurities characterized by a relative retention time of about 0.88 for HPLC (impurity B). For example, in some embodiments, impurity B may be present in an amount of less than 0.5%, less than 0.2%, or less than 0.1%, as measured by HPLC. In some embodiments, impurity B is not detected by HPLC (or is below the detection limit). In some embodiments, substantially pure compound 4 may be substantially free of impurities a and B, e.g., less than 0.5%, less than 0.2%, or less than 0.1% of the total content of impurities a and B as measured by HPLC. In some embodiments, impurities a and B are not detected (or are below the detection limit) by HPLC. In some embodiments, the term "measuring by HPLC" refers to using the HPLC method shown in example 1, e.g., using 220nm as the detection wavelength. Pharmaceutically acceptable salts prepared by the methods herein can generally have an HPLC purity of greater than about 90% (e.g., about 95%, about 97%, about 98%, about 99%, or any range between the specified values).

In some embodiments, substantially pure compound 4 is converted to its monomethanesulfonate salt compound 5. In some embodiments, compound 5 is also substantially pure. For example, in some embodiments, compound 5 is characterized by a purity (by weight and/or HPLC area) of at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or at least 99%). In some embodiments, substantially pure compound 5 has an HPLC purity of at least 95% (e.g., about 96%, about 97%, about 98%, about 98.5%, about 99%, or greater than 99%). In some embodiments, substantially pure compound 5 may be substantially free of impurity a (as defined herein). For example, in some embodiments, impurity a may be present in an amount of less than 1%, less than 0.8%, less than 0.5%, less than 0.2%, less than 0.1%, as measured by HPLC. In some embodiments, impurity a is not detected by HPLC (or is below the detection limit). In some embodiments, substantially pure compound 5 may be substantially free of impurity B (as defined herein). For example, in some embodiments, impurity B may be present in an amount of less than 0.5%, less than 0.2%, or less than 0.1%, as measured by HPLC. In some embodiments, impurity B is not detected by HPLC (or is below the detection limit). In some embodiments, substantially pure compound 5 may be substantially free of impurity a and impurity B, e.g., less than 0.5%, less than 0.2%, or less than 0.1% of the total content of impurities a and B as measured by HPLC. In some embodiments, impurities a and B are not detected (or are below the detection limit) by HPLC. In some embodiments, the term "measuring by HPLC" refers to using the HPLC method shown in example 1, e.g., using 220nm as the detection wavelength.

Compound 1 can be prepared by various methods. In some embodiments, compound 1 for use in the synthetic methods described herein can be prepared by a process comprising reducing compound a9 or a salt thereof with hydrogen in the presence of a catalyst (e.g., a heterogeneous catalyst, such as palladium on carbon) in an ether solvent (e.g., tetrahydrofuran, t-butyl methyl ether, and the like):

it has been unexpectedly found that the reduction of the nitro group of a9 with hydrogen can be performed more efficiently in ether solvents such as Tetrahydrofuran (THF) rather than in more commonly used alcohol solvents such as ethanol. For example, the hydrogenation reaction can be carried out to completion (or near completion) in a shorter time, thereby improving yield. Various hydrogenation reaction conditions may be used. For example, various hydrogenation catalysts, such as heterogeneous catalysts, are suitable and may be selected by one of skill in the art in light of this disclosure. Other ethereal solvents similar to tetrahydrofuran may also be used. In light of the present disclosure, one skilled in the art can adjust the pressure of hydrogen and the reaction temperature, the concentration of compound a9 in the solvent. Exemplary hydrogenation conditions are shown in the examples section. Compound 1 prepared by the methods herein can also have a high purity, e.g., an HPLC purity of at least 85% (e.g., about 90%, about 95%, about 96%, about 97%, about 98%, about 98.5%, about 99%, or greater than 99%). In some embodiments, compound 1 may also be in a crystalline form, for example, in some embodiments, compound 1 may be recrystallized from a suitable solvent such as ethyl acetate and the like. In some embodiments, crystalline compound 1 may be used as a starting material for reacting with an acid of formula IV-acid or an activated form thereof, such as an acid chloride of formula IV, to form a compound of formula III under amide forming conditions.

Compound a9 can be prepared by various methods. In some embodiments, compound a9 for use in the synthetic methods described herein can be prepared by including in the presence of a base (e.g., a carbonate base, such as potassium carbonate) in a solvent (e.g., CH)₃CN) with compound A8 or a salt thereof:

various reaction conditions may be used. For example, various bases and amounts thereof can be selected by one skilled in the art in light of this disclosure. Typically, it has been found that an amount of base carbonate, e.g., potassium carbonate, of less than 3 equivalents, e.g., about 1.2 equivalents, can result in an increase in yield. Such as CH₃Solvents for CN may be used for this conversion. In light of this disclosure, one skilled in the art can adjust the reaction temperature, the concentration of compound a7 in the solvent, and the like. Exemplary reaction conditions are shown in the examples section.

Compound a7 can also be prepared by various suitable methods. In some embodiments, compound a7 for use in the synthetic methods described herein may be prepared by a process comprising reacting compound a5 with compound a6 or a salt thereof in the presence of a catalytic acid, such as p-toluenesulfonic acid (PTSA), in a solvent, such as isopropanol:

various reaction conditions for analogous metathesis reactions are known in the art and may be applied to the methods herein. It was found that only catalytic amounts of acid were required for the reaction. For example, as shown in the examples section, using as low as 0.2 equivalents of PTSA in the reaction of compounds a5 and a6, compound a7 is obtained. Typically, compound a7 is produced in the form of the hydrochloride salt, which can optionally be neutralized before the next step, e.g. before reaction with compound A8. Solvents such as isopropanol may be used for this conversion. In light of this disclosure, one skilled in the art can adjust the reaction temperature, the concentration of compound a5 in the solvent, and the like. Exemplary reaction conditions are shown in the examples section.

Compound a5 can also be prepared by various methods. Compound a5 for use in the synthetic methods described herein can be prepared by a method comprising reacting compound A3 with compound a4 in the presence of a base (e.g., a carbonate base, such as potassium carbonate) in a solvent (e.g., N-dimethylacetamide (DMAc)):

it has been found that the use of weak bases such as carbonate bases is sufficient to carry out the conversion and is more advantageous. For example, earlier methods for this conversion used strong bases (such as NaH), which limited their applicability in large scale preparation. In the present disclosure, it was found that similar yields were obtained using carbonate bases such as sodium carbonate as with NaH. Thus, the reactions of A3 and a4 to provide a5 can be advantageously carried out with carbonate bases, which is much easier for large scale synthesis than prior methods. The solvent for this conversion with carbonate can generally be an aprotic polar solvent, such as DMAc. In light of this disclosure, one skilled in the art can adjust the reaction temperature, the concentration of compound a3 in the solvent, and the like. Exemplary reaction conditions are shown in the examples section.

Compound a3 can also be prepared by various methods. In some embodiments, compound A3 for use in the synthetic methods described herein can be prepared by a method comprising reacting compound a1 with an indole in the presence of a base (e.g., MeMgBr) in a solvent (e.g., 2-methyltetrahydrofuran (2-MeTHF)):

it was found that the Lewis acid catalyzed Friedel-Crafts reaction of A1 with indole also did not work. In contrast, if a base, such as MeMgBr, is used, typically in excess of 1 equivalent, e.g., about 2 equivalents, the reaction proceeds smoothly with high yield. The solvent used for this transformation may typically be an ether solvent, such as 2-MeTHF. In light of this disclosure, one skilled in the art can adjust the reaction temperature, the concentration of compound a1 in the solvent, and the like. Exemplary reaction conditions are shown in the examples section.

In some embodiments, a method of preparing compound 4, or a pharmaceutically acceptable salt thereof, may comprise a reaction scheme according to:

suitable conditions and/or reagents for transformation in the protocol include any of those described herein. Exemplary conditions are described in the examples section.

In some embodiments, the present disclosure also provides compound 4, or a pharmaceutically acceptable salt thereof, produced by any of the methods herein.

It will be apparent to those skilled in the art that conventional protecting groups may be necessary to prevent certain functional groups from undergoing undesirable reactions. Suitable protecting groups for various functional groups and suitable conditions for protecting and deprotecting particular functional groups are well known in the art. For example, in "protecting group in organic synthesis" of 4 th edition, p.g.m.wuts; numerous protecting groups are described in t.w. greene, John Wiley, 2007, and the references cited therein. The reagents for the reactions described herein are generally known compounds or may be prepared by known methods or obvious modifications thereof. For example, many reagents are available from commercial suppliers, such as Aldrich Chemical Co (Milwaukee, Wis.), Sigma (St. Louis, Mo.). Other reagents can be prepared by procedures described in standard references or obvious modifications thereof, such as organic synthetic reagents of Fieser and Fieser, volumes 1-15 (John Wiley and Sons, 1991), Rodd's "carbon compound chemistry", volumes 1-5 and supplementations (Ex. Weissel scientific Press, 1989), organic reactions, volumes 1-40 (John Wiley and Sons, 1991), 3 months of "advanced organic chemistry" (7 th edition, Weili), and Larock's "comprehensive organic transformation" (Wiley-VCH, 1999), and any available updates in this document.

Pharmaceutical composition

In some embodiments, the present invention provides a pharmaceutical composition comprising one or more compounds described herein (e.g., substantially pure compound 4 or a pharmaceutically acceptable salt thereof, such as compound 5). Typically, the pharmaceutical composition comprises a therapeutically effective amount of one or more compounds described herein (e.g., substantially pure compound 4 or a pharmaceutically acceptable salt thereof, such as compound 5) and optionally a pharmaceutically acceptable excipient or carrier. In some embodiments, the pharmaceutical composition comprises substantially pure compound 4, or a pharmaceutically acceptable salt thereof, as described herein. In some embodiments, the pharmaceutical composition comprises compound 4, or a pharmaceutically acceptable salt thereof, produced by any of the synthetic methods herein. In some embodiments, the pharmaceutical composition comprises substantially pure compound 5 described herein. In some embodiments, the pharmaceutical composition comprises compound 5 in crystalline form I and/or amorphous form. The pharmaceutical compositions may be formulated for any route of administration, for example oral administration.

Certain particular embodiments of the present invention relate to pharmaceutical compositions comprising a therapeutically effective amount of substantially pure compound 4, or a pharmaceutically acceptable salt thereof, e.g., as described herein, and optionally a pharmaceutically acceptable excipient or carrier. In some embodiments, a pharmaceutical composition comprising substantially pure compound 4 or a pharmaceutically acceptable salt thereof, e.g., as described herein, can be formulated for oral, parenteral, nasal, pulmonary, buccal, topical, or transdermal administration. In some embodiments, a pharmaceutical composition comprising substantially pure compound 4 or a pharmaceutically acceptable salt thereof, e.g., as described herein, can be formulated as a solid dosage form. In some embodiments, the solid dosage form is an oral solid dosage form. In some embodiments, the solid dosage form is a capsule or tablet. In some embodiments, the solid dosage form is enterically coated. However, in some embodiments, the solid dosage form is not enterically coated.

Certain embodiments of the present invention relate to pharmaceutical compositions comprising a therapeutically effective amount of compound 5, and optionally a pharmaceutically acceptable excipient or carrier. In some embodiments, a pharmaceutical composition comprising compound 5 can be formulated for oral, parenteral, nasal, pulmonary, buccal, topical, or transdermal administration. In some embodiments, a pharmaceutical composition comprising compound 5 can be formulated as a solid dosage form. In some embodiments, the solid dosage form is an oral solid dosage form. In some embodiments, the solid dosage form is a capsule or tablet. In some embodiments, the solid dosage form is enterically coated. However, in some embodiments, the solid dosage form is not enterically coated. In some embodiments, the pharmaceutical composition comprises substantially pure compound 5 described herein. In some embodiments, compound 5 is present in crystalline form I. In some embodiments, the pharmaceutical composition is free or substantially free of any solid form of compound 5 other than form I. For example, in some embodiments, no solid form of compound 5 is detectable in the pharmaceutical composition other than form I. In some embodiments, form I of compound 5 in the pharmaceutical composition can be characterized by its particle size distribution, such as those described herein. For example, in some embodiments, form I of compound 5 in the pharmaceutical composition is characterized by a particle size distribution of D90: about 80 μm to about 120 μm; and D50: about 30 μm to about 60 μm. Other suitable particle size distributions for form I of compound 5 are described herein. In some embodiments, the pharmaceutical composition comprises compound 5 in a mixture of form I and amorphous form.

Pharmaceutical compositions comprising compound 5 are generally storage stable. For example, in one embodiment, the pharmaceutical composition contains substantially the same amount of compound 5 as the HPLC assay, and a similar level (e.g., no increase) of impurities or degradants as the HPLC assay, and has substantially the same dissolution profile as the dissolution method described herein, when tested for stability by storage in an environment of 40 ℃, 75% relative humidity for up to 6 months. In general, the solid dosage forms herein can be formulated as immediate release formulations, e.g., when using the general rule 40931 of method 2 in the 2015 edition "chinese pharmacopoeia" and a paddle rotation speed of 50rpm, at least 70% (e.g., at least 80%, at least 85%, at least 90% or substantially all) of compound 5 is released within 30 minutes.

The pharmaceutical compositions herein can comprise compound 5 in various amounts, e.g., compound 5 in an amount effective to treat a disease or disorder described herein, e.g., cancer (e.g., non-small cell lung cancer). In some embodiments, the active ingredient in the pharmaceutical composition can consist essentially of compound 5 or consist of compound 5. Typically, compound 5 can be included in an amount of about 5% to about 25% by weight of the pharmaceutical composition.

In a typical dosage form, a pharmaceutically acceptable salt of compound 4 (e.g., as described herein) can be used as the Active Pharmaceutical Ingredient (API). However, in some embodiments, the API of the dosage form can include compound 4 (e.g., as described herein) as the free base. In some embodiments, the API can comprise two or more different pharmaceutically acceptable salts of compound 4 (e.g., as described herein). In some embodiments, the API can include compound 4 (e.g., as described herein) as a free base in a mixture with one or more pharmaceutically acceptable salts of compound 4 (e.g., as described herein).

Various excipients or carriers may be included in the pharmaceutical compositions described herein. Generally, the pharmaceutical compositions herein may comprise one or more excipients or carriers selected from fillers (e.g., lactose, microcrystalline cellulose, mannitol, and the like), disintegrants (e.g., croscarmellose sodium), glidants (e.g., colloidal silicon dioxide), lubricants (e.g., sodium stearyl fumarate), antioxidants, stabilizers, preservatives, diluents, solvents, sweeteners, viscosity increasing agents, chelating agents, coloring agents, surfactants, flavoring agents, coatings, gelling agents, binders, and mold release agents. One skilled in the art knows that other excipients/carriers may also be used, and knows how to select the appropriate excipient/carrier when formulating the compounds herein according to the intended use. In some particular embodiments, the pharmaceutical composition comprises one or more (e.g., 1,2, 3, 4, 5, or 6) of: lactose, microcrystalline cellulose, mannitol, croscarmellose sodium, colloidal silicon dioxide and sodium stearyl fumarate. Any suitable amount of such excipients and carriers may be used. The amount of excipient and/or carrier can also be adjusted, for example, to achieve the desired immediate release dissolution profile described herein. In some embodiments, the excipients and carriers are used in amounts equal to or below the upper limit of excipients or carriers that the U.S. food and drug administration or other corresponding regulatory authority determines to be safe for human use. Other suitable examples of excipients or carriers can be found in "remington pharmaceutical science" by Mack pub. Science and practice of pharmacy, Lippincott Williams & Wilkins, philadelphia, 20 th edition (2003) and 21 th edition (2005), the contents of which are incorporated herein by reference in their entirety.

Method of treatment

The compounds and pharmaceutical compositions described herein are useful for treating various diseases and disorders. As described in U.S. publication No. 2017/0355696a1, compound 4 and pharmaceutically acceptable salts thereof can be potent and selective inhibitors against EGFR variants or mutants, such as activating or resistant mutant forms of EGFR, e.g., L858R mutants, Exon19 deletion mutants, and/or T790M mutants.

In some embodiments, methods of treating a disease or disorder in a subject in need thereof are provided. In some embodiments, the method comprises administering to the subject substantially pure compound 4 or a pharmaceutically acceptable salt thereof, e.g., as described herein. In some embodiments, the method comprises administering to the subject compound 4, or a pharmaceutically acceptable salt thereof, produced by any of the synthetic methods herein. In some embodiments, the method comprises administering compound 5 to the subject. In some embodiments, the disease or disorder is mediated by EGFR (e.g., an activated or resistant mutant form of EGFR). In some embodiments, the disease or disorder is mediated by an L858R mutant, an Exon19 deletion mutant, and/or a T790M mutant. In some embodiments, the disease or disorder can be, but is not limited to, ovarian cancer, cervical cancer, colorectal cancer (e.g., colon adenocarcinoma), breast cancer, pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer, leukemia, lymphoma, non-hodgkin lymphoma, gastric cancer, lung cancer (e.g., non-small cell lung cancer), liver cancer, gastrointestinal stromal tumor (GIST), thyroid cancer, bile duct cancer, endometrial cancer, renal cancer, anaplastic large cell lymphoma, Acute Myelogenous Leukemia (AML), multiple myeloma, and/or mesothelioma.

In some embodiments, the method is for treating cancer. In some embodiments, the method is for treating ovarian cancer, cervical cancer, colorectal cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer, leukemia, lymphoma, non-hodgkin lymphoma, gastric cancer, lung cancer, liver cancer, gastrointestinal stromal tumor, thyroid cancer, bile duct cancer, endometrial cancer, renal cancer, anaplastic large cell lymphoma, acute myelogenous leukemia, multiple myeloma, mesothelioma, or any combination thereof. In some embodiments, the method is for treating lung cancer. In some embodiments, the cancer is non-small cell lung cancer.

In any of the methods described herein, the subject may be resistant to one or more EGFR inhibitors other than compound 4 or a pharmaceutically acceptable salt thereof (e.g., compound 5). In some embodiments, the subject may be resistant to one or more EGFR inhibitors selected from gefitinib, erlotinib and inctinib. In some embodiments, the subject may experience the side effects of wild-type EGFR inhibition.

In general, the methods described herein comprise administering to the subject a therapeutically effective amount of compound 4 or a pharmaceutically acceptable salt thereof, e.g., compound 5 or a pharmaceutical composition described herein. In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising substantially pure compound 4 or a pharmaceutically acceptable salt thereof, e.g., as described herein. In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising compound 4, or a pharmaceutically acceptable salt thereof, produced by any of the synthetic methods herein. The compounds and pharmaceutical compositions can be administered to a subject by any route of administration. For example, in some embodiments, the compounds and pharmaceutical compositions can be administered orally to a subject. In some specific embodiments, the method comprises orally administering to the subject a therapeutically effective amount of compound 5.

In any of the embodiments described herein, compound 4, e.g., substantially pure compound 4, or a pharmaceutically acceptable salt thereof, e.g., compound 5, can be used in a monotherapy. For example, in some embodiments, the active agent for use in the methods or pharmaceutical compositions herein can consist of or consist essentially of compound 4 or a pharmaceutically acceptable salt thereof, e.g., substantially pure compound 4 or a pharmaceutically acceptable salt thereof, e.g., compound 5, can be in crystalline form I, amorphous form, or a combination thereof. The methods described herein may also be used in combination with other therapies. For example, the methods herein may be used in combination with one or more selected from the group consisting of surgical procedures (e.g., conventional anti-cancer surgical treatments), radiation therapy, chemotherapy, and anti-tumor immunotherapy.

In some embodiments, the method can include, for example, orally administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising substantially pure compound 4 or a pharmaceutically acceptable salt thereof, e.g., concurrently, sequentially or separately administered with chemotherapy or anti-tumor immunotherapy, as described herein. In some embodiments, the method can include, e.g., orally administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising compound 4 or a pharmaceutically acceptable salt thereof produced by any of the synthetic methods herein, e.g., as described herein, in parallel, simultaneously, sequentially or separately with chemotherapy or anti-tumor immunotherapy. In some embodiments, the method can comprise, e.g., orally administering to the subject a therapeutically effective amount of compound 5, simultaneously, sequentially or separately, in parallel with the chemotherapy, anti-tumor immunotherapy. Chemotherapy or immunotherapy includes, but is not limited to, one or more antineoplastic agents of the following types: alkylating agents (e.g., carboplatin, oxaliplatin, cisplatin, cyclophosphamide, nitrosoureas, nitrogen mustards, melphalan), antimetabolites (e.g., gemcitabine) and antifolates (e.g., 5-fluorouracil and tegafur, raltitrexed, methotrexate, cytarabine, hydroxyurea), topoisomerase inhibitors (e.g., etoposide, topotecan, camptothecin), antimitotics (e.g., vincristine, vinblastine, vinorelbine, paclitaxel), antitumor antibiotics (e.g., doxorubicin, bleomycin, doxorubicin, daunomycin, mitomycin C, actinomycin), antiestrogens (e.g., tamoxifen, fulvestrant, toremifene, raloxifene, droloxifene), antiandrogenic drugs (e.g., bicalutamide, flutamide, nilutamide), LHRH antagonists or LHRH agonists (e.g., goserelin, leuprolide, and brenserin), aromatase inhibitors (e.g., anastrozole, letrozole), CYP17 lyase inhibitors (e.g., abiraterone), the anti-erbB 2 antibody trastuzumab [ Herceptin ], the anti-EGFR antibody cetuximab [ Erbitux ]; tyrosine kinases, serine/threonine kinase inhibitors (e.g., imatinib, nilotinib, sorafenib, trametinib, crizotinib); cyclin-dependent kinase inhibitors (e.g., CDK4 inhibitor, palbociclib), bevacizumab anti-human vascular endothelial growth factor antibody (Avastin) and VEGF receptor tyrosine kinase inhibitors (apatinib); anti-tumor immunotherapy, such as anti-PD-1 antibody (pembrolizumab, nivolumab), anti-PD-L1 antibody, anti-LAG-3 antibody, anti-CTLA-4 antibody, anti-4-1 BB antibody, anti-GITR antibody, anti-ICOS antibody, interleukin 2.

Definition of

As used herein, the term "about" modifying amounts relevant to the present invention refers to a change in numerical value that may occur, for example, by conventional testing and handling; pass inadvertent errors in such testing and handling; by differences in the manufacture, source or purity of the ingredients used in the present invention; and so on. As used herein, "about" certain values also include the certain value, e.g., about 10% includes 10%. The claims, whether or not modified by the term "about," include equivalents to the amounts recited. In one embodiment, the term "about" means within 20% of the reported numerical value.

For the purpose of identifying a certain impurity of compound 4 (or a pharmaceutically acceptable salt thereof), the impurity can be described using Relative Retention Time (RRT). Retention Time (RT) measured by specific impurities_Impurities) (e.g., in minutes) divided by the Retention Time (RT) measured for Compound 4_{Compound 4}) (e.g., in minutes), the relative retention time of a particular impurity in a particular HPLC method (e.g., example 1) can be determined, i.e., according to the following equation: RRT ═ RT_Impurities/RT_{Compound 4}. Thus, impurities having an RRT < 1 (e.g., 0.88) elute (e.g., from an HPLC column) before compound 4 and impurities having an RRT > 1 (e.g., 1.04) elute after compound 4. One skilled in the art will appreciate that the retention time of compound 4 or a pharmaceutically acceptable salt thereof will be the same using the HPLC method described in example 1, regardless of the retention time of the counterion. Similarly, when using the HPLC method described in example 1, an impurity as defined herein, e.g. impurity a or B, whether present in basic or protonated form, will have the same retention time. For purposes of this disclosure, impurities a or B include any form that may be present in a test sample.

The terms "purity" and "impurity" are used according to their respective art-recognized meanings. The term "purity by HPLC area", "purity by HPLC" or "purity as measured by HPLC" refers to the purity of the corresponding compound as measured using an HPLC method, such as the HPLC method described in example 1, expressed as percentage of HPLC area. For the purposes of this disclosure, the purity of a compound herein as measured by the HPLC method described in example 1 can be expressed as the percentage of the area of the peak representing the compound using 220nm or 254nm as the detection wavelength. For example, in some embodiments, unless otherwise specified or contrary to the context, when a compound herein has a purity of about 95% as determined by HPLC, this means that when measured by the HPLC method described in example 1, using one or both of 220nm and 254nm as the detection wavelength, the peak representing the compound has an area percentage of about 95%. In any of the embodiments described herein, unless otherwise indicated or contrary to the context, the HPLC purity of a compound can be determined by the HPLC method described in example 1 and expressed as the area percentage of the peak representing the compound in the HPLC curve using 220nm as the detection method. The relative amounts of impurities should be understood similarly. See HPLC in fig. 4A, 4B, 5A and 5B for purity examples. For example, in fig. 4A, the HPLC purity of compound 4 was 97.5% (peak area with retention time of 10.976, representing compound 4), the content of impurity a was 1.26% (peak area with retention time of 11.638, representing impurity a), and the content of impurity B was 0.23% (peak area with retention time of 9.680, representing impurity B). The term "purity" as used herein, unless specifically referring to weight purity, should not be construed to mean purity by weight, such as purity measured by HPLC or by HPLC, or similar terms, although the weight percent purity of a test sample can also be determined by HPLC methods.

As used herein, the terms "treat", "treating", "treatment", and the like refer to the elimination, alleviation, or amelioration of a disease or disorder and/or symptoms associated therewith. Although not excluded, treating a disease or condition does not require complete elimination of the disease, condition or symptom associated therewith. As used herein, the terms "treat, treating," and the like, can include "prophylactic treatment," which refers to reducing the likelihood of recurrence of a disease or disorder or of prior control of a disease or disorder in a subject who is not, but at risk of or susceptible to recurrence of a disease or disorder. The terms "treatment" and synonyms contemplate administration of a therapeutically effective amount of a compound described herein to a subject in need of such treatment.

The term "leaving group" has its usual meaning in the field of synthetic organic chemistry, and may for example refer to an atom or a group capable of being substituted by a nucleophile. See, for example, Smith, March Advanced Organic Chemistry 6 th edition (501-502). Examples of suitable leaving groups include, but are not limited to, halogen (e.g., F, Cl, Br or I (iodo)), alkoxycarbonyloxy, aryloxycarbonyloxy, alkylsulfonyloxy, arenesulfonyloxy, alkyl-carbonyloxy (e.g., acetoxy), arylcarbonyloxy, aryloxy, methoxy, N, O-dimethylhydroxylamine, 9-phenylxanthine-9-yl (pixyl), and haloformates. In some embodiments, the hydroxy or alkoxy group may also be a leaving group herein.

The term "pharmaceutically acceptable salts" refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic response, and the like, and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art.

As used herein, the term "therapeutically effective amount" refers to an amount of a therapeutic agent (e.g., compound 4 or 5) sufficient to alleviate one or more disorders or conditions (e.g., lung cancer), or prevent the appearance or development of a disease or disorder, or cause the regression or cure of a disease or disorder.

As used herein, the term "subject" (alternatively referred to herein as "patient") refers to an animal, preferably a mammal, most preferably a human, who has been the subject of treatment, observation or experiment. In any of the embodiments described herein, the subject may be a human.

Examples

Example 1 general procedure

Materials: starting materials, reagents, solvents, and the like are generally commercially available.

Powder X-ray diffraction (XRPD): the solid samples were examined using an X-ray diffractometer (Bruker D8 advance). The system is equipped with a LynxEye detector. The sample was scanned at an angle of 3 to 40 degrees 2 theta with a step size of 0.02 degrees 2 theta. The tube voltage and current were 40KV and 40mA, respectively. The sample was transferred from the sample container to a zero background XRPD holder and lightly grounded.

TGA analysis: TGA analysis was performed on a TA Instruments TGA Q500. The samples were placed in decoked platinum or aluminum pans, automatically weighed, and then inserted into a TGA furnace. The sample was heated to the final temperature at a rate of 10 ℃/min. The purging gas is nitrogen, the equilibrium speed is 40mL/min, and the sample detection time is 60 mL/min.

DSC analysis: DSC analysis was performed on TA Instruments Q200. The calibration standard is indium. The weight samples were placed in a TA DSC pan and the weight was accurately recorded. The coil edge was compressed for analysis and the sample was heated to the final temperature under nitrogen (50mL/min) at a rate of 10 deg.C/min.

Particle size analysis:

the particle size distribution was measured by a laser particle size analyzer.

HPLC analysis: representative HPLC methods are shown below, which can be used, for example, to analyze the purity of the compounds herein.

The instrument comprises the following steps: agilent

Flow rate: 1.0mL/min

Mobile phase: a: 0.1% TFA in water

B：CH₃CN

Sample introduction amount: 5 μ L

A chromatographic column: agilent Zorbax Box RP, 3.5um, 4.6 x 150mm

Column temperature: 25 deg.C

And (3) detection: 220 or 254nm

Operating time: 30 minutes

Gradient (T/B%): 0.0/10, 15.0/40, 20.0/90, 30.0/90, 31.0/10 and 35.0/10

Example 2 preparation of (N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (1- (2,2, 2-trifluoroethyl) -1H-indol-3-yl) pyrimidin-2-yl) amino) phenyl) acrylamide) monomethanesulfonate

Compound 1 can be prepared according to the same method as described in U.S. publication No. 2017/0355696a1, published 12/14/2017, which is incorporated herein by reference in its entirety. In U.S. publication No. 2017/0355696a1, compound 1 was prepared by reduction of the corresponding nitro precursor using iron/ammonium chloride. It has been found that the conversion can also be carried out under hydrogen using palladium on carbon.

Compound 1(1 equivalent) was mixed with tetrahydrofuran (about 10mL/g compound 1) and water (about 1mL/g compound 1) under nitrogen. The mixture was cooled to-10 to-5 ℃. Compound 2(1.2 equivalents) was then added to the mixture, with the temperature controlled between-10 and 0 ℃ during the addition. Thereafter, the mixture was stirred at this temperature for about 30 minutes. Then, potassium hydroxide (3.8 equivalents) was added in portions to the mixture, and the temperature was controlled below 20 ℃ during the addition of potassium hydroxide. Thereafter, the reaction mixture is heated to 55-65 ℃ and held at that temperature for about 16 hours, and then cooled to 25 ℃ or less. After work-up and purification, compound 4 was obtained with an HPLC purity of greater than 98.5%. Alternatively, compound 4 can be dissolved in isopropanol/water (5:1, total volume of about 8mL/g compound 4) at 75-85 deg.C, then slowly cooled to 40-50 deg.C, stirred and held for 1-2 hours, then slowly cooled to 10-20 deg.C, stirred for 2 hours. The precipitated solid was then collected, washed and dried to provide compound 4 with an HPLC purity of greater than 98.5%.

Analytical data for compound 3: LCMS (ESI, m/z): [ M + H ]]⁺＝604.0；m/z。HNMR(400MHz，DMSO-d₆，ppm)：10.39(brs，1H)，9.83(s，1H)，8.63(s，1H)，8.58(s，1H)，8.37(d，J＝5.6Hz，1H)，8.35(m，1H)，8.06(s，1H)，7.72(d，J＝8.0Hz，1H)，7.29(m，1H)，7.26(d，J＝5.6Hz，1H)，7.26(m，1H)，6.95(s，1H)，5.34(q，J＝9.2Hz，2H)，3.90(t，J＝6.4Hz，2H)，3.88(s，3H)，3.31(m，2H)，3.28(m，2H)，3.11(t，J＝6.4Hz，2H)，2.75(s，6H)，2.63(s，3H)。CNMR(100MHz，DMSO-d₆，ppm)：168.90，161.68，160.59，158.38，147.62，139.42，138.02，133.39，125.79，125.04，124.88，123.30，122.33，122.03，117.80，115.07，111.28，107.85，10468 56.51、53.95、49.30、46.98、43.47、42.65、41.59、39.46。FNMR(376MHz，DMSO-d₆，ppm)：-69.92(3F)。

Compound 4(1 eq) was then dissolved in ethanol and ethyl acetate (1: 1, combined volume about 10mL/g of compound 4). The mixture was heated to 55-65 ℃ and methanesulfonic acid (0.95 eq) was added under nitrogen. The reaction is held at this temperature for about 20-30 minutes. The reaction mixture is then slowly cooled to provide the mesylate compound 5 of form I, which serves as a seed crystal. Representative of Compound 5¹H NMR is shown in figure 1.

Such cooling can be carried out in the presence of seed crystals, for example, after the seed crystals are added to the reaction mixture (e.g., about 3% to about 15% w/w). The seeded mixture may be slowly cooled to 45-55 deg.C, stirred and held for about 2 hours, then slowly cooled to 35-45 deg.C, stirred and held for 1-2 hours, then slowly cooled to 25-35 deg.C, stirred and held for 1-2 hours, and finally slowly cooled to 15-25 deg.C and stirred for at least 2 hours. The resulting solid was then filtered and collected to provide pure compound 5. In one embodiment, 3 wt.% of seed crystals are added to the reaction mixture prior to the gradual cooling step. The resulting crystals were analyzed for particle size distribution as described herein. The D90 for this batch of crystals was found to be: about 82.4 μm; d50: about 34.3 μm; and D10 was about 11.4 μm.

Compound 5 can also be prepared by adding 25 to 50 mole% methanesulfonic acid to a solution of compound 4 in ethyl acetate and ethanol and seed crystals (e.g., about 3% to about 15% w/w) at 50-60 ℃ under nitrogen. The reaction was held at this temperature for about 30 minutes and the remaining 50 to 75 mol% of methanesulfonic acid was added. The mixture with the seed crystals may be slowly cooled to 45-55 deg.C, stirred and held for about 2-3 hours, then slowly cooled to 40-50 deg.C, stirred and held for 1-2 hours, then slowly cooled to 30-40 deg.C, stirred and held for 1-2 hours, and finally slowly cooled to 20-30 deg.C and stirred for about 2-32 hours. The resulting solid was then filtered and collected to provide pure compound 5. The solid can be further treated in isopropanol and water, stirred at 55-65 deg.C for 10-12 hours, slowly cooled to 40-50 deg.C, stirred and held for 1-2 hours, then slowly cooled to 30-40 deg.C, stirred and held for 1-2 hours, then slowly cooled to 20-30 deg.C, and stirred for 1-2 hours.

Typically, compound 5 obtained by this method has an HPLC purity of greater than 98.5%.

Example 2 A.alternative preparation of (N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (1- (2,2, 2-trifluoroethyl) -1H-indol-3-yl) pyrimidin-2-yl) amino) phenyl) acrylamide) monomethanesulfonate

Compound A1(1 equivalent) was mixed with 2-methyltetrahydrofuran (about 10mL/g Compound A1) under nitrogen. The mixture was cooled to-5 to 5 ℃. MeMgBr (0.85 eq) was added to the mixture and the temperature was controlled between-5 and 5 ℃. Thereafter, compound A2(1.1 equiv.) was added between-5 and 5 ℃. After stirring for 1 hour, the mixture was heated to 78 to 83 ℃ and stirred for another 1 hour. MeMgBr (0.44 eq) was added to the mixture, the temperature was controlled between 78 and 83 ℃ and the mixture was stirred for a further 0.5-1 hour. Two portions of MeMgBr (0.44 equivalents per portion) were added to the mixture following a similar procedure as above. The mixture was heated at 78 to 83 ℃ for 19-21 hours under nitrogen and then cooled to 0-5 ℃. Aqueous HOAc (12%, 3.3 equivalents) was added to quench the reaction and the temperature was controlled below 40 ℃. The mixture was separated and the aqueous phase was extracted with EtOAc (2g/g Compound A1). The combined organic layers were washed with brine (5g/g of Compound A1) and then concentrated at 45-50 ℃ to a volume of 2-3mL/g of Compound 1. EtOH (4g/g Compound A1) was added to the residue and the mixture was concentrated to a volume of 3-4mL/g Compound A1. The residue was heated at 70-75 ℃ for 1 hour and then cooled to 20-25 ℃. Water (3g/g of Compound A1) was added and the mixture was stirred for 2 hours. Filtering, and drying the filter cake at 60-65 ℃ under vacuum to obtain the compound A3.

Compound A3 was mixed with DMAc (10mL/g Compound A3) under nitrogen. Potassium carbonate (1.5 equiv.) is added and the mixture is stirred at 15-20 ℃ for 1.5-2 hours. Compound a4(1.15 eq) was added and the mixture was heated at 40-45 ℃ for 16-20 hours. Water (10g/g Compound A3) was added to control the temperature below 20 ℃. The mixture was cooled to 0-5 ℃ and stirred for 1.5-2 hours. The mixture was filtered and the filter cake was washed with water and EtOAc respectively and dried under vacuum at 60-65 ℃ to give compound a 5.

Compound A5 was mixed with Compound A6(1.1 equiv.), p-TSA (0.2 equiv.) and isopropanol (9g/g Compound A5) under nitrogen. The mixture was heated at 77-87 ℃ for 16-20 hours and then cooled to 30-40 ℃. Filtration, washing of the filter cake with isopropanol and water respectively, and drying at 60-65 ℃ under vacuum gave compound a7 hydrochloride. The hydrochloride was dissolved in DMF (11mL/g hydrochloride) and Et was added₃N (1.2 eq), followed by the addition of water (11mL/g hydrochloride). The mixture was stirred at 25-30 ℃ for 1-2 hours and filtered. The filter cake was washed with water and dried under vacuum at 60-65 ℃ to give compound a 7.

Compound a7 was mixed with potassium carbonate (1.3 eq) and MeCN (10mL/g compound a7) under nitrogen. Compound A8(1.4 equivalents) was added to the mixture and the temperature was controlled at 15-30 ℃. The mixture was heated at 77-87 ℃ for 2-4 hours, then water (15mL/g Compound A7) was added. The mixture was stirred at 10-20 ℃ for 2-4 hours and filtered. The filter cake was washed with water and dried under vacuum at 45-50 ℃ to give compound a 9.

Compound A9 was combined under nitrogen with palladium on carbon (0.1g/g Compound A9) and tetrahydrofuran (10mL/g Compound A9). The mixture was changed from nitrogen to hydrogen atmosphere and heated at 30-40 ℃. The mixture was stirred under a hydrogen atmosphere (0.1-0.2MPa) for 9-11 hours, then changed to a nitrogen atmosphere, cooled to 15-25 ℃ and filtered through Celite (Celite). For the filtrate, the solvent was converted from tetrahydrofuran to EtOAc by distillation with an additional amount of EtOAc (3X 10mL/g Compound 9). The mixture was heated at 50-60 ℃ to a clear solution, then slowly cooled to 0-10 ℃ and a solid precipitated. The suspension is stirred at 0-10 ℃ for 4-5 hours and then filtered. The filter cake was washed with EtOAc and dried under vacuum at 40-45 ℃ to give compound 1.

Compound 1(1 equivalent) was mixed with tetrahydrofuran (about 10mL/g of compound 1) and water (about 1mL/g of compound 1) under nitrogen. The mixture was cooled to-10 to-5 ℃ and the temperature was controlled at-10 to 0 ℃ while adding compound 2(1.2 equivalents). Thereafter, the mixture was stirred at-10 to 0 ℃ for about 30 minutes. The reaction was sampled for HPLC to ensure complete conversion of compound 1 (compound 1. ltoreq.0.5%). Potassium hydroxide (4.8 equivalents) was then added to the mixture, and the temperature was controlled below 20 ℃ during the addition. Thereafter, the reaction mixture was heated to 55-65 ℃ and held at that temperature for about 16 hours. Samples were taken from the reaction and subjected to HPLC, and the end of the reaction (Compound 3. ltoreq. 0.2%) was ensured before cooling to 15-25 ℃. Water (9g/g Compound 1) was added and the mixture was separated. The organic layer was diluted with EtOAc (10mL/g of Compound 1) and washed twice with brine and water, respectively. The solvent was converted from EtOAc to isopropanol by distillation with an additional amount of isopropanol (2x12g/g compound 1). Water (1.4g/g Compound 1) was added and the mixture was heated at 75-85 ℃ to a clear solution and then slowly cooled to 10-20 ℃ to precipitate a solid. The suspension was stirred at 10-20 ℃ for 2-3 hours and filtered. The filter cake was washed with isopropanol/water (5: 1) and dried under vacuum at 45-50 ℃ to give compound 4. It was found that by using an additional crystallization process, the purity of compound 4 could be further improved, based on the area% calculated for 220nm detection, with the reduction of impurity a (relative retention time, about 1.02) and impurity B (relative retention time, about 0.88) to below 0.2% as tested by the HPLC method in example 1. See fig. 4A (before crystallization in IPA/water) and fig. 4B (after crystallization in IPA/water). The retention time for compound 4 was 10.976 minutes in fig. 4A and 10.452 minutes in fig. 4B. The total yield from aniline 1 to compound 4 is about 70%.

Compound 4(1 eq) was dissolved in ethanol and ethyl acetate (1: 1, combined volume about 12mL/g compound 4). The mixture was heated to 55-60 ℃ and methanesulfonic acid (0.24 eq) was added under nitrogen. After the seed crystals are added to the reaction mixture (e.g., about 3% to about 15% w/w), additional methanesulfonic acid (0.71 equiv.) is added at 55-60 ℃. The mixture was stirred at 55-60 ℃ for 2-3 hours, then slowly cooled to 45-55 ℃, stirred and held for about 2 hours. The mixture was then slowly cooled to 35-45 ℃, stirred and held for 1-2 hours. The mixture was then slowly cooled to 25-35 ℃, stirred and held for 1-2 hours. The mixture was then slowly cooled to 15-25 ℃ and stirred for at least 2 hours. The mixture was then filtered and the solids collected to give wet compound 5. Wet Compound 5 was slurried in a mixed solvent of isopropanol and water (7:0.4, 7.4mL/g Compound 4) and the suspension was wet-milled to control particle size (D50: 30-60 μm, D90: 80-120 μm). After milling, the suspension is filtered and the filter cake is dried under vacuum at 40-50 ℃ to give compound 5. The HPLC purity of compound 5 obtained was greater than 99% (HPLC method of example 1, area% based on 220nm detection). Note that when impurity B was not removed in the process of preparing compound 4, the formation of the methanesulfonate salt did not remove the impurity, as shown in fig. 5A and 5B. Figure 5B is an HPLC profile obtained by formation of a salt of compound 4, which was further purified by recrystallization in IPA/water as described above; figure 5A is an HPLC profile obtained by forming a salt of compound 4 without further purification by recrystallization in IPA/water as described above. As understood by the person skilled in the art, for the determination of HPLC purity, it is not important whether the impurity a or B is present in salt form after the salt formation step, as under the HPLC conditions of the method of example 1, the protonation state of the impurity a or B in the sample is not important for the retention time observed in HPLC. The retention time of compound 5 was 11.079 minutes in fig. 5A and 10.478 minutes in fig. 5B.

EXAMPLE 3 solid State analysis of Compound 5 form I

The crystal obtained from example 2 was analyzed by XRPD and DSC and designated as form I. A representative XRPD spectrum is shown in fig. 2A. (see also Table 1 below for a list of peaks of relative intensity). A representative DSC profile is shown in figure 2B.

TABLE 1 XRPD peaks

Angle of rotation	Strength of	Angle of rotation	Strength of	Angle of rotation	Strength of
						2-θ/°	％	2-θ/°	％	2-θ/°	％
6.134	10.2	18.903	32.2	27.337	7
						6.565	9.9	19.212	31.1	27.756	17.7
7.802	100	19.476	11.5	28.17	3.4
						8.476	9.9	19.776	41.9	28.583	7.9
10.189	26.9	20.572	25.3	29.294	3.6
						10.773	37.2	20.85	17.3	30.313	3.3
11.239	18.9	21.13	14.9	30.62	2.5
						11.871	8.5	21.356	9.9	31.046	5
12.119	10	22.17	6.4	31.613	3.6
						13.213	8.7	22.591	15.3	31.967	4.3
13.605	23.5	22.873	7.3	33.189	5.3
						13.883	28.8	23.267	5.9	33.521	4.1
14.628	6.4	23.463	11.5	34.088	3.9
						14.977	12.5	23.684	36.2	35.375	4.1
15.712	9	23.924	54	36.025	3.1
						16.053	9.2	24.453	7.5	37.588	2.5
16.409	34.7	24.919	19.4	38.066	2.1
						17.729	65	25.314	7.2	38.748	3.1
18.376	27.2	25.902	11.5	39.249	2.5
						18.651	18.7	26.541	11.4
18.903	32.2	26.97	4.3

It was also found that form I can be converted to amorphous form after heat treatment by heating the crystals in the DSC cell to 220 ℃ and then immediately placing them in ice ("quenching"). See, for example, fig. 3. In addition, when form I was mechanically milled, some loss of crystallinity was observed for form I, although no new form was found.

The TGA curve shows a weight loss of 0.1935% before decomposition. The DSC curve shows that an endothermic peak appears due to melting, with an onset and maximum temperature of 208.64 ℃ and 210.12 ℃ respectively, and an enthalpy of 96.87J/g.

Form I was also found to be non-hygroscopic. Only a negligible weight increase was observed upon storage at 25 ℃ with a relative humidity of 80%.

It should be understood that the "detailed description" section, rather than the "abstract" and "brief description of the invention" sections, is intended to be used to interpret the claims. The brief description of the invention and abstract sections may set forth one or more, but not all exemplary embodiments of the invention contemplated by the inventors, and are therefore not intended to limit the invention and the appended claims in any way.

The present invention has been described above with the aid of functional constructs illustrating specific functions and relationships thereof. For convenience of description, the boundaries of these functional configurations have been arbitrarily defined herein. Other boundaries may be defined so long as the specified functions and relationships thereof are appropriately performed.

The foregoing description of the specific embodiments will so fully reveal the general nature of the invention that others can, by applying knowledge within the skill of the art, readily modify and/or adapt for various applications such specific embodiments, without undue experimentation, without departing from the general concept of the present invention. Therefore, such modifications and adaptations are intended to be within the meaning and range of equivalents of the disclosed embodiments, based on the teaching and guidance presented herein. It is to be understood that the phraseology or terminology herein is for the purpose of description and not of limitation, such that the terminology or phraseology of the present specification is to be interpreted by the skilled artisan in light of the teachings and guidance.

Claims

1. A process for preparing Compound 4 or a pharmaceutically acceptable salt thereof,

the method comprises the following steps:

1) converting compound 1 or a salt thereof to a compound of formula III or a salt thereof under amide forming conditions, wherein Lg in formula III is a leaving group;

2) converting a compound of formula III or a salt thereof to compound 4 under elimination reaction conditions; and in the alternative,

3) reacting compound 4 with a suitable acid to form a pharmaceutically acceptable salt of compound 4.

2. The method of claim 1, wherein Lg in formula III is a halogen or an oxygen-containing leaving group.

3. The method according to claim 1, wherein Lg in formula III is chloro.

4. The process of any one of claims 1-3, wherein converting compound 1 to a compound of formula III, or a salt thereof, comprises reacting compound 1 with an acid chloride reagent having formula IV, wherein Lg is a leaving group:

5. the method according to claim 4, wherein Lg in formula IV is chloro.

6. The process of any one of claims 1 to 5, which comprises contacting the compound of formula III with a base (e.g. a base, such as potassium hydroxide) to convert the compound of formula III or a salt thereof to compound 4.

7. The process of any one of claims 1-6, wherein compound 1 is prepared by a process comprising reducing compound a9 with hydrogen in the presence of a catalyst (e.g., palladium on carbon) in an ether solvent (e.g., tetrahydrofuran):

8. compound 4, or a pharmaceutically acceptable salt thereof, produced by the process of any one of claims 1-7.

9. Compound 4 or a pharmaceutically acceptable salt thereof,

its HPLC purity was at least 98.5%.

10. Compound 4 or a pharmaceutically acceptable salt thereof, according to claim 9, substantially free of impurities, characterized by a relative retention time on HPLC of about 1.02 (impurity a).

11. Compound 4 or a pharmaceutically acceptable salt thereof according to claim 9 or 10, substantially free of impurities, characterized by a relative retention time on HPLC of about 0.88 (impurity B).

12. A pharmaceutical composition comprising compound 4 or a pharmaceutically acceptable salt thereof according to any one of claims 8-11, and optionally a pharmaceutically acceptable excipient or carrier.

13. The pharmaceutical composition of claim 12, in the form of a tablet or capsule.

14. A method of treating one or more diseases or disorders mediated by an EGFR variant, comprising administering to a subject in need thereof a therapeutically effective amount of compound 4, or a pharmaceutically acceptable salt thereof, according to any one of claims 8-11, or a pharmaceutical composition according to any one of claims 12-13.

15. The method of claim 14, wherein the EGFR variant is a L858R mutant, an Exon19 deletion mutant, and/or a T790M mutant.

16. The method of claim 14 or 15, wherein the one or more diseases or disorders are selected from ovarian cancer, cervical cancer, colorectal cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer, leukemia, lymphoma, non-hodgkin lymphoma, gastric cancer, lung cancer, liver cancer, gastrointestinal stromal tumors, thyroid cancer, cholangiocarcinoma, endometrial cancer, renal cancer, anaplastic large cell lymphoma, acute myelogenous leukemia, multiple myeloma, and mesothelioma.

17. A method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of compound 4 or a pharmaceutically acceptable salt thereof according to any one of claims 8-11, or a pharmaceutical composition according to any one of claims 12-13.

18. The method of claim 17, wherein the cancer is ovarian cancer, cervical cancer, colorectal cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer, leukemia, lymphoma, non-hodgkin lymphoma, gastric cancer, lung cancer, liver cancer, gastrointestinal stromal tumor, thyroid cancer, bile duct cancer, endometrial cancer, renal cancer, anaplastic large cell lymphoma, acute myelogenous leukemia, multiple myeloma, and/or mesothelioma.

19. The method of claim 17, wherein the cancer is lung cancer.

20. The method of claim 17, wherein the cancer is non-small cell lung cancer.

21. The method of any one of claims 14-20, wherein the subject is resistant to one or more EGFR inhibitors selected from gefitinib, erlotinib and inctinib.

22. A compound of formula III:

wherein Lg is a leaving group.

23. The compound or salt according to claim 22, wherein Lg is chloro or hydroxy.