Crystalline forms, salt crystal forms, preparation methods and uses of substituted 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-C]quinolin-2-one derivatives

Abstract

The present invention provides crystalline forms of free base and salt of substituted 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivatives (represented by formula (I)), their preparation method and use, specifically the crystalline forms of free base and fumarate of 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivatives represented by formula (I). The crystalline forms of free base and salt provided by the present invention have good stability, can be used for the treatment of cancer, cancer with CNS metastasis, meningeal metastasis, primary brain cancer or glioma, etc., especially when combined with DNA double strand break inducer, and have good bioavailability and half-life, all of which are very important for further research on the efficacy of such solid drugs. [Formula 1] JPEG2025514643000015.jpg5364

Description

[Technical field]

[0001] The present invention relates to a crystal form, a salt form and a crystal form thereof of a substituted 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative, specifically, to a free base crystal form, a fumarate crystal form and a method for preparing the same and a method for using the same of (R)-1-(3,3-difluoro-1-methylpiperidin-4-yl)-8-(6-methoxypyridin-3-yl)-3-methyl-1,3-dihydro-2H-imidazo[4,5-c]quinolin-2-one (I). [Background technology]

[0002] Glioblastoma, or glioblastoma multiforme (GBM), is an advanced WHO grade IV malignant brain tumor. Due to the blood-brain barrier (BBB), no targeted therapy that can achieve effective doses in the brain has been approved for the treatment of patients with GBM or diffuse intrinsic pontine glioma (DIPG). Radiation therapy and surgery are the main therapeutic approaches along with temozolomide, but the therapeutic effect is limited. For DIPG patients in particular, radiation therapy remains the only treatment option for disease that recurs within 5.5 months. (See, for example, Hamer et al., Neuro-Oncology 12(3):304-316, 2010.) GBM is a whole-brain disease, and all GBMs have clinically significant tumor areas with intact BBB. If effective treatment does not reach all areas of GBM, treatment will fail and recurrence is inevitable (SaRkaRIa et al. Neuro-Oncology 20(2):184-191,2018). GBM is inherently resistant to radiation therapy (RT), and the development of brain-penetrating radiosensitizers has been one strategy to overcome this limitation. Repair of DNA double-strand breaks induced by RT is mediated by the protein kinase ataxia telangiectasia mutated (ATM). The role of ATM in radiation sensitivity and the availability of brain-penetrant ATM inhibitors as radiosensitizers could be highly effective in the treatment of brain cancer or cancers with central nervous system metastases, especially for conditions where radiation is the only treatment option, such as DIPG.

[0003] Therefore, it would be particularly useful to develop drugs that have both biological activity through ATM kinase and the ability to cross the blood-brain barrier as radiosensitizers in combination with radiotherapy for the treatment or prevention of glioma.

[0004] As is well known, the stability, solubility and bioavailability of the crystal form, salt form and salt crystal form of the same drug are very different, which may affect the efficacy of the drug.Therefore, it is very useful to develop new salt forms and new crystal forms of substituted 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-C]quinolin-2-one derivatives, which are more useful for drug processing and pharmaceutical composition, and provide more qualitative and quantitative information on the efficacy and safety of solid drugs, which is very important. Summary of the Invention Problem that the invention aims to solve

[0005] The problem that the present invention aims to solve is that existing ATM inhibitors cannot effectively cross the blood-brain barrier and cannot reach effective drug concentrations in the brain, and the properties of the free base amorphous 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivatives (I) of the present invention do not promote problems occurring during pharmaceutical processing and use of pharmaceutical compositions, and provide free base crystalline forms and salt crystalline forms of 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivatives that promote pharmaceutical processing and use of pharmaceutical compositions. The free base crystalline forms, salt crystalline forms, preparation methods and uses provide more qualitative and quantitative information for the study of efficacy and safety of solid drugs. [Means for solving the problem]

[0006] The 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivatives provided by the present invention have biological activity by ATM kinase and effectively cross the blood-brain barrier. The 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivatives are represented by formula (I): [ka]

[0007] The object of the present invention is achieved by the following technical solutions:

[0008] The technical solution adopted by the present invention is to provide new 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivatives (I) with chirality of R. The chiral purity is preferably ee>90%, more preferably ee>97%.

[0009] The technical solution adopted by the present invention is to provide new 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-C]quinolin-2-one derivatives (I) with R chirality, which have the biological activity of ATM kinase.

[0010] The technical solution adopted by the present invention is to provide a novel 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) with the biological activity of ATM kinase and at the same time with high blood-brain barrier passing ability and R chirality.

[0011] The present invention also provides a synthetic method for preparing the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivatives (I) of the present invention.

[0012] The technical solution adopted by the present invention is to provide a free base crystalline form of 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative, whose XRPD pattern has the following peaks at 2θ:

[0013] Main characteristic peaks: 10.2, 16.9, 20.3, 25.6 Secondary characteristic peaks: 10.8, 12.6, 13.5, 14.8, 16.1, 20.6, 21.2, 23.3 The error range for 2θ values ​​is ±0.2.

[0014] The XRPD pattern of the free base crystal form of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) provided by the present invention is preferably 2θ=10.19, 10.76, 12.56, 13.37, 14.84, 16.08, 16.87, 20.30, 20.51, 21.22, 23.26, 25.56. The differential peaks are at 5.08, 15.33, 16.53, 19.12, 19.51, 22.65, 23.72, 24.77, 26.67, 27.04, 28.38, 28.77, 29.26, 29.60, 30.09, 31.05, 32.61, 32.97, 34.90, 36.96, 38.21, and 38.50, and the error range of the above 2θ values ​​is ±0.2.

[0015] Another technical solution adopted by the present invention is to provide a crystalline form of 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-C]quinolin-2-one derivative fumarate salt, which has an XRPD pattern with peaks at the following 2θ:

[0016] Main characteristic peaks: 6.8, 8.6, 12.2, 13.7, 17.3, 19.4, 26.1 Secondary characteristic peaks: 6.1, 11.0, 14.6, 16.1, 16.5, 18.0, 20.2, 22.1, 26.8 The error range for 2θ values ​​is ±0.2.

[0017] The XRPD pattern of the fumarate crystal form of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) provided by the present invention is preferably at 2θ=6.07, 6.78, 8.60, 10.99, 12.21, 13.66, 14.55, 16.09, 16.49, 17.32, 18.04, 19.41, 20.24, 22.15, 26.06, 26.81. Differential peaks are at 12.57, 18.37, 18.64, 20.96, 23.54, 24.82, 25.40, 28.23, 30.19, 33.40, 36.25, and the error range of the above 2θ values ​​is ±0.2.

[0018] The free base crystalline form of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative of the present invention preferably has an XRPD pattern substantially as shown in FIG.

[0019] The fumarate salt crystalline form of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative of the present invention preferably has an XRPD pattern substantially as shown in FIG.

[0020] The present invention also provides a method for preparing the free base crystalline form of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) of the present invention, which comprises adding 100-120 mg of an amorphous form of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative sample of formula (I) to 1-1.5 mL of an organic solvent mixture and stirring at 50° C. or higher, whereupon the lower layer solid is separated by centrifugation to obtain the free base crystalline form.

[0021] The present invention also provides a method for preparing the fumarate crystalline form of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) of the present invention, which comprises adding the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative of formula (I) and 2-2.5 equivalents of fumaric acid to an organic solvent, stirring the mixture at 22-28° C., and collecting the solid by centrifugation, in which 10-200 mg of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative of formula (I) is added per ml of organic solvent.

[0022] In the above method, the organic solvent is preferably a mixture of dichloromethane and methanol (1:1 to 1:1.7, v / v).

[0023] Furthermore, in the method for preparing the free base crystalline form of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I), the organic solvent is preferably an alcohol, an ether, an ester, a ketone, an aliphatic alkane, an aromatic solvent, and more preferably methyl tert-butyl ether / water (1:10, v / v).

[0024] The present invention also provides a pharmaceutical composition comprising a crystalline free base or crystalline fumarate salt of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative of formula (I), or a combination thereof, and a pharma- ceutically acceptable excipient or auxiliary ingredient.

[0025] The adjuvants or auxiliary components preferably include carriers, excipients, diluents, vehicles and adjuvants.

[0026] The present invention also provides the use of a free base crystalline form, a fumarate crystalline form, or the above pharmaceutical composition of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative represented by formula (I) in the manufacture of a medicament for the treatment or prevention of a disease mediated by ATM kinase.

[0027] The drug is preferably a drug for the treatment or prevention of a disease mediated by ATM kinase and caused by ATM activation following DNA double-strand breaks.

[0028] More preferably, the medicament is manufactured for the treatment or prevention of non-small cell lung cancer brain metastasis, pancreatic cancer, meningeal metastasis, head and neck squamous cell carcinoma, squamous cell carcinoma, brain stem tumor, DIPG, primary brain cancer, or glioma. Effect of the Invention

[0029] Those skilled in the art can adjust the dosage and method of the reagents used in the present invention, including increasing or decreasing the amount of raw materials or solvents, based on their knowledge and experience, and these adjustment schemes are also included in the method of the present invention.Compared with the prior art, the present invention has the following beneficial effects:(1) The novel chiral (R)-1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) and its pharmaceutical free base and salt of the present invention have an unexpected blood-brain barrier passing ability, and can be used as a protein kinase inhibitor, especially for pathologies caused by ATM activation, and can be used for the treatment or prevention of disorders related to abnormal protein kinase activity, such as cancer, cancer with brain metastasis, cancer with meningeal metastasis, and central nervous system diseases, especially when combined with a DNA double-strand break inducer. (2) The 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) and its pharmaceutical free base and salt of the present invention have a low efflux rate, are not P-glycoprotein efflux enzyme substrates, breast cancer drug resistance efflux enzyme substrates, or aldehyde oxidase substrates, and can reduce efflux resistance and improve absorption. (3) The free base crystal form of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative of the present invention and its pharmaceutical salt have good pharmacokinetics, long half-life, and high biological activity, which can reduce the burden of tablet intake on patients and improve tablet intake cone pliance. (4) The free base crystal form and fumarate crystal form of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) of the present invention provided by the present invention have good stability and water solubility, which are beneficial for use in pharmaceutical processing and pharmaceutical compositions, and can treat ATM kinase-mediated cancers such as non-small cell lung cancer brain metastasis, meningeal metastasis, head and neck squamous cell carcinoma, pancreatic cancer, squamous cell carcinoma, brain stem tumor, primary brain cancer or glioma, and have good bioavailability and half-life, providing qualitative and quantitative information on efficacy and safety, which is of great significance for further research on the efficacy of such solid drugs. [Brief description of the drawings]

[0030] Other features, objects and advantages of the present invention will become more apparent upon reading the detailed description of the non-limiting embodiments with reference to the following drawings, in which: [Figure 1] 1 is an XRPD pattern of the free base crystalline form of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative of the present invention. [Diagram 2] 1 is an XRPD pattern of the fumarate salt crystal form of a 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative of the present invention. [Diagram 3] FIG. 1 is a schematic diagram of an efficacy study of 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivatives of the present invention in a humanized glioma mouse intracranial model. [Figure 4A] Figure 4A shows the TGA and DSC spectra of the free base crystal form of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative of the present invention, where the TGA spectrum shows no significant weight loss, and Figure 4B shows the DSC spectrum, where the crystal form is stable even at high temperatures, the crystal form does not change, and the melting point is 206°C. [Figure 4B] Figure 4A shows the TGA and DSC spectra of the free base crystal form of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative of the present invention, where the TGA spectrum shows no significant weight loss, and Figure 4B shows the DSC spectrum, where the crystal form is stable even at high temperatures, the crystal form does not change, and the melting point is 206°C. [Figure 5A] Figure 5A shows the TGA and DSC spectra of the fumarate salt of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative of the present invention, where Figure 5A shows the TGA spectrum without significant weight loss, and Figure 5B shows the DSC spectrum, where the crystal form is stable even at high temperatures, the crystal form does not change, and the melting point is 220°C. [Figure 5B]Figure 5A shows the TGA and DSC spectra of the fumarate salt of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative of the present invention, where Figure 5A shows the TGA spectrum without significant weight loss, and Figure 5B shows the DSC spectrum, where the crystal form is stable even at high temperatures, the crystal form does not change, and the melting point is 220°C. [Figure 6] 1 is a synthesis scheme for intermediate 3,3-difluoro-1-methylpiperidin-4-amine A6. [Figure 7] 1 is a synthesis scheme of compound (I). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0031] (Explanation of terms used in the present invention) The terms "ameliorate" and "treat" are used interchangeably to mean, but are not limited to, a therapeutic benefit and / or a prophylactic benefit, as well as reducing, inhibiting, preventing, or stabilizing the onset or progression of a condition (e.g., a disease or disorder described herein).

[0032] "Disease" refers to a specific disorder or disease that damages or interferes with the normal function of a cell, organ, or tissue.

[0033] "Marker" refers to any change associated with a disease or disorder, for example a protein or polynucleic acid that has an altered expression level or activity associated with a disease or disorder.

[0034] In this context, the terms "comprise," "contain," and "are," and other similar terms, have the meaning given to them in the patent law, and "consisting substantially of" or "consisting essentially of" have the same meaning given to them in the patent law, and the term is open to the presence of matter other than the cited subject matter, so long as the presence of such matter does not change the basis or novel characteristics of the referenced subject matter, but does not include prior art implementations.

[0035] As used herein, the terms "antagonist" and "inhibitor" are used interchangeably and refer to the ability of a compound or agent to inhibit the biological function of a target protein or polypeptide, for example, by inhibiting the activity or expression of the protein or polypeptide. Some of the antagonists herein interact with a particular target protein or polypeptide (e.g., bind to ATM kinase), but the inhibitors also, within the definition, inhibit the biological activity of the target protein or polypeptide by interacting with other members of a signaling pathway that targets the protein or polypeptide, including those that inhibit the development of tumors that develop, grow, or spread, or those associated with undesirable immune responses exhibited by autoimmune diseases.

[0036] As used herein, the terms "anti-cancer agent", "anti-tumor agent", or "chemotherapeutic agent" refer to any agent useful in the treatment of a neoplastic disorder. The class of anti-cancer agents includes chemotherapeutic agents. "Chemotherapy" refers to one or more chemotherapeutic agents and / or other agents administered in a variety of ways, including intravenously, orally, subcutaneously, intramuscularly, intraperitoneally, intravesically, transdermally, bucally, or by inhalation.

[0037] As used herein, "cell proliferation" refers to an increase in cell number as a result of cell division and cell growth (eg, increase in size) consistent with proliferation signals by cell morphology.

[0038] As used herein, the term "co-administration" refers to the use of two or more agents simultaneously, the use of co-present compositions using two or more agents, and the administration of two or more agents and / or their metabolites at different times, or the administration of two or more agents and / or their metabolites alone.

[0039] As used herein, the term "effective amount" or "therapeutically effective amount" refers to an amount of a compound or pharmaceutical composition described herein that is sufficient to achieve the intended use, including, but not limited to, the treatment of a disease. In some embodiments, the amount is detected as effective in killing cancer cells, inhibiting the growth or spread of tumors, tumor size or number, or the severity, stage, and progression of cancer. An effective therapeutic amount may vary depending on the intended application, such as in vitro or in vivo, disease state and severity, subject age, weight, or method of administration. The term also refers to a dose that induces target cells to respond in a particular way, such as, for example, reducing cell migration. Specific doses will vary, for example, depending on the particular compound selected, the subject's species (including age and pre-existing health conditions), route of administration, severity of disease, combination with other drugs, time of administration, tissue to be administered, and administration device.

[0040] As used herein, the term "therapeutic benefit" includes therapeutic benefit and / or prophylactic benefit. A prophylactic benefit includes delaying or eliminating the onset of a disease or condition, delaying or eliminating the onset of symptoms or disorders of a disease, slowing, halting or reversing a disease or condition, or a combination thereof.

[0041] As used herein, the term "signal transduction" refers to the process by which a stimulatory or inhibitory signal is transmitted to a cell to initiate an intracellular response. A "modulator" of a signal transduction pathway refers to a compound that regulates one or more activities of a cellular protein mediated by a particular signal transduction pathway. A "modulator" can either enhance (agonist) or inhibit (antagonist) the activity of a signal transduction molecule.

[0042] As used herein, the term "selective inhibition" refers to the ability of a compound to selectively reduce a target signaling activity relative to an off-target target activity through direct or indirect interaction. For example, the activity of a compound that selectively inhibits ATM is at least about 2-fold, about 3-fold, about 5-fold, about 10-fold, about 20-fold, about 50-fold, about 100-fold or more than the activity of ATR or DNA-PK kinase.

[0043] As used herein, the term "radiation therapy" refers to the exposure of a subject to radiation emitters such as, but not limited to, alpha-particle emitting radionuclides (e.g., actinium and thorium radionuclides) (e.g., beta emitters), conversion electron emitters (e.g., strontium-89 and samarium-153-EDTMP), or high-energy radiation, including, but not limited to, x-rays, gamma rays, and neutrons.

[0044] As used herein, the term "subject" includes, but is not limited to, humans (e.g., of any age group and sex) and / or other primates (e.g., cynomolgus monkeys, rhesus monkeys), mammals, including commercially relevant mammals such as cows, sheep, goats, pigs, horses, cats and / or dogs, and / or birds, including commercially relevant birds such as chickens, geese, quail, ducks and / or turkeys.

[0045] As used herein, the term "in vivo" refers to activities that take place within a subject's body, including occurring in rodents such as rats, mice, guinea pigs, etc.

[0046] As used herein, the term "in vitro" refers to events that occur outside the body. For example, in vitro testing includes any detection that is performed outside the body. In vitro assays include cell determinations based on live or dead cells, and cell-free assays that are used with non-intact cells.

[0047] The term "compound" as used herein also includes salts of compounds of the general formula herein. This term also includes any solvates, hydrates, and polymorphs of the foregoing. In certain aspects of the invention described in this application, specific reference to "solvates," "hydrates," or "polymorphs" is to be taken as such. In other aspects of the invention where the term "compound" is used without reference to these other forms, such forms are not excluded.

[0048] The salts of the compounds of the invention are formed between an acid and a basic group of the compound, such as an amino function. According to another preferred embodiment, it is a pharma-ceutically acceptable acid addition salt.

[0049] As used herein, the term "pharmaceutical acceptable" refers to a pharmaceutical composition that is suitable for use in contact with the tissues of humans and other mammals without undue toxicity, irritation, allergic response, etc., and has reasonable benefits / risks rather than ingredients. "Pharmaceutically acceptable salt" refers to a non-toxic salt that, upon administration to a recipient, can directly or indirectly provide a prodrug or compound of the present invention.

[0050] Acids commonly used to form pharma- ceutically acceptable salts include inorganic acids such as hydrochloric, hydrobromic, sulfuric, hydroiodic, and phosphoric acids, trifluoroacetic acid, citric acid, camphorsylthiocyanate, maleic acid, oxalic acid, picric acid, acetic acid, adipic acid, alginic acid, aspartic acid, sulfuric acid, boric acid, butyric acid, valeric acid, camphoric acid, camphorsylthiocyanate, digluconic acid, dodecylsulfuric acid, pivalic acid, formic acid, fumaric acid, hydroiodic acid, benzoic acid, 2-hydroxyethanesulfonic acid, fumaric acid, stearic acid, lactobionic acid, protic acid, and the like. These include inorganic acids such as pionic, lauric, oleic, nicotinic, lactic, cinnamic, amber, mandelic, malic, tartaric, lactic acid, pyruvic, pectinic, methanesulfonic, pamoic, benzenesulfonic, persulfuric, palmitic, malonic, glycerophosphoric, 2-naphthalenesulfonic, p-toluenesulfonic, salicylic, ascorbic, 2-phenylpropionic, gluconic, glucuronic, phosphoric, glutamic, ethanesulfonic, p-bromobenzenesulfonic, and carbonic acid, as well as related inorganic and organic acids.

[0051] As used herein, the term "hydrate" refers to a compound that contains a stoichiometric or non-stoichiometric amount of water bound by non-covalent intermolecular forces. As used herein, the term "solvate" refers to a compound that contains a stoichiometric or non-stoichiometric amount of a solvent bound by non-covalent intermolecular forces, such as water, dichloromethane, 2-propanol, acetone, methanol, ethanol, etc. Pharmaceutically acceptable solvates and hydrates are complexes that may contain, for example, 1 to about 100, 1 to about 10, 1 to about 4, about 3, or about 2 solvent or water molecules. As used herein, the term "compound" is understood to include the compounds and solvates, hydrates, and mixtures thereof of the compounds described.

[0052] As used herein, "polymorph" refers to a solid crystalline form of a compound or a complex thereof. Different polymorphs of the same compound may exhibit different physical, chemical, and / or spectral properties. Different physical properties include, but are not limited to, stability (e.g., heat, light, or moisture), density, hygroscopicity, solubility, compressibility, and dissolution rate.

[0053] The term "isomers" as used herein refers to different compounds that have the same molecular formula. "Stereoisomers" are only isomers in which atoms are arranged in a different way. The term "isomers" as used herein includes all geometric and stereoisomers. For example, "isomers" including geometric double bond cis- and trans-isomers, also referred to as E- and Z-isomers, R- and S-enantiomers, diastereomers, (D)-isomers and (L)-isomers, racemic mixtures thereof, and other mixtures thereof are disclosed herein.

[0054] Double bonds surrounding carbon-carbon substituents are designated as being in the "Z" or "E" configuration, where the terms "Z" and "E" are used in accordance with IUPAC nomenclature. Unless otherwise stated, structures represent both the "E" and "Z" isomers.

[0055] Substitutable substituents surrounding a carbon-carbon double bond may also be designated "cis" or "trans", where "cis" refers to substituents on the same side of the double bond and "trans" refers to substituents on both sides. The arrangement of the carbocyclic rings around the substituents may also be designated "cis" or "trans", where "cis" refers to substituents on the same side in the plane of the ring and "trans" refers to substituents on both sides of the plane of the ring. A mixture of substituents on the same and opposite sides of the plane of the two rings is herein designated as "cis / trans".

[0056] The term "enantiomer" as used herein refers to a pair of non-superimposable stereoisomers that are mirror images of one another. A mixture of enantiomers in specific ratios may be referred to as a "racemic" mixture. "(±)" is used to designate a racemic mixture when necessary. "Diastereomers" refer to mirror images that have at least two asymmetric atoms but are not stereoisomers of each other. Absolute stereochemistry is specified according to the Cahn-Ingold-PRelog RS system. If the compound is an enantiomer, the stereochemistry of each chiral carbon may be designated R or S. Because the absolute configuration of a compound is unknown, it may be designated (+) or (-) depending on the direction of rotation (right or left) of polarized light at the wavelength of the sodium D line. Some of the substances described herein contain one or more asymmetric centers and can therefore give rise to enantiomers, diastereomers, and other stereoisomeric forms that can be defined by the absolute configuration of each asymmetric atom (R) or (S), and the pharmaceutical compositions and methods include all these possible isomers, including racemic mixtures, optically pure forms, and intermediate mixtures. Optically active (R)- and (S)- can also be prepared by chiral synthetic methods or chiral reagents, or by conventional techniques.

[0057] As used herein, the term "enantiomeric excess" or "enantiomeric excess" can be calculated using the formula shown below: In the example shown below, the composition contains one enantiomer, e.g., 90% S enantiomer, and the other enantiomer, e.g., 10% R enantiomer. ee value = (90-10) / 100 = 80%

[0058] Thus, a composition that contains 90% of one enantiomer and 10% of the other enantiomer has an enantiomeric excess of 80%. Some of the compositions described herein contain at least about 50% enantiomeric excess, about 75%, about 90%, about 95%, or about 99% S enantiomer. In other words, the composition has an enantiomeric excess of the S enantiomer in the R enantiomer. In other embodiments, some of the compositions described herein contain at least about 50% enantiomeric excess, about 75%, about 90%, about 95%, or about 99% R enantiomer. In other words, the composition has an enantiomeric excess of the R enantiomer in the S enantiomer. For example, in some embodiments, an isomer / enantiomer may provide the ee value of the corresponding enantiomer and may be referred to as "optically enriched," "enantiomerically enriched," "enantiomerically pure," and "non-racemic," which are used interchangeably herein. These terms mean that the weight percentage of one enantiomer is greater than the amount of a control mixture than a racemic composition in one enantiomer (e.g., greater than 1:1 by weight). For example, the enantiomeric enantiomer of the S enantiomer is present at about 75% of the weight of the enantiomer (e.g., greater than about 50% by weight of the compound or at least about 80% by weight). In some embodiments, enrichment is greater than about 80% by weight, providing "substantially enantiomerically enriched," "substantially enantiomerically pure," or "substantially non-racemic," meaning that the weight of an enantiomer is at least 85% by weight of the composition, e.g., at least about 90% by weight of the formulation, and even, e.g., at least about 95% by weight relative to one of the other enantiomers. In certain embodiments, the compounds provided herein may be present in an amount of about 90% by weight of at least one enantiomer. In other embodiments, the compounds may be present in an amount of at least about 95%, about 98%, or about 100% by weight of the enantiomer. In some embodiments, the compounds are (S)- and (R)-racemic mixtures.In other embodiments, processes are provided in which the individual compounds of the mixture (S) are predominantly mixtures of compounds or are predominantly mixtures of compounds in which (R) is present. For example, the compound mixture has greater than about 55%, greater than about 60%, greater than about 65%, greater than about 70%, greater than about 75%, greater than about 80%, greater than about 85%, greater than about 90%, greater than about 95%, greater than about 96%, greater than about 97%, greater than about 98%, greater than about 99%, greater than about 99.5%, or more. In other embodiments, the mixture of compounds has an (S)-enantiomeric excess of greater than about 55% to about 99.5%, greater than about 60% to about 99.5%, greater than about 65% to about 99.5%, greater than about 70% to about 99.5%, greater than about 75% to about 99.5%, greater than about 80% to about 99.5%, greater than about 85% to about 99.5%, greater than about 96% to about 99.5%, greater than about 97% to about 99.5%, greater than about 98% to about 99.5%, greater than about 99% to about 99.5%, or more. In other embodiments, the (R)-enantiomer purity of the compound mixture is greater than about 55%, greater than about 60%, greater than about 65%, greater than about 70%, greater than about 75%, greater than about 80%, greater than about 85%, greater than about 90%, greater than about 95%, greater than about 96%, greater than about 97%, greater than about 98%, greater than about 99%, greater than about 99.5%, or more. In other embodiments, the mixture of compounds has an (R)-enantiomeric excess of greater than about 55% to about 99.5%, greater than about 60% to about 99.5%, greater than about 65% to about 99.5%, greater than about 70% to about 99.5%, greater than about 75% to about 99.5%, greater than about 95% to about 99.5%, greater than about 85% to about 99.5%, greater than about 96% to about 99.5%, greater than about 97% to about 99.5%, greater than about 98% to about 99.5%, greater than about 99% to about 99.5% or greater.

[0059] In other embodiments, the compound mixtures contain the same chemical entities in addition to their stereochemical orientation, i.e., (S)- or (R)-. For example, if a compound has a -CH(R)- unit and R is not hydrogen, then -CH(R)- is the same chemical entity as either the (S)- or (R)-stereochemical orientation. In some embodiments, the (S)-isomer in the mixture of the same chemical is present in an (S) enantiomeric excess of greater than about 55% to about 99.5%, greater than about 60% to about 99.5%, greater than about 65% to about 99.5%, greater than about 90% to about 99.5%, greater than about 90% to about 99.5%, greater than about 90% to about 99.5%, greater than about 90% to about 99.5%, greater than about 95% to about 99.5%, greater than about 96% to about 99.5%, greater than about 97% to about 99.5%, greater than about 98% to about 99.5%, greater than about 99% to about 99.5% or more.

[0060] In another embodiment, the (R) isomer is present at about 55%, about 60%, about 65%, about 90%, about 95%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, about 99.5% or more of the same chemical entity (excluding stereochemical orientation) as the (S) isomer. In some embodiments, the (R)-enantiomeric excess in a mixture of the same chemical entity (excluding its stereochemical orientation) is greater than about 55% to about 99.5%, greater than about 60% to about 99.5%, greater than about 75% to about 99.5%, greater than about 75% to about 99.5%, greater than about 75% to about 99.5%, greater than about 90% to about 99.5%, greater than about 95% to about 99.5%, greater than about 96% to about 99.5%, greater than about 97% to about 99.5%, greater than about 98% to about 99.5%, greater than about 99% to about 99.5%, or greater.

[0061] Enantiomers can be isolated from a racemic mixture by any method known to those skilled in the art, including chiral high performance liquid chromatography (HPLC), the formation and crystallization of chiral salts, or asymmetric synthesis.

[0062] Optical isomers can also be obtained by conventional resolution of racemic mixtures with optically active acids or bases, for example by forming diastereomeric salts. Examples of suitable acids include, but are not limited to, tartaric acid, diacetyl, dibenzoyl, dibenzoyltartaric acid, camphorsulfonic acid. Separation of isomers from mixtures of optically active bases of these salts can be achieved by diastereomeric crystallization. Alternatively, covalent diastereomeric molecules are synthesized when the open compound is reacted with an activated optically pure acid or an optically pure isocyanate. The synthesized enantiomers can be isolated by conventional methods such as chromatography, distillation, crystallization, or sublimation, and then hydrolyzed to obtain enantiomerically enriched compounds. Optically active compounds can also be obtained by using active substances. In some embodiments, these isomers can be in the form of free acids, free bases, esters, or salts.

[0063] In certain embodiments, the pharma- ceutically acceptable forms are tautomers. As used herein, "tautomer" refers to a type of isomer that includes at least one form of change of two or more interconverting compounds with a hydrogen atom and a covalent bond (e.g., from a single bond to a double bond, a triple bond to a single bond, or vice versa). "Tautomerism" includes proton or proton-transfer tautomerism, which is considered a subset of acid-base chemistry. "Proton-transfer tautomerism" involves a proton transfer with a bond change. The exact ratio of tautomers depends on various factors, including temperature, solvent, and pH. Tautomerism between them is possible (e.g., in solution), and a chemical equilibrium of tautomerism can be reached. Tautomerism (i.e., a reaction that provides a tautomeric pair) can occur with acid or base catalysis, or in the presence or absence of external factors. Such tautomeric additions include, but are not limited to, ketones to enols, amides to imides, enamines to imines, and the addition of one form of an enamine to a different enamine. Specific examples of ketone to enol tautomers are pentane-2,4-dione and 4-hydroxypent-3-en-2-one tautomers. Another example of tautomerism is the tautomerism of phenols and ketones. Specific examples of phenol and ketone tautomers are pyridine-4-phenol and pyridin-4-(1H)-one tautomers.

[0064] Unless otherwise stated, structures depicted herein are meant to include compounds that are present only at one or more isotopically enriched atoms, for example, compounds having structures where a hydrogen is replaced with deuterium or tritium, or carbon-13 or carbon-14 enriched structures within the disclosed ranges.

[0065] The present disclosure also includes "isotopically labeled derivatives," which are pharma- ceutically acceptable forms of the compounds recited herein except for one or more atoms of a different atomic mass commonly found in nature. Examples of isotopes that can be incorporated into the disclosed compounds include: 2 H, 3 H, 13 C. 14 C.15 N, 18 O. 17 O. 18 F and 36 and isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as Cl. Certain isotopically labeled disclosed compounds (e.g., 3 H and 14 C) are useful for determining the tissue distribution of compounds and / or substrates. 3 H) and carbon 14 (i.e. 14 C) isotopes can be easily produced and tested. In addition, deuterium (i.e. 2 Substitution with heavier isotopes such as H or D can provide certain therapeutic advantages (e.g., increased half-life in vivo and reduced dose requirements) resulting from higher metabolic stability. Isotopically labeled disclosed compounds can generally be produced by substituting a non-isotopically labeled reagent for an isotopically labeled reagent. Some embodiments provided herein may also include one or more non-natural atomic isotopes to produce such compounds. All isotopic variants of the disclosed compounds, whether radioactive or not, are used herein within the scope of the present disclosure. In some embodiments, radiolabeled compounds may be used to study the metabolism and tissue distribution of compounds by altering metabolic pathways or rates or other biological functions.

[0066] The term "CDCl3" refers to deuterated chloroform.

[0067] The term "DMSO-d6" refers to deuterated dimethylsulfoxide.

[0068] The term "LC-MS: (ESI)" refers to electrospray-ionization liquid chromatography mass spectrometry.

[0069] The term "alteration" as used herein refers to a change in relative physiological state. Examples of alterations include mutations, deletions, fusions with other proteins, overexpression, or underexpression.

[0070] Compounds of the invention: In one aspect, the present invention provides a compound of formula (I), or a salt thereof, or a hydrate, solvate or polymorph thereof: [ka]

[0071] The synthesis of the novel 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivatives (I) with the chirality of R of the present invention can be easily achieved by the general synthesis researcher. For example, the relevant methods and intermediates are disclosed herein. All patents, patent applications, and publications referred to herein, whether published in conventional journals or available only on the Internet, are hereby incorporated by reference in their entirety.

[0072] Other methods for synthesizing compound (I) described herein can be readily adapted from the references cited herein. Modification of these procedures and their optimization are within the capabilities of one of ordinary skill in the art.

[0073] The present invention also provides compositions comprising an effective amount of the compounds described herein, or, if applicable, pharma- ceutically acceptable salts, solvates, hydrates, or polymorphs of the compounds.The compositions of the present invention are preferably formulated for pharmaceutical use ("pharmaceutical compositions"). Here, the carrier is a pharma- ceutically acceptable carrier.In consideration of compatibility with other components of the formulation, in the case of a pharma- ceutical acceptable carrier, the carrier must be "acceptable" and not harmful to the recipient in the amount normally used in medicines.

[0074] A "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" includes any solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. A pharmaceutically acceptable carrier or excipient does not interfere with the pharmacological activity of the disclosed compound and is non-toxic when administered in a dosage of the compound sufficient to deliver. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions disclosed herein is contemplated.Examples of pharma- ceutically acceptable carriers and excipients include sugars such as lactose, sucrose, and glucose; starches such as potato starch and corn starch; celluloses and derivatives thereof such as sodium carboxymethylcellulose, cellulose acetate, and ethylcellulose; gelatin, powdered tragacanth, talc, malt, cacao butter and suppository wax; oils such as peanut oil, safflower oil, cottonseed oil, olive oil, sesame oil, corn oil, and soybean oil; diols such as polyethylene glycol and propylene glycol; esters such as ethyl oleate and ethyl laurate; agar, buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid, aldehydes, phosphates, phosphate buffers, sodium lauryl sulfate, and Non-toxic compatible lubricants such as magnesium stearate, colorants, coating agents, release agents, sweeteners, flavors and fragrances, surfactants for pharmaceutical dosage forms such as (SEDDS) such as vitamin E polyethylene glycol 1000 succinate, Tween or other similar polymeric delivery matrices, serum proteins such as human serum albumin, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or protamine sulfate, electrolytes such as potassium hydrogen phosphate, disodium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulosic materials, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, and other solubilizing derivatives to improve delivery of the compound, such as cyclodextrins, such as alpha-, beta- and gamma-cyclodextrin, chemically modified derivatives such as hydroxyalkyl cyclodextrins including 2- and 3-hydroxypropyl cyclodextrin.

[0075] The pharmaceutical composition of the present invention can be administered in solid or liquid form, for example, by oral administration such as irrigation (aqueous or non-aqueous solution or suspension), tablet (for oral subcutaneous and systemic absorption), hard or soft capsule, pill, syrup, powder, granule, tongue paste, duodenal route, parenteral administration including intravenous, intraarterial, subcutaneous, intramuscular, topical administration, sublingual, catheter or stent, for example, as fine powder or via inhalation (for example, as fine powder or liquid aerosol).

[0076] Examples of suitable aqueous and non-aqueous carriers in the pharmaceutical compositions to be injected include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils such as olive oil, and organic esters such as ethyl oleate. The desired particle size of the dispersion can be maintained by using coating materials such as lecithin, and the proper fluidity can be maintained by using surfactants. These compositions can include adjuvants such as preservatives, wetting agents, emulsifying agents, dispersing agents, lubricants, and / or antioxidants. The antimicrobial action of the compounds described herein can be ensured by including various antibacterial and antifungal agents, such as, for example, p-hydroxybenzoic acid, chlorobutanol, phenol sorbic acid, etc. The compositions can also include isotonic agents such as sugars, sodium chloride, etc. In addition, the absorption of the injectable dosage form can be prolonged by including absorption-delaying agents such as aluminum monostearate and gelatin.

[0077] Methods for preparing such formulations or compositions include the steps of selecting a compound and / or chemotherapeutic vehicle described herein and one or more accessory ingredients. In general, the formulations are produced by uniformly structuring a compound disclosed herein with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product. Excipients are also within the scope of this disclosure insofar as any conventional excipient medium is not incompatible with the compounds provided herein, e.g., by interacting with any other components of the pharma-ceutically acceptable composition to produce undesirable biological or deleterious effects.

[0078] In some embodiments, the concentration of one or more disclosed compounds is about 100%, about 90%, about 80%, about 70%, about 60%, about 50%, about 40%, about 14%, about 13%, about 12%, about 11%, about 10%, about 5%, about 4%, about 3%, about 2%, about 1%, about 0.5%, about 0.4%, about 0.3%, about 0.2%, about 0.09%, about 0.08%, about 0.07%, about 0.06%, about 0.05%, about 0.04%, about 0.03%, about 0.02%, about 0.04%, about 0.05%, about 0.06%, about 0.07%, about 0.08%, about 0.09%, about 0.08%, about 0.07%, about 0.06%, about 0.05%, about 0.04%, about 0.03%, about 0.02%, about 0.04 ... It may be less than 0.01%, about 0.009%, about 0.008%, about 0.007%, about 0.006%, about 0.005%, about 0.004%, about 0.003%, about 0.002%, about 0.001%, about 0.0009%, about 0.0008%, about 0.0007%, about 0.0006%, about 0.0005%, about 0.0004%, about 0.0003%, about 0.0002% or about 0.0001% weight / weight, weight / volume, or volume / volume.

[0079] In some embodiments, the concentration of one or more compounds disclosed herein is about 90%, about 80%, about 70%, about 60%, about 50%, about 40%, about 30%, about 20%, about 18.5%, about 18.25%, about 17.5%, about 17.25%, about 17%, about 16.5%, about 16.25%, about 16%, about 15.5%, about 15.25%, about 15%, about 14.5%, about 14.25%, about 14%, about 13.5%, about 13.25%, about 13%, about 12.5%, about 12.25%, about 12%, about 11.5%, about 11.25%, about 11%, about 10.75%, about 10.5%, about 10%, about 9.75%, about 9.5%, about 9.25%, about 9%, about 8.75%, about 8.5%, about 8.25%, about 8%, about 7.75%, about 7.5%, about 7.25%, about 7%, about 6.75%, about 6.5%, about 6.25%, about 6%, about 5.75%, about 5.5%, about 5.25%, about 5%, about 4.75%, about 4.5%, 4.25%, 4%, 3.75%, 3.5%, 3%, 2.75%, 2.50%, 2.25%, 2%, 1.75%, 1.50%, 1.25%, 1%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.0 It may exceed 1%, about 0.009%, about 0.008%, about 0.007%, about 0.006%, about 0.005%, about 0.004%, about 0.003%, about 0.002%, about 0.001%, about 0.0009%, about 0.0008%, about 0.0007%, about 0.0006%, about 0.0005%, about 0.0004%, about 0.0003%, about 0.0002%, or about 0.0001% weight / weight, weight / volume, or volume / volume.In some embodiments, the concentration of one or more compounds disclosed herein ranges from about 0.0001% to about 50%, from about 0.001% to about 40%, from about 0.01% to about 30%, from about 0.02% to about 20%, from about 0.09% to about 24%, from about 0.08% to about 23%, from about 0.07% to about 22%, from about 0.06% to about 24%, from about 0.2% to about 2 The weight / weight, weight / volume, or volume / volume ratio may be 0%, about 0.1% to about 21%, about 0.2% to about 20%, about 0.3% to about 19%, about 0.4% to about 18%, about 0.5% to about 17%, about 0.6% to about 16%, about 0.7% to about 15%, about 0.8% to about 14%, about 0.9% to about 12%, or about 1% to about 10%. In some embodiments, the concentration of one or more compounds disclosed herein may range from about 0.001% to about 10%, about 0.01% to about 5%, about 0.02% to about 4.5%, about 0.03% to about 4%, about 0.04% to about 3.5%, about 0.05% to about 3%, about 0.06% to about 2.5%, about 0.07% to about 2%, about 0.08% to about 1.5%, about 0.09% to about 1%, or about 0.1% to about 0.9% weight / weight, weight / volume, or volume / volume.

[0080] In some embodiments, the amount of one or more compounds disclosed herein is about 10 grams, about 9.5 grams, about 9.0 grams, about 8.5 grams, about 8.0 grams, about 7.5 grams, about 7.0 grams, about 6.5 grams, about 6 grams, about 5.5 grams, about 5 grams, about 4.5 grams, about 4 grams, about 3.5 grams, about 3 grams, about 2.5 grams, about 2 grams, about 1.5 grams, about 1.0 grams, about 0.95, about 0.9 grams, about 0.85 grams, about 0.8 grams, about 0.75 grams, about 0.7 grams, about 0.65 grams, about 0.6 grams, about 0.55 grams, about 0.5 grams, about 0.45 grams, about 0.4 grams, about 0.35 grams, about 0.3 grams, about 0.25 grams, about 0.2 grams, about 0.15 grams, about The amount may be less than or equal to 0.1 grams, about 0.09 grams, about 0.08 grams, about 0.07 grams, about 0.06 grams, about 0.05 grams, about 0.04 grams, about 0.03 grams, about 0.02 grams, about 0.01 grams, about 0.009 grams, about 0.008 grams, about 0.007 grams, about 0.006 grams, about 0.005 grams, about 0.004 grams, about 0.003 grams, about 0.002 grams, about 0.001 grams, about 0.0009 grams, about 0.0008 grams, about 0.0007 grams, about 0.0006 grams, about 0.0005 grams, about 0.0004 grams, about 0.0003 grams, about 0.0002 grams, or about 0.0001 grams.In some embodiments, the amount of one or more compounds disclosed herein is about 0.0001 grams, about 0.0002 grams, about 0.0003 grams, about 0.0004 grams, about 0.0005 grams, about 0.0006 grams, about 0.0007 grams, about 0.0008 grams, about 0.0009 grams, about 0.001 grams, about 0.0015 grams, about 0.002 grams, about 0.0025 grams, about 0.003 grams, about 0.0035 grams, about 0.004 grams, about 0.0045 grams, about 0.005 grams, about 0.0055 grams, about 0.006 grams, about 0.0065 grams, about 0.007 grams, about 0.0075 grams, about 0.008 grams, about 0.0085 grams, about 0.009 grams, about 0.0095 grams, about 0.01 grams, about 0.015 grams, about 0.02 grams, about 0.025 grams, about 0.03 grams, about 0.035 grams, about 0.04 grams, about 0.045 grams rum, about 0.05 grams, about 0.055 grams, about 0.06 grams, about 0.065 grams, about 0.07 grams, about 0.075 grams, about 0.08 grams, about 0.085 grams, about 0.09 grams, about 0.095 grams, about 0.1 grams, about 0.15 grams, about 0.2 grams, about 0.25 grams, about 0.3 grams, about 0.35 grams, about 0.4 grams, about 0.45 grams, about 0.5 grams, about 0.55 grams, about 0.6 grams, about 0.65 grams In some embodiments, the amount of the glycerol in the composition may be greater than about 0.7 grams, about 0.75 grams, about 0.8 grams, about 0.85 grams, about 0.9 grams, about 0.95 grams, about 1 gram, about 1.5 grams, about 2 grams, about 2.5 grams, about 3 grams, about 3.5 grams, about 4 grams, about 4.5 grams, about 5 grams, about 5.5 grams, about 6 grams, about 6.5 grams, about 7 grams, about 7.5 grams, about 8 grams, about 8.5 grams, about 9 grams, about 9.5 grams, or about 10 grams.

[0081] In some embodiments, the amount of one or more compounds disclosed herein may range from about 0.0001 grams to about 10 grams, from about 0.0005 grams to about 9 grams, from about 0.001 grams to about 0.5 grams, from about 0.001 grams to about 8 grams, from about 0.005 grams to about 7 grams, from 0.01 grams to about 6 grams, from about 0.05 grams to about 5 grams, from about 0.1 grams to about 4 grams, from about 0.5 grams to about 4 grams, or from about 1 gram to about 3 grams.

[0082] Certain preferred embodiments are pharmaceutical compositions comprising an oral administration of the compounds disclosed herein and pharmaceutical excipients suitable for oral administration. In some embodiments, provided herein is a pharmaceutical composition for oral administration comprising (1) an effective amount of the compounds disclosed herein, (2) an effective amount of one or more second agents, and (3) one or more pharma- ceutically acceptable excipients for oral administration. In some embodiments, the pharmaceutical composition further comprises (4) an effective amount of a third agent.

[0083] In some embodiments, the pharmaceutical composition may be a liquid pharmaceutical composition suitable for oral administration. Pharmaceutical compositions suitable for oral administration may be available as separate dosage forms such as capsules, cachets, tablets, or liquids, solutions, aerosol sprays or suspensions of a predetermined amount of active ingredient, including powder or granules, water or non-aqueous liquids, or liquid emulsions in water or water in liquid emulsions. Such dosage forms may be produced by certain pharmaceutical methods, but all methods include the step of producing the composition by uniformly and intimately combining the active ingredient with liquid carriers, liposomes, or finely divided solid carriers, or both. In general, pharmaceutical compositions are formed by uniformly and intimately mixing the active ingredient with liquid carriers or finely divided solid carriers, or both, and, if necessary, shaping the product into the desired shape. For example, a tablet may be one or more components that are compressed or molded. Tablets may be produced by mixing a free-flowing form, such as the active ingredient in powder or granules, with selected excipients, such as, but not limited to, binders, lubricants, inert diluents, and / or surfactants or dispersing agent mixtures, and compressing in specialized equipment. Molded tablets can be produced by molding a mixture of the powdered compound moistened with an inert liquid diluent in a specialized device. The tablets, whether coated, uncoated, or nicked, can be formulated to provide release or controlled release of the active ingredient therein, thereby providing a sustained effect over an extended period of time, such as with glyceryl monostearate or glyceryl distearate. Oral preparations can be produced as hard gelatin capsules in which the active ingredient can be mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate, or kaolin, or as soft gelatin capsules in which the active ingredient can be mixed with water or an oil medium, such as peanut oil, liquid paraffin, olive oil, or mixtures thereof.

[0084] The active ingredient can be intimately combined with a pharma- ceutically acceptable carrier by conventional drug mixing techniques. The carrier can be in various forms, depending on the desired form of administration of the formulation. In preparing pharmaceutical compositions for oral dosage forms, any of the usual pharmaceutical media, such as water, glycols, oils, ethanol, flavoring agents, preservatives, coloring agents, and oral liquid formulations (e.g., liquids, solutions, elixirs) or aerosols, can be used as carriers, or carriers such as starch, sugar, microcrystalline cellulose, diluents, granulators, lubricants, binders, disintegrants, etc., can be used for oral solid formulations. In some embodiments, lactose is not used. In some embodiments, the compound can be mixed with lactose, sucrose, starch powder, cellulose esters of alkanoic acid, cellulose alkyl esters, talc, stearic acid, magnesium stearate, magnesium oxide, calcium phosphate, sodium phosphate, calcium sulfate, sodium sulfate, gelatin, gum arabic, sodium alginate, polyvinylpyrrolidone, and / or polyvinyl alcohol for further formulation. For example, suitable carriers for preparing solid oral dosage forms include powder, capsule, and tablet forms, hi some embodiments, tablets can be coated by standard aqueous or nonaqueous techniques.

[0085] Suitable binders for use in pharmaceutical compositions and dosage forms include, but are not limited to, corn starch, potato starch or other starches, gelatin, natural binders, synthetic gums such as gum arabic, sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum, cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate, calcium carboxymethylcellulose, sodium carboxymethylcellulose), polyvinylpyrrolidone, cellulose, pregelatinized starch, hydroxypropyl methylcellulose, microcrystalline cellulose, and mixtures thereof.

[0086] Examples of fillers suitable for use in pharmaceutical compositions and dosage forms include, but are not limited to, talc, calcium carbonate (e.g., granules or powder), microcrystalline cellulose, powdered cellulose, glucose binders, kaolin, mannitol, silicic acid, sorbitol, starch, pregelatinized starch, and mixtures thereof.

[0087] Disintegrants are described herein as being useful in pharmaceutical compositions to provide tablets that disintegrate when exposed to an aqueous environment. Too much disintegrant may cause the tablet to disintegrate in the bottle. Too little may not be enough to disintegrate, but may alter the rate and extent of release of the active ingredient in the dosage form. Thus, the amount of disintegrant must be sufficient, but not so small or large that it impairs the release of the active ingredient. The amount of disintegrant depends on the formulation and method of administration, but can be easily adjusted by one of skill in the art. About 0.5 to about 15% by weight or about 1 to about 5% by weight of disintegrant may be used in pharmaceutical compositions. Disintegrants for pharmaceutical compositions and for forming dosage forms include, but are not limited to, agar, alginic acid, calcium carbonate, microcrystalline cellulose closcar mellow sucrose povidone, sodium acetate, potato or tapioca starch, other starches, preformed starches, clays, other algae, other celluloses, gums, or mixtures thereof.

[0088] Lubricants can be used to form pharmaceutical compositions. They include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerol, sorbitol, mannitol, polyethylene glycol, other diols, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oils (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, soybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, or mixtures thereof. Lubricants can also include silica gel, solidified aerosol, or mixtures thereof. They can also be added in an amount of less than about 1% by weight of the pharmaceutical composition.

[0089] When aqueous suspensions and / or elixirs are used for oral administration, the active ingredient may be combined with various sweetening or flavoring agents, coloring agents or dyes, diluents such as emulsifying and / or suspending agents, water, ethanol, propylene glycol, glycerol, and combinations thereof.

[0090] Surfactants that can be used to form pharmaceutical compositions and dosage forms include, but are not limited to, hydrophilic surfactants, lipophilic surfactants, and mixtures thereof. Suitable hydrophilic surfactants will usually have an HLB value of at least about 10, and suitable lipophilic surfactants will usually have an HLB value of less than about 10. An empirical parameter that can be used to characterize relative hydrophilicity and hydrophobicity is the hydrophilic lipophilic balance value (HLB value). The lower the HLB value of a surfactant, the more lipophilic or hydrophobic it is and the higher its solubility in oil. On the other hand, active agents with higher HLB values ​​are more hydrophilic and have higher solubility in aqueous solutions. Hydrophilic surfactants are generally considered to have HLB values ​​greater than about 10, although in the case of anionic, cationic, or zwitterionic compounds, HLB values ​​are generally not applicable. Similarly, lipophilic (i.e., hydrophobic) surfactants have HLB values ​​of about 10 or less. However, the HLB value of a surfactant is only a rough marker for the general use of industrial, pharmaceutical, and cosmetic emulsions.

[0091] Hydrophilic surfactants can be either ionic or non-ionic.Suitable oligopeptide and polypeptide surfactants include, but are not limited to, alkyl ammonium salts, fusidate salts, fatty acid derivatives of amino acids, oligopeptides and polypeptides, derivatives of glycerol esters of amino acids, oligopeptides and polypeptides, lecithin and phospholipids and their derivatives, carnitine fatty acid ester salts, alkyl sulfate salts, fatty acid salts, docusate sodium, acyl lactates, tartrate esters of mono / diacetylated mono / diglycerides, succinylated mono / diglycerides, citrate esters of mono / diglycerides, and mixtures thereof. Ionic surfactants include, but are not limited to, lecithin, lysolecithin, phospholipids, lysophospholipids and their derivatives, carnitine fatty acid ester salts, alkyl sulfate salts, fatty acid salts, acyl lactate salts, diacetyl tartaric acid esters of mono / mono / diglycerides, succinylated mono / diglycerides, citrate esters of mono / diglycerides, and mixtures thereof. Hydrophilic nonionic surfactants include, but are not limited to, alkyl glycosides, alkyl maltose, alkyl thioglycosides, lauroyl polyethylene glycol glycerides, polyethylene glycols such as polyethylene glycol alkylphenols, polyoxyalkylene alkyl ethers such as polyoxyalkylene alkylphenols, polyoxyalkylene alkylphenol fatty acid esters such as polyethylene glycol fatty acid monoesters and polyethylene glycol fatty acid diesters, diol glycerol fatty acid esters, polyglycerin fatty acid esters, polyoxyalkylene sorbitol fatty acid esters, glycerol esters, vegetable oils, hydrogenated vegetable oils, fatty acids and sterols, polyoxyethylene sterols, derivatives thereof, polyoxyethylenated vitamins and derivatives thereof, polyoxyethylene-polyoxypropylene block copolymers, and mixtures thereof, hydrophilic transesterification products of polyethylene glycol sorbitan fatty acid esters with at least one polyol selected from the group consisting of triglycerides, vegetable oils, and hydrogenated vegetable oils.The polyol can be glycerol, ethylene glycol, polyethylene glycol, sorbitol, propylene glycol, pentaerythritol, or a carbohydrate. Other hydrophilic nonionic surfactants include, but are not limited to, PEG-10 laurate, PEG-12 laurate, PEG-20 laurate, PEG-32 laurate, PEG-32 dilaurate, PEG-12 oleate, PEG-15 oleate, PEG-20 oleate, PEG-20 dioleate, PEG-32 oleate, PEG-200 oleate, PEG-40 oleate, PEG-15 stearate, PEG-32 distearate lactone, PEG-40 stearate, PEG-100 stearate, PEG-20 dilaurate, PEG-25 glycerol trioleate, PEG-32 dioleate, PEG-20 glyceryl lauret lactone, PEG-30 glyceryl laureate, PEG-20 glycate, and the like.PEG-20 Glyceryl Oleate, PEG-30 Glycerol, PEG-30 Glyceryl, PEG-40 Castor Oil, PEG-40 Castor Oil, PEG-40 Castor Oil, PEG-40 Castor Oil, PEG-40 Castor Oil, PEG-40 Castor Oil, PEG-40 Castor Oil, PEG-40 Castor Oil, PEG-40 Castor Oil, PEG-40 Castor Oil, PEG-40 Castor Oil, PEG-40 Castor Oil, PEG-40 Castor Oil, PEG-40 Castor Oil, PEG-40 Castor Oil, PEG-40 Castor Oil, PEG-40 Castor Oil, PEG-60 Corn Oil, PEG-6 Glyceryl / Capric Glyceride, PEG-8 Capric / Capric Glyceride, Polyglyceryl 1-10 Laurate, PEG-30 Cholesterol, PEG-25 Phytosterol, PEG-30 Also included are, but are not limited to, soy sterols, PEG-20 trioleate, PEG-40 sorbitol oleate, PEG-80 sorbitan laurate, polysorbate 20, polysorbate 80, POE-9 dodecyl ether, POE-23 lauryl ether, POE-10 oleyl ether, POE-20 oleyl ether, POE-20 stearin, PEG-100 tocopherol succinate, PEG-24 cholesterol, Tween 40, Tween 60, sucrose monostearate, sucrose monolaurate, sucrose monopalmitate, PEG 10-100 nonylphenol series, PEG 15-100 octylphenol series, and poloxamers. Suitable lipophilic surfactants include, but are not limited to, fatty alcohols, glycerin fatty acid esters, acetylated glycerol fatty acid esters, lower alcohol fatty acid esters, propylene glycol fatty acid esters, sorbitol fatty acid esters, diol sorbitan fatty acid esters, sterols and sterol derivatives, polyoxyethylenated sterols and sterol derivatives, polyethylene glycol alkyl ethers, sugar esters, sugar ethers, lactic acid derivatives of mono / diglycerides, and the like.

[0092] The pharmaceutical composition may include a solubilizing agent to ensure good solubilization and / or dissolution of the compound and minimize precipitation of the compound. Solubilizing agents may be particularly useful in pharmaceutical compositions for parenteral applications, such as injectable pharmaceutical compositions. Solubilizing agents may be added to enhance the solubility of other components, such as hydrophilic drugs and / or surfactants, or to maintain the pharmaceutical composition as a stable or homogeneous solution or dispersion. Examples of suitable solubilizing agents include, but are not limited to, alcohols and polyols, such as ethanol, isopropanol, butanol, benzyl alcohol, ethylene glycol, propylene glycol, butylene glycol and its isomers, glycerol, pentaerythritol, sorbitol, mannitol, dimethylisosorbide, polyethylene glycol, polypropylene glycol, polyvinyl alcohol, and other cellulose derivatives, cyclodextrins and cyclodextrin derivatives, ethers of polyethylene glycol with molecular weights of about 200 to about 6000, such as tetrahydrofurfuryl alcohol PEG ether (tetrahydrofuran polyglycol ether) or methoxy PEG. These include, but are not limited to, amides and other nitrogen-containing compounds such as 2-pyrrolidone, 2-piperidone, ε-caprolactam, N-alkylpyrrolidone, N-hydroxyalkylpyrrolidone, N-alkylpiperidine, N-alkylcaprolactam, dimethylacetamide, polyvinylpyrrolidone, ethyl propionate, esters, acetyl triethyl citrate, triethyl citrate, triethyl citrate, ethyl oleate, ethyl octanoate, ethyl butyrate, glycerol triacetate, propylene glycol monoacetate, propylene glycol diacetate, ε-caprolactone and its isomers, delta-valine esters and its isomers, butyrolactone and its isomers, and other known solubilizing agents such as dimethylacetamide, dimethyl isosorbide, N-methylpyrrolidone, diethylene glycol monoethyl ether, water, etc. Mixtures of solubilizing agents may also be used.

[0093] The amount of a given solubilizer is limited to a biologically acceptable amount and can be easily determined by those skilled in the art. The solubilizer can be determined in a weight ratio of about 10%, about 25%, about 50%, about 100%, or up to about 200% based on the total weight of the drug and other excipients. It can also be about 5%, 2%, 1% or less, if necessary. Usually, the solubilizer can be present in a weight ratio of about 1% to about 100%, about 5% to about 25%.

[0094] The described pharmaceutical compositions may also include one or more pharma- ceutically acceptable additives and excipients, flavoring agents, coloring agents, suspending agents, binders, fillers, plasticizers, lubricants, and mixtures thereof. Preservatives include, but are not limited to, antioxidants, chelating agents, antibacterial preservatives, antifungal preservatives, alcohol preservatives, acidic preservatives, other preservatives, and the like. Antioxidants include, but are not limited to, alpha-tocopherol, ascorbic acid, butyl hydroxyanisole, butyl hydroxytoluene, monothioglycerol, potassium pyrosulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium sulfite. Chelating agents include, but are not limited to, ethylenediaminetetraacetic acid (EDTA), citrate monohydrate, disodium ethylenediaminetetraacetate, dipotassium ethylenediaminetetraacetate, edetic acid, fumaric acid, malic acid, phosphoric acid, sodium edetate, tartaric acid, and triethylenediaminetetraethyl citrate. Antimicrobial preservatives include, but are not limited to, for example, benzalkonium chloride, benzethonium chloride, benzyl alcohol, bromonitropyrene glycol, cetrimonium bromide, cetylpyridinium chloride, chlorocresol, cresol, ethanol, glycerol, heptacidine, imidazolidine, phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate, and propylene glycol. Examples of antifungal agents include, but are not limited to, butyl parahydroxybenzoate, methyl parahydroxybenzoate, ethyl parahydroxybenzoate, propyl parahydroxybenzoate, benzoic acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium benzoate, sodium propionate, sorbic acid, etc. Examples of preservatives include, but are not limited to, ethanol, polyethylene glycol, phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoate, phenylethanol, etc. Examples of acidic preservatives include, but are not limited to, vitamin A, vitamin C, vitamin E, beta carotene, citric acid, acetic acid, dehydroacetic acid, ascorbic acid, sorbic acid, phytic acid, etc.Other preservatives include, but are not limited to, for example, tocopherol acetate, cetrimonium bromide, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium bisulfate, sodium metabisulfite, potassium metabisulfite, and methyl p-hydroxybenzoate. In certain embodiments, the preservative can be an antioxidant. In other embodiments, the preservative can be a chelating agent.

[0095] In some embodiments, provided herein are pharmaceutical compositions for parenteral administration comprising (1) an effective amount of a disclosed compound, optionally (2) an effective amount of one or more second reagents, (3) one or more pharmaceutical excipients suitable for parenteral administration, and (4) an effective amount of a third reagent.

[0096] The pharmaceutical compositions may be administered in the form of aqueous or oily suspensions or emulsions, sesame oil, corn oil, cottonseed oil or peanut oil, and elixirs, mannitol, glucose, or sterile aqueous solutions, and similar pharmaceutical carriers may be used. Saline solutions are also commonly used for injection. Ethanol, glycerol, propylene glycol, liquid polyethylene glycol, benzyl alcohol, and the like (and suitable mixtures thereof), cyclodextrin derivatives, sodium chloride, tragacanth, buffers, and vegetable oils may also be used. Proper fluidity can be maintained by the use of coating agents such as lecithin, and by maintaining the desired particle size in the case of dispersions using surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, p-hydroxybenzoic acid esters, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. The pharmaceutical composition may be injected with a suitable carrier including saline, glucose or water, or may be solubilized with cyclodextrins, cosolvents (eg propylene glycol) or micelles (eg Tween 80).

[0097] Sterile injectable solutions can be prepared by filtration and sterilization using the desired amount of the compounds disclosed herein with the appropriate solvent for the various other ingredients listed above. Typically, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile carrier containing a basic dispersion medium and the appropriate other ingredients listed above. Sterile injectable solutions are prepared from sterile powders, some of which are carried out by vacuum drying and freeze-drying techniques to produce the active ingredient and the other sterile-filtered ingredients listed above. Sterile injectable preparations can also be prepared by a solution of a non-toxic parenterally acceptable diluent or solvent, such as a solution in 1,3-butanediol or a sterile injectable solution. Acceptable carriers and solvents that can be used include, but are not limited to, water, Ringer's solution, isotonic sodium chloride solution, and the like. Sterile, fixed oils are also commonly used as solvents or suspending media, including, but not limited to, synthetic monoglycerides or diglycerides. In addition, fatty acids, such as oleic acid, can also be used in the preparation of injectables. Injectable formulations can be sterilized, for example, by a bacterial-retaining filter or by adding sterilizing agents incorporated into sterile solid compositions that can be dissolved or dispersed in sterile water or other sterile injectable medium. Injectable compositions can be in a weight ratio of about 0.1% to about 5% of the compounds disclosed herein.

[0098] The pharmaceutical compositions provided herein can be formulated in solid, semi-solid or liquid forms suitable for topical or external use, such as gelling agents, water-soluble gels, liniments, creams, lotions, suspensions, foams, powders, ointments, solutions, oils, pastes, suppositories, sprays, emulsions, saline, dimethylsulfoxide (DMSO)-based solutions, etc. Generally, carriers with higher density can provide areas with prolonged exposure to the active ingredient. In contrast, solution formulations result in more direct contact with the selected area by the active ingredient. For example, ointment formulations can be paraffinic or water-miscible. Alternatively, the active ingredient is formulated as an oil-in-water cream-based cream. The aqueous phase of the cream matrix can include at least about 30% by weight of a polyol, such as propylene glycol, butane-1,3-diol, mannitol, sorbitol, glycerol, polyethylene glycol, and mixtures thereof. The pharmaceutical compositions may also include suitable solid or gel phase carriers or excipients that enhance or aid in the delivery of the compound through the stratum corneum skin barrier layer. Examples include urea (e.g., urea), (e.g., menthol), amines, amides, alkanes, alkanols, water, and polymers such as isopropyl myristate and sodium sulfate, pyrrolidone, glycerol monolaurate, sulfoxides, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polyethylene glycol.

[0099] The pharmaceutical composition of the present invention can be administered in the form of suppositories for rectal administration. These compositions can be prepared by mixing the compound of the present invention with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature and melts in the rectum to release the active ingredient. Such materials include, but are not limited to, polyethylene glycol, beeswax, and cocoa butter.

[0100] The pharmaceutical compositions of the present invention can be administered by nasal aerosol or inhalation. The compositions can be prepared as saline solutions using known pharmaceutical preparation techniques and can include the use of benzyl alcohol or other suitable preservatives to enhance bioavailability, absorption enhancers, fluorocarbons, and other solubilizing or dispersing agents common in the art.

[0101] When applying a therapeutic agent to a subject, it can be administered locally at a target site. A variety of techniques can be used to apply the host composition to the target site, such as by injection, catheter, gel, stent, trocar, propellant, drug-releasing polymer, or other device to provide internal access.

[0102] In another embodiment, the present invention provides an implantable medical device that comprises a compound of the present invention or a composition containing a compound of the present invention, such that the compound of the present invention provides an effective treatment.

[0103] In another embodiment, the present invention provides a method of injecting an implantable drug delivery device, comprising the step of contacting the drug delivery device with a compound or composition of the present invention, the implantable drug delivery device including, but not limited to, biodegradable polymer capsules or pills, non-degradable, dispersible polymer capsules, and biodegradable polymer flakes.

[0104] In another embodiment, the composition of the present invention also includes a second therapeutic agent. The second therapeutic agent includes a specific compound or therapeutic agent that is known to exert or exerts beneficial properties when administered alone or in combination with any of the compounds of the general formula herein. Drugs that can be combined with these compounds to obtain beneficial properties include other kinase inhibitors and / or other chemotherapeutic agents for treating the diseases and disorders listed above. Such agents are described in detail in the art. It is preferred if the second therapeutic agent is an agent that can be used to treat or prevent a disease or condition selected from cancer.

[0105] In another embodiment, the present invention provides separate dosage forms of a compound of the present invention and a second therapeutic agent associated with each other. As used herein, the term "associated with each other" means that the individual dosage forms are packaged together or linked together such that the individual dosage forms are provided or administered at the same time (less than 24 hours, consecutively or simultaneously).

[0106] The pharmaceutical compositions of the invention contain an effective amount of a compound of the invention. As used herein, the term "effective amount" refers to an amount that, when administered in a proper dosing regimen, is sufficient to reduce or ameliorate the severity, duration, or onset of the disorder being treated, prevent the progression of the disorder, halt the disorder being treated, or enhance or improve the prophylactic or therapeutic effects of another therapy.

[0107] An effective amount of the compounds of the present invention may range from about 0.001-1 mg / kg to about 500 mg / kg, about 0.01 mg / kg to about 50 mg / kg, about 0.1 mg / kg to about 2.5 mg / kg. As will be appreciated by those of skill in the art, the effective dosage will vary depending on the condition being treated, the severity of the condition, the route of administration, the age, sex, and general health of the patient, the use of excipients, and other treatment methods commonly used (e.g., the use of other drugs), and the judgment of the treating physician.

[0108] In the case of pharmaceutical compositions containing a second therapeutic agent, the effective amount of the second therapeutic agent is between about 20% and 100% of the dose normally used in a monotherapy regimen using only the agent, and preferably between about 70% and 100% of the normal monotherapy dose.

[0109] Some of the second therapeutic agents mentioned herein are expected to act synergistically with the compounds of the invention. If so, the effective dose of the second therapeutic agent and / or the compounds of the invention will be lower than that required for monotherapy. This has the advantage of minimizing secondary side effects of the second therapeutic agent or the compounds of the invention, improving efficacy, ease of administration or use, and / or reducing the overall cost of manufacturing or formulating the compounds.

[0110] Treatment includes the following:

[0111] According to another embodiment, the present invention provides a method of treating a subject suffering from or susceptible to a disease or disorder or a symptom thereof (e.g., as described herein), comprising administering to said subject an effective amount of a compound or composition of the present invention. These diseases are well known in the art and disclosed herein.

[0112] This includes the treatment of diseases mediated by protein kinases, such as ATM kinase.

[0113] In another aspect, the invention provides a method of treating a disease in a subject comprising administering to the subject a composition comprising any of the compounds of the general formulae herein.

[0114] In certain embodiments, the disease is mediated by ATM kinase.

[0115] In another embodiment, the disease is cancer or a proliferative disease.

[0116] In another embodiment, the compounds of formula (I) and pharmaceutically acceptable salts as inhibitors against ATM kinase are expected to be present or partially mediated by the activity of ATM kinase, such as the treatment of cancer or disease conditions, which may be used to treat the types of cancers treated with the compounds of formula (I) or pharmaceutically acceptable salts, including, but not limited to, ovarian cancer, cervical cancer, colon cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer, leukemia, lymphoma, non-Hodgkin's lymphoma, gastric cancer, lung cancer, liver cancer, bone cancer, gastrointestinal stromal tumor (GIST), thyroid cancer, cholangiocarcinoma, endometrial cancer, renal cell carcinoma, anaplastic large cell lymphoma, acute myeloid leukemia (AML), multiple myeloma, melanoma, mesothelioma, brain cancer, adenocarcinoma, skin cancer, or head and neck squamous cell carcinoma.

[0117] In another embodiment, the disease is glioma.

[0118] In another embodiment, the disease is non-small cell lung cancer (NSCLC) with central nervous system metastases.

[0119] In another embodiment, the disease is a central nervous system disease.

[0120] In one embodiment, the method of the present invention is used to treat a subject suffering from or susceptible to a disease or condition. These diseases, disorders, or symptoms thereof include, for example, those controlled by protein kinases (e.g., ATM protein kinase). The disease or disease symptoms may be, for example, cancer or a proliferative disease or disorder. The disease or disease symptoms may be ovarian cancer, cervical cancer, colon cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer, leukemia, lymphoma (GIST), thyroid cancer, cholangiocarcinoma, endometrial cancer, kidney cancer, anaplastic large cell lymphoma, acute myeloid leukemia (AML), gastric cancer, lung cancer, liver cancer, multiple myeloma, melanoma, mesothelioma, brain tumor, membranous adenocarcinoma, skin cancer, or head and neck squamous cell carcinoma. The methods described herein include subjects identified as in need of the specifically described treatment. Subject identification requires that treatment is within the judgment of the subject or a medical professional and may be subjective (eg, opinion) or objective (eg, measurable by testing or diagnostic methods).

[0121] In another embodiment, the compounds of the general formulas herein (and compositions thereof) are useful for treating diseases or disorders that have been treated with other therapeutic agents (e.g., anti-cancer agents, neurotrophic agents, psychotropic agents, cardiovascular disease therapeutic agents, anti-obesity agents, or anti-diabetic agents) and make the subject resistant. In one aspect, the methods herein include administering a compound of the formulas of the methods (or compositions thereof) to a subject that is resistant to the treatment (or has been identified as resistant to treatment with gefitinib, erlotinib). In another aspect, the subject thus responds to the treatment, resulting in the disorder being regulated or improved prior to treatment with the compounds of the formulas of the present invention.

[0122] In another embodiment, the invention provides a method for modulating the activity of a protein kinase (e.g., a protein kinase, a kinase described herein) in a cell comprising contacting the cell with one or more compounds of the general formulae herein.

[0123] The above anti-cancer treatments can be carried out as monotherapy or in combination with conventional compounds, or as radiotherapy, chemotherapy or immunotherapy using the compounds of the invention.Such chemotherapy may be administered simultaneously, sequentially or separately in combination with the compounds of the present invention and may include antiproliferative / antitumor agents, alkylating agents (e.g., cisplatin, oxaliplatin, carboplatin, cyclophosphamide, nitrogen mustard, melphalan, chlorambucil, busulfan, temozolomide and nitrosoureas), antimetabolites (e.g., antifungal acids such as gemcitabine, 5-fluorouracil and tegafur, raltitrexed, methotrexate, cytarabine and hydroxyurea), antitumor antibiotics (e.g., doxorubicin, bleomycin, adriamycin, daunorubicin, epirubicin, idarubicin, mitomycin, C, gentamicin, grilamycin, and other anthracyclines), antimitotics (e.g., vincal kaloids such as vincristine, alkaloids such as paclitaxel and tacrolimus, and polo kinase inhibitors), and topoisomerase inhibitors (e.g., the epipodophyllotoxins etoposide, pyridine glycosides, acridines, topotecan, and camptothecin), cell proliferation inhibitors such as antihormones (e.g., tamoxifen, fluvatrant, toremifene, raloxifene, droloxifene, and difoxifene), antiandrogens (e.g., amylamines, flutamide, nilutamide acetate, and cyclopropanone), LHRH antagonists or LHRH agonists (e.g., These may include, but are not limited to, one or more of the following categories of anti-tumor agents: 5-alpha reductase inhibitors such as goserelin, leuprolide, and bucorin, progesterones (e.g., megestrol acetate), aromatase inhibitors (e.g., anastrozole, letrozole, buoxazole, and exemestane) and finasteride, anti-invasion agents (e.g., c-Src kinase family inhibitors such as cetatinib, dasatinib, and bosutinib, and bosustifen), and metalloprotease inhibitors such as inhibitors of horse, urokinase plasminogen activator receptor, or antibody heparinase.Inhibitors of growth factor function, such as, for example, such inhibitors, include growth factor antibodies and growth factor receptor antibodies (e.g., the anti-erbB2 antibody trastuzumab [Herceptin brand], the anti-EGFR antibody panitumumab, the anti-ErbB antibody cetuximab (Erbito, C225) and those by Stem et al.) A critical review in Oncology / Hematology disclosed growth factor receptors or growth factor receptor antibodies (2005, Vol. 54, pp. 11-29). Aurora kinase inhibitors. Such inhibitors also include tyrosine kinase inhibitors such as epidermal growth factor family inhibitors (e.g., EGFR family inhibitors such as gefitinib, erlotinib, icotinib, afatinib, dacomitinib and tagrisso, erbB2 tyrosine kinase inhibitors such as lapatinib, neratinib), hepatocyte growth factor family inhibitors, platelet derived growth factor family inhibitors such as imatinib and / or nilotinib, inhibitors of serine / threonine kinases (e.g., RAS / RAF signaling inhibitors, e.g., sorafenib, phenyltransferase inhibitors such as rifabutinib, tipifarnib, lonafarnib), MEK and / or AKT kinase cell signaling inhibitors, c-kit inhibitors, abl fusion kinase inhibitors, PI3 kinase inhibitors, PLT3 kinase inhibitors, CSF-1R kinase inhibitors, IGF receptor (insulin-like growth factor) kinase inhibitors, Aurora kinase inhibitors, cyclin dependent kinase inhibitors such as CDK2 and / or CDK4 inhibitors, angiogenesis inhibitors, for example the antibody bevacizumab (Avastin), which inhibit the effects of vascular endothelial growth factor. 商標), as well as VEGF receptor tyrosine kinase inhibitors, such as vandetanib, vatalanib, sunitinib, axitinib, pazopanib and cediranib, compounds acting via other mechanisms (e.g. tricarboxyaminoquinolines, integrin αV3 function inhibitors and angiogenesis inhibitors), antisense (nucleic acid) therapy including replacement of abnormal genes, such as the above-mentioned abnormal p53 and abnormal BRCA1 or BRCA2, such as ISIS 2503, anti-ras gene antisense (nucleic acid) (e.g. olaparnib, niraparib, rucaparib, talazoparib), immunogenicity of T cells, using cytokines such as interleukin 2, 4, granulocyte macrophage stimulating factor, transfected immune cells such as cytokine-transfected dendritic cells, cytokine-transfected anti-idiotypic antibodies, regulatory T cells, medullary inhibitory cells, or IDO, TDO methods of improving a patient's resistance to chemotherapy or radiation therapy, such as enzymes using cytosine deaminase, thymidine kinase, bacterial nitroreductase, and multidrug resistance gene therapy immunotherapy, and the use of antibodies derived from tumor associated antigens, such as proteins or peptides derived from NY-ES0-1, MAGE-3, WTI or HER2 / neu, or other drugs commonly used as foundational drugs or adjuvants in cancer treatment regimens (e.g., antiemetics, anti-anemic drugs, etc.).

[0124] As used herein, the term "co-administration" means that the second therapeutic agent may be administered in combination with the compound of the present invention, either partially as a single dosage form (e.g., a composition comprising the compound of the present invention and the second therapeutic agent described above) or as an independent multiple dosage form. Alternatively, the additional agent may be administered before, in conjunction with, or after administration of the compound of the present invention. In such combination therapy, the compound of the present invention and the second therapeutic agent are administered by conventional methods. The administration of the composition of the present invention comprising the compound of the present invention and the second therapeutic agent to a subject does not preclude either the same therapeutic agent, another second therapeutic agent, or the compound of the present invention being administered independently to the subject at other times during the course of treatment. The administration of the second component sequentially or separately, or the administration is delayed, should not eliminate the benefits of the effect obtained from the use of the combination.

[0125] In one embodiment of the present invention, when a second therapeutic agent is administered to a subject, the effective amount of the compound of the present invention is lower than the effective amount of the second therapeutic agent when the second therapeutic agent is not administered. In another embodiment, the effective amount of the second therapeutic agent is lower than the effective amount of the second therapeutic agent when the compound of the present invention is not administered. In this way, undesirable side effects associated with any of the doses of the drugs can be minimized. The potential advantages to one of ordinary skill in the art are clear (e.g., including, but not limited to, improved dosing schedules and / or reduced drug costs).

[0126] In another aspect, the invention provides the use of any of the compounds of the general formula herein, alone or in combination with one or more of the second therapeutic agents described herein, either as a single composition or as separate dosage forms, in the manufacture of a medicament for the treatment or prevention of a disease, disorder or condition described herein in a subject. Another aspect of the invention is the use of a compound of the general formula herein for the treatment or prevention of a disease, disorder or condition described herein in a subject.

[0127] In other embodiments, the methods herein further include a method of monitoring the subject's response to the therapeutic administration. Such monitoring may include periodically sampling the subject's tissues, bodily fluids, cerebrospinal fluid, samples, cells, proteins, chemical markers, genetic material, etc., as markers or indicators of the treatment regimen. In other methods, subjects are pre-screened or identified as in need of such treatment by assessing the suitability of relevant markers or indicators for such treatment.

[0128] In one embodiment, the present invention provides a method for monitoring the progress of a treatment. The method includes determining a diagnostic marker (marker) in a subject suffering from or susceptible to or diagnosed (e.g., screened, assayed) a disorder or condition described herein (e.g., any target or cell type described herein modulated by a compound described herein), and the subject is administered a therapeutic amount of a compound of the present invention sufficient to treat the disease or condition. The level of the marker determined in this manner can be compared to well-known levels in healthy normal controls or other affected patients to establish the disease state of the subject. In a preferred embodiment, a second level of the marker in the subject is measured at a later time point than the measurement of the first level, and the two levels are compared to monitor the progression of the disease or the effectiveness of the treatment. In certain preferred embodiments, the pre-treatment level of the marker in the subject is measured before starting treatment according to the present invention, and the pre-treatment level of the marker can be the same as the level of the marker in the subject after starting treatment to determine the effectiveness of the treatment.

[0129] In certain method embodiments, the level of the marker or marker activity in the subject is determined at least once. The marker level can be compared, for example, with another measurement from the same patient, another patient, or another subject obtained previously or subsequently from said subject, to determine whether the treatment according to the invention has the desired effect, and, if necessary, to adjust the dose level. The determination of the level of the marker can be performed using any suitable sampling / expression assay method known in the art or described herein. Preferably, a tissue or liquid sample is first removed from the subject. Examples of suitable samples include blood, urine, cerebrospinal fluid, tissue, mouth or cheek cells, and hair samples including roots. Other suitable samples are known to those skilled in the art. The measurement of protein levels, ctDNA, cfDNA and / or mRNA levels (e.g., marker levels) in the sample can utilize any suitable technique known in the art, including, but not limited to, enzyme immunoassay, ELISA, radiolabeling techniques, Western blot / chemiluminescence, real-time PCR, electrochemical signals, and the like.

[0130] The present invention also provides kits for treating diseases, disorders, or conditions described herein. Such kits include (1) a pharmaceutical composition comprising any of the compounds of the general formulas herein or salts thereof, or a prodrug thereof, or a salt thereof, or a hydrate, solvate, or polymorph thereof in a container, and (2) a description of a method for treating a disease, disorder, or condition described herein using the pharmaceutical composition. The container can be any container or other sealed or sealable device that can contain the pharmaceutical composition. Examples include bottles, reservoir bottles with separate or multiple chambers, each compartment or section containing a single dose of the composition, and separate foil packages, each compartment containing a single dose of the composition, dispensing the single dose of the composition. The container can be of any conventional shape or form known in the art, made of pharma- ceutically acceptable materials, such as paper or cardboard boxes, glass or plastic bottles or cans, resealable bags (e.g., tablet "refills" used to place into another container), or single dose blister packs used to push the package out of the treatment schedule. The container used may depend on the exact dosage form involved, for example a conventional cardboard box would not normally be used to contain a liquid suspension. It is possible for multiple containers to be used together in one package to sell a single dosage form. For example tablets may be contained in a bottle which is then housed in a box. Preferably the container is a blister pack.

[0131] The kit may further include information and / or instructions from a physician, pharmacist, or the subject. These memory aids include numbers (corresponding to the order or number of days that the capsules should be taken) printed on each compartment or divider containing the medication, number of weeks printed on each compartment or divider, or cards containing the same type of information.

[0132] When the present invention is used with ATM kinase, the enantiomeric purity of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) of the present invention is greater than about 55% to about 99.5%, greater than about 60% to about 99.5%, greater than about 65% to about 99.5%, greater than about 70% to about 99.5%, greater than about 75% to about 99.5%, greater than about 80% to about 99.5%, greater than about 85% to about 99.5%, greater than about 90% to about 99.5%, greater than about 95% to about 99.5%, greater than about 96% to about 99.5%, greater than about 97% to about 99.5%, greater than about 100% to about 100%, greater than about 98% to about 99.5%, greater than about 99% to about 99.5%, or more.

[0133] The compounds described herein can be evaluated for their biological activity using protocols known in the art, including, for example, those described herein. Some of the compounds herein exhibit unexpectedly superior properties (e.g., metabolic stability, high selectivity, low excretion rate, high permeability, non-P-glycoprotein efflux substrates, etc.), making them excellent candidates as potential therapeutic agents.

[0134] All references cited herein, whether electronic, printed, computer readable, or otherwise, are expressly incorporated herein by reference in their entirety, and include, but are not limited to, abstracts, articles, journals, publications, textbooks, papers, technical data sheets, internet sites, databases, patents, patent applications, and patent publications.

[0135] The present invention will be described in detail below with reference to examples. The following examples will help those skilled in the art to further understand the present invention without limiting the present invention. It should be noted that many modifications and improvements can be made by those skilled in the art without departing from the spirit of the present invention. All of these are within the scope of the present invention. EXAMPLES

[0136] Example 1. Preparation of 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivatives (I)

[0137] 1.1 The intermediate 3,3-difluoro-1-methylpiperidin-4-amine A6 was synthesized as follows. [ka] Step 1: At 25° C., a solution of 2-methylpropane-2-sulfinamide (404 g, 3.33 mol) and TI(OEt)4 (1003 g, 4.44 mol) was added to THF (2.5 L) and added dropwise to a solution of A1 (500 g, 2.22 mol). The reaction mixture was stirred at 70° C. for 1 h and then cooled to 0° C. The reaction mixture was then poured into NaBH4 (166 g, 2.0 equiv.) in THF (2.5 L) in a separate reactor at 0° C. The reaction mixture was stirred at 0° C. for 0.5 h and then slowly warmed to 25° C. and continued to stir for 0.5 h. Methanol (2.5 L) was added and then stirred for 0.5 h. The mixture was added to a vessel containing saturated NaCl solution (2.5 L) and stirred at 25° C. for 1 h. The reaction mixture was extracted twice with DCM (2.5 L). The organic layer was dried over anhydrous sodium sulfate, filtered and evaporated in vacuo to give product A2.

[0138] Step 2: A solution of crude A2 in ethyl acetate (1 L) at 25° C. was added to HCl (5.0 equiv., 4 M in ethyl acetate), stirred at 25° C. for 1 h, and filtered to give A3.

[0139] Step 3: Water (1.5 L) was added to the filter cake A3, and then the pH was adjusted to 8-9 with 20% aqueous solution. NaOH solution was added at 25 °C. The mixture was extracted with DCM (1.5 L), and the organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Triethylamine (267 g, 1.2 equiv.) was added to the crude product, followed by (Boc)2O (577 g, 1.2 equiv.), and the mixture was stirred at 25 °C for 1 h. Then 10% citric acid solution (2 L) was added to the mixture, and then the organic layer was separated. The aqueous layer was extracted with DCM (1 L). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give A4 (430 g, 59.4%).

[0140] Step 4: To a solution of A2 (380 g, 1.0 equiv) in MeOH (3.8 L) was added Pd(OH)2 / C (20%, w / w) and the mixture was stirred at 20 atm for at least 1 h at 25° C. The mixture was filtered and the filtrate was concentrated to give A5 (250 g, 91%).

[0141] Step 5: To a solution of A5 (236 g, 1.0 equiv.) in MeOH (2.4 L) at 5° C. was added HCHO (37% in water, w / w). The reaction mixture was stirred at 5° C. for 1 h, then NaBH(OAc)3 was added portionwise until LC-MS showed disappearance of A5. 10% aqueous NaOH solution was added to the reaction mixture. The pH of the NaOH solution was adjusted to 8-9 at 25° C. and stirred at 25° C. for 0.5 h. After evaporating the methanol under vacuum, the residue was extracted with ethyl acetate (1.2 L). The organic layer was separated and concentrated. HCl (5.0 equiv., 4 M in ethyl acetate) was added followed by filtration. The filter cake was collected and dried under vacuum at 45° C. to give the HCl salt of A6 (125.1 g, 53.7%).

[0142] 1.2 Synthesis of 1-(3,3-difluoro-1-methylpiperidin-4-yl)-8-(6-methoxypyridin-3-yl)-3-methyl-1,3-dihydro-2H-imidazo[4,5-c]quinolin-2-one derivatives (I):

[0143] The synthetic route is as follows: [ka]

[0144] Step 1: To a mixture of B1 (25 g, 79.48 mmol) in DMA (500 mL) was added 2 (14.92 g, 99.34 mmol) and DIEA (30 g, 238.744 mmol). The mixture was heated at 60 °C for 5 h. The mixture was cooled to room temperature and poured into a mixture of DCM (500 mL) and H2O (250 mL). The organic layer was separated, washed with brine, dried over anhydrous Na2SO4, filtered and concentrated under vacuum to give B2 (15 g, 44.09%) as an off-white solid. LC-MS: (ESI) m / z=404[M+H]+.

[0145] Step 2: To a mixture of B2 (15 g, 33.35 mmol) in MeOH / H2O / THF (300 mL / 150 mL / 150 mL) at room temperature was added NaOH (8 g, 100 mmol) and heated at 50° C. for 2 h. The mixture was cooled to room temperature and concentrated in vacuo to remove the organic layer. The residue was added to HO (150 mL), adjusted to pH = 4 with 10% HCl, and filtered to give B3 (11 g, 78.57%) as an off-white solid: 1H NMR (400 MHz, DMSO-d6) δ 12 -11.85(m, 1H), 8.80-8.65 (m, 1H), 8.01-7.67 (m, 2H), 5.35-5.01 (m, 1H), 3.68-3.51(m, 1H), 3.30-3.22 (m, 1H), 3.17 (d, J = 8.0 Hz, 1H), 2.92 (d, J = 12.0 Hz, 1H), 2.63-2.53(m, 1H), 2.47-2.29(m, 4H), 2.07-1.92(m, 1H).

[0146] Step 3: To a solution of B3 (11 g, 27.5 mmol) and DPPA (7.12 mL, 33 mmol) in DMF (350 mL) was added TEA (28.2 mL, 82.5 mmol). The mixture was stirred at 60 °C for 2 h. The reaction was cooled to room temperature and then poured into H2O (500 mL). The precipitate was filtered, washed with H2O (50 mL), and the filter cake was dried to give B4 (10 g, 91.82%) as a brown solid.

[0147] Step 4: To a mixture of B4 (10 g, 25.2 mmol) in DMF (250 mL) at room temperature was added DMF-DMA (15.15 g, 126 mmol). The mixture was then heated to 80° C. for 2 h, and the mixture was cooled to room temperature and then filtered. The filtrate was washed with HO and dried to give B5 (9 g, 86.95%) as an off-white solid. 1H NMR(400MHz,DMSO-d6) δ 9.05-8.95(m,1H),8.75-8.36(m,1H),8.04-7.71(m,2H),5.37-5.16(m,1H),3.67-3.55(m,1H),3.50(s,2H),3 .30-3.23(m,1H),2.93(d,J=8.0Hz,1H),2.74-2.53(m,1H),2.48-2.32(m,4H),2.07-1.93(m,1H).LC-MS:(ESI) m / z=411,413[M+H]+.

[0148] Step 5: A mixture of B5 (2 g, 4.87 mmol), (6-methoxypyridin-3-yl)boronic acid (888 mg, 5.844 mmol), PdCl2(dtdppf) (318 mg, 4.87 mmol) and K2CO3 (2 g, 14.61 mmol) in 1,4-dioxane / HO (100 mL / 40 mL) was degassed twice with N2 and then heated at 80 °C for 3 h. The mixture was poured into a mixture of DCM (500 mL) and HO (500 mL), and the organic layer was washed with brine, dried over anhydrous Na2SO4 and purified by column chromatography (DCM:MeOH = 30:1) to give product 1 (1.2 g, 56.07%) as a white solid. 1H NMR(400MHz,DMSO) δ 8.94 (d,J =25.6 Hz,1H),8.80 (s,1H),8.68 (dd,J =39.6,2.0Hz,1H),8.38 (s,1H),8.29-8.10(m,2H),8.05-7.95 (m,1H),7.01 (dd,J=18.8,8.8Hz,1H),5.45-5.25(m,1H),3.94(d,J=4.8Hz,3H),3.73-3.61(m,1H),3.56(d,J=33.2Hz,3H),3.44- 3.35(m,1H),3.15-2.87(m,1H),2.72-2.53(m,1H),2.48-2.39(m,2H),2.35(s,2H),2.10-1.91(m,1H).LC-MS:(ESI) m / z=440.1 [M+H]+.

[0149] The sample was dissolved in approximately 150 mL of EtOH and injected every 7 mL using a chiral Pre-SFC column (WateRs SFC150, 250×25 mm, 10 m DAI-CEL CHIRALPAK® AS SFC column) to separate the enantiomers (55:45 supercritical CO2:EtOH mobile phase at a flow rate of 70 g / min and monitoring wavelength of 214 nm). The R enantiomer was dissolved in 2,2,2-trifluoroethanol and concentrated to give the amorphous free base product.

[0150] The above method is amorphous, and the crystal form changes during different solvent systems and heating processes. It is difficult to prepare solid formulations under pharmaceutical processing conditions. Therefore, it is necessary to further prepare the preferred free base crystal form, salt form and salt crystal form of 1-(3,3-difluoro-1-methylpiperidin-4-yl)-8-(6-methoxypyridin-3-yl)-3-methyl-1,3-dihydro-2H-imidazo[4,5-c]quinolin-2-one derivative with excellent physicochemical properties, which can be advantageously used in pharmaceutical processing and pharmaceutical compositions.

[0151] Example 2. Preparation of the free base crystalline form of the (R)-1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) of the present invention 100-120 mg of amorphous form of (R)-1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative sample of formula (I) is added to 1-1.5 mL of methyl tert-butyl ether / water (1:10, v / v) mixture, stirred at 50°C for 6 hours, and the bottom solid is separated by centrifugation to obtain the free base crystalline form. According to XRPD detection, the solid is the free base crystalline form.

[0152] Example 3. Preparation of the fumarate salt crystalline form of the (R)-1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) of the present invention Approximately 500 mg of 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative represented by formula (I) (i.e., (R)-1-(3,3-difluoro-1-methylpiperidin-4-yl)-8-(6-methoxypyridin-3-yl)-3-methyl-1,3-dihydro-2H-imidazo[4,5-c]quinolin-2-one (I)) sample was weighed and placed in a 100 ml vial, and 8 ml of chloromethane / methanol (1:11:1.7, v / v) and 132.5 mg of fumaric acid were added, and the mixture was stirred at room temperature for about 4 hours, and then the lower wet solid was separated by centrifuge. According to XRPD detection, the solid was in the form of fumarate salt crystals.

[0153] Example 4. The crystalline forms of the free base and salts of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) of the present invention are characterized by XRPD patterns.

[0154] XRPD instrument information is as follows:

[0155] XRPD spectra were collected on a Bruker X-ray powder diffraction analyzer, and the XRPD parameters are shown in Table 1.

[0156] [Table 1] The XRPD (X-ray powder diffraction) pattern of the free base crystalline form of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) prepared according to the method described in this example is shown in FIG. 1, and the specific properties are shown in Table 2 below.

[0157] [Table 2]

[0158] The XRPD (X-ray powder diffraction) pattern of the fumarate salt crystal form of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) prepared according to the method described in this example is shown in FIG. 2, and its specific properties are shown in Table 3 below. [Table 3]

[0159] "d value" is the distance between two adjacent crystal planes in a crystal lattice in Angstroms, and "I%" is the relative intensity.

[0160] As can be seen from FIG. 1, the XRPD pattern of the free base crystal form of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) provided by the present invention is as follows: 2θ=5.08, 10.19, 10.76, 12.56, 13.37, 14.84, 15.33, 16.08, 16.53, 16.87, 19.12, The positions are 19.51, 20.30, 20.51, 21.22, 22.65, 23.26, 23.72, 24.77, 25.56, 26.67, 27.04, 28.38, 28.77, 29.26, 29.60, 30.09, 31.05, 32.61, 32.97, 34.90, 36.96, 38.21, and 38.50, and the error range of the above 2θ values ​​is ±0.2.

[0161] As can be seen from FIG. 2, the XRPD pattern of the fumarate crystal form of 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) provided by the present invention is at 2θ=6.07, 6.78, 8.60, 10.99, 12.21, 12.57, 13.66, 14.55, 16.09, 16.49, 17.32, 18.04, 18.37, 18.64, 19.41, 20.24, 20.96, 22.15, 23.54, 24.82, 25.40, 26.06, 26.81, 28.23, 30.19, 33.40, 36.25, and the error range of the above 2θ values ​​is ±0.2.

[0162] After testing, the error range of the 2θ values ​​may also be ±0.2. Those skilled in the art should understand that these diffraction peaks do not represent the details of the diffraction peaks of the free base crystal form and the fumarate salt crystal form of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I). The 2θ values ​​of the X-ray powder diffraction pattern may vary slightly due to machine changes, sample preparation, and batch-to-batch changes, and the quoted values ​​are not considered absolute values. It should also be understood that the absolute intensity of the peaks may also vary due to orientation effects. Therefore, the intensities shown in the present invention are illustrative and are not used for absolute comparison.

[0163] Example 5. Biological activity of 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivatives (I) ODYSSEY CLx (LI-COR) was used to detect the phosphorylation of ATM protein using MCF-7 cells, about 25 μl of cells were seeded into each blank 384-well plate, and after 24 hours, compounds and etoposide diluted proportionally were added using a pinzer, and after 1 hour of incubation at 37°C, the cells were fixed for 20 minutes at room temperature by adding 25 μl of 8% paraformaldehyde. The cells were permeabilized with 1XPBS containing 0.1% Triton X-100, blocked by adding 50 μl of Odyssey blocking buffer, and shaken at room temperature for 1.5 hours. The blocking buffer was removed, and 20 μl of anti-pKAP1 antibody was added, and the anti-pKAP1 antibody was incubated overnight at 4°C on a Gentol shaker. 20 μl of secondary antibody (IRDye 800CW Goat anti-Rabbit IgG antibody) solution containing DNA stain DRAQ5 (1 / 4,000) was added to each plate well and diluted in blocking buffer containing 0.1% Tween-20 (1 / 5000), and the secondary antibody was incubated for 1 h. The solution was washed away. To block the phosphorylation of ATM protein, the plates were immediately scanned using ODYSSEY CLx (LI-COR). In cell-based inhibition of ATM phosphorylation, compounds of the present invention are effective (<1 nM) in inhibiting pATM signaling on MCF-7 and therefore have the potential to overcome radiotherapy or chemotherapy resistance by DNA damage repair of ATM protein.

[0164] Example 6. Blood-brain barrier permeability of 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivatives (I) To determine whether the quinazoline derivatives (I) can cross the blood-brain barrier (BBB), the test compounds were orally administered to rats. Four hours after administration, the rats were killed, and blood and brain tissue were collected to analyze the concentration of the test compounds. Brain permeability is defined as the ratio of the concentration of the compound in the brain tissue to the concentration in the plasma. The passage of the blood-brain barrier is the ratio of the free concentration of the drug in the brain tissue to the free concentration of the drug in the plasma. P-glycoprotein is an efflux protein of the blood-brain barrier, which effluxes P-glycoprotein substrates into the skull. Breast cancer resistance protein is an efflux protein from the blood-brain barrier, which effluxes breast cancer resistance protein into the brain. Table 4 shows the ability of 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivatives (I) to cross the blood-brain barrier.

[0165] [Table 4]

[0166] In the detection of the rate of passing through the blood-brain barrier, the ratio of the free concentration of compound (I) in brain tissue to the free concentration in plasma is higher than 30%, and it is not a substrate of P-glycoprotein or BCRP. Because it can pass through the blood-brain barrier, it can achieve an effective blood drug concentration in the brain, and can be used in the treatment and prevention of central nervous system diseases such as the treatment of glioma, cancer brain metastasis, meningeal metastasis, and brain cancer, and has the potential to reduce the risk of dose-limiting toxicity outside the skull.

[0167] Example 7. Pharmacological efficacy of 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivatives (I) in combination with radiotherapy against glioma in an intracranial pdx mouse model The patient's primary tumor tissue (brain tumor, about 100,000 cells) was implanted intracranially in mice, and one week later, 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivatives (I) (5mg / kg or 10mg / kg, QD) were orally administered 15 minutes before radiation (2Gy) for 5 days. The first group was the control group and did not contain any drugs. The second group received radiation therapy (2Gy) for only 5 days. The third group received 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivatives (I) (5mg / kg, orally, once a day) 15 minutes before radiation therapy (2Gy) for 5 days. The fourth group of 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivatives (I) (10 mg / kg, orally, once a day) was administered 15 minutes before radiation therapy (2 Gy) for 5 days. As shown in Figure 3, the drug combined with radiation groups (third and fourth groups) showed longer survival with statistically significant efficacy (P<0.01) compared with the first group (control group) and the second group (radiation only, 2 Gy for 5 days), and showed a dose-dependent manner in the animal model.

[0168] Example 8. Equilibrium solubility of 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) and its salt crystalline forms Experimental method: Approximately 10 mg of solid sample (free base and various salt forms, crystalline forms) was weighed, 1.5 mL of water was added, and after equilibration at room temperature for 24 hours, 0.3 mL of the cloudy solution was taken and the bottom solid layer and the supernatant were separated by centrifugation. The supernatant was filtered through a 0.45 μm (PTFE) filter, and the concentration of free alkali was tested.

[0169] [Table 5]

[0170] As can be seen from Table 5, the free base crystal form and fumarate crystal form of 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) of the present invention have good equilibrium solubility in water.Preferably, the fumarate crystal form of 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) of the present invention has an equilibrium solubility in water of more than 5mg / mL.This is very beneficial for the absorption of the drug.

[0171] Example 9. Stability Studies The physical and chemical stability of the free base crystal form and the fumarate crystal form of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) of the present invention for one week is stable in physical and chemical properties when stored at 25°C / 60%RH and 40°C / 75%RH for one week. During the heating process, the TGA chart shows that there is no significant change in the crystal form of Example 4. The DSC chart shows that the free base crystal form and the fumarate crystal form show good stability. As shown in Figures 4A and 4B, the free base crystal form has no significant weight loss during the heating process and has a melting point of about 206°C. As shown in Figures 5A and 5B, the fumarate crystal form has no significant weight loss during the heating process and has a melting point of about 220°C. Such properties are useful for the preparation and processing of tablets.

[0172] Example 10. Good bioavailability of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) of the present invention In the rat pharmacokinetic study, the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) of the present invention was intravenously injected at 1-2 mg / kg to a group of rats (3 rats) at seven time points (0.25, 0.5, 1, 2, 4, 8, 16 hours) and orally administered at 5-10 mg / kg to a second group of rats (3 rats). Blood was collected, the concentration of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) of the present invention in the blood was measured, and the peak area and half-life were calculated. The calculation method of bioavailability is (drug oral / peak area of ​​oral dose) / (drug intravenous injection / peak area of ​​intravenous dose)×100%. The free base crystal form and fumarate crystal form of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative (I) of the present invention have good bioavailability, both of which are >35% compared to the amorphous form (oral bioavailability <35% and half-life <10 hours), and have a long half-life (>10 hours), providing better confluence and lower pill burden (half-life in rats of the free base crystal form: 22 hours, fumarate crystal form: 11.2 hours). Furthermore, the compound of the present invention is not a substrate of human aldehyde oxidase, which has a long half-life (>20 hours), and therefore has the potential to achieve effective efficacy in the brain with low metabolic clearance for the treatment and / or prevention of cancer brain metastasis, meningeal metastasis, glioma, glioblastoma, DIPG and other central nervous system diseases.

[0173] Human aldehyde oxidase (hAOX) is a cytosolic drug metabolizing enzyme expressed in human liver that, like CYPs, contributes significantly to the oxidation of a significant amount of quinoline derivatives, but acts in the absence of the NADPH cofactor. Drugs that are substrates of AOX often exhibit high metabolic clearance, resulting in low exposure and therefore reduced efficacy in humans (Lepri et al. PNAS, 2017, pp. E3178-E3187).

[0174] The free base crystalline form and the fumarate crystalline form of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative of the present invention have good solubility in water, do not undergo any change in crystal form during heating, and have good stability.

[0175] The free base crystal form and fumarate crystal form of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative of the present invention have unexpectedly excellent physicochemical properties, which are beneficial for use in pharmaceutical processing and pharmaceutical compositions.This can be applied to the treatment of cancer, cancer with CNS metastasis, brainstem tumor, primary brain cancer, DIPG or glioma, etc., especially when combined with DNA double strand break inducer, and at the same time, provide qualitative and quantitative information on efficacy and safety, which is very important for further research on the efficacy and safety of such solid drugs.

[0176] The specific embodiments of the present invention have been described above. The present invention is not limited to the above specific embodiments, and it should be understood that those skilled in the art can make various modifications and alterations without affecting the essence of the present invention within the scope of the claims.

Claims

1. The crystalline form of the free base of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative of the present invention, wherein the derivative is represented by formula (I): 【Chemistry 1】 ， wherein the XRPD pattern of said crystalline form has major characteristic peaks at 10.2, 16.9, 20.3, 25.6 2θ and secondary characteristic peaks at 10.8, 12.6, 13.5, 14.8, 16.1, 20.6, 21.2, 23.3 2θ, the error range of said 2θ values ​​being ±0.

2.

2. A process for preparing a crystalline form of the free base of a 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative, comprising the steps of: adding an amorphous form of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative sample of formula (I) to an organic solvent (50-150 mg / mL), wherein formula (I) is represented as follows: 【Chemistry 2】 and, and agitating at 50° C. and centrifuging to separate the bottom solid to obtain the free base crystalline form, wherein 50-150 mg of the amorphous derivative of formula (I) is added per ml of the organic solvent, the first organic solvent being methyl tert-butyl ether / water (1:10, v / v).

3. A crystalline form of a fumarate salt of a 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative represented by formula (I): 【Chemistry 3】 wherein the XRPD pattern of said crystalline form has major characteristic peaks at 2θ of 6.8, 8.6, 12.2, 13.7, 17.3, 19.4, 26.1, and said secondary characteristic peaks are at 6.1, 11.0, 14.6, 16.1, 16.5, 18.0, 20.2, 22.1, 26.8, and the error range of said 2θ values ​​is ±0.

2.

4. A method for producing a crystalline form of the fumarate salt of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative according to claim 3, comprising the steps of: 【Chemistry 4】 and 2-2.5 equivalents of fumaric acid to the organic solvent, stirring the mixture at 22-28° C. and collecting the solid by centrifugation, wherein 10-200 mg of the 1-(3,3-difluoropiperidin-4-yl)-imidazo[4,5-c]quinolin-2-one derivative of formula (I) is added per ml of organic solvent, and the first organic solvent is dichloromethane / methanol (1:1, v / v).

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