Methods of treating solid tumors using heteroaromatic macrocyclic ether compounds

Abstract

Provided herein are methods of treating, preventing or managing solid tumors using a heteroaromatic macrocyclic ether compound (e.g., Compound 1), or a stereoisomer thereof, or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof.

Description

[0001] This application claims the benefit of priority to U.S. application serial number 63 / 251,536 filed on October 1, 2021 and U.S. application serial number 63 / 357,309 filed on June 30, 2022, each of which is incorporated herein by reference in its entirety. Background Art

[0002] Receptor tyrosine kinases (RTKs) are enzymes on the surface of cells that receive external signals, such as whether to grow and divide, and transmit those signals inside the cell via kinase activity. Many RTKs are proto-oncogenes; abnormal RTK activity can drive cell survival, growth, and proliferation, leading to cancer and related conditions. This abnormal kinase activity may be caused by mutations such as activating mutations in the kinase domain, gene rearrangements that produce fusion proteins containing the complete kinase domain, amplification, and other means. RTK proto-oncogenes include ROS1, anaplastic lymphoma kinase (ALK), NTRK1 (encoding TRKA), NTRK2 (encoding TRKB), and NTRK3 (encoding TRKC).

[0003] ROS1 is a RTK proto-oncogene, and ROS1 rearrangement is detected in non-small cell lung cancer (NSCLC), glioblastoma, inflammatory myofibroblastic tumor (IMT), cholangiocarcinoma, ovarian cancer, gastric cancer, colorectal cancer, angiosarcoma and Spitzoid melanoma. Carcinogenic ROS1 gene fusions contain the kinase domain (3' region) of ROS1 fused with the 5' region of multiple partner genes. The examples of ROS1 fusion partner genes observed in NSCLC include SLC34A2, CD74, TPM3, SDC4, EZR, LRIG3, KDELR2, CEP72, CLTL, CTNND2, GOPC, GPRC6A, LIMA1, LRIG3, MSN, MYO5C, OPRM1, SLC6A17 (presumed), SLMAP, SRSF6, TFG, TMEM106B, TPD52L1, ZCCHC8 and CCDC6. Other fusion partners include CAPRIN1, CEP85L, CHCHD3, CLIP1 (putative), EEF1G, KIF21A (putative), KLC1, SART3, ST13 (putative), TRIM24 (putative), ERC1, FIP1L1, HLAA, KIAA1598, MYO5A, PPFIBP1, PWWP2A, FN1, YWHAE, CCDC30, NCOR2, NFKB2, APOB, PLG, RBP4, and GOLGB1.

[0004] NTRK1, NTRK2, and NTRK3 are RTK proto-oncogenes encoding TRK family kinases, and chromosomal rearrangements of NTRK1, NTRK2, and NTRK3 are detected at low frequencies in many cancers. However, for the treatment of ROS1-positive or ALK-positive patients, TRK inhibition, especially in the central nervous system (CNS), is associated with adverse reactions, including dizziness / ataxia / gait disturbance, paresthesia, weight gain, and cognitive changes.

[0005] Agents used to treat oncogenic ROS1 and ALK have significant drawbacks. These drawbacks may represent one or more of the following: associated TRK inhibition, limited CNS activity, and insufficient activity against resistance mutations. Treatment of ROS1-positive or ALK-positive patients with TRK inhibition is associated with adverse reactions, especially in the CNS, including dizziness / ataxia / gait disturbance, paresthesia, weight gain, and cognitive changes. In addition, there is a need for CNS penetrating and TRK sparing inhibitors of wild-type ROS1 kinase domain and ROS1 with acquired resistance mutations occurring alone or in combination, including G2032R, D2033N, S1986F, S1986Y, L2026M, L1951R, E1935G, L1947R, G1971E, E1974K, L1982F, F2004C, F2004V, E2020K, C2060G, F2075V, V2089M, V2098I, G2101A, D2113N, D2113G, L2155S, L2032K and L2086F. Likewise, there is a need for CNS penetrating and TRK sparing inhibitors of ALK with acquired resistance mutations. A variety of ALK resistance mutations have been reported, either alone or in combination, including G1202R, L1196M, G1269A, C1156Y, I1171T, I1171N, I1171S, F1174L, V1180L, S1206Y, E1210K, 1151Tins, F1174C, G1202del, D1203N, S1206Y, S1206C, L1152R, L1196Q, L1198P, L1198F, R1275Q, L1152P, C1156T, and F1245V. Summary of the invention

[0006] Provided herein are methods for treating, preventing, or controlling solid tumors using a heteroaromatic macrocyclic ether compound (eg, Compound 1), or a stereoisomer thereof, or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof.

[0007] In one embodiment, provided herein is a method of treating a solid tumor comprising administering to a patient in need thereof a therapeutically effective amount of Compound 1:

[0008]

[0009] or a stereoisomer thereof, or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is Compound 1.

[0010] In one embodiment, the solid tumor is an advanced ROS1-positive solid tumor. In one embodiment, the solid tumor is a locally advanced ROS1-positive solid tumor. In one embodiment, the solid tumor is an advanced ROS1-positive non-small cell lung cancer (NSCLC).

[0011] In one embodiment, the solid tumor is a metastatic ROS1 positive solid tumor. In one embodiment, the solid tumor is a metastatic ROS1 positive solid tumor of the central nervous system (CNS) (e.g., brain). In one embodiment, the solid tumor is a metastatic ROS1 positive NSCLC. In one embodiment, the solid tumor CNS (e.g., brain) is a metastatic ROS1 positive NSCLC.

[0012] In one embodiment, the patient is not treated with tyrosine kinase inhibitor (TKI) therapy. In one embodiment, the patient has been treated with one or more previous TKI therapies. In one embodiment, the patient has been treated with a previous ROS1 TKI therapy (e.g., crizotinib or entrectinib). In one embodiment, the patient has been treated with two or more previous ROS1 TKI therapies. In one embodiment, the patient's tumor has no known oncogene-driven gene changes except ROS1.

[0013] In one embodiment, the compound is administered once a day (QD) or twice a day (BID) in an amount from about 5 mg to about 400 mg. In one embodiment, the compound is administered once a day (QD) or twice a day (BID) in an amount from about 5 mg to about 200 mg (by weight of Compound 1). In one embodiment, the compound is administered to the patient on an empty stomach (e.g., at least 1 hour before and not less than 2 hours after the intake of food and / or beverages other than water). In one embodiment, the compound is administered to the patient after the intake of food and / or beverages. In one embodiment, the patient is not taking any of a potent inhibitor of CYP3A4, a potent inducer of CYP3A4, a sensitive substrate of CYP3A4, a substrate of P-gp / multidrug resistance protein (MDR1), a substrate of BCRP / breast cancer resistance protein (ABCG2), a substrate of MATE1, or a gastric acid reducing agent. In one embodiment, the patient is taking any of a strong inhibitor of CYP3A4, a strong inducer of CYP3A4, a sensitive substrate of CYP3A4, a substrate of P-gp / multidrug resistance protein (MDR1), a substrate of BCRP / breast cancer resistance protein (ABCG2), a substrate of MATE1, or a gastric acid reducing agent. In one embodiment, the compound is administered in the absence of a strong inhibitor of CYP3A4, a strong inducer of CYP3A4, or a sensitive substrate of CYP3A4. In one embodiment, the compound is administered in the presence of a strong inhibitor of CYP3A4, a strong inducer of CYP3A4, or a sensitive substrate of CYP3A4. In one embodiment, the compound is administered in the absence of a strong inhibitor of CYP3A4. In one embodiment, the compound is administered in the absence of a strong inducer of CYP3A4. In one embodiment, the compound is administered in the absence of a sensitive substrate of CYP3A4. In one embodiment, the compound is administered in the absence of a substrate for P-gp / multidrug resistance protein 1 (MDR1), a substrate for BCRP / breast cancer resistance protein (ABCG2), or MATE1. In one embodiment, the compound is administered in the absence of a substrate for P-gp / multidrug resistance protein 1 (MDR1), a substrate for BCRP / breast cancer resistance protein (ABCG2), or MATE1. In one embodiment, the compound is administered in the absence of a gastric acid reducing agent (e.g., a proton pump inhibitor, such as lansoprazole). In one embodiment, the compound is administered in the presence of a gastric acid reducing agent (e.g., a proton pump inhibitor, such as lansoprazole). In one embodiment, the compound is administered in the absence of a strong inducer of CYP3A4. In one embodiment, the compound is administered in the presence of a strong inducer of CYP3A4.

[0014] In one embodiment, the compound is administered in the absence of a sensitive substrate of CYP3A4. In one embodiment, the compound and the sensitive substrate of CYP3A4 are not administered simultaneously. In one embodiment, the sensitive substrate of CYP3A4 includes one or more of buspirone, everolimus, lovastatin, midazolam, simvastatin, triazolam, maraviroc, conivaptan, and darifenacin. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 Phase 1 and Phase 2 study designs are shown.

[0016] Figure 2A , Figure 2B and Figure 2C Compound 1-induced regression in Lu01-0414, CTG-0848, and CTG-2532 patient-derived xenograft models (PDXs) are shown, respectively. Activity of Compound 1 in various ROS1 PDX models implanted subcutaneously in Balb / c mice. The vehicle was 20% HP-β-CD (hydroxypropyl-β-cyclodextrin). Mean ± SEM plots. No significant changes in body weight were observed (data not shown). Compound 1-induced regression regardless of fusion partner (SDC4 or CD74).

[0017] Figure 3 Shown is ROS1 fusion signaling through the PI3K and MAP kinase pathways to effect proliferation and other transcriptional changes.

[0018] Figure 4A Representative Western blots of Lu01-0414 PDX (BID×5PO) treated with Compound 1 are shown, showing inhibition of ROS1-dependent cellular signaling through reduced pROS1, ROS1, pAKT, and pERK levels. Pharmacokinetic analysis indicated dose-dependent differences in unbound plasma concentrations of Compound 1. In this model, the 0.04 mg / kg dose achieved stagnation and the 5 mg / kg dose achieved regression. Figure 4B , Figure 4C and Figure 4D : Western blot quantification of three PDX models (BID×5PO) treated with compound 1 confirms reduced ROS1, pROS1 and pERK levels. The Y axis represents the fold change normalized to vehicle treatment, indicated by an open black box and a horizontal gray line. Mean ± SEM plotted.

[0019] Figure 5A , Figure 5B and Figure 5CShown is Compound 1 inhibition of ROS1 signaling in tumors by quantitative digital image analysis of immunohistochemical staining. Mean ± SEM plotted. Effects on pERK, ERK, and pAKT were model dependent.

[0020] Fig. 6A , Figure 6B and Figure 6C Compound 1 reduces expression levels of MAP kinase pathway genes in tumors. Gene expression profiling of tumor samples was performed using the NanoString 770 gene nCounter PanCancer Pathways panel. Expression levels of MAP kinase pathway genes DUSP6, FOS, IL1R1, and SPRY4 were reduced in a dose-dependent manner in tumors following Compound 1 treatment. The Y axis represents relative expression normalized to vehicle treatment, indicated by black dots and horizontal gray lines. Mean ± SEM plotted.

[0021] Figure 7 An additional Phase 1 study design is shown.

[0022] Fig. 8A Representative computed tomography (CT) images are shown showing a confirmed partial response (PR) to treatment with Compound 1 in a patient with CD74-ROS1 fusion-positive lung adenocarcinoma harboring the ROS1 G2032R resistance mutation after prior crizotinib, lorlatinib, and chemotherapy. The arrow indicates a left lung nodule that decreased in size during treatment.

[0023] Figure 8B Representative CT (upper panel) and magnetic resonance imaging (lower panel) images are shown, showing confirmed PR to Compound 1 treatment in a patient with EZR-ROS1 fusion-positive lung adenocarcinoma carrying the ROS1 G2032R resistance mutation after prior entrectinib, lopretinib, and chemotherapy. Arrows in the upper panel indicate segments 5 / 6 and 3 liver metastases with continuous regression during treatment. Arrows in the lower panel indicate right occipital lobe metastases, which decreased in size at week 4 and became barely noticeable at week 16.

[0024] Figure 8C Representative computed tomography images are shown showing a confirmed partial response to Compound 1 in a patient with EZR-ROS1 fusion-positive lung adenocarcinoma without a known ROS1 resistance mutation after prior crizotinib treatment. The arrow indicates a left pleural-based lung nodule that decreased in size at week 4 and became barely noticeable at week 24.

[0025] Fig. 9Compound 1 was shown to inhibit signaling through the MAPK and PI3K / AKT pathways and induce apoptosis in SDC4-ROS1-bearing PDX tumors.

[0026] Fig. 10A is a representative X-ray powder diffraction (XRPD) pattern of Form 1, the free base of Compound 1. Fig. 10B is a representative differential scanning calorimetry (DSC) thermogram of Form 1 of the free base of Compound 1.

[0027] Fig.11 ctDNA data are shown, indicating that Compound 1 is active against both ROS1 fusions and ROS1G2032R solvent front mutations at all dose levels tested.

[0028] Fig.12 Radiographic tumor regression is shown for patients dosed with Compound 1. PD = progressive disease; PR = confirmed partial response; QD = once daily; SD = stable disease; TKI = tyrosine kinase inhibitor. a Single time point PR was not confirmed. b Sustained partial responses are pending confirmation. c Best response PR due to residual non-target disease. d Other prior ROS1 TKIs are ceritinib or cabozantinib. e Including immunotherapy, bevacizumab, and investigational therapies.

[0029] Fig.13 The duration of treatment is shown for response-evaluable NSCLC patients given Compound 1. PD = progressive disease; PR = partial response; TKI = tyrosine kinase inhibitor.

[0030] Fig.14 Compound 1-induced reduction in tumor burden in patients with ROS1 G2032R (left panel) and Compound 1-induced reduction in the ROS1 G2032R allele (right panel) are shown. b Bars represent week 2 results; week 8 results are pending.

[0031] Fig.15 Shown are radiological images indicating intracranial response in a 65-year-old woman with CD74-ROS1 fusion NSCLC who had previously been treated with chemotherapy, crizotinib, and lorlatinib with CNS progression and no known ROS1 resistance mutation.

[0032] Fig.16Shown are radiological images indicating intracranial and extracranial activity in TKI-refractory ROS1 G2032R+ NSCLC in a patient diagnosed with EZR-ROS1 fusion NSCLC.

[0033] Fig.17 Shown are EZR-ROS1 (left y-axis) and ROS1 G2032R (right y-axis) variant allele frequencies in the plasma of a 65-year-old patient with stage IV lung adenocarcinoma and multiple brain metastases, as determined by the Guardant 360 assay, with EZR-ROS1 identified by plasma testing before (C1D1) and during treatment (C1D15 and C3D1).

[0034] Fig.18A , Fig.18B and Fig.18C Unbound plasma concentrations of Compound 1 during 24 hours on Day 15 of Treatment Cycle 1 are shown for three patients (see the Results section of Example 5). The dashed line indicates the minimum effective concentration threshold of 5.3 ng / mL = 13 nM, which is based on the effective threshold in the murine CTG-2532CD74-ROS1 G2032R PDX study.

[0035] Fig.19 Compound 1 exposure in clinical trials was shown to exceed target levels that provided resolution in preclinical models (see Example 5). Compound 1 exhibited favorable pharmacokinetics, including low intra-cohort patient PK variability, increasing exposure with increasing dose levels, and a half-life of approximately 20 hours supporting QD dosing. For patients treated through September 1, 2022, data are as of September 13, 2022. Pharmacokinetic data for the 125 mg QD cohort are not shown due to immaturity. QD = once a day. a,b Based on tumor regression in in vivo models carrying SDC4-ROS1 and CD74-ROS1 G2032R, respectively. c,d The values ​​of a and b were divided by the predicted human CNS Kp, respectively.

[0036] Fig. 20 The Phase 1 population in Example 5 is shown to have heavily pre-treated ROS1-positive solid tumors according to data on September 13, 2022 for patients treated until September 1, 2022. Unless otherwise specified, all data are shown as n (%). CNS, central nervous system; ECOGPS, Eastern Cooperative Oncology Group performance status; NSCLC, non-small cell lung cancer; RECIST 1.1, solid tumor response evaluation criteria version 1.1; TKI, tyrosine kinase inhibitor. a Patients with untreated CNS lesions were included. bTherapies given for early-stage disease are not included. c The categories are not mutually exclusive.

[0037] Fig.21 Treatment-related adverse events (TRAEs) occurring in >1 patient in the Phase 1 trial (Example 5) based on data as of September 13, 2022 are shown for patients treated through September 1, 2022. The safety profile of Compound 1 was favorable and consistent with the highly ROS1-selective, TRK-sparing Compound 1. AE, adverse event; ALT, alanine aminotransferase; AST, aspartate aminotransferase; DLT, dose-limiting toxicity; SAE, serious adverse event. a Includes edema and peripheral edema. DETAILED DESCRIPTION

[0038] definition

[0039] Unless otherwise defined, all technical and scientific terms used herein have the meanings commonly understood by those skilled in the art of the present disclosure. The following references provide general definitions of many terms used in the present disclosure for those skilled in the art: Singleton et al., Dictionary of Microbiology and Molecular Biology (2nd edition 1994); The Cambridge Dictionary of Science and Technology (Walker edition, 1988); The Glossary of Genetics, 5th edition, R. Rieger et al. (eds.), Springer Verlag (1991); and Hale and Marham, The Harper Collins Dictionary of Biology (1991). Unless otherwise indicated, the following terms as used herein have the following meanings assigned to them.

[0040] In some embodiments, chemical structures are disclosed with the corresponding chemical names. In case of conflict, the chemical structure controls the meaning, not the name.

[0041] As used herein, the terms "comprising" and "including" can be used interchangeably. The terms "comprising" and "including" should be interpreted as specifying the presence of the features or components mentioned, but do not exclude the presence or addition of one or more features or components or groups thereof. In addition, the terms "comprising" and "including" are intended to include examples covered by the term "consisting of". Therefore, the term "consisting of" can be used instead of the terms "comprising" and "including" to provide more specific embodiments of the present invention.

[0042] The term "consisting of" means that the subject matter has at least 90%, 95%, 97%, 98% or 99% of the features or components that constitute the subject matter. In another embodiment, the term "consisting of" excludes any other features or components from the scope of any subsequent description, except those that are not necessary for the technical effect to be achieved.

[0043] Unless otherwise specified or obvious from the context, as used herein, the term "or" is understood to be inclusive. Unless otherwise specified or obvious from the context, as used herein, the terms "a / an" and "the" are understood to be singular or plural. For example, when a compound provided herein is administered to a "patient", it includes administering the compound to an individual patient or a patient population.

[0044] As used herein and unless otherwise indicated, "stereoisomer" refers to various stereoisomeric forms of compounds containing one or more asymmetric centers or steric hindrances in the structure. In some embodiments, a stereoisomer is an enantiomer, a mixture of enantiomers, atropisomers, or a tautomer thereof. For example, the compounds described herein may be in the form of a single enantiomer, a diastereomer, or a geometric isomer (e.g., atropisomers), or may be in the form of a mixture of stereoisomers, including a racemic mixture and a mixture enriched in one or more stereoisomers. In some embodiments, the compounds provided herein may be atropisomers. In certain embodiments, atropisomers are stereoisomers produced due to hindered rotation around a single bond, wherein energy differences due to steric strain or other contributing factors produce rotational barriers high enough to allow separation of individual conformers. Stereoisomers can be separated from a mixture by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferably isomers can be prepared by asymmetric synthesis. See, e.g., Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, EL Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, SH Tables of Resolving Agents and Optical Resolutions, p. 268 (EL Eliel, ed., Univ. of Notre Dame Press, Notre Dame, IN 1972). The present invention further includes compounds as individual isomers substantially free of other isomers, or as compounds as mixtures of various isomers.

[0045] In certain embodiments, provided herein is a compound that can be racemic. In certain embodiments, provided herein is a compound that can be enriched in one enantiomer. For example, provided herein is a compound that can have greater than about 30%ee, about 40%ee, about 50%ee, about 60%ee, about 70%ee, about 80%ee, about 90%ee or even about 95% or greater ee. In certain embodiments, provided herein is a compound that can have more than one stereocenter. In certain such embodiments, provided herein is a compound that can be enriched in one or more diastereomers. For example, provided herein is a compound that can have greater than about 30%de, about 40%de, about 50%de, about 60%de, about 70%de, about 80%de, about 90%de or even about 95% or greater de.

[0046] In certain embodiments, therapeutic preparations can be enriched to mainly provide one enantiomer of compound.Enantiomerically enriched mixtures can include, for example, at least about 60mol% of one enantiomer, or more specifically at least about 75, about 90, about 95 or even about 99mol%.In certain embodiments, the compound enriched in one enantiomer is substantially free of other enantiomers, wherein substantially free of means that compared with the amount of other enantiomers in, for example, a composition or a mixture of compounds, the material in question accounts for less than about 10%, or less than about 5%, or less than about 4%, or less than about 3%, or less than about 2%, or less than about 1%.For example, if a composition or a mixture of compounds contains about 98 grams of the first enantiomer and about 2 grams of the second enantiomer, it is considered that it contains about 98mol% of the first enantiomer and only about 2% of the second enantiomer.

[0047] In certain embodiments, the therapeutic preparation can be enriched to provide primarily one diastereomer of the compound. A diastereomerically enriched mixture can contain, for example, at least about 60 mol% of one diastereomer, or more specifically at least about 75, about 90, about 95, or even about 99 mol%.

[0048] In some embodiments, a moiety in a compound exists as a mixture of tautomers. A "tautomer" is a structural isomer of a moiety or compound that is easily interconverted with another structural isomer. For example, a pyrazole ring has two tautomers:

[0049]

[0050] They differ in the position of pi bonds and hydrogen atoms. Unless specifically stated otherwise, a drawing of one tautomer of a moiety or compound includes all possible tautomers.

[0051] The term "subject" contemplated for administration includes, but is not limited to, humans (i.e., males or females of any age group, such as pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young adults, middle-aged adults, or elderly people)) and / or other primates (e.g., cynomolgus monkeys, rhesus monkeys); mammals, including commercially relevant mammals, such as cattle, pigs, horses, sheep, goats, cats, and / or dogs; and / or birds, including commercially relevant birds, such as chickens, ducks, geese, quail, and / or turkeys. In certain embodiments, the subject is a human. In certain embodiments, the subject is an adult of at least 40 years of age. In certain embodiments, the subject is an adult of at least 50 years of age. In certain embodiments, the subject is an adult of at least 60 years of age. In certain embodiments, the subject is an adult of at least 70 years of age. In certain embodiments, the subject is an adult of at least 18 years of age or at least 12 years of age. As used herein and unless otherwise indicated, a human subject contemplated for administration of a therapeutic agent (e.g., a compound described herein) to treat, prevent, or manage a disease, condition, or disorder, or a symptom thereof, is also referred to as a "patient."

[0052] As used herein, a therapeutic agent for "preventing" a disease or illness refers to a compound that reduces the occurrence of a disease or illness in a treated sample, or delays the onset of, or reduces the severity of one or more symptoms of a disease or illness, relative to an untreated control sample, in a statistical sample. These effects are also referred to as "preventive" effects. Therefore, as used herein and unless otherwise indicated, the terms "prevention" and "preventing" refer to methods for obtaining beneficial or desired results, including but not limited to preventive benefits. For preventive benefits, a therapeutic agent can be administered to a patient at risk of developing a particular disease, or to a patient who reports one or more physiological symptoms of a disease, even if a diagnosis of the disease may not yet be made. In one embodiment, a therapeutic agent is administered before the clinical manifestation of an undesirable disease (e.g., a disease or other undesirable state of a subject) to obtain a preventive benefit (e.g., it protects the subject from developing an undesirable disease).

[0053] As used herein and unless otherwise indicated, the terms "treatment" and "treating" refer to treatment or palliative measures. Beneficial or desired clinical results include, but are not limited to, complete or partial alleviation of symptoms associated with a disease or disorder or illness, reduction in the extent of the disease, stabilization of the disease state (i.e., no worsening), delay or slowing of the disease process, improvement or alleviation of the disease state (e.g., one or more symptoms of the disease), and relief (whether partial or total relief), whether detectable or undetectable. "Treatment" may also mean prolonged survival compared to survival expected when not receiving treatment. In one embodiment, "treatment" includes administering a therapeutic agent after the occurrence of an unwanted disorder (i.e., it is intended to reduce, improve, or stabilize an existing unwanted disorder or its side effects).

[0054] As used herein and unless otherwise indicated, the term "managing" includes preventing the recurrence of a particular disease or condition in a patient suffering from the particular disease or condition, prolonging the time that a patient suffering from the disease or condition remains in remission, reducing the mortality rate of a patient, and / or keeping the severity of the disease or condition under control reduced or avoiding symptoms associated with the disease or condition.

[0055] As used herein, an "effective amount" refers to an amount sufficient to achieve a desired biological effect. As used herein, a "therapeutically effective amount" refers to an amount sufficient to achieve a desired therapeutic effect. For example, a therapeutically effective amount may refer to an amount sufficient to ameliorate at least one sign or symptom of cancer.

[0056] A "response" to a treatment approach can include a decrease or improvement in negative symptoms, a decrease in the progression of the disease or its symptoms, an increase in beneficial symptoms or clinical outcomes, a decrease in side effects, stabilization of the disease, partial or complete treatment of the disease, etc.

[0057] As used herein and unless otherwise indicated, the term "relapse" refers to a condition, disease or disorder that responded to (eg, achieved a complete response) a previous treatment and then progressed. The previous treatment may include one or more therapies.

[0058] As used herein and unless otherwise indicated, the term "refractory" refers to a condition, disease or disorder that has not responded to previous treatment, which may include one or more therapies.

[0059] As used herein and unless otherwise specified, the terms "about" and "approximately" when used in conjunction with the dosage, amount or weight percentage of the ingredients of the composition or dosage form, mean a dosage, amount or weight percentage that is recognized by one of ordinary skill in the art as providing a pharmacological effect equivalent to that obtained with the specified dosage, amount or weight percentage. In certain embodiments, the terms "about" and "approximately" when used in this context, encompass dosages, amounts or weight percentages within 30%, within 20%, within 15%, within 10% or within 5% of the specified dosage, amount or weight percentage.

[0060] The term "between" includes the endpoint numbers on both ends of the range. For example, the range described by "between 3 and 5" includes the numbers "3" and "5".

[0061] As used herein and unless otherwise specified, the term "pharmaceutically acceptable salt" refers to those salts that are suitable for use in contact with the tissues of a subject without undue toxicity, irritation, allergic response, and the like, within the scope of sound medical judgment, and commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66: 1-19. In certain embodiments, pharmaceutically acceptable salts include, but are not limited to, alkyl, dialkyl, trialkyl, or tetraalkyl ammonium salts. In certain embodiments, pharmaceutically acceptable salts include, but are not limited to, L-arginine, phenethylbenzylamine, benzophenone, betaine, calcium hydroxide, choline, diarno, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucamine, hydrabamine, 1H-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, 1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine, and zinc salts. In certain embodiments, pharmaceutically acceptable salts include, but are not limited to, Na, Ca, K, Mg, Zn, or other metal salts.

[0062] Pharmaceutically acceptable acid addition salts may also exist as various solvates, such as with water, methanol, ethanol, dimethylformamide, and the like. Mixtures of such solvates may also be prepared. The source of the solvate may be from the crystallization solvent, inherent in the preparation or crystallization solvent, or exogenous to the solvent.

[0063] Pharmaceutically acceptable anionic salts include, but are not limited to, acetate, aspartate, benzenesulfonate, benzoate, benzenesulfonate, bicarbonate, bitartrate, bromide, camphorsulfonate, carbonate, chloride, citrate, decanoate, edetate, essulfonate, fumarate, glucoheptonate, gluconate, glutamate, glycolate, hexanoate, hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, malate, maleate, mandelate, methanesulfonate, methylsulfate, mucate, naphthenate, nitrate, octanoate, oleate, pamoate, pantothenate, phosphate, polygalacturonate, propionate, salicylate, stearate, acetate, succinate, sulfate, tartrate, theoclorate, and toluenesulfonate.

[0064] Compound

[0065] In one embodiment, the compound used in the methods provided herein is a compound of the formula, also referred to as "Compound 1":

[0066]

[0067] or a stereoisomer thereof, or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof. The chemical name of compound 1 is (19R)-3-ethyl-16-fluoro-10,19-dimethyl-20-oxa-3,4,9,10,11,23-hexaazapentacyclo[19.3.1.0 2 , 6 .0 8 , 12 .0 13 , 18 ] pentacosa-1(24),2(6),4,8,11,13,15,17,21(25),22-decene-22-amine, and described in International Application No. PCT / US2021 / 030842, the entirety of which is incorporated herein by reference. As used herein, "Compound 1", "Compound 1 free base", "Compound 1 (free base)" and "free base Compound 1" are used interchangeably.

[0068] It should be understood that, unless otherwise specified, when a compound as described herein is provided as a pharmaceutically acceptable salt thereof, the moiety referenced by weight does not include the salt moiety (ie, as Compound 1 free base).

[0069] In one embodiment, compound 1 (free base) is used in the methods provided herein. In one embodiment, tautomers of compound 1 are used in the methods provided herein. In one embodiment, compound 1 substantially free of (S)-enantiomers is used in the methods provided herein. As used herein, "substantially free" means that the amount of (S)-enantiomer present is less than about 10% by weight of compound 1 (e.g., less than about 5% by weight of compound 1 or less than about 0.05% by weight). In one embodiment, substantially pure compound 1 has an enantiomeric purity of at least about 98% (e.g., 99% or 99.5%). In one embodiment, a pharmaceutically acceptable salt of compound 1 is used in the methods provided herein. In one embodiment, a pharmaceutically acceptable salt is a benzenesulfonate. In one embodiment, a pharmaceutically acceptable salt is a phosphate.

[0070] In one embodiment, the compound used in the methods provided herein is a solid form of Compound 1. In one embodiment, the solid form is Form 1 of Compound 1. A representative XRPD pattern of Form 1 of Compound 1 is provided at Fig. 10A .

[0071] In one embodiment, the solid form of Compound 1 is characterized by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22 or all XRPD peaks at about 10.7, 12.0, 12.2, 13.9, 15.0, 17.4, 18.4, 18.6, 20.8, 21.2, 21.3, 21.6, 21.7, 23.3, 23.5, 24.0, 25.2, 26.0, 26.2, 26.7, 27.7, 28.0, and 29.6 ° 2θ when measured using Cu Ka radiation. In one embodiment, the solid form is characterized by 3 peaks. In one embodiment, the solid form is characterized by 5 peaks. In one embodiment, the solid form is characterized by 7 peaks. In one embodiment, the solid form is characterized by 9 peaks. In one embodiment, the solid form is characterized by 11 peaks. In one embodiment, the solid form is characterized by all peaks.

[0072] In one embodiment, the solid form (e.g., crystalline form) of Compound 1 is characterized by an XRPD pattern comprising at least three peaks selected from the group consisting of approximately (e.g., ± 0.2°) 10.7, 15.0, 17.4, 20.8, 21.2, 21.3, 21.6, 24.0, and 25.2 ° 2θ when measured using Cu Ka radiation. In one embodiment, the solid form is characterized by an XRPD pattern comprising at least four peaks selected from the group consisting of approximately (e.g., ± 0.2°) 10.7, 15.0, 17.4, 20.8, 21.2, 21.3, 21.6, 24.0, and 25.2 ° 2θ. In one embodiment, the solid form is characterized by an XRPD pattern comprising at least five peaks selected from the group consisting of approximately (e.g., ± 0.2°) 10.7, 15.0, 17.4, 20.8, 21.2, 21.3, 21.6, 24.0, and 25.2° 2θ.

[0073] In one embodiment, the solid form (e.g., crystalline form) of Compound 1 is characterized by an XRPD pattern comprising at least three peaks selected from the group consisting of approximately (e.g., ± 0.2°) 10.7, 12.0, 12.2, 13.9, 15.0, 17.4, 18.4, 20.8, 21.2, 21.3, 21.6, 24.0, and 25.2 ° 2θ when measured using Cu Ka radiation. In one embodiment, the solid form is characterized by an XRPD pattern comprising at least four peaks selected from the group consisting of approximately (e.g., ± 0.2°) 10.7, 12.0, 12.2, 13.9, 15.0, 17.4, 18.4, 20.8, 21.2, 21.3, 21.6, 24.0, and 25.2 ° 2θ. In one embodiment, the solid form is characterized by an XRPD pattern comprising at least five peaks selected from the group consisting of approximately (e.g., ± 0.2°) 10.7, 12.0, 12.2, 13.9, 15.0, 17.4, 18.4, 20.8, 21.2, 21.3, 21.6, 24.0, and 25.2° 2θ.

[0074] In one embodiment, the solid form (e.g., crystalline form) of Compound 1 is characterized by an XRPD pattern comprising peaks at approximately (e.g., ± 0.2°) 10.7, 15.0, and 21.2° 2θ. In one embodiment, the XRPD pattern further comprises peaks at approximately (e.g., ± 0.2°) 17.4 and 21.3° 2θ. In one embodiment, the XRPD pattern further comprises peaks at approximately (e.g., ± 0.2°) 12.0, 12.2, and 13.9° 2θ. In another embodiment, the XRPD pattern further comprises peaks at approximately (e.g., ± 0.2°) 21.6 and 24.0° 2θ. In one embodiment, the XRPD pattern includes peaks at approximately (e.g., ± 0.2°) 10.7, 15.0, 17.4, 20.8, 21.2, 21.3, 21.6, 24.0, and 25.2° 2θ. In one embodiment, the XRPD pattern includes peaks at approximately (e.g., ± 0.2°) 10.7, 12.0, 12.2, 13.9, 15.0, 17.4, 18.4, 20.8, 21.2, 21.3, 21.6, 24.0, and 25.2° 2θ.

[0075] In one embodiment, the solid form is characterized by Fig. 10A The XRPD pattern matches the XRPD pattern described in.

[0076] In one embodiment, the XRPD patterns described herein are obtained using Cu Ka radiation. In one embodiment, the XRPD patterns are obtained by XRPD using a The wavelength of Kα1 radiation and The Kα2 radiation of the wavelength is measured by Cu Kα radiation, where the Kα1:Kα2 ratio is 0.5.

[0077] A representative DSC thermogram of Form 1 is provided at Fig. 10B In one embodiment, the solid form exhibits a thermal event (endotherm) as characterized by DSC having an onset temperature of about 265°C (e.g., ±2°). In one embodiment, the thermal event also has a peak temperature of about 267°C (e.g., ±2°). In one embodiment, without being bound by a particular theory, the thermal event corresponds to melting. In one embodiment, the solid form is characterized by Fig. 10B The DSC thermograph described matches the DSC thermograph. In one embodiment, the DSC thermograph is as measured by DSC using a scan rate of about 10°C / minute.

[0078] In another embodiment, the solid form has a melting point of about 270°C (eg, ±2°).

[0079] In one embodiment, the solid form (e.g., crystalline form) of Compound 1 has the following approximate unit cell dimensions: α = 90°, β = 106°, and γ = 90°. In one embodiment, the solid form has the following approximate unit cell dimensions: α = 90°, β = 105.6°, and γ = 90°. In one embodiment, the solid form has the following approximate unit cell dimensions: α = 90°, β = 105.55°, and γ = 90°. In one embodiment, Form 1 has a unit cell with a space group of P21. In one embodiment, the solid form has about / unit cell volume. In one embodiment, Form 1 has a Z value of 2. In one embodiment, Form 1 has a volume of about 1.362 g / cm 3 density.

[0080] In one embodiment, the (S)-enantiomer of Compound 1 is used in the methods provided herein. In one embodiment, the tautomer of the (S)-enantiomer of Compound 1 is used in the methods provided herein. In one embodiment, a pharmaceutically acceptable salt of the (S)-enantiomer of Compound 1 is used in the methods provided herein.

[0081] In one embodiment, the racemic mixture of Compound 1 is used in the methods provided herein. In one embodiment, a tautomer of the racemic mixture of Compound 1 is used in the methods provided herein. In one embodiment, a pharmaceutically acceptable salt of the racemic mixture of Compound 1 is used in the methods provided herein.

[0082] How to use

[0083] In one embodiment, provided herein is a method of treating cancer comprising administering a heteroaromatic macrocyclic ether compound (such as Compound 1), or a stereoisomer thereof, or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof.

[0084] In one embodiment, provided herein is a method for treating, preventing or controlling solid tumors using a heteroaromatic macrocyclic ether compound (eg, Compound 1), or a stereoisomer thereof, or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof.

[0085] In one embodiment, provided herein is a method of treating a patient with a solid tumor, comprising administering to the patient a therapeutically effective amount of Compound 1:

[0086]

[0087] or a stereoisomer thereof, or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof.

[0088] In one embodiment, the cancer is anaplastic large cell lymphoma (ALCL), atypical meningioma, breast cancer, cholangiocarcinoma, gastric cancer, glioblastoma, inflammatory myofibroblastic tumor (IMT), inflammatory hepatocellular adenoma (HCA), melanoma, pancreatic cancer, papillary thyroid carcinoma, salivary gland cancer, serous ovarian cancer, or Spitz nevus.

[0089] In one embodiment, the solid tumor is an advanced solid tumor. In one embodiment, the solid tumor is a locally advanced solid tumor. In one embodiment, the advanced solid tumor relapses, is refractory or resists after previous treatment with a tyrosine kinase inhibitor (TKI). In one embodiment, the solid tumor is non-small cell lung cancer (NSCLC). In one embodiment, the solid tumor is ROS1 positive (eg, ROS1 fusion) NSCLC. In one embodiment, the solid tumor is ALK positive (eg, ALK fusion) NSCLC. In one embodiment, the solid tumor is LTK positive (eg, LTK fusion) NSCLC. In one embodiment, the solid tumor is advanced NSCLC. In one embodiment, the solid tumor is locally advanced NSCLC. In one embodiment, the solid tumor is metastatic. In one embodiment, the solid tumor is CNS metastatic (also referred to herein as "CNS metastatic" or "metastatic CNS"). In one embodiment, the solid tumor is metastatic NSCLC. In one embodiment, the solid tumor is CNS metastatic NSCLC. As used herein and unless otherwise indicated, "advanced tumor" refers to a tumor that cannot be cured or grows beyond the initial site of origin, whether locally advanced or metastatic.

[0090] In one embodiment, the solid tumor (or cancer) is ROS1 positive. In one embodiment, the solid tumor is ROS1 positive NSCLC. In one embodiment, the solid tumor is an advanced ROS1 positive solid tumor. In one embodiment, the solid tumor is a locally advanced ROS1 positive solid tumor. In one embodiment, the solid tumor is an advanced ROS1 positive NSCLC. In one embodiment, the solid tumor is a locally advanced ROS1 positive NSCLC. In one embodiment, the solid tumor is a metastatic ROS1 positive solid tumor. In one embodiment, the solid tumor is a CNS metastatic ROS1 positive solid tumor. In one embodiment, the solid tumor is a metastatic ROS1 positive NSCLC. In one embodiment, the solid tumor is a CNS metastatic ROS1 positive NSCLC. In one embodiment, the solid tumor (or cancer) has a ROS1 mutation. In one embodiment, the ROS1 mutation is G2032R. In one embodiment, the ROS1 mutation comprises G2032R and one or more of S1986F, S1986Y, F2004C, F2004V, L2026M, D2033N, or G2101 A. In one embodiment, the solid tumor (or cancer) has a ROS1 fusion.

[0091] In one embodiment, the solid tumor (or cancer) is ALK positive. As used herein and unless otherwise specified, "ALK positive" (ALK+) cancer, disease or condition refers to a cancer, disease or condition characterized by abnormally high expression of the ALK gene and / or the presence of mutations in the ALK gene and / or the presence of partial deletion of ALK protein. In one embodiment, the solid tumor is ALK positive NSCLC. In one embodiment, the solid tumor is an advanced ALK positive solid tumor. In one embodiment, the solid tumor is a locally advanced ALK positive solid tumor. In one embodiment, the solid tumor is an advanced ALK positive NSCLC. In one embodiment, the solid tumor is a locally advanced ALK positive NSCLC. In one embodiment, the solid tumor is a metastatic ALK positive solid tumor. In one embodiment, the solid tumor is a CNS metastatic ALK positive solid tumor. In one embodiment, the solid tumor is a metastatic ALK positive NSCLC. In one embodiment, the solid tumor (or cancer) has an ALK mutation.

[0092] In one embodiment, the ALK mutation comprises one or more ALK rearrangements (in one embodiment, one rearrangement). In one embodiment, the ALK mutation comprises one or more ALK fusions (in one embodiment, one fusion). In some embodiments, the cancer treated by the methods of the present disclosure comprises an ALK fusion. In one embodiment, ALK is fused to one of the fusion partners described in Ou et al., JTO Clinical and Research Reports, 1(1): 1-10, which is incorporated herein by reference in its entirety. In one embodiment, ALK is fused to one of the fusion partners selected from the group consisting of EML4, TFG, KIF5B, KLC1, STRN, HIP1, TPR, BIRC6, DCTN1, SQSTM1, SOCS5, SEC31A, CLTC, PRKAR1A, PPM1B, EIF2AK3, CRIM1, CEBPZ, PICALM, CLIP1, BCL11A, GCC2, LMO7, PHACTR1, CMTR1, VIT, DYSF, ITGAV, PLEKHA7, CUX1, VKORC1L1, FBXO36, SPTBN1, EML6, FBXO11, CLIP4, CAMKMT, NCOA1, MYT1L, SRBD1, SRD5A2, NYAP2, MPRIP, ADAM17, ALK, L PIN1, WDPCP, CEP55, ERC1, SLC16A7, TNIP2, ATAD2B, SLMAP, FBN1, SWAP70, TCF12, TRIM66, WNK3 , AKAP8L, SPECC1L, PRKCB, CDK15, LCLAT1, YAP1, PLEKHM2, DCHS1, PPFIBP1, ATP13A4, C12orf75 , EPAS1, FAM179A, FUT8, LIMD1, LINC00327, LOC349160, LYPD1, RBM20, TACR1, TANC1, TTC27, TU BBB, SMPD4, SORCS1, LINC00211, SOS1, C9orf3, CYBRD1, MTA3, THADA, TSPYL6, WDR37 and PLEKHH2.In one embodiment, ALK is fused to one of the fusion partners selected from the group consisting of EML4, TMP1, WDCP, GTF2IRD1, TPM3, TPM4, CLTC, LMNA, PRKAR1A, RANBP2, TFG, FN1, KLC1, VCL, STRN, HIP1, NPM1, DCTN1, SQSTM1, TPR, CRIM1, PTPN3, FBXO36, ATIC, and KIF5B. In one embodiment, ALK is mutated to EML4-ALK, a fusion between the echinoderm microtubule-associated protein-like 4 (EML4) gene and the ALK tyrosine kinase domain. Many variants of EML4-ALK vary depending on the breakpoint connection, with variants 1 (v1) and 3 (v3) being the most common clinically. In one embodiment, ALK is mutated to NPM1-ALK. In one embodiment, ALK is mutated to STRN-ALK.

[0093] In one embodiment, solid tumors (or cancers) are leukocyte receptor tyrosine kinase (LTK) positive. As used herein and unless otherwise indicated, "LTK positive" (LTK+) cancers, diseases or conditions refer to abnormally high expression of LTK genes and / or mutations in LTK genes, including LTK gene rearrangements that lead to LTK fusion proteins, characterized by cancers, diseases or conditions. In one embodiment, solid tumors are LTK positive breast invasive ductal carcinoma, prostate adenocarcinoma, pancreatic adenocarcinoma, unknown primary adenocarcinoma or bladder urothelial carcinoma. In one embodiment, cancer is LTK positive leukemia. In one embodiment, solid tumors are LTK positive lung cancer. In one embodiment, solid tumors are LTK positive NSCLC. In one embodiment, solid tumors (or cancers) have LTK mutations. In one embodiment, LTK mutations are G269A, F218I, N257T, A13fs or A214fs. In one embodiment, solid tumors (or cancers) have LTK fusions. In one embodiment, LTK fusions are CLIP1-LTK. See Cooper AJ, Sequist LV, Johnson TW, Lin JJ. LTK fusions: A new target emerges in non-small cell lung cancer. Cancer Cell. 2022 Jan 10; 40(1): 23-25; and Izumi, H., Matsumoto, S., Liu, J. et al. The CLIP1–LTK fusion is an oncogenic driver in non-small-cell lung cancer. Nature 600, 319–323 (2021), each of which is incorporated herein by reference.

[0094] In one embodiment, the patient has not been treated with a prior therapy. In one embodiment, the patient is not treated with (ie, has not received) any tyrosine kinase inhibitor (TKI) therapy.

[0095] In one embodiment, the patient has been treated with one or more previous therapies. In one embodiment, the patient has been treated with at least one previous TKI therapy. In one embodiment, the patient has been treated with at least two previous TKI therapies. In one embodiment, the patient has been treated with one previous TKI therapy. In one embodiment, the patient has been treated with two previous TKI therapies. In one embodiment, the TKI is a ROS1 TKI (e.g., crizotinib or entrectinib). In one embodiment, the previous TKI therapy is selected from one or more of the group consisting of: crizotinib, entrectinib, repotrectinib, taletrectinib, and lorlatinib.

[0096] In one embodiment, the patient has not been treated with a previous platinum-based chemotherapy. In one embodiment, the patient has been treated with at most one previous platinum-based chemotherapy. In one embodiment, the patient has been treated with at least one previous platinum-based chemotherapy. In one embodiment, the patient has been treated with at least two previous platinum-based chemotherapy. In one embodiment, the patient has been treated with one previous platinum-based chemotherapy. In one embodiment, the patient has been treated with two previous platinum-based chemotherapy.

[0097] As used herein, "platinum-based chemotherapy" refers to a chemotherapeutic agent that is a coordination complex of platinum. Exemplary platinum-based chemotherapy include, but are not limited to, cisplatin, oxaliplatin, nedaplatin, or carboplatin.

[0098] In one embodiment, the patient has not been treated with immunotherapy. In one embodiment, the patient has been treated with immunotherapy. In one embodiment, the patient has been treated with at least one previous immunotherapy. In one embodiment, the patient has been treated with at least two previous immunotherapies. In one embodiment, the patient has been treated with one previous immunotherapy. In one embodiment, the patient has been treated with two immunotherapies.

[0099] As used herein, "immunotherapy" refers to treating a disease by activating or suppressing the immune system. Immunotherapy designed to trigger or amplify immune responses is classified as activated immunotherapy, while reduced or suppressed immunotherapy is classified as suppressive immunotherapy. Immunotherapy regulates immune effector cells (such as lymphocytes, macrophages, dendritic cells, natural killer cells (NK cells), cytotoxic T lymphocytes (CTLs), etc.) to cooperate against cancer cells by targeting abnormal antigens expressed on the surface of tumors. Exemplary immunotherapy includes but is not limited to checkpoint inhibitors (such as anti-cytotoxic T lymphocyte-associated protein 4 (CTLA-4) and anti-programmed cell death protein 1 (PD-1) antibodies). Exemplary PD-1 inhibitors include but are not limited to pembrolizumab (Keytruda), nivolumab (Opdivo) and cemilimab (Libtayo). Exemplary PD-L1 inhibitors include but are not limited to atezolizumab monoclonal antibody (Tecentriq), avelumab (avelumab) (Bavencio), and durvalumab (Imfinzi). Exemplary CTLA-4 inhibitors include, but are not limited to, ipilimumab (Yervoy).

[0100] In one embodiment, the patient has not been treated with chemotherapy. In one embodiment, the patient has been treated with at least one prior line of chemotherapy. In one embodiment, the patient has been treated with at least two prior lines of chemotherapy.

[0101] In one embodiment, the patient has been treated with at least three prior lines of anticancer therapy. In one embodiment, the patient has been treated with at least two prior lines of anticancer therapy selected from the group consisting of: ROS1 TKI (e.g., investigational ROS1 TKI, crizotinib, lorlatinib, entrectinib, tarentinib, lopretinib) and chemotherapy.

[0102] In one embodiment, the patient has been treated with at least one line of ROS1 TKI and one line of chemotherapy. In one embodiment, the patient has been treated with at least two lines of ROS1 TKI and one line of chemotherapy. In one embodiment, the patient has been treated with at least three lines of ROS1 TKI and one line of chemotherapy. In one embodiment, the patient has been treated with at least two lines of chemotherapy. In one embodiment, the patient has been treated with at least one line of ROS1 TKI and two lines of chemotherapy. In one embodiment, the patient has been treated with at least two lines of ROS1 TKI and two lines of chemotherapy. In one embodiment, the patient has been treated with at least three lines of ROS1 TKI and two lines of chemotherapy.

[0103] In one embodiment, the ROS1 TKI is crizotinib. In one embodiment, the ROS1 TKI is entrectinib. In one embodiment, the ROS1 TKI is lorlatinib. In one embodiment, the ROS1 TKI is lopatinib. In one embodiment, the patient has been treated with lorlatinib and lopatinib. In one embodiment, the ROS1 TKI is talatinib.

[0104] In one embodiment, the solid tumor is a metastatic ROS1 -positive solid tumor, and the patient has been treated with at least one prior ROS1 TKI therapy.

[0105] In one embodiment, the solid tumor is metastatic ROS1 -positive NSCLC, and the patient has not been treated with TKI therapy and has been treated with at most one prior platinum-based chemotherapy, with or without immunotherapy.

[0106] In one embodiment, the solid tumor is metastatic ROS1 -positive NSCLC, and the patient has been treated with one prior ROS1 TKI therapy and has not been treated with prior platinum-based chemotherapy or immunotherapy.

[0107] In one embodiment, the solid tumor is metastatic ROS1 -positive NSCLC, and the patient has been treated with one prior ROS1 TKI therapy and one prior platinum-based chemotherapy, with or without immunotherapy.

[0108] In one embodiment, the solid tumor is metastatic ROS1 -positive NSCLC, and the patient has been treated with at least two prior ROS1 TKI therapies and at most one prior platinum-based chemotherapy, with or without immunotherapy.

[0109] In one embodiment, the solid tumor is a metastatic ROS1 positive solid tumor, and the patient progresses during the previous therapy. In one embodiment, the previous therapy is a previous ROS1TKI therapy. In one embodiment, the previous therapy is a previous chemotherapy (e.g., platinum-based chemotherapy). In one embodiment, the previous therapy is a previous immunotherapy.

[0110] In one embodiment, the solid tumor is advanced ROS1 -positive NSCLC and the patient has not been treated with TKI therapy and has been treated with at most one prior platinum-based chemotherapy, with or without immunotherapy.

[0111] In one embodiment, the solid tumor is advanced ROS1 -positive NSCLC, and the patient has been treated with one prior ROS1 TKI therapy and has not been treated with prior platinum-based chemotherapy or immunotherapy.

[0112] In one embodiment, the solid tumor is advanced ROS1 -positive NSCLC, and the patient has been treated with one prior ROS1 TKI therapy and one prior platinum-based chemotherapy, with or without immunotherapy.

[0113] In one embodiment, the solid tumor is advanced ROS1-positive NSCLC, and the patient has been treated with at least two prior ROS1 TKI therapies and at most one prior platinum-based chemotherapy, with or without immunotherapy.

[0114] In one embodiment, the solid tumor is an advanced ROS1-positive solid tumor and the patient has progressed on a prior therapy. In one embodiment, the prior therapy is a prior ROS1 TKI therapy.

[0115] In one embodiment, the ROS1 TKI is crizotinib. In one embodiment, the ROS1 TKI is entrectinib.

[0116] In one embodiment, Compound 1 is administered to a patient for one or more days. In one embodiment, Compound 1 is administered to a patient for at least one treatment cycle. In one embodiment, a treatment cycle is at least 7 days. In one embodiment, a treatment cycle is at least 14 days. In one embodiment, a treatment cycle is at least 21 days. In one embodiment, a treatment cycle is at least 28 days.

[0117] In one embodiment, the patient does not experience a Grade 4 adverse event after administration of Compound 1. In one embodiment, the patient does not experience a Grade 3 adverse event after administration of Compound 1. In one embodiment, the patient does not experience a Grade 2 adverse event after administration of Compound 1. In one embodiment, the patient does not experience a Grade 1 adverse event after administration of Compound 1. As used herein and unless otherwise specified, the grade of adverse events follows the grade of the National Cancer Institute Common Terminology Criteria for Adverse Events (CTCAE).

[0118] In one embodiment, the patient experiences only nausea after administration of Compound 1. In one embodiment, the patient does not experience neurological adverse events after administration of Compound 1. In one embodiment, the patient does not experience one or more CNS adverse events selected from the group consisting of dizziness, ataxia, gait disturbance, paresthesia, weight gain, hyperphagia, paresthesia, movement abnormalities, cognitive changes, seizures, hallucinations, speech effects (e.g., aphasia, dysarthria, slow speech or speech disorders), mood disorders (e.g., irritability, anxiety, depression, affective disorders, mood swings, affective disorders, aggression, agitation, mood changes, depressed mood, euphoria, suicidal ideation or mania), mental state, sleep disorders and cognitive disorders (e.g., memory impairment, cognitive disorders, amnesia, confusion, attention disorders, delirium, mental disorders, attention deficit / hyperactivity disorder, dementia, sleep disorders, disorientation or reading disorders). In one embodiment, the neurological adverse event is one or more selected from the group consisting of cognitive impairment, mood disorders, sleep disorders, dizziness and ataxia. In one embodiment, the patient does not have adverse events of weight gain and / or glucose metabolism disorders. In one embodiment, the glucose metabolism disorder is hyperglycemia (e.g., diabetes). In one embodiment, the glucose metabolism disorder is hypoglycemia. In one embodiment, a grade 1 treatment-related adverse event occurs after the patient is administered compound 1. In one embodiment, a grade 1 treatment-related adverse event occurs at most after the patient is administered compound 1. In one embodiment, a grade 1 treatment-related adverse event is fatigue, myalgia, edema (e.g., edema and peripheral edema), alanine aminotransferase (ALT) increases, aspartate aminotransferase (AST) increases, or nausea.

[0119] In one embodiment, the patient has no treatment-related serious adverse events (SAEs). In one embodiment, the patient has no treatment-related CNS adverse events. In one embodiment, the patient has no treatment-related dizziness.

[0120] In one embodiment, less than about 40% of the patient population experience at most a Grade 1 adverse event (e.g., TRAE) after administration of Compound 1. In one embodiment, less than about 30% of the patient population experience at most a Grade 1 adverse event (e.g., TRAE) after administration of Compound 1. In one embodiment, less than about 20% of the patient population experience at most a Grade 1 adverse event (e.g., TRAE) after administration of Compound 1. In one embodiment, less than about 15% of the patient population experience at most a Grade 1 adverse event (e.g., TRAE) after administration of Compound 1. In one embodiment, a Grade 1 adverse event (e.g., TRAE) is fatigue, myalgia, edema (edema and peripheral edema), increased alanine aminotransferase (ALT), increased aspartate aminotransferase (AST), or nausea.

[0121] In one embodiment, less than about 30% of the patient population experience Grade 2 adverse events (e.g., TRAEs) after administration of Compound 1. In one embodiment, less than about 20% of the patient population experience Grade 2 adverse events (e.g., TRAEs) after administration of Compound 1. In one embodiment, less than about 15% of the patient population experience Grade 2 adverse events (e.g., TRAEs) after administration of Compound 1. In one embodiment, less than about 10% of the patient population experience Grade 2 adverse events (e.g., TRAEs) after administration of Compound 1. In one embodiment, less than about 5% of the patient population experience Grade 2 adverse events (e.g., TRAEs) after administration of Compound 1. In one embodiment, no Grade 2 adverse events (e.g., TRAEs) occur in the patient population after administration of Compound 1.

[0122] In one embodiment, less than about 20% of the patient population experience Grade 3 adverse events (e.g., TRAEs) after administration of Compound 1. In one embodiment, less than about 15% of the patient population experience Grade 3 adverse events (e.g., TRAEs) after administration of Compound 1. In one embodiment, less than about 10% of the patient population experience Grade 3 adverse events (e.g., TRAEs) after administration of Compound 1. In one embodiment, less than about 5% of the patient population experience Grade 3 adverse events (e.g., TRAEs) after administration of Compound 1. In one embodiment, no Grade 3 adverse events (e.g., TRAEs) occur in the patient population after administration of Compound 1.

[0123] In one embodiment, less than about 10% of the patient population experience a Grade 4 adverse event (e.g., TRAE) after administration of Compound 1. In one embodiment, less than about 5% of the patient population experience a Grade 4 adverse event (e.g., TRAE) after administration of Compound 1. In one embodiment, less than about 1% of the patient population experience a Grade 4 adverse event (e.g., TRAE) after administration of Compound 1. In one embodiment, no Grade 4 adverse event (e.g., TRAE) occurs in the patient population after administration of Compound 1.

[0124] In one embodiment, the patient has a complete response after one or more cycles of treatment. In one embodiment, the patient has a partial response after one or more cycles of treatment. In one embodiment, the patient reaches stable disease after one or more cycles of treatment. In one embodiment, the patient does not have progressive disease after one or more cycles of treatment.

[0125] As used herein and unless otherwise specified, complete response (CR) refers to the disappearance of all target lesions. In some embodiments of CR, the short axis of any pathological lymph node (whether targeted or non-targeted) must be reduced to <10mm. Partial response (PR) refers to the sum of the diameters of one or more target lesions being reduced by at least 30%. Progressive disease (PD) refers to the sum of the diameters of one or more target lesions being increased by at least 20%. In some embodiments of PD, in addition to a relative increase of 20%, the sum also indicates an increase of at least 5mm. In some embodiments, the occurrence of one or more new lesions is also considered to be progress. Stable disease (SD) refers to neither sufficient reduction to prove PR nor sufficient increase to prove PD, with the minimum diameter sum as a reference during the study. According to the additional description of these terms in RECIST 1.1, it can be consulted in EUROPEAN JOURNAL OF CANCER 45 (2009) 228–247.

[0126] In one embodiment, the patient has brain metastases. In one embodiment, the patient has brain metastases and has not experienced intracranial progression after at least one treatment cycle.

[0127] In one embodiment, the patient has a reduction of at least about 5% to about 100% in the ROS1 allele variant in the circulating tumor DNA after at least one treatment cycle. In one embodiment, the patient has a reduction of at least about 35% in the ROS1 allele variant in the circulating tumor DNA after at least one treatment cycle. In one embodiment, the patient has a reduction of at least about 45% in the ROS1 allele variant in the circulating tumor DNA after at least one treatment cycle. In one embodiment, the patient has a reduction of at least about 55% in the ROS1 allele variant in the circulating tumor DNA after at least one treatment cycle. In one embodiment, the patient has a reduction of at least about 65% in the ROS1 allele variant in the circulating tumor DNA after at least one treatment cycle. In one embodiment, the patient has a reduction of at least about 75% in the ROS1 allele variant in the circulating tumor DNA after at least one treatment cycle. In one embodiment, the patient has a reduction of at least about 85% in the ROS1 allele variant in the circulating tumor DNA after at least one treatment cycle. In one embodiment, the patient has a reduction of at least about 100% in the ROS1 allele variant in the circulating tumor DNA after at least one treatment cycle. In one embodiment, the patient has undetectable ROS1 allelic variant in circulating tumor DNA after at least one treatment cycle. In one embodiment, the ROS1 allelic variant is G2032R.

[0128] In one embodiment, the prior therapy is prior ROS1 TKI therapy. In one embodiment, the ROS1 TKI is crizotinib, entrectinib, lorlatinib, ceritinib, cabozantinib, taceratinib, or lopatinib.

[0129] In one embodiment, administration of Compound 1 provides an area under the curve (AUC) of the compound from 0 to 24 hours. 0-24 ) is in the range of (80% to 125% of 500 ng*h / mL) to (80% to 125% of 30000 ng*h / mL). In one embodiment, administration of Compound 1 provides an area under the curve (AUC 0-24 ) is in the range of about 500 to about 30000 ng*h / mL. In one embodiment, after about half a treatment cycle (e.g., 15 days), the area under the curve (AUC) of the compound from 0 to 24 hours is 0-24 ) is in the range of about 1000 to about 13000 ng*h / mL. In one embodiment, after about half a treatment cycle (e.g., 15 days), the area under the curve (AUC) of the compound from 0 to 24 hours is 0-24) is in the range of about 1500 to about 10000 ng*h / mL. In one embodiment, after about half a treatment cycle (e.g., 15 days), the area under the curve (AUC) of the compound from 0 to 24 hours is 0-24 ) is in the range of about 2000 to about 8000 ng*h / mL.

[0130] In one embodiment, such administration provides, for each mg of Compound 1 administered, an area under the curve (AUC 0-24 ) is in the range of (80% to 125% of 20 ng*h / mL) to (80% to 125% of 500 ng*h / mL). In one embodiment, for each mg of Compound 1 administered, such administration provides an area under the curve (AUC) of the compound from 0 to 24 hours after administration. 0-24 ) is in the range of about 20 to about 500 ng*h / mL. In one embodiment, after about half a treatment cycle (e.g., 15 days), such administration provides an area under the curve (AUC) of the compound from 0 to 24 hours after administration. 0-24 ) is in the range of about 50 to about 200 ng*h / mL.

[0131] In one embodiment, administration of Compound 1 provides an area under the curve of unbound Compound 1 from 0 hours to the last measurable plasma concentration (AUC of Compound 1). 最后,未结合 In the range of (80% to 125% of 400 ng*h / mL) to (80% to 125% of 7000 ng*h / mL). In one embodiment, administration of Compound 1 provides an AUC of 最后,未结合 In the range of about 400 to about 7000 ng*h / mL. In one embodiment, the AUC of the compound is 最后,未结合 In the range of about 1000 to about 6200 ng*h / mL. In one embodiment, after administration of about 25 mg of the compound, the AUC of the compound is 最后,未结合 In the range of about 1100 to about 1500 ng*h / mL. In one embodiment, after administration of about 50 mg of the compound, the AUC of the compound is 最后,未结合 In the range of about 2800 to about 3200 ng*h / mL. In one embodiment, after administration of about 75 mg of the compound, the AUC of the compound is 最后,未结合 In the range of about 3800 to about 4200 ng*h / mL. In one embodiment, after administration of about 100 mg of the compound, the AUC of the compound is 最后,未结合 In the range of about 5500 to about 5900 ng*h / mL. In one embodiment, after administration of about 125 mg of the compound, the AUC of the compound is 最后,未结合In the range of about 4800 to about 5200 ng*h / mL. In one embodiment, after administration of about 150 mg of the compound, the AUC of the compound is 最后,未结合 In the range of about 2600 to about 3100 ng*h / mL. In one embodiment, after administration of about 150 mg of the compound, the AUC of the compound is 最后,未结合 In some embodiments, the last measurable plasma concentration is at 24 hours.

[0132] In one embodiment, such administration provides an area under the curve (AUC) of unbound compound from 0 hours to the last measurable plasma concentration of that compound for each mg of Compound 1 administered. 最后,未结合 ) is in the range of (80% to 125% of 10 ng*h / mL) to (80% to 125% of 80 ng*h / mL). In one embodiment, for each mg of Compound 1 administered, such administration provides a Compound AUC 最后,未结合 In the range of from about 10 to about 80 ng*h / mL. In one embodiment, for each mg of Compound 1 administered, such administration provides a Compound AUC 最后,未结合 In the range of from about 30 to about 70 ng*h / mL. In one embodiment, for each mg of Compound 1 administered, such administration provides a Compound AUC 最后,未结合 In the range of from about 35 to about 65 ng*h / mL. In one embodiment, for each mg of Compound 1 administered, the AUC of the compound is 最后,未结合 In the range of from about 15 to about 25 ng*h / mL. In one embodiment, for each mg of Compound 1 administered, the AUC of the compound is 最后,未结合 In some embodiments, the last measurable plasma concentration is at 24 hours.

[0133] In one embodiment, administration of Compound 1 provides an area under the curve (AUC) of that compound from 0 to the end of the dosing interval. tau ) is in the range of 2000 hr*ng / mL to 10000 hr*ng / mL. In one embodiment, administration of Compound 1 provides an AUC of the compound tau In the range of about 2000 to about 8000 ng*h / mL. In one embodiment, after about half a treatment cycle (e.g., 15 days), administration of Compound 1 provides an AUC of the compound of tau In the range of about 2000 to about 8000 ng*h / mL.

[0134] In one embodiment, administration of Compound 1 provides an area under the curve (AUC) of that compound from 0 to the end of the dosing interval for each mg of Compound 1 administered.tau ) is in the range of 30 hr*ng / mL to 200 hr*ng / mL. In one embodiment, for each mg of Compound 1 administered, the compound AUC provided by administration of Compound 1 is tau In the range of from about 50 to about 150 ng*h / mL. In one embodiment, for each mg of Compound 1 administered after about half a treatment cycle (e.g., 15 days), administration of Compound 1 provides a Compound AUC of tau In the range of about 50 to about 150 ng*h / mL.

[0135] In one embodiment, administration of Compound 1 provides an area under the curve (AUC) of the compound from 0 to infinity. inf ) is in the range of 3500 hr*ng / mL to 20000 hr*ng / mL. In one embodiment, administration of Compound 1 provides an AUC of the compound inf In the range of about 4000 to about 15000 ng*h / mL. In one embodiment, after about half a treatment cycle (e.g., 15 days), administration of Compound 1 provides an AUC of the compound of inf In the range of about 4000 to about 15000 ng*h / mL.

[0136] In one embodiment, administration of Compound 1 provides an area under the curve (AUC) of Compound 1 from 0 to infinity for each mg of Compound 1 administered. inf ) is in the range of 50 hr*ng / mL to 300 hr*ng / mL. In one embodiment, for each mg of Compound 1 administered, administration of Compound 1 provides a Compound AUC of inf In the range of from about 50 to about 250 ng*h / mL. In one embodiment, for each mg of Compound 1 administered after about half a treatment cycle (e.g., 15 days), the administration of Compound 1 provides a Compound AUC of inf In the range of about 50 to about 250 ng*h / mL.

[0137] In one embodiment, such administration provides a maximum plasma concentration (C max ) ranges from (80% to 125% of 100 ng / mL) to (80% to 125% of 1500 ng / mL). In one embodiment, such administration provides a maximum plasma concentration (C max ) is in the range of about 100 to about 1500 ng / mL. In one embodiment, such administration provides a maximum plasma concentration (C max) is in the range of about 200 to about 1000 ng / mL. In one embodiment, after about half a treatment cycle (e.g., 15 days), the C of the compound provided by Compound 1 is administered. max In the range of about 200 to about 1000 ng*h / mL.

[0138] In one embodiment, for each mg of Compound 1 administered, such administration provides a maximum plasma concentration (C max ) is in the range of about 2 to about 50 ng / mL. In one embodiment, for each mg of Compound 1 administered, such administration provides a maximum plasma concentration (C max ) is in the range of about 5 to about 20 ng / mL. In one embodiment, for each mg of Compound 1 administered after about half a treatment cycle (e.g., 15 days), the administration of Compound 1 provides Compound C max In the range of about 5 to about 20 ng / mL.

[0139] In one embodiment, such administration provides a minimum plasma concentration (C min ) is in the range of about 50 to about 400 ng / mL. In one embodiment, such administration provides a minimum plasma concentration (C min ) is in the range of about 50 to about 350 ng / mL. In one embodiment, after about half a treatment cycle (e.g., 15 days), such administration provides a minimum plasma concentration (C ) of Compound 1 achieved during the time interval between administrations of two doses. min ) is in the range of about 50 to about 350 ng / mL.

[0140] In one embodiment, such administration provides a maximum plasma concentration (C max,未结合 ) is in the range of (80% to 125% of 30 ng / mL) to (80% to 125% of 400 ng / mL). In one embodiment, such administration provides an unbound maximum plasma concentration (C max,未结合 ) is in the range of about 30 ng / mL to about 400 ng / mL. In one embodiment, such administration provides an unbound maximum plasma concentration (C max,未结合 ) is in the range of about 50 ng / mL to about 350 ng / mL. In one embodiment, after administration of about 50 mg of the compound, the C of the compound is max,未结合 In the range of about 90 to about 140 ng / mL. In one embodiment, after administration of about 75 mg of the compound, the C of the compound is max,未结合In the range of about 200 to about 250 ng / mL. In one embodiment, after administration of about 100 mg of the compound, the C of the compound max,未结合 In the range of about 300 to about 400 ng / mL. In one embodiment, after administration of about 150 mg of the compound, the C of the compound is max,未结合 In the range of about 200 to about 300 ng / mL. In one embodiment, after administration of about 150 mg of the compound, the C of the compound is max,未结合 In the range of about 200 to about 300 ng / mL. In one embodiment, after administration of about 150 mg of the compound, the C of the compound is max,未结合 is about 258 ng / mL (about 0.62 μM). In one embodiment, after administration of about 100 mg of the compound, the C of the compound is about 258 ng / mL (about 0.62 μM). max,未结合 In the range of about 150 to about 200 ng / mL. In one embodiment, after administration of about 100 mg of the compound, the C of the compound max,未结合 is about 172 ng / mL (about 0.41 μM). In one embodiment, for each mg of Compound 1 administered, such administration provides a C of the compound of max,未结合 In the range of about 1 to about 4 ng / mL. In one embodiment, for each mg of Compound 1 administered, the C of the compound is max,未结合 It is about 1.7ng / mL.

[0141] In one embodiment, without being bound by a particular theory, Compound 1 has a brain penetration (brain-to-plasma ratio) of about 0.26 (about 26%).

[0142] In one embodiment, such administration provides a T of the compound in the range of about 0.25 h to about 5 h after administration. max In one embodiment, such administration provides a T of the compound in the range of about 0.25 h to about 4 h after administration. max In one embodiment, such administration provides a T of the compound in the range of about 0.5 h to about 5 h after administration. max In one embodiment, such administration provides a T of the compound in the range of about 0.5 h to about 2 h after administration. max In one embodiment, such administration provides a T of the compound in the range of about 0.5 h to about 1.0 h after administration. max In one embodiment, such administration provides a T of the compound at about 1 hour after administration. max In one embodiment, such administration provides a T of the compound at about 0.5 h after administration. max .

[0143] In one embodiment, such administration provides a t of the compound in the range of about 2 hours to about 50 hours after administration.1 / 2 In one embodiment, such administration provides a t of the compound within the range of about 8 hours to about 25 hours after administration. 1 / 2 In one embodiment, such administration provides a t of the compound within the range of about 10 hours to about 24 hours after administration. 1 / 2 In one embodiment, such administration provides a t of the compound within the range of about 10 hours to about 20 hours after administration. 1 / 2 In one embodiment, such administration provides a t of the compound within the range of about 16 hours to about 25 hours after administration. 1 / 2 In one embodiment, such administration provides a t of the compound at about 10 h, about 11 h, about 12 h, about 13 h, about 14 h, about 15 h, about 16 h, about 17 h, about 18 h, about 19 h, about 20 h, about 21 h, about 22 h, about 23 h, or about 24 h after administration. 1 / 2 In one embodiment, such administration provides a t of the compound about 20 hours after administration. 1 / 2 .

[0144] In one embodiment, the administration provides a dose plasma concentration of the compound that is at least 70%, 80%, 90%, 95%, 97%, 98% or 99% higher than the IC of the compound against the ROS1 G2032R mutant during about 24 hours immediately following administration. 50 At least 10%, 20%, 30%, 40% or 50% higher.

[0145] A method of reducing lesions in a subject having a ROS1-positive solid tumor (e.g., NSCLC), comprising

[0146] (i) obtaining a first radiological measurement of the size of the lesion;

[0147] (ii) administering a pharmaceutically effective amount of Compound 1 once a day for one or more days; and

[0148] (iii) obtaining a second radiological measurement of the size of the lesion;

[0149] Thereby the second measurement is at most 100% of the first measurement.

[0150] In one embodiment, the second measurement is at most about 90% of the first measurement. In one embodiment, the second measurement is at most about 80% of the first measurement. In one embodiment, the second measurement is at most about 70% of the first measurement. In one embodiment, the second measurement is at most about 60% of the first measurement. In one embodiment, the second measurement is at most about 50% of the first measurement. In one embodiment, the second measurement is about 0.01% to about 90% of the first measurement. In one embodiment, the second measurement shows no detectable lesions.

[0151] In one embodiment, the compound used herein (Compound 1 or its stereoisomer, or mixture of stereoisomers, or pharmaceutically acceptable salt thereof) is administered once a day (QD). In one embodiment, the compound is administered twice a day (BID). In certain embodiments, the compound used herein is Compound 1.

[0152] In one embodiment, the compound is administered in an amount such that the area under the curve (AUC 0-24 ) is in the range of from about 500 to about 30000 ng*h / mL. In one embodiment, AUC 0-24 In the range of from about 750 to about 20000 ng*h / mL. In one embodiment, AUC 0-24 In the range of from about 750 to about 15000 ng*h / mL. In one embodiment, AUC 0-24 In the range of from about 1000 to about 13000 ng*h / mL. In one embodiment, AUC 0-24 In the range of from about 1500 to about 10000 ng*h / mL. In one embodiment, AUC 0-24 In the range of from about 750 to about 4500 ng*h / mL. In one embodiment, AUC 0-24 In the range of from about 1000 to about 3000 ng*h / mL. In one embodiment, AUC 0-24 In the range of from about 1250 to about 3000 ng*h / mL. In one embodiment, AUC 0-24 In the range of from about 1500 to about 2500 ng*h / mL. In one embodiment, AUC 0-24 In the range of from about 1750 to about 7500 ng*h / mL. In one embodiment, AUC 0-24 In the range of from about 2000 to about 6000 ng*h / mL. In one embodiment, AUC 0-24 In the range of from about 3000 to about 5000 ng*h / mL. In one embodiment, AUC 0-24 In the range of from about 3000 to about 14000 ng*h / mL. In one embodiment, AUC 0-24 In the range of from about 3000 to about 9000 ng*h / mL. In one embodiment, AUC 0-24 In the range of from about 4500 to about 7500 ng*h / mL. In one embodiment, AUC 0-24 In the range of from about 4000 to about 13000 ng*h / mL. In one embodiment, AUC0-24 In the range from about 6000 to about 10000 ng*h / mL.

[0153] In one embodiment, the compound is administered in an amount such that the maximum plasma concentration (C max ) is in the range of from about 100 to about 1500 ng / mL. In one embodiment, C max In the range of from about 150 to about 1400 ng / mL. In one embodiment, C max In the range of from about 150 to about 350 ng / mL. In one embodiment, C max In the range of from about 300 to about 900 ng / mL. In one embodiment, C max In the range of from about 300 to about 700 ng / mL. In one embodiment, C max In the range of from about 450 to about 1050 ng / mL. In one embodiment, C max In the range of from about 600 to about 1400 ng / mL.

[0154] In one embodiment, the compound is administered in an amount such that the plasma concentration of the compound is above (e.g., at least 10%, 20%, 30%, 40%, or 50% higher) a predetermined value (e.g., the IC of the compound against the ROS1 G2032R mutant) for at least 70%, 80%, 90%, 95%, 97%, 98%, or 99% of the time following about 24 hours of administration. 50 ).

[0155] In one embodiment, the compound has a half-life in a patient of about 2 to about 50 hours. In one embodiment, a half-life of about 10 to about 20 hours. In one embodiment, a half-life of about 15 to about 25 hours. In one embodiment, a half-life of about 20 hours.

[0156] In one embodiment, the pharmacokinetic parameters provided herein (e.g., AUC 0-24 , AUC 最后,未结合 , C max , T max and half-life) refer to the arithmetic mean of the patient population. In one embodiment, the pharmacokinetic parameters provided herein (e.g., AUC 0-24 , C max ) refers to the arithmetic mean of the patient population. In certain embodiments, t 1 / 2 Refers to the arithmetic mean of a patient population. In certain embodiments, T max Refers to the median range of a patient population. In certain embodiments, AUC 0-24 , AUC 最后未结合, AUC tau , AUC tau DN , AUC inf , AUC inf_DN , C max , C max_DN , C max,未结合 and C min Refers to the geometric mean of the patient population.

[0157] In one embodiment, the pharmacokinetic parameters provided herein (e.g., AUC 0-24 , C max , T max and half-life) are measured after the first administration of the compound to the patient in a treatment cycle (e.g., day 1 of a treatment cycle, e.g., day 1 of cycle 1 of a 28-day treatment cycle). In one embodiment, the pharmacokinetic parameters provided herein (e.g., AUC 0-24 , C max and half-life) are measured after multiple administrations of the compound to a patient during a treatment cycle (eg, the compound may not have reached a steady state in the patient) (eg, Day 15 of Cycle 1 of a 28-day treatment cycle).

[0158] In one embodiment, the compound used herein (Compound 1 or its stereoisomer, or mixture of stereoisomers or pharmaceutically acceptable salt thereof) is administered in an amount of about 5 mg to about 500 mg per day (based on the weight of free base Compound 1). In one embodiment, the compound is administered in an amount of about 25 mg to about 250 mg per day. In one embodiment, the compound is administered in an amount of about 25 mg to about 200 mg per day. In one embodiment, the compound is administered in an amount of about 50 mg to about 200 mg per day. In one embodiment, the compound is administered in an amount of about 5 mg to about 150 mg per day. In one embodiment, the compound is administered in an amount of about 25 mg to about 150 mg per day. In one embodiment, the compound is administered in an amount of about 25 mg to about 125 mg per day. In one embodiment, the compound is administered in an amount of about 25 mg to about 100 mg per day. In one embodiment, the compound is administered in an amount of about 50 mg to about 125 mg per day. In one embodiment, the compound is administered in an amount of about 50 mg to about 100 mg per day. In one embodiment, the compound is administered in an amount of about 5, about 10, about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, about 95, about 100, about 105, about 110, about 115, about 120, about 125, about 130, about 135, about 140, about 145, about 150, about 155, about 160, about 165, about 170, about 175, about 180, about 185, about 190, about 195, or about 200 mg per day. In one embodiment, the amount is about 5 mg per day. In one embodiment, the amount is about 10 mg per day. In one embodiment, the amount is about 15 mg per day. In one embodiment, the amount is about 20 mg per day. In one embodiment, the amount is about 25 mg per day. In one embodiment, the amount is about 30 mg per day. In one embodiment, the amount is about 35 mg per day. In one embodiment, the amount is about 40 mg per day. In one embodiment, the amount is about 45 mg per day. In one embodiment, the amount is about 50 mg per day. In one embodiment, the amount is about 75 mg per day. In one embodiment, the amount is about 100 mg per day. In one embodiment, the amount is about 125 mg per day. In one embodiment, the amount is about 150 mg per day. As used herein, weight refers to the weight of free base compound 1. In certain embodiments, the compound used herein is compound 1.

[0159] In one embodiment, the compound used herein (Compound 1 or its stereoisomer, or mixture of stereoisomers or pharmaceutically acceptable salt thereof) is administered once in an amount of about 5 mg to about 500 mg per day (based on the weight of the free base Compound 1). In one embodiment, the compound is administered once in an amount of about 25 mg to about 250 mg per day. In one embodiment, the compound is administered once in an amount of about 25 mg to about 200 mg per day. In one embodiment, the compound is administered once in an amount of about 50 mg to about 200 mg per day. In one embodiment, the compound is administered once in an amount of about 5 mg to about 150 mg per day. In one embodiment, the compound is administered once in an amount of about 25 mg to about 150 mg per day. In one embodiment, the compound is administered once in an amount of about 25 mg to about 125 mg per day. In one embodiment, the compound is administered once in an amount of about 25 mg to about 100 mg per day. In one embodiment, the compound is administered once in an amount of about 50 mg to about 125 mg per day. In one embodiment, the compound is administered once in an amount of about 50 mg to about 100 mg per day. In one embodiment, the compound is administered once a day in an amount of about 5, about 10, about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, about 95, about 100, about 105, about 110, about 115, about 120, about 125, about 130, about 135, about 140, about 145, about 150, about 155, about 160, about 165, about 170, about 175, about 180, about 185, about 190, about 195, or about 200 mg. In one embodiment, the amount is about 5 mg once a day. In one embodiment, the amount is about 10 mg once a day. In one embodiment, the amount is about 15 mg once a day. In one embodiment, the amount is about 20 mg once a day. In one embodiment, the amount is about 25 mg once a day. In one embodiment, the amount is about 30 mg once a day. In one embodiment, the amount is about 35 mg once a day. In one embodiment, the amount is about 40 mg once a day. In one embodiment, the amount is about 45 mg once a day. In one embodiment, the amount is about 50 mg once a day. In one embodiment, the amount is about 75 mg once a day. In one embodiment, the amount is about 100 mg once a day. In one embodiment, the amount is about 125 mg once a day. In one embodiment, the amount is about 150 mg once a day. As used herein, weight refers to the weight of free base compound 1. In certain embodiments, the compound used herein is compound 1.

[0160] In one embodiment, the compound used herein (Compound 1 or its stereoisomer, or mixture of stereoisomers or pharmaceutically acceptable salt thereof) is administered twice in an amount of about 5 mg to about 500 mg per day (based on the weight of the free base Compound 1). In one embodiment, the compound is administered twice in an amount of about 5 mg to about 250 mg per day. In one embodiment, the compound is administered twice in an amount of about 25 mg to about 250 mg per day. In one embodiment, the compound is administered twice in an amount of about 25 mg to about 200 mg per day. In one embodiment, the compound is administered twice in an amount of about 5 mg to about 100 mg per day. In one embodiment, the compound is administered twice in an amount of about 10 mg to about 50 mg per day. In one embodiment, the compound is administered twice in an amount of about 10 mg to about 100 mg per day. In one embodiment, the compound is administered twice in an amount of about 25 mg to about 100 mg per day. In one embodiment, the compound is administered twice in an amount of about 50 mg to about 100 mg per day. In one embodiment, the compound is administered twice in an amount of about 50 mg to about 100 mg per day. In one embodiment, the compound is administered twice in an amount of about 50 mg to about 100 mg per day. In one embodiment, the compound is administered twice in an amount of about 50 mg to about 150 mg per day. In one embodiment, the compound is administered twice in an amount of about 25 mg to about 150 mg per day. In one embodiment, the compound is administered twice in an amount of about 5, about 10, about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, about 95, about 100, about 105, about 110, about 115, about 120, about 125, about 130, about 135, about 140, about 145, about 150, about 155, about 160, about 165, about 170, about 175, about 180, about 185, about 190, about 195, or about 200 mg per day. In one embodiment, the amount is about 5 mg twice a day. In one embodiment, the amount is about 10 mg twice a day. In one embodiment, the amount is about 15 mg twice a day. In one embodiment, the amount is about 20 mg twice a day. In one embodiment, the amount is about 25 mg twice a day. In one embodiment, the amount is about 30 mg twice a day. In one embodiment, the amount is about 35 mg twice a day. In one embodiment, the amount is about 40 mg twice a day. In one embodiment, the amount is about 45 mg twice a day. In one embodiment, the amount is about 50 mg twice a day. In one embodiment, the amount is about 75 mg twice a day. In one embodiment, the amount is about 100 mg twice a day. In one embodiment, the amount is about 125 mg twice a day. In one embodiment, the amount is about 150 mg twice a day. As used herein, weight refers to the weight of free base compound 1. In certain embodiments, the compound used herein is compound 1.

[0161] In one embodiment, the compound is administered orally.

[0162] In one embodiment, the compound is administered in the form of one or more tablets. In one embodiment, the tablet has a unit dosage strength of about 5 mg based on the weight of free base compound 1. In one embodiment, the tablet has a unit dosage strength of about 25 mg based on the weight of free base compound 1.

[0163] In one embodiment, the compound is administered to a fasting subject. In one embodiment, the compound is administered to a subject in a fasting state. In one embodiment, the compound is administered to a subject who has not taken in food. In one embodiment, the compound is administered to a subject at least 1 hour before taking in a beverage other than food and / or water and no earlier than 2 hours after taking in a beverage other than food and / or water. In one embodiment, the compound is administered to a subject in a full stomach state. In one embodiment, the compound is administered to a subject in a fed state. In one embodiment, the compound is administered to a subject who has taken in food. In one embodiment, the compound is administered together with food and / or beverage intake.

[0164] In one embodiment, the patient experiences an improvement in one or more symptoms selected from the group consisting of cognitive impairment, mood disorders, sleep disorders, dizziness, ataxia, and weight gain after administration of the compound. In one embodiment, the patient does not experience one or more symptoms selected from the group consisting of cognitive impairment, mood disorders, sleep disorders, dizziness, ataxia, and weight gain after administration of the compound. In one embodiment, after administration of the compound, the patient experiences a decrease in the level of one or more of pROS1, ROS1, pAKT, and pERK. In one embodiment, after administration of the compound, the patient experiences a decrease in the expression level of one or more MAP kinase pathway genes in the tumor. In one embodiment, after administration of the compound, the patient experiences a decrease in the expression level of one or more MAP kinase pathway genes in a solid tumor. In one embodiment, one or more MAP kinase pathway genes are selected from the group consisting of: DUSP6, FOS, IL1R1, and SPRY4.

[0165] Cancer is an uncontrolled cell proliferation disease caused by changes in certain genes. Some of these changes occur in genes encoding receptor tyrosine kinases (RTKs), which are a family of membrane-bound proteins that can transmit signals from outside the cell to promote cell survival, growth and proliferation. Abnormal RTK activation can cause excessive cell growth, leading to cancer. Typically, RTKs include an N-terminal domain that binds to an extracellular ligand, a transmembrane domain, and a C-terminal kinase domain that catalyzes intracellular signal transduction.

[0166] In some embodiments, the compound of formula (I) is a human ROS1 inhibitor. ROS1 is an RTK encoded by the ROS1 gene. The ligands and biological functions of human ROS1 are not yet clear, but it has been shown that its homologs in some other species can bind extracellular ligands and stimulate cell differentiation. For example, mouse ROS1 is essential for male gamete maturation and reproduction. In humans, ROS1 chromosomal rearrangements are a well-documented cause of cancer, accounting for 1-2% of non-small cell lung cancer (NSCLC) and a subset of many other cancers. These rearrangements result in the fusion of the C-terminus of ROS1 with the N-terminus of various partner proteins, the most common of which is CD74. ROS1 fusion has constitutive kinase activity, driving tumor growth via MAPK, PI3K and JAK / STAT signal transduction pathways. Small molecule tyrosine kinase inhibitors (TKIs) have been used to target ROS1 fusions in cancer, including crizotinib and entrectinib. Crizotinib is the first FDA-approved TKI for the treatment of ROS1-positive NSCLC. Despite initial responses, most patients develop resistance to crizotinib and relapse. The main mechanism of resistance is the G2032R mutation in the solvent front, which greatly reduces crizotinib affinity. The FDA has not approved any inhibitors with activity against the ROS1-G2032R fusion, indicating a need in the art.

[0167] In one embodiment, the compounds provided herein selectively inhibit ROS1. In one embodiment, the compounds selectively inhibit ROS1 compared to ALK. As non-limiting examples, the selectivity ratio can be greater than about 1.5 times, greater than about 2 times, greater than about 3 times, greater than about 4 times, greater than about 5 times, greater than about 10 times, greater than about 20 times, greater than about 30 times, greater than about 50 or greater than about 100 times, wherein the selectivity can be measured by IC 50 In one embodiment, the selectivity of ROS1 for ALK is measured by the IC 50 Values ​​and IC against ROS1 50 Measured as a ratio of values.

[0168] In one embodiment, the compound selectively inhibits ROS1 (e.g., TRKA, TRKB, and / or TRBC) over TRK. As non-limiting examples, the selectivity ratio may be greater than about 5 times, greater than about 10 times, greater than about 50 times, greater than about 100 times, greater than about 200 times, greater than about 400 times, greater than about 600 times, greater than about 800 times, greater than about 1000 times, greater than about 1500 times, greater than about 2000 times, greater than about 5000 times, greater than about 10,000 times, or greater than about 20,000 times, wherein the selectivity may be measured by IC 50In one embodiment, the selectivity of ROS1 for TRK is measured by IC values ​​for TRK. 50 Values ​​and IC against ROS1 50 Measured as a ratio of values.

[0169] In one embodiment, a method for selectively inhibiting ROS1 compared to ALK is provided herein, wherein the inhibition occurs in a cell. In one embodiment, a method for selectively inhibiting ROS1 (e.g., TRKA, TRKB, and / or TRBC) compared to TRK is provided herein, wherein the inhibition occurs in a cell. In one embodiment, the method includes contacting ROS1 with an effective amount of a compound provided herein. In one embodiment, this contact occurs in a cell. In one embodiment, this contact occurs in a mammal such as a human cell. In one embodiment, this contact occurs in a human patient suffering from a cancer provided herein.

[0170] In one embodiment, provided herein is a method of selectively inhibiting ROS1 compared to ALK, wherein the inhibition occurs in a subject suffering from cancer, the method comprising administering to the subject an effective amount of a compound or pharmaceutical composition provided herein. In certain embodiments, provided herein is a method of treating a subject suffering from a cancer associated with ROS1, the method comprising selectively inhibiting ROS1 compared to ALK by administering to the subject an amount of a compound or pharmaceutical composition provided herein, wherein the amount is sufficient to selectively inhibit ROS1 compared to ALK.

[0171] In one embodiment, provided herein is a method for selectively inhibiting ROS1 (e.g., TRKA, TRKB, and / or TRBC) compared to TRK, wherein the inhibition occurs in a subject suffering from cancer, the method comprising administering to the subject an effective amount of a compound or pharmaceutical composition provided herein. In certain embodiments, provided herein is a method for treating a subject suffering from a cancer associated with ROS1, the method comprising selectively inhibiting ROS1 (e.g., TRKA, TRKB, and / or TRBC) compared to TRK by administering to the subject an amount of a compound or pharmaceutical composition provided herein, wherein the amount is sufficient to selectively inhibit ROS1 (e.g., TRKA, TRKB, and / or TRBC) compared to TRK.

[0172] As used herein and unless otherwise specified, inhibiting ROS1 includes inhibiting wild-type ROS1 or a mutation thereof; inhibiting ALK includes inhibiting wild-type ALK or a mutation thereof; and inhibiting TRK includes inhibiting wild-type TRK or a mutation thereof.

[0173] Cancers treated by the methods provided herein include, but are not limited to, lung cancer, such as non-small cell lung cancer, inflammatory myofibroblastic tumor, ovarian cancer, such as serous ovarian cancer, melanoma, such as spitzoid melanoma, glioblastoma, bile duct cancer, such as cholangioepithelial carcinoma, gastric cancer, colorectal cancer, angiosarcoma, anaplastic large cell lymphoma, diffuse large B-cell lymphoma, large B-cell lymphoma, esophageal cancer, such as esophageal squamous cell carcinoma, kidney cancer, such as renal medullary carcinoma or renal cell carcinoma, breast cancer, such as triple-negative breast cancer, thyroid cancer, such as papillary thyroid carcinoma, neuroblastoma, epithelioid hemangioendothelioma, colon cancer, and Spitz nevus tumor.

[0174] Cancers treated by the methods provided herein include cancers derived from one or more oncoproteins selected from ROS1, ALK, TRKA, TRKB and TRKC. In certain embodiments, cancers treated by the methods provided herein include cancers resistant to treatment for one or more oncoproteins selected from ROS1, ALK, TRKA, TRKB and TRKC.

[0175] In one embodiment, the cancer in the method provided herein is ROS1 positive (ROS1+). As used herein and unless otherwise specified, "ROS1 positive" (ROS1+) cancer, disease or condition refers to a cancer, disease or condition characterized by abnormally high expression of the ROS1 gene and / or the presence of a mutation in the ROS1 gene. In one embodiment, the mutation changes the biological activity of the ROS1 nucleic acid molecule or polypeptide. As used herein and unless otherwise specified, "mutation" or "mutant" of ROS1 includes one or more deletions, substitutions, insertions, inversions, repetitions, translocations or amplifications in the amino acid or nucleotide sequence of ROS1 or its fragments. As used herein and unless otherwise specified, ROS1 "rearrangement" refers to a genetic translocation involving the ROS1 gene, which can produce a ROS1 fusion gene and / or a ROS1 fusion protein. ROS1 fusions can also include one or more deletions, substitutions, insertions, inversions, repetitions, translocations or amplifications or their fragments, as long as the mutant retains kinase phosphorylation activity.

[0176] In one embodiment, ROS1 mutations include one or more of ROS1 point mutations. In some embodiments, the cancer treated by the methods provided herein includes one or more mutations in ROS1 kinases. In one embodiment, one or more ROS1 point mutations are selected from E1935, L1947, L1951, G1971, E1974, L1982, S1986, F2004, E2020, L2026, G2032, D2033, C2060, F2075, L2086, V2089, V2098, G2101, D2113, 1981Tins, M2001T and L2155. In one embodiment, one or more ROS1 point mutations are selected from G2032R, G2032K, D2033N, S1986F, S1986Y, L2026M, L1951R, E1935G, L1947R, G1971E, E1974K, L1982F, F2004C, F2004V, E2020K, C2060G, F2075V, V2089M, V2098I, G2101A, D2113N, D2113G, L2155S and L2086F. In one embodiment, ROS1 mutates to G2032R. In one embodiment, ROS1 mutates to S1986F. In one embodiment, ROS1 mutates to S1986Y. In one embodiment, ROS1 mutates to L2026M. In one embodiment, ROS1 mutates to D2033N. In one embodiment, ROS1 mutates to L2086F. In one embodiment, ROS1 mutates to F2004C. In one embodiment, ROS1 mutates to F2004V. In one embodiment, ROS1 mutates to G2101A. In one embodiment, ROS1 mutates to L1982F. In one embodiment, ROS1 mutates to a co-mutation of one or more of G2032R and S1986F, S1986Y, F2004C, F2004V, L2026M or D2033N.

[0177] In one embodiment, a ROS1 mutation comprises one or more ROS1 rearrangements (in one embodiment, one rearrangement). In one embodiment, a ROS1 mutation comprises one or more ROS1 fusions (in one embodiment, one fusion). In some embodiments, a cancer treated by the methods provided herein comprises a ROS1 fusion. In one embodiment, ROS1 is fused to one of the fusion partners selected from the group consisting of SLC34A2, CD74, TPM3, SDC4, EZR, LRIG3, KDELR2, CEP72, CLTL, CTNND2, GOPC (e.g., GOPC-S, GOPC-L), GPRC6A, LIMA1, LRIG3, MSN, MYO5C, OPRM1, SLC6A17 SLMAP, SRSF6, TFG, TMEM106B, TPD52L1, ZCCHC8, CCDC6, CAPRIN1, CEP85L, CHCHD3, CLIP1, EEF1G, KIF21A, KLC1, SART3, ST13, TRIM24, ERC1, FIP1L1, HLAA, KIAA1598, MYO5A, PPFIBP1, PWWP2A, FN1, YWHAE, CCDC30, NCOR2, NFKB2, APOB, PLG, RBP4, and GOLGB1. In one embodiment, the ROS1 fusion is a CD74-ROS1 fusion. In one embodiment, the ROS1 fusion is a SDC4-ROS1 fusion. In one embodiment, the ROS1 fusion is an EZR-ROS1 fusion. In one embodiment, the ROS1 fusion is a SLC34A2-ROS1 fusion. In one embodiment, the ROS1 fusion is a GOPC-ROS1 fusion (eg, GOPC-ROS1-S, GOPC-ROS1-L). In one embodiment, the ROS1 fusion is a CEP85L-ROS1 fusion.

[0178] In one embodiment, ROS1 mutations include a ROS1 rearrangement and one or more ROS1 point mutations. In one embodiment, ROS1 mutations include one or more ROS1 rearrangements from CD74-ROS1, EZR-ROS1, SLC34A2-ROS1, GOPC-ROS1 (e.g., GOPC-ROS1-S, GOPC-ROS1-L) and CEP85L-ROS1, and one or more ROS1 point mutations selected from F2004C, F2004V and G2032R. In one embodiment, ROS1 mutations include one or more ROS1 rearrangements from CD74-ROS1, EZR-ROS1 and SLC34A2-ROS1, and a ROS1 point mutation of G2101A.

[0179] In one embodiment, ROS1 mutates to CD74-ROS1 F2004C. In one embodiment, ROS1 mutates to CD74-ROS1 F2004V. In one embodiment, ROS1 mutates to CD74-ROS1 G2101A. In one embodiment, ROS1 mutates to CD74-ROS1 G2032R. In one embodiment, ROS1 mutates to CD74-ROS1 S1986F. In one embodiment, ROS1 mutates to CD74-ROS1 L2026M. In one embodiment, ROS1 mutates to CD74-ROS1 D2033N. In one embodiment, ROS1 mutates to EZR-ROS1 F2004C. In one embodiment, ROS1 mutates to EZR-ROS1 F2004V. In one embodiment, ROS1 mutates to EZR-ROS1 G2101A. In one embodiment, ROS1 mutates to EZR-ROS1 G2032R. In one embodiment, ROS1 is mutated to SLC34A2-ROS1F2004C. In one embodiment, ROS1 is mutated to SLC34A2-ROS1 F2004V. In one embodiment, ROS1 is mutated to SLC34A2-ROS1 G2101A. In one embodiment, ROS1 is mutated to SLC34A2-ROS1 G2032R. In one embodiment, ROS1 is mutated to GOPC-ROS1 F2004C (e.g., GOPC-ROS1-S F2004C, GOPC-ROS1-LF2004C). In one embodiment, ROS1 is mutated to GOPC-ROS1 F2004V (e.g., GOPC-ROS1-S F2004V, GOPC-ROS1-L F2004V). In one embodiment, ROS1 is mutated to GOPC-ROS1 G2032R (e.g., GOPC-ROS1-SG2032R, GOPC-ROS1-L G2032R). In one embodiment, ROS1 is mutated to CEP85L-ROS1F2004C. In one embodiment, ROS1 is mutated to CEP85L-ROS1 F2004V. In one embodiment, ROS1 is mutated to CEP85L-ROS1G2032R. In one embodiment, ROS1 is mutated to GOPC-ROS1 L1982F (e.g., GOPC-ROS1-S L1982F, GOPC-ROS1-L L1982F). In one embodiment, ROS1 is mutated to CD74-ROS1L1982F.

[0180] In some embodiments, provided herein is a method for treating a patient population with ROS1-positive solid tumors, comprising administering to each of the patient population an effective amount of compound 1 or a pharmaceutical composition provided herein, wherein the objective response rate (ORR) after one treatment cycle of compound 1 is at least about 10% (RECIST 1.1). In one embodiment, the ORR after one treatment cycle of compound 1 is at least about 20% (RECIST 1.1). In one embodiment, the ORR after one treatment cycle of compound 1 is at least about 30% (RECIST 1.1). In one embodiment, the ORR after one treatment cycle of compound 1 is at least about 40% (RECIST 1.1). In one embodiment, the ORR after one treatment cycle of compound 1 is at least about 50% (RECIST 1.1). In one embodiment, the ORR after one treatment cycle of compound 1 is at least about 60% (RECIST 1.1). In one embodiment, the ORR after one treatment cycle of compound 1 is at least about 70% (RECIST 1.1). In one embodiment, the ORR is at least about 80% (RECIST 1.1) after one treatment cycle of Compound 1. In one embodiment, the ORR is at least about 90% (RECIST 1.1) after one treatment cycle of Compound 1. In one embodiment, at least about 20% of the patient population achieves a partial response. In one embodiment, at least about 30% of the patient population achieves a partial response. In one embodiment, at least about 40% of the patient population achieves a partial response. In one embodiment, at least about 50% of the patient population achieves a partial response. In one embodiment, at least about 60% of the patient population achieves a partial response. In one embodiment, at least about 70% of the patient population achieves a partial response. In one embodiment, at least about 80% of the patient population achieves a partial response. In one embodiment, at least about 10% of the patient population achieves stable disease. In one embodiment, at least about 20% of the patient population achieves stable disease. In one embodiment, at least about 30% of the patient population achieves stable disease. In one embodiment, at least about 40% of the patient population achieves stable disease. In one embodiment, at least about 50% of the patient population achieves stable disease. In one embodiment, at least about 60% of the patient population achieves stable disease. In one embodiment, at least about 70% of the patient population achieves stable disease.

[0181] In some embodiments, provided herein is a method for treating a patient population having a solid tumor with a ROS1 mutation (e.g., G2032R), comprising administering to each of the patient population an effective amount of Compound 1 or a pharmaceutical composition provided herein, wherein the ORR is at least about 30% (RECIST 1.1) after one treatment cycle of Compound 1. In one embodiment, the ORR is at least about 40% (RECIST 1.1) after one treatment cycle of Compound 1. In one embodiment, the ORR is at least about 50% (RECIST 1.1) after one treatment cycle of Compound 1. In one embodiment, the ORR is at least about 60% (RECIST 1.1) after one treatment cycle of Compound 1. In one embodiment, the ORR is at least about 70% (RECIST 1.1) after one treatment cycle of Compound 1. In one embodiment, the ORR is at least about 80% (RECIST 1.1) after one treatment cycle of Compound 1. In one embodiment, the ORR is at least about 90% (RECIST 1.1) after one treatment cycle of Compound 1. In one embodiment, at least about 20% of the patient population achieves a partial response. In one embodiment, at least about 30% of the patient population achieves a partial response. In one embodiment, at least about 40% of the patient population achieves a partial response. In one embodiment, at least about 50% of the patient population achieves a partial response. In one embodiment, at least about 60% of the patient population achieves a partial response. In one embodiment, at least about 70% of the patient population achieves a partial response. In one embodiment, at least about 80% of the patient population achieves a partial response. In one embodiment, at least about 90% of the patient population achieves a partial response. In one embodiment, at least about 5% of the patient population achieves stable disease. In one embodiment, at least about 10% of the patient population achieves stable disease. In one embodiment, at least about 15% of the patient population achieves stable disease. In one embodiment, at least about 20% of the patient population achieves stable disease. In one embodiment, at least about 30% of the patient population achieves stable disease. In one embodiment, at least about 40% of the patient population achieves stable disease. In one embodiment, at least about 50% of the patient population achieves stable disease. In one embodiment, at least about 60% of the patient population achieves stable disease.

[0182] In some embodiments, provided herein is a method for treating a patient population having ROS1-positive solid tumors and CNS diseases at baseline, comprising administering to each of the patient population an effective amount of compound 1 or a pharmaceutical composition provided herein, wherein the ORR is at least about 30% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 40% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 50% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 60% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 70% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 80% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 90% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, at least about 30% of the patient population achieves a partial response. In one embodiment, at least about 40% of the patient population achieves a partial response. In one embodiment, at least about 50% of the patient population achieves a partial response. In one embodiment, at least about 60% of the patient population achieves a partial response. In one embodiment, at least about 70% of the patient population achieves a partial response. In one embodiment, at least about 80% of the patient population achieves a partial response. In one embodiment, at least about 90% of the patient population achieves a partial response. In one embodiment, at least about 5% of the patient population achieves stable disease. In one embodiment, at least about 10% of the patient population achieves stable disease. In one embodiment, at least about 15% of the patient population achieves stable disease. In one embodiment, at least about 20% of the patient population achieves stable disease. In one embodiment, at least about 30% of the patient population achieves stable disease. In one embodiment, at least about 40% of the patient population achieves stable disease. In one embodiment, at least about 50% of the patient population achieves stable disease. In one embodiment, at least about 60% of the patient population achieves stable disease.

[0183] In some embodiments, provided herein is a method for treating a patient population with ROS1-positive solid tumors after receiving at least two prior ROS1 TKI therapies and at least one chemotherapy, comprising administering to each of the patient population an effective amount of compound 1 or a pharmaceutical composition provided herein, wherein the ORR is at least about 10% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 20% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 30% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 40% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 50% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 60% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 70% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 80% (RECIST 1.1) after one treatment cycle of Compound 1. In one embodiment, the ORR is at least about 90% (RECIST 1.1) after one treatment cycle of Compound 1. In one embodiment, at least about 20% of the patient population achieves a partial response. In one embodiment, at least about 30% of the patient population achieves a partial response. In one embodiment, at least about 40% of the patient population achieves a partial response. In one embodiment, at least about 50% of the patient population achieves a partial response. In one embodiment, at least about 60% of the patient population achieves a partial response. In one embodiment, at least about 70% of the patient population achieves a partial response. In one embodiment, at least about 80% of the patient population achieves a partial response. In one embodiment, at least about 90% of the patient population achieves a partial response. In one embodiment, at least 10% of the patient population achieves stable disease. Stable disease is achieved. In one embodiment, at least about 20% of the patient population achieves stable disease. In one embodiment, at least about 30% of the patient population achieves stable disease. In one embodiment, at least about 40% of the patient population achieves stable disease. In one embodiment, at least about 50% of the patient population achieves stable disease. In one embodiment, at least about 60% of the patient population achieves stable disease. In one embodiment, at least about 70% of the patient population achieves stable disease. In one embodiment, at least about 80% of the patient population achieves stable disease.

[0184] In some embodiments, provided herein is a method for treating a patient population with ROS1-positive solid tumors after receiving at least one prior ROS1 TKI therapy and at least one chemotherapy, comprising administering to each of the patient population an effective amount of compound 1 or a pharmaceutical composition provided herein, wherein the ORR is at least about 10% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 20% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 30% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 40% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 50% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 60% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 70% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 80% (RECIST 1.1) after one treatment cycle of Compound 1. In one embodiment, the ORR is at least about 90% (RECIST 1.1) after one treatment cycle of Compound 1. In one embodiment, at least about 20% of the patient population achieves a partial response. In one embodiment, at least about 30% of the patient population achieves a partial response. In one embodiment, at least about 40% of the patient population achieves a partial response. In one embodiment, at least about 50% of the patient population achieves a partial response. In one embodiment, at least about 60% of the patient population achieves a partial response. In one embodiment, at least about 70% of the patient population achieves a partial response. In one embodiment, at least about 80% of the patient population achieves a partial response. In one embodiment, at least about 90% of the patient population achieves a partial response. In one embodiment, at least 10% of the patient population achieves stable disease. Stable disease is achieved. In one embodiment, at least about 20% of the patient population achieves stable disease. In one embodiment, at least about 30% of the patient population achieves stable disease. In one embodiment, at least about 40% of the patient population achieves stable disease. In one embodiment, at least about 50% of the patient population achieves stable disease. In one embodiment, at least about 60% of the patient population achieves stable disease. In one embodiment, at least about 70% of the patient population achieves stable disease. In one embodiment, at least about 80% of the patient population achieves stable disease.

[0185] In some embodiments, provided herein is a method for treating a patient population having ROS1-positive solid tumors after receiving prior treatment with lorlatinib and / or lopatinib, comprising administering to each of the patient population an effective amount of compound 1 or a pharmaceutical composition provided herein, wherein the ORR is at least about 10% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 20% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 30% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 40% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 50% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 60% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 70% (RECIST 1.1) after one treatment cycle of compound 1. In one embodiment, the ORR is at least about 80% (RECIST 1.1) after one treatment cycle of Compound 1. In one embodiment, the ORR is at least about 90% (RECIST 1.1) after one treatment cycle of Compound 1. In one embodiment, at least about 20% of the patient population achieves a partial response. In one embodiment, at least about 30% of the patient population achieves a partial response. In one embodiment, at least about 40% of the patient population achieves a partial response. In one embodiment, at least about 50% of the patient population achieves a partial response. In one embodiment, at least about 60% of the patient population achieves a partial response. In one embodiment, at least about 70% of the patient population achieves a partial response. In one embodiment, at least about 80% of the patient population achieves a partial response. In one embodiment, at least about 90% of the patient population achieves a partial response. In one embodiment, at least 10% of the patient population achieves stable disease. Achieved stable disease. In one embodiment, at least about 10% of the patient population achieves stable disease. In one embodiment, at least about 20% of the patient population achieves stable disease. In one embodiment, at least about 30% of the patient population achieves stable disease. In one embodiment, at least about 40% of the patient population achieves stable disease. In one embodiment, at least about 50% of the patient population achieves stable disease. In one embodiment, at least about 60% of the patient population achieves stable disease. In one embodiment, at least about 70% of the patient population achieves stable disease. In one embodiment, at least about 80% of the patient population achieves stable disease.

[0186] In one embodiment, ROS1+ cancer is determined by an FDA approved test or other test known in the art. Tests that can be used include, for example, Thermo Fisher Scientific's Oncomine TM Dx Target Test. (a qualitative in vitro diagnostic test that uses targeted high-throughput, parallel sequencing technology to detect sequence changes in 23 genes in DNA and RNA isolated from formalin-fixed, paraffin-embedded (FFPE) tumor tissue specimens from patients with non-small cell lung cancer (NSCLC) using the Ion PGMDx System); Vysis ROS1 BreakApart FISH Probe Kit (a qualitative test that detects rearrangements involving the ROS1 gene rearrangement at 6q22 in formalin-fixed, paraffin-embedded (FFPE) non-small cell lung cancer (NSCLC) tissue specimens via fluorescence in situ hybridization (FISH)) or reverse transcription-polymerase chain reaction (RT-PCR) or next-generation sequencing (NGS) via a local diagnostic test.

[0187] Also provided is a method for treating a subject with cancer (e.g., a ROS1-positive cancer), comprising: determining whether a cancer cell sample obtained from a subject with cancer and previously administered a first ROS1 inhibitor has one or more ROS1 inhibitor resistance mutations; and if the subject has cancer cells with one or more ROS1 inhibitor resistance mutations, administering a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof as a monotherapy or in combination with another anticancer agent to the subject. In some embodiments, the one or more ROS1 inhibitor resistance mutations confer increased resistance to cancer cells or tumors to treatment with a first ROS1 inhibitor. In some embodiments, the one or more ROS1 inhibitor resistance mutations include one or more ROS1 inhibitor resistance mutations. For example, the one or more ROS1 inhibitor resistance mutations may include substitutions at one or more of amino acid positions 2032, 2033, 1986, 2026, 1951, 1935, 1947, 1971, 1974, 1982, 2004, 2020, 2060, 2075, 2089, 2098, 2101, 2113, 2155, 2032, and 2086, such as G2032R, D2032R, D2033 3N, S1986F, S1986Y, L2026M, L1951R, E1935G, L1947R, G1971E, E1974K, L1982F, F2004C, F2004V, E2020K, C2060G, F2075V, V2089M, V2098I, G2101A, D2113N, D2113G, L2155S, L2032K and L2086F. In some embodiments, the another anticancer agent is any anticancer agent known in the art. For example, the another anticancer agent may be another ROS1 inhibitor (e.g., a second ROS1 inhibitor).

[0188] In one embodiment, provided herein is a compound that penetrates the CNS. In one embodiment, after administering an effective amount of a compound provided herein (e.g., orally or intravenously), the compound is able to penetrate the CNS (e.g., the blood-brain barrier) and reach a concentration in the CNS (e.g., the brain) that is still sufficient to inhibit (e.g., selectively inhibit) ROS1.

[0189] In one embodiment, provided herein is a method for treating CNS metastasis of cancer, comprising administering to a subject in need thereof an effective amount of a compound provided herein, e.g., a compound of formula (I) or a stereoisomer thereof, or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof. In one embodiment, CNS metastasis is brain metastasis. In one embodiment, cancer is ROS1+ cancer.

[0190] In some embodiments, the compound is an inhibitor of human tropomyosin receptor kinase A, B or C. In certain embodiments, the compound inhibits the IC of mutant or non-mutant ROS1 or ALK. 50 Does not exceed the IC of the compound for inhibition of wild-type tropomyosin receptor kinase A, B, or C 50 TRK inhibition, especially in the central nervous system (CNS), is associated with adverse reactions including dizziness / ataxia / gait disturbance, paresthesia, weight gain, and cognitive changes.

[0191] In some embodiments, a method for minimizing treatment-related adverse events in a subject in need of treatment of cancer (e.g., ROS1-positive cancer) is provided, the method comprising administering to the subject a therapeutically effective amount of a compound provided herein, for example, a compound of formula (I), a stereoisomer thereof, or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, and wherein the method minimizes treatment-related adverse events associated with a TRK inhibitor. In some embodiments, cancer is a ROS1-related cancer. In some embodiments, the adverse event is one or more of a TRK-related CNS adverse event.

[0192] As used herein, "minimizing" adverse events refers to reducing the incidence of adverse events in subjects or patient populations compared to the example incidence of adverse events in subjects or patient populations treated with TRK inhibitors (e.g., entrectinib, lopatinib or lorlatinib). In some embodiments, the incidence of adverse events refers to the frequency or percentage of specific adverse events in subjects or patient populations. In some embodiments, the incidence of adverse events refers to the total number of adverse events experienced by individual subjects. In some embodiments, minimizing adverse events refers to minimizing TRK-related CNS adverse events. In some embodiments, minimizing TRK-related CNS adverse events means that less than 40% of the patient population has TRK-related CNS adverse events. In some embodiments, minimizing TRK-related CNS adverse events means that less than 35%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10% or less than 5% of the patient population has TRK-related CNS adverse events. In some embodiments, minimizing TRK-related CNS adverse events means that less than 12% of the patient population has more than one TRK-related CNS adverse event. In some embodiments, minimizing TRK-related CNS adverse events means less than 11%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, or less than 3% of the patient population has more than one TRK-related CNS adverse event.

[0193] In some embodiments, a TRK-related CNS treatment-related adverse event refers to one or more of: dizziness, ataxia, gait disturbance, paresthesia, weight gain, increased appetite, paresthesia, movement abnormalities, cognitive changes, speech effects (e.g., dysarthria, slow speech, or speech disorders), mood disorders (e.g., irritability, anxiety, depression, mood swings, personality changes, mood swings, affective disorders, aggression, restlessness, mood changes, depressed mood, euphoria, or mania), and cognitive disorders (e.g., memory impairment, cognitive disorders, amnesia, confusion, attention disorders, delirium, mental disorders, attention-deficit / hyperactivity disorder, dementia, sleep disorders, or reading disorders).

[0194] In one embodiment, provided herein is a method for preventing or limiting CNS side effects or adverse events related to TRK-related treatment in cancer treatment, including administering an effective amount of a compound provided herein to a subject in need, for example, a compound of formula (I) or its stereoisomer, or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof. In one embodiment, the method prevents the occurrence of TRK-related CNS adverse events. In one embodiment, the method limits the frequency of occurrence of TRK-related CNS adverse events. In one embodiment, the method limits the severity of TRK-related side effects. In one embodiment, provided herein is a method for treating CNS metastasis of cancer by reducing TRK-related side effects, including administering an effective amount of a compound provided herein to a subject in need, for example, a compound of formula (I) or its stereoisomer, or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof. In one embodiment, the reduction / limitation / prevention of CNS side effects or adverse events is determined in a statistical sample as compared to standard of care treatment for ROS1+ and / or ALK+ cancers, e.g., approved ROS1 and / or ALK inhibitors (e.g., crizotinib, entrectinib, lorlatinib, or lopatinib). In one embodiment, TRK-related side effects are TRKB-related CNS side effects. In one embodiment, TRK-related CNS side effects or adverse events are dizziness, ataxia, gait disturbance, paresthesia, weight gain, cognitive impairment, mood disorders, or sleep disorders.

[0195] In one embodiment, provided herein is a method for treating cancer, comprising administering to a subject in need thereof a therapeutically effective amount of a compound provided herein, e.g., a compound of formula (I) or a stereoisomer thereof, or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof. In one embodiment, the cancer is a ROS1-related cancer. In one embodiment, the cancer is a ROS1+ cancer. In one embodiment, the cancer is a ROS1-associated cancer.

[0196] In one embodiment, provided herein is a method for treating ROS1+ cancer, comprising administering to a subject in need thereof a therapeutically effective amount of a compound provided herein, e.g., a compound of formula (I) or a stereoisomer thereof, or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof.

[0197] In one embodiment, provided herein is a method for treating cancer in a subject, comprising: (i) identifying the subject's cancer as ROS1+, and (ii) administering to the subject a therapeutically effective amount of a compound provided herein, e.g., a compound of formula (I) or a stereoisomer thereof, or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof.

[0198] In one embodiment, cancer (or ROS1+ cancer) is a solid tumor. In one embodiment, cancer (or ROS1+ cancer) is an advanced solid tumor. In one embodiment, cancer (or ROS1+ cancer) is a locally advanced solid tumor. In one embodiment, cancer (or ROS1+ cancer) is lung cancer, such as non-small cell lung cancer (NSCLC) or bronchial cancer, glioblastoma, inflammatory myofibroblastic tumor (IMT), bile duct cancer such as bile duct cancer, ovarian cancer such as serous ovarian cancer, gastric cancer, colorectal cancer, angiosarcoma, melanoma such as spitz-like melanoma, epithelial hemangioendothelioma, esophageal cancer such as esophageal squamous cell carcinoma (ESCC), renal cancer such as renal medullary carcinoma or renal cell carcinoma, breast cancer such as triple-negative breast cancer, colon cancer, thyroid cancer such as papillary thyroid cancer, Spitz nevus-like tumor, pancreatic cancer, inflammatory hepatocellular adenoma or neuroblastoma.

[0199] In one embodiment, the cancer is lung cancer. In one embodiment, the cancer is non-small cell lung cancer. In one embodiment, the cancer is ROS1+ non-small cell lung cancer. In one embodiment, the cancer is relapsed or refractory non-small cell lung cancer. In one embodiment, the cancer is relapsed or refractory ROS1+ non-small cell lung cancer. In one embodiment, the cancer is newly diagnosed non-small cell lung cancer. In one embodiment, the cancer is newly diagnosed ROS1+ non-small cell lung cancer. In one embodiment, the cancer is bronchial carcinoma. In one embodiment, the cancer is ROS1+ bronchial carcinoma. In one embodiment, the cancer is relapsed or refractory bronchial carcinoma. In one embodiment, the cancer is relapsed or refractory ROS1+ bronchial carcinoma. In one embodiment, the cancer is newly diagnosed bronchial carcinoma. In one embodiment, the cancer is newly diagnosed ROS1+ bronchial carcinoma.

[0200] In one embodiment, the cancer is glioblastoma. In one embodiment, the cancer is relapsed or refractory glioblastoma. In one embodiment, the cancer is relapsed or refractory ROS1+ glioblastoma. In one embodiment, the cancer is newly diagnosed glioblastoma. In one embodiment, the cancer is newly diagnosed ROS1+ glioblastoma.

[0201] In one embodiment, the cancer is IMT. In one embodiment, the cancer is ROS1+IMT. In one embodiment, the cancer is relapsed or refractory IMT. In one embodiment, the cancer is relapsed or refractory ROS1+IMT. In one embodiment, the cancer is newly diagnosed IMT. In one embodiment, the cancer is newly diagnosed ROS1+IMT.

[0202] In one embodiment, the cancer is cholangiocarcinoma. In one embodiment, the cancer is cholangiocarcinoma. In one embodiment, the cancer is ROS1+ cholangiocarcinoma. In one embodiment, the cancer is relapsed or refractory cholangiocarcinoma. In one embodiment, the cancer is relapsed or refractory ROS1+ cholangiocarcinoma. In one embodiment, the cancer is newly diagnosed cholangiocarcinoma. In one embodiment, the cancer is newly diagnosed ROS1+ cholangiocarcinoma.

[0203] In one embodiment, the cancer is ovarian cancer. In one embodiment, the cancer is ROS1+ ovarian cancer. In one embodiment, the cancer is relapsed or refractory ovarian cancer. In one embodiment, the cancer is relapsed or refractory ROS1+ ovarian cancer. In one embodiment, the cancer is newly diagnosed ovarian cancer. In one embodiment, the cancer is newly diagnosed ROS1+ ovarian cancer. In one embodiment, the ovarian cancer is serous ovarian cancer. In one embodiment, the ovarian cancer is high-grade serous ovarian cancer.

[0204] In one embodiment, the cancer is gastric cancer. In one embodiment, the cancer is ROS1+ gastric cancer. In one embodiment, the cancer is recurrent or refractory gastric cancer. In one embodiment, the cancer is recurrent or refractory ROS1+ gastric cancer. In one embodiment, the cancer is newly diagnosed gastric cancer. In one embodiment, the cancer is newly diagnosed ROS1+ gastric cancer.

[0205] In one embodiment, the cancer is colorectal cancer. In one embodiment, the cancer is ROS1+ colorectal cancer. In one embodiment, the cancer is recurrent or refractory colorectal cancer. In one embodiment, the cancer is recurrent or refractory ROS1+ colorectal cancer. In one embodiment, the cancer is newly diagnosed colorectal cancer. In one embodiment, the cancer is newly diagnosed ROS1+ colorectal cancer.

[0206] In one embodiment, the cancer is angiosarcoma. In one embodiment, the cancer is ROS1+ angiosarcoma. In one embodiment, the cancer is relapsed or refractory angiosarcoma. In one embodiment, the cancer is relapsed or refractory ROS1+ angiosarcoma. In one embodiment, the cancer is newly diagnosed angiosarcoma. In one embodiment, the cancer is newly diagnosed ROS1+ angiosarcoma.

[0207] In one embodiment, the cancer is melanoma. In one embodiment, the cancer is a Spitz tumor. In one embodiment, the cancer is a Spitz melanoma. In one embodiment, the cancer is a ROS1+ Spitz melanoma. In one embodiment, the cancer is a relapsed or refractory Spitz melanoma. In one embodiment, the cancer is a relapsed or refractory ROS1+ Spitz melanoma. In one embodiment, the cancer is a newly diagnosed Spitz melanoma. In one embodiment, the cancer is a newly diagnosed ROS1+ Spitz melanoma.

[0208] In one embodiment, the cancer is epithelial hemangioendothelioma. In one embodiment, the cancer is ROS1+ epithelial hemangioendothelioma. In one embodiment, the cancer is relapsed or refractory epithelial hemangioendothelioma. In one embodiment, the cancer is relapsed or refractory ROS1+ epithelial hemangioendothelioma. In one embodiment, the cancer is newly diagnosed epithelial hemangioendothelioma. In one embodiment, the cancer is newly diagnosed ROS1+ epithelial hemangioendothelioma.

[0209] In one embodiment, the cancer is esophageal cancer. In one embodiment, the cancer is ESCC. In one embodiment, the cancer is ROS1+ESCC. In one embodiment, the cancer is relapsed or refractory ESCC. In one embodiment, the cancer is relapsed or refractory ROS1+ESCC. In one embodiment, the cancer is newly diagnosed ESCC. In one embodiment, the cancer is newly diagnosed ROS1+ESCC.

[0210] In one embodiment, the cancer is renal cancer. In one embodiment, the cancer is renal medullary carcinoma. In one embodiment, the cancer is ROS1+ renal medullary carcinoma. In one embodiment, the cancer is recurrent or refractory renal medullary carcinoma. In one embodiment, the cancer is recurrent or refractory ROS1+ renal medullary carcinoma. In one embodiment, the cancer is newly diagnosed renal medullary carcinoma. In one embodiment, the cancer is newly diagnosed ROS1+ renal medullary carcinoma. In one embodiment, the cancer is renal cell carcinoma. In one embodiment, the cancer is ROS1+ renal cell carcinoma. In one embodiment, the cancer is recurrent or refractory renal cell carcinoma. In one embodiment, the cancer is recurrent or refractory ROS1+ renal cell carcinoma. In one embodiment, the cancer is newly diagnosed renal cell carcinoma. In one embodiment, the cancer is newly diagnosed ROS1+ renal cell carcinoma.

[0211] In one embodiment, the cancer is breast cancer. In one embodiment, the cancer is ROS1+ breast cancer. In one embodiment, the cancer is relapsed or refractory breast cancer. In one embodiment, the cancer is relapsed or refractory ROS1+ breast cancer. In one embodiment, the cancer is newly diagnosed breast cancer. In one embodiment, the cancer is newly diagnosed ROS1+ breast cancer. In one embodiment, breast cancer is triple negative breast cancer.

[0212] In one embodiment, the cancer is colon cancer. In one embodiment, the cancer is ROS1+ colon cancer. In one embodiment, the cancer is relapsed or refractory colon cancer. In one embodiment, the cancer is relapsed or refractory ROS1+ colon cancer. In one embodiment, the cancer is newly diagnosed colon cancer. In one embodiment, the cancer is newly diagnosed ROS1+ colon cancer.

[0213] In one embodiment, the cancer is thyroid cancer. In one embodiment, the cancer is papillary thyroid cancer. In one embodiment, the cancer is ROS1+ papillary thyroid cancer. In one embodiment, the cancer is relapsed or refractory papillary thyroid cancer. In one embodiment, the cancer is relapsed or refractory ROS1+ papillary thyroid cancer. In one embodiment, the cancer is newly diagnosed papillary thyroid cancer. In one embodiment, the cancer is newly diagnosed ROS1+ papillary thyroid cancer.

[0214] In one embodiment, the cancer is a ROS1+ glioma (e.g., grade 1, grade 2, grade 3, or grade 4). In one embodiment, the cancer is a recurrent or refractory glioma. In one embodiment, the cancer is a recurrent or refractory ROS1+ glioma. In one embodiment, the cancer is a newly diagnosed ROS1+ glioma. In one embodiment, the cancer is a ROS1+ glioblastoma. In one embodiment, the cancer is a newly diagnosed ROS1+ glioblastoma. In one embodiment, the cancer is a recurrent or refractory glioblastoma. In one embodiment, the cancer is a recurrent or refractory ROS1+ glioblastoma. In one embodiment, the cancer is a neuroblastoma. In one embodiment, the cancer is a ROS1+ neuroblastoma. In one embodiment, the cancer is a recurrent or refractory neuroblastoma. In one embodiment, the cancer is a recurrent or refractory ROS1+ neuroblastoma. In one embodiment, the cancer is a newly diagnosed neuroblastoma. In one embodiment, the cancer is a newly diagnosed ROS1+ neuroblastoma.

[0215] In one embodiment, the cancer is ROS1+ pancreatic cancer. In one embodiment, the cancer is relapsed or refractory pancreatic cancer. In one embodiment, the cancer is relapsed or refractory ROS1+ pancreatic cancer. In one embodiment, the cancer is newly diagnosed neuroblastoma. In one embodiment, the cancer is newly diagnosed ROS1+ pancreatic cancer.

[0216] In one embodiment, the cancer is ROS1+ inflammatory hepatocellular adenoma. In one embodiment, the cancer is recurrent or refractory inflammatory hepatocellular adenoma. In one embodiment, the cancer is recurrent or refractory ROS1+ inflammatory hepatocellular adenoma. In one embodiment, the cancer is newly diagnosed neuroblastoma. In one embodiment, the cancer is newly diagnosed ROS1+ inflammatory hepatocellular adenoma.

[0217] In one embodiment, the cancer (or ROS1+ cancer, or ALK+ cancer) is a hematological cancer. In one embodiment, the cancer (or ROS1+ cancer or ALK+ cancer) is a lymphoma. In one embodiment, the lymphoma is a non-Hodgkin lymphoma. In one embodiment, the lymphoma is an anaplastic large cell lymphoma (ALCL), a diffuse large B-cell lymphoma (DLBCL), or a large B-cell lymphoma. In addition to hematological cancers, methods for treating other blood disorders or hematological malignancies of ROS1+ or ALK+ are also provided herein.

[0218] In one embodiment, the cancer is ALCL. In one embodiment, the cancer is ROS1+ALCL. In one embodiment, the cancer is ALK+ALCL. In one embodiment, the cancer is relapsed or refractory ALCL. In one embodiment, the cancer is relapsed or refractory ROS1+ALCL. In one embodiment, the cancer is relapsed or refractory ALK+ALCL. In one embodiment, the cancer is newly diagnosed ALCL. In one embodiment, the cancer is newly diagnosed ROS1+ALCL. In one embodiment, the cancer is newly diagnosed ALK+ALCL.

[0219] In one embodiment, the cancer is DLBCL. In one embodiment, the cancer is ROS1+DLBCL. In one embodiment, the cancer is ALK+DLBCL. In one embodiment, the cancer is relapsed or refractory DLBCL. In one embodiment, the cancer is relapsed or refractory ROS1+DLBCL. In one embodiment, the cancer is relapsed or refractory ALK+DLBCL. In one embodiment, the cancer is newly diagnosed DLBCL. In one embodiment, the cancer is newly diagnosed ROS1+DLBCL. In one embodiment, the cancer is newly diagnosed ALK+DLBCL.

[0220] In one embodiment, the cancer is large B cell lymphoma. In one embodiment, the cancer is ROS1+ large B cell lymphoma. In one embodiment, the cancer is ALK+ large B cell lymphoma. In one embodiment, the cancer is relapsed or refractory large B cell lymphoma. In one embodiment, the cancer is relapsed or refractory ROS1+ large B cell lymphoma. In one embodiment, the cancer is relapsed or refractory ALK+ large B cell lymphoma. In one embodiment, the cancer is newly diagnosed large B cell lymphoma. In one embodiment, the cancer is newly diagnosed ROS1+ large B cell lymphoma. In one embodiment, the cancer is newly diagnosed ALK+ large B cell lymphoma. In one embodiment, cancer or (ROS1+ cancer) is newly diagnosed. In one embodiment, cancer or (ROS1+ cancer) is previously untreated.

[0221] In one embodiment, the cancer or (ROS1+ cancer) is relapsed or refractory. In one embodiment, the cancer is relapsed. In one embodiment, the cancer or (ROS1+ cancer) is refractory.

[0222] In one embodiment, the subject is previously untreated. In one embodiment, the subject is not treated with tyrosine kinase inhibitor (TKI) therapy. In one embodiment, the subject receives one or more previous therapies. In one embodiment, the subject receives two or more previous therapies. In one embodiment, the subject produces resistance to one or more previous therapies. In one embodiment, previous therapy includes tyrosine kinase inhibitors (TKI). In one embodiment, previous TKI therapy includes treatment with one or more of crizotinib, ceritinib, alectinib, brigatinib, lorlatinib, entrectinib, lopatinib, cabozantinib, forritinib, tarentinib, merestinib, masitinib and ensartinib. In one embodiment, previous therapy includes one or more chemotherapy. In one embodiment, one or more chemotherapy is attached to TKI therapy.

[0223] In one embodiment, the cancer is an advanced cancer, eg, relapsed, refractory, or resistant after prior treatment with a TKI.

[0224] In one embodiment, the cancer (or ROS1+ cancer) is resistant to a tyrosine kinase inhibitor (TKI).

[0225] In one embodiment, the cancer is resistant lung cancer. In one embodiment, the cancer is resistant bronchial cancer. In one embodiment, the cancer is resistant non-small cell lung cancer. In one embodiment, the cancer is TKI-resistant non-small cell lung cancer. In one embodiment, the cancer is ROS1+ non-small cell lung cancer that is resistant to TKI.

[0226] In one embodiment, the cancer is lung cancer (eg, NSCLC, bronchial carcinoma). In one embodiment, the cancer is advanced lung cancer, eg, relapsed or refractory after prior treatment with TKIs.

[0227] In one embodiment, the compounds provided herein are administered as first-line therapeutic agents. In one embodiment, the compounds provided herein are administered as second-line therapeutic agents. In one embodiment, the compounds provided herein are administered as third or fourth-line therapeutic agents.

[0228] In one embodiment, the cancer or (ROS1+cancer) is metastatic. In one embodiment, the cancer has CNS metastases. In one embodiment, the cancer has brain metastases. In one embodiment, the cancer is metastatic non-small cell lung cancer (NSCLC). In one embodiment, the cancer is metastatic ROS1+NSCLC.

[0229] In one embodiment, provided herein is a method for treating a patient with metastatic ROS1+ non-small cell lung cancer (NSCLC), comprising administering to the patient a therapeutically effective amount of a compound provided herein, e.g., a compound of formula (I) or a stereoisomer thereof, or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof.

[0230] In one embodiment, the patient is an adult patient. In one embodiment, the patient is a pediatric patient.

[0231] In one embodiment, provided herein is a method for treating an adult patient with metastatic ROS1+ NSCLC, comprising administering to the patient a therapeutically effective amount of a compound provided herein, e.g., a compound of formula (I) or a stereoisomer thereof, or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof.

[0232] In one embodiment, provided herein is a method for treating an adult patient with metastatic ROS1+ NSCLC, comprising administering to the patient a therapeutically effective amount of a compound provided herein, e.g., a compound of formula (I) or a stereoisomer thereof, or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein the patient has progressed on or is intolerant to at least one prior TKI therapy.

[0233] In one embodiment, provided herein is a method for treating an adult patient with metastatic NSCLC that is ROS1+ due to the solvent front mutation G2032R, comprising administering to the patient a therapeutically effective amount of a compound provided herein, e.g., a compound of Formula (I) or a stereoisomer thereof, or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein the patient has progressed on or is intolerant to at least one prior TKI therapy.

[0234] In one embodiment, provided herein is a method for treating a ROS1-associated (or ROS1+) cancer in a subject in need thereof, wherein the cancer develops resistance to a tyrosine kinase inhibitor (TKI), the method comprising administering to the subject a therapeutically effective amount of a compound provided herein, e.g., a compound of formula (I) or a stereoisomer thereof, or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof.

[0235] Provided herein is a method for treating a ROS1-related (or ROS1+) cancer in a subject in need thereof, wherein the cancer produces resistance to a tyrosine kinase inhibitor (TKI), and wherein the cancer is identified as having one or more ROS1 inhibitor resistance mutations, the method comprising administering to the subject a therapeutically effective amount of a compound provided herein, for example, a compound of formula (I) or a stereoisomer thereof, or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof. In one embodiment, one or more ROS1 inhibitor resistance mutations include one or more amino acid substitutions at amino acid positions selected from 1986, 2004, 2026, 2032, and 2033. In one embodiment, one or more ROS1 inhibitor resistance mutations include one or more amino acid substitutions selected from S1986F, S1986Y, F2004C, F2004V, L2026M, G2032R, D2033N, L2086F, and G2101A. In one embodiment, one or more ROS1 inhibitor resistance mutations are G2032R. In one embodiment, the one or more ROS1 inhibitor resistance mutations include G2032R and one or more of S1986F, S1986Y, F2004C, F2004V, L2026M, D2033N, or G2101 A. In one embodiment, the ROS1 inhibitor resistance mutation is L2086F.

[0236] In one embodiment, the TKI is a ROS1 inhibitor. In one embodiment, the TKI is an ALK inhibitor. In one embodiment, the TKI is crizotinib, ceritinib, alectinib, brigatinib, lorlatinib, entrectinib, lopretinib, cabozantinib, foreitinib, mesartinib, tarentinib, masitinib or ensartinib. In one embodiment, the TKI is crizotinib. In one embodiment, the TKI is entrectinib. In one embodiment, the TKI is lorlatinib. In one embodiment, the TKI is lopretinib.

[0237] In certain embodiments, the subject relapses after a first-line treatment for cancer. In other embodiments, the subject relapses after a second-line treatment for cancer.

[0238] In certain embodiments, methods of treating or preventing cancer can be demonstrated by one or more responses, such as increased apoptosis, inhibition of tumor growth, reduction of tumor metastasis, inhibition of tumor metastasis, reduction of microvessel density, reduction of neovascularization, inhibition of tumor migration, tumor regression, and increased survival of the subject.

[0239] Combination therapy

[0240] In some embodiments, methods of treating or preventing cancer may include administering a compound of Formula (I) in combination with one or more other chemotherapeutic agents.

[0241] As used herein and unless otherwise indicated, "combination" or "with ... combination" does not mean that other medicaments and formula (I) compounds must be administered simultaneously and / or they are formulated for delivery together, although these delivery methods are within the scope of the present disclosure. Compounds provided herein can be administered simultaneously with one or more other medicaments (e.g., one or more other additional medicaments), before (e.g., before 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, 12 weeks or 16 weeks) or after (e.g., after 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, 12 weeks or 16 weeks). Typically, each therapeutic agent is administered at a dose and / or schedule determined for that particular agent. Other therapeutic agents may be administered with the compounds provided herein in a single composition or separately in different compositions. Triple therapy is also contemplated herein.

[0242] Chemotherapeutic agents that can be administered in combination with the compounds provided herein include: 1-amino-4-phenylamino-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate (Acid Blue 25), 1-amino-4-[4-hydroxyphenyl-amino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, 1-amino-4-[4-aminophenylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, 1-amino-4-[1-naphthylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, 1-amino-4-[4-fluoro-2-carboxyphenylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, 1-amino-4-[4-fluoro-2-carboxyphenylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, 1-amino-4-[ 2-Anthracenylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, ABT-263, afatinib maleate, axitinib, aminoglutethimide, amsacrine, anastrozole, APCP, asparaginase, AZD5363, bacillus Calmette-Guérin (bcg), bicalutamide, bleomycin, bortezomib, β-methylene-ADP (AOPCP), buserelin, busulfan, cabazitaxel, cabozantinib, camptothecin, capecitabine, carboplatin, carfilzomib, carmustine, ceritinib, chlorambucil, chloroquine, cisplatin, cladribine, clodronate, cobimetinib ), colchicine, crizotinib, cyclophosphamide, cyproterone, cytarabine, dacarbazine, actinomycin D, daunorubicin, demethoxychloroquine, dexamethasone, dichloroacetate, diethylstilbestrol, diethylstilbestrol, docetaxel, doxorubicin, epirubicin, eribulin, erlotinib, estradiol, estramustine, etoposide, everolimus, exemestane, filgrastim, fludarabine, fludrocortisone, fluorouracil, fluoxymesterone, flutamide, gefitinib, gemcitabine, genistein, goserelin, GSK1120212, hydroxyurea, idarubicin, ifosfamide, imatinib, dry interferon, irinotecan, ixabepilone, lenalidomide, letrozole, folinic acid, leuprolide, levamisole, lomustine, lonidamine, mechlorethamine, medroxyprogesterone, megestrol, melphalan, mercaptopurine, mesna, metformin, methotrexate, miltefosine, mitomycin, mitotane, mitoxantrone, MK-2206, mutamycin, N-(4-sulfamoylphenylthiocarbamoyl)trimethylacetamide, NF279, NF449, nilutamide,Nocodazole, octreotide, olaparib, oxaliplatin, paclitaxel, pamidronate, pazopanib, pemexetred, pentostatin, perifosine, PF-04691502, plicamycin, pomalidomide, porfimer, PPADS, procarbazine, quercetin, Tetrexed, ramucirumab, reactive blue 2, rituximab, rolofylline, romidepsin, rucaparib, selumetinib, sirolimus, sodium 2,4-dinitrobenzenesulfonate, sorafenib, streptozocin, sunitinib, suramin, talazoparib, tamoxifen, temozolomide, temsirolimus, teniposide, testosterone, thalidomide, thioguanine, thiotepa, titanocene dichloride dichloride), tonaphthol, tonaphthol, topotecan, trametinib, trastuzumab, retinoic acid, veliparib, vinblastine, vincristine, vindesine, vinorelbine and vorinostat (SAHA). In other embodiments, chemotherapeutic agents that can be administered in combination with the compounds provided herein include: ABT-263, dexamethasone, 5-fluorouracil, PF-04691502, romidepsin and vorinostat (SAHA). In other embodiments, chemotherapeutic agents that can be administered in combination with the compounds provided herein include: 1-amino-4-phenylamino-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate (Acid Blue 25), 1-amino-4-[4-hydroxyphenyl-amino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, 1-amino-4-[4-aminophenylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, 1-amino-4-[4-aminophenylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, 1-amino-4- [1-naphthylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, 1-amino-4-[4-fluoro-2-carboxyphenylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, 1-amino-4-[2-anthrylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, APCP, β-methylene-ADP (AOPCP), capecitabine, cladribine, cytarabine, fludarabine, doxorubicin,Gemcitabine, N-(4-sulfamoylphenylthiocarbamoyl)trimethylacetamide, NF279, NF449, PPADS, quercetin, reactive blue 2, rolofylline 2,4-dinitrobenzenesulfonate sodium, sumarin and topofylline. In some embodiments, the chemotherapeutic agent is a biologic such as an ADC or a MET antibody. In some embodiments, the chemotherapeutic agent is a MET inhibitor, a MEK inhibitor, a RET inhibitor, another ALK inhibitor, or a ROS1 inhibitor. In some embodiments, the compounds as described herein are used with one or more platinum-based chemotherapies and / or immunotherapies (e.g., checkpoint inhibitors). ,

[0243] Many combination therapies for treating cancer have been developed. In certain embodiments, the compounds provided herein (e.g., compounds of formula (I)) can be co-administered with one or more combination therapies. Examples of combination therapies that can be co-administered with the compounds provided herein are included in Table 1.

[0244] Table 1: Exemplary combination therapies for treating cancer

[0245]

[0246]

[0247]

[0248]

[0249]

[0250]

[0251] In certain embodiments, the combination therapies provided herein include co-administration with other types of chemotherapeutic agents, such as immuno-oncology agents. Cancer cells typically have specific cell surface antigens that can be recognized by the immune system. Therefore, immuno-oncology agents, such as monoclonal antibodies, can selectively bind to cancer cell antigens and cause cell death. Other immuno-oncology agents can inhibit tumor-mediated natural immune responses, inhibit or otherwise activate immune responses and thereby promote recognition of tumors by the immune system. Exemplary antibody immuno-oncology agents include, but are not limited to, abagovomab, adecatumumab, afutuzumab, alemtuzumab, anatumomab mafenatox), apolizumab, belintoumab, BMS-936559, catumaxomab, durvalumab, epacadostat, epratuzumab, indoximod, okiatuzumab, intelumumab, ipilimumab, isatuximab, lamblizumab (lambrolizumab), MED14736, MPDL3280A, nivolumab, obotuzumab, ocaratuzumab, ofatumumab, olatatumab, pembrolizumab, pidilizumab, rituximab, ticilimumab, samalizumab and tremelimumab. In some embodiments, the antibody immuno-oncology agent is selected from anti-CD73 monoclonal antibody (mAb), anti-CD39 mAb, anti-PD-1 mAb and anti-CTLA4 mAb. Therefore, in some embodiments, the methods provided herein include the combined administration of one or more immuno-oncology agents, such as the agents mentioned above.

[0252] In some embodiments, combination therapy includes administering a compound provided herein, such as a compound of Formula (I), in combination with an SH2 inhibitor, such as CGP78850, CPG85793, C90, C126, G7-18NATE, G7-B1, and NSC642056.

[0253] In some embodiments, combination therapy comprises administering a solid form or pharmaceutical composition provided herein, such as a compound of Formula (I), in combination with an ERK1 / 2 inhibitor such as ASN007, GDC-0994, KO-947, LTT462, LY3214996, MK-8353, ulixertinib.

[0254] In some embodiments, combination therapy includes administering a compound provided herein, such as a compound of Formula (I), in combination with a MEK inhibitor, such as trametinib, cobimetinib, binimetinib, selumetinib, PD-325901, CI-1040, and TAK-733.

[0255] In some embodiments, the combination therapy comprises administering a compound provided herein, such as a compound of Formula (I), in combination with a MET inhibitor selected from JNJ-38877605, PF-04217903, foreitinib, AMG 458, tivantinib, cabozantinib, crizotinib, capmatinib hydrochloride, tepotinib hydrochloride, and savotinib.

[0256] In some embodiments, the combination therapy comprises administering a compound of the present disclosure, such as Formula (I), in combination with a SHP2 inhibitor selected from: TNO-155, RMC-4630, JAB-3068, or RLY-1971.

[0257] In some embodiments, combination therapy includes administering a compound provided herein, such as a compound of Formula (I), in combination with a RAS inhibitor selected from the group consisting of aliskiren, captopril, losartan, irbesartan, olmesartan, candesartan, valsartan, fimasartan, azilsartan, telmisartan, eprosartan, benazepril, enalapril, lisinopril, perindopril, quinapril, ramipril, and trandolapril.

[0258] In some embodiments, combination therapy includes administering a compound provided herein, such as a compound of formula (I), in combination with a TKI. In one embodiment, the TKI is a ROS1 inhibitor. In one embodiment, the TKI is an ALK inhibitor. In one embodiment, the TKI is crizotinib, ceritinib, alectinib, brigatinib, lorlatinib, entrectinib, lopretinib, cabozantinib, foretinib, mesartinib, tarentinib, masitinib or ensartinib. In one embodiment, the TKI is crizotinib. In one embodiment, the TKI is entrectinib. In one embodiment, the TKI is alectinib. In one embodiment, the TKI is brigatinib. In one embodiment, the TKI is lorlatinib.

[0259] In some embodiments, combination therapy includes the co-administration of a compound provided herein, such as a compound of formula (I) and an anti-PD-1 therapy. In certain embodiments, combination therapy includes the co-administration of a compound provided herein, such as a compound of formula (I), and oxaliplatin. In other embodiments, combination therapy includes the co-administration of a compound provided herein, such as a compound of formula (I), and doxorubicin.

[0260] In certain embodiments, the compounds provided herein can be co-administered with non-chemotherapy for cancer treatment. In certain embodiments, the compounds provided herein can be co-administered with radiotherapy. In certain embodiments, the compounds provided herein can be co-administered with surgery, thermal ablation, focused ultrasound therapy, cryotherapy, or any combination of these therapies.

[0261] In certain embodiments, the compounds provided herein can be co-administered with one or more other compounds provided herein. In addition, these combinations can be co-administered with other therapeutic agents, such as other agents suitable for treating cancer, immune or neurological diseases, such as the agents identified above. In certain embodiments, one or more additional chemotherapeutic agents are co-administered with the compounds provided herein to provide a synergistic effect. In certain embodiments, one or more additional chemotherapeutic agents are co-administered to provide an additive effect.

[0262] Pharmaceutical composition

[0263] In certain embodiments, provided herein are pharmaceutical preparations suitable for use in human patients, comprising compounds provided herein (e.g., compounds of formula (I)), and one or more pharmaceutically acceptable excipients. In certain embodiments, pharmaceutical preparations can be used to treat or prevent conditions or diseases described herein. Compounds provided herein can be used to manufacture drugs for treating any disease or illness disclosed herein.

[0264] In one embodiment, the pharmaceutical composition comprises a compound of formula (I):

[0265]

[0266] Or its stereoisomer, or mixture of stereoisomers, or pharmaceutically acceptable salts thereof, and diluents, binders, disintegrants and lubricants. In one embodiment, compound 1, or its stereoisomer, or mixture of stereoisomers, or pharmaceutically acceptable salts thereof in the pharmaceutical composition is the free base of compound 1. In one embodiment, the free base of compound 1 is amorphous. In one embodiment, the free base of compound 1 is one of the solid forms of the free base of compound 1 provided herein. In one embodiment, the free base of compound 1 is Form 1 of the free base of compound 1.

[0267] In one embodiment, the diluent is mannitol. In one embodiment, the diluent is Pearlitol 100SD, Pearlitol 110C, Pearlitol 160C, Pearlitol 25C, Pearlitol 300DC, Pearlitol 400DC, Pearlitol 500DC, Pearlitol SW-F200, Parteck M 100, Parteck M 200 or lactose monohydrate. In one embodiment, the diluent is mannitol Pearlitol 50C. In one embodiment, the diluent is mannitol Pearlitol 200SD.

[0268] In one embodiment, the binder is hydroxypropyl cellulose (HPC), silicified microcrystalline cellulose (SMCC) or a mixture thereof. In one embodiment, the binder is HPC. In one embodiment, the binder is SMCC. In one embodiment, the binder is a mixture of HPC and SMCC. In certain embodiments, the binder is carboxymethyl cellulose, alginate, gelatin, polyvinyl pyrrolidone, sucrose and / or gum arabic or a mixture thereof.

[0269] In one embodiment, the disintegrant is sodium starch glycolate (SSG). In one embodiment, the disintegrant is croscarmellose sodium.

[0270] In one embodiment, the lubricant is magnesium stearate. In another embodiment, the lubricant is sodium lauryl sulfate.

[0271] In one embodiment, the diluent is mannitol, the binder is a mixture of HPC and SMCC, the disintegrant is sodium starch glycolate (SSG), and the lubricant is magnesium stearate.

[0272] In one embodiment, the tablets are coated with a non-functional film coating (eg, Opadry II pink).

[0273] In one embodiment, the pharmaceutical composition provided herein is an oral dosage form. In one embodiment, the oral dosage form is a tablet. In one embodiment, the oral dosage form is an immediate release tablet.

[0274] The compositions and methods provided herein can be used to treat subjects in need. In certain embodiments, the subject is a mammal such as a human, or a non-human mammal. When applied to a subject such as a human, the composition or compound is preferably administered as a pharmaceutical composition comprising, for example, a compound provided herein and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions such as water or physiologically buffered saline or other solvents or vehicles such as glycols, glycerol, oils such as olive oil or injectable organic esters. In a preferred embodiment, when these pharmaceutical compositions are used for human administration, especially for invasive administration routes (i.e., avoiding routes of transport or diffusion via epithelial barriers, such as injection or implantation), the aqueous solution is pyrogen-free, or substantially pyrogen-free. Excipients may be selected, for example, to achieve delayed release of an agent or selective targeting of one or more cells, tissues or organs. The pharmaceutical composition may be in the form of dosage units such as tablets, capsules (including scattered capsules and gelatin capsules), particles, lyophilized products for reconstruction, powders, solutions, syrups, suppositories, injections or the like. The composition may also be present in a transdermal delivery system such as a skin patch. The composition may also be presented in a solution suitable for topical administration such as eye drops.

[0275] A pharmaceutically acceptable carrier may contain a physiologically acceptable agent, which is used, for example, to stabilize a compound such as provided herein, to increase its solubility or to increase its absorption. Such physiologically acceptable agents include, for example, carbohydrates such as glucose, sucrose or dextran, antioxidants such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins or other stabilizers or excipients. The selection of a pharmaceutically acceptable carrier including a physiologically acceptable agent depends on, for example, the route of administration of the composition. The preparation or pharmaceutical composition may be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (preparation) may also be a liposome or other polymer matrix, which may incorporate, for example, a compound provided herein. For example, a liposome including a phospholipid or other lipid is a non-toxic, physiologically acceptable and metabolizable carrier that is relatively easy to manufacture and administer.

[0276] The phrase "pharmaceutically acceptable" is used herein to refer to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for contact with the tissues of subjects without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0277] As used herein, the phrase "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of its formulation and not deleterious to the subject. Some examples of materials that can serve as pharmaceutically acceptable carriers include: (1) sugars such as lactose, glucose, and sucrose; (2) starches such as corn starch and potato starch; (3) cellulose and its derivatives such as sodium carboxymethylcellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients such as cocoa butter and suppository waxes; (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn starch, and cellulose acetate. oil and soybean oil; (10) glycols such as propylene glycol; (11) polyols such as glycerol, sorbitol, mannitol and polyethylene glycol; (12) esters such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffer solution; and (21) other non-toxic compatible substances used in pharmaceutical preparations.

[0278] The pharmaceutical composition (formulation) can be administered to a subject by any of a number of routes of administration, including, for example, oral (e.g., large doses of liquid medicine such as in aqueous or non-aqueous solutions or suspensions, tablets, capsules (including sprinkle capsules and gelatin capsules), pills, powders, granules, pastes for the tongue); absorption through the oral mucosa (e.g., sublingual); anal, rectal or vaginal (e.g., as a pessary, cream or foam); parenteral (including intramuscular, intravenous, subcutaneous or intrathecal, such as sterile solutions or suspensions); nasal; intraperitoneal; subcutaneous; transdermal (e.g., as a patch applied to the skin); and topical (e.g., as a cream, ointment or spray applied to the skin, or as eye drops). The compound can also be formulated for inhalation. In certain embodiments, the compound can be simply dissolved or suspended in sterile water. Details of suitable routes of administration and compositions appropriate for such routes can be found, for example, in U.S. Pat. Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970, and 4,172,896, and patents cited therein.

[0279] The preparation can be conveniently presented in unit dosage form and can be prepared by any method well known in pharmaceutical technology. In certain embodiments, the unit dosage form is a tablet. In certain embodiments, the unit dosage form is a tablet of 5mg (by weight of free base compound 1) dosage strength. In certain embodiments, the unit dosage form is a tablet of 25mg (by weight of free base compound 1) dosage strength. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will vary depending on the subject being treated and the specific mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form is generally the amount of the compound that produces a therapeutic effect. Typically, in one hundred percent, this amount is in the range of about 1% to about 99% of the active ingredient, preferably about 5% to about 70%, and most preferably about 10% to about 30%.

[0280] The method for preparing these preparations or compositions includes the step of associating an active compound, such as a compound provided herein, with a carrier and optionally one or more auxiliary ingredients. Generally, the preparation is prepared by uniformly and intimately associating a compound provided herein with a liquid carrier, or a finely divided solid carrier, or both, and then, if necessary, shaping the product.

[0281] Formulations provided herein suitable for oral administration can be in the form of capsules (including sprinkle capsules and gelatin capsules), cachets, pills, tablets, lozenges (using flavoring agents, usually sucrose and gum arabic or tragacanth), lyophilizates, powders, granules, or as solutions or suspensions in aqueous or non-aqueous liquids, or as oil-in-water or water-in-oil liquid emulsions, or as elixirs or syrups, or as soft tablets (using inert bases such as gelatin and glycerin or sucrose and gum arabic) and / or as mouthwashes and the like, each containing a predetermined amount of a compound provided herein as an active ingredient. The composition or compound can also be administered as a pill, dry medicine electuary or paste.

[0282] Solid dosage forms (capsules (including sprinkle capsules and gelatin capsules), tablets, pills, sugar tablets, powders, granules and the like) for oral administration are prepared by mixing the active ingredient with one or more pharmaceutically acceptable carriers, such as sodium citrate or calcium hydrogen phosphate, and / or any of the following: (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginate, gelatin, polyvinyl pyrrolidone, sucrose and / or gum arabic; (3) humectants , such as glycerol; (4) disintegrants, such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate; (5) solution retardants, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as, for example, cetyl alcohol and glyceryl monostearate; (8) absorbents, such as kaolin and bentonite; (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate and mixtures thereof; (10) complexing agents, such as modified and unmodified cyclodextrins; and (11) coloring agents. In the case of capsules (including sprinkle capsules and gelatin capsules), tablets and pills, the pharmaceutical composition may also include a buffer. Solid compositions of a similar type may also be used as fillers in soft and hard-filled gelatin capsules using excipients such as lactose or milk sugar, as well as high molecular weight polyethylene glycols and their analogs.

[0283] Tablets can be made by compression or molding, optionally with one or more auxiliary ingredients. Compressed tablets can be prepared using a binder (e.g., gelatin or hydroxypropyl methylcellulose), a lubricant, an inert diluent, a preservative, a disintegrant (e.g., sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), a surface active or dispersant. Molded tablets can be produced by molding a mixture of powdered compounds moistened with an inert liquid diluent in a suitable machine.

[0284] Tablets and other solid dosage forms of pharmaceutical compositions, such as sugar tablets, capsules (including sprinkle capsules and gelatin capsules), pills and granules, may be optionally scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in pharmaceutical formulation technology. The dosage form may also be formulated to provide a slow or controlled release of the active ingredient therein by using, for example, hydroxypropylmethylcellulose, other polymer matrices, liposomes and / or microspheres in different proportions to provide the desired release profile. It may be sterilized by filtering, for example, through a bacteria-retaining filter, or by incorporating a sterilizing agent in the form of a sterile solid composition, which may be dissolved in sterile water or some other sterile injectable medium directly before use. These compositions may also optionally contain an emulsifier and may have a certain composition so that it may optionally release the active ingredient in a delayed manner, only or preferentially in a certain part of the gastrointestinal tract. Examples of embedding compositions that can be used include polymers and waxes. Where appropriate, the active ingredient may also be in a microencapsulated form with one or more of the above-mentioned excipients.

[0285] Liquid dosage forms suitable for oral administration include pharmaceutically acceptable emulsions, lyophilisates for reconstitution, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage form may contain an inert diluent commonly used in the art, such as, for example, water or other solvents, cyclodextrins and their derivatives, solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofuran methanol, polyethylene glycol and fatty acid esters sorbitan and mixtures thereof.

[0286] Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

[0287] Suspensions, in addition to the active compounds, may contain suspending agents such as ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

[0288] Formulations of pharmaceutical compositions for rectal, vaginal or urethral administration may be presented in the form of suppositories which may be prepared by mixing one or more active compounds with one or more suitable non-irritating excipients or carriers including, for example, cocoa butter, polyethylene glycol, suppository wax or salicylates and which are solid at room temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.

[0289] The formulation of the pharmaceutical composition for administration to the oral cavity may be presented in the form of a mouthwash or oral spray or oral ointment.

[0290] Alternatively or additionally, the composition can be formulated to be delivered via a catheter, stent, wire or other intraluminal device. Delivery via these devices can be particularly suitable for delivery to the bladder, urethra, ureter, rectum or intestine.

[0291] Formulations suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or spray formulations containing such carriers as are known in the art to be appropriate.

[0292] Dosage forms for topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active compound may be mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers or propellants that may be required.

[0293] Ointments, pastes, creams and gels may contain, in addition to the active compounds, excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

[0294] In addition to the active compounds, powders and sprays may contain excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays may additionally contain customary propellants, such as chlorofluorocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

[0295] Transdermal patches have the additional advantage of controlled delivery of compounds provided herein to the body. These dosage forms can be prepared by dissolving or dispersing the active compound in a suitable medium. Absorption enhancers can also be used to increase the flux of the compound through the skin. The rate of this flux can be controlled by providing a rate-controlling membrane or by dispersing the compound in a polymer matrix or gel.

[0296] Ophthalmic preparations, eye ointments, powders, solutions and the like are also contemplated to be within the scope of the present disclosure. Exemplary ophthalmic preparations are described in U.S. Publication Nos. 2005 / 0080056, 2005 / 0059744, 2005 / 0031697 and 2005 / 004074 and U.S. Patent No. 6,583,124, the contents of which are incorporated herein by reference. If desired, liquid ophthalmic preparations have properties similar to tears, aqueous humor or vitreous humor or are compatible with these fluids. Preferably, the route of administration is topical (e.g., topical, such as eye drops, or administered via implantation).

[0297] As used herein, the phrases "parenteral administration" and "administered parenterally" mean modes of administration other than enteral and topical administration, usually by injection, and include, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion.

[0298] Pharmaceutical compositions suitable for parenteral administration include one or more active compounds and one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just before use and which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents.

[0299] Examples of suitable aqueous and nonaqueous carriers that can be used in the pharmaceutical compositions provided herein include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol and the like) and suitable mixtures thereof, vegetable oils such as olive oil and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials such as lecithin, by maintaining the required particle size in the case of dispersions, and by the use of surfactants.

[0300] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifiers and dispersants. Various antibacterial and antifungal agents may be included to prevent the action of microorganisms, for example, parabens, chlorobutanol, phenol sorbic acid and the like. Isotonic agents such as sugar, sodium chloride and the like may also need to be included in the composition. In addition, the long-term absorption of injectable pharmaceutical forms may be achieved by including agents that delay absorption such as aluminum monostearate and gelatin.

[0301] In some cases, in order to prolong the effect of the drug, it is necessary to slow down the absorption of the drug from subcutaneous or intramuscular injection. This can be accomplished by using a liquid suspension of a crystalline or amorphous material with poor water solubility. The absorption rate of the drug then depends on its dissolution rate, which in turn can depend on crystal size and crystalline form. Alternatively, delayed absorption of parenteral drug forms is achieved by dissolving or suspending the drug in an oil vehicle.

[0302] Injectable depot forms are made by forming a microencapsulation matrix of the target compound in a biodegradable polymer such as polylactic acid-polyglycolide. Depending on the ratio of drug to polymer, and the nature of the specific polymer used, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissues.

[0303] For use in the methods provided herein, the active compound can be administered per se or as a pharmaceutical composition containing, for example, 0.1 to 99.5% (more preferably, 0.5 to 90%) of active ingredient and a pharmaceutically acceptable carrier.

[0304] The introduction method can also be provided by a rechargeable or biodegradable device. In recent years, various sustained-release polymer devices have been developed and tested in vivo for the controlled delivery of drugs, including protein biopharmaceuticals. A variety of biocompatible polymers (including hydrogels) including biodegradable and non-degradable polymers can be used to form implants that continuously release compounds at specific target sites.

[0305] Actual dosage levels of the active ingredients in the pharmaceutical compositions may be varied to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without causing toxicity to the patient.

[0306] The selected dosage level will depend upon a variety of factors including the activity of the particular compound or combination of compounds employed, or the esters, salts or amides thereof, the route of administration, the time of administration, the rate of excretion of the particular compound employed, the duration of the treatment, other drugs, compounds and / or materials used in combination with the particular compound employed, the age, sex, weight, condition, general health and prior medical history of the subject being treated, and like factors well known in the medical arts.

[0307] A physician or veterinarian with ordinary skills in the art can easily determine and prescribe the therapeutically effective amount of the desired pharmaceutical composition. For example, a physician or veterinarian can start the dosage of the pharmaceutical composition or compound at a level lower than the level required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. "Therapeutically effective amount" refers to a concentration of the compound sufficient to cause the desired therapeutic effect. It is generally understood that the effective amount of the compound will vary according to the subject's weight, sex, age and medical history. Other factors affecting the effective amount may include, but are not limited to, the severity of the subject's condition, the condition being treated, the stability of the compound, and, if necessary, another type of therapeutic agent administered with the compound provided herein. A larger total dose can be delivered by multiple administrations of the agent. Methods for determining efficacy and dosage are known to those skilled in the art (Isselbacher et al. (1996) Harrison's Principles of Internal Medicine 13th Edition, 1814-1882, which is incorporated by reference).

[0308] If desired, the effective daily dose of the active compound can be administered as one, two, three, four, five, six or more sub-doses at appropriate intervals throughout the day, optionally in unit dosage form. In certain embodiments provided herein, the active compound can be administered twice or three times a day. In a preferred embodiment, the active compound is administered once a day.

[0309] In certain embodiments, provided herein are compounds that can be used alone or in combination with another type of therapeutic agent. As used herein, the term "combined administration" refers to any form of administration of two or more different therapeutic compounds, so that the previously administered therapeutic compound is still effective in vivo while the second compound is administered (for example, two compounds are effective in the subject at the same time, which may include the synergistic effect of the two compounds). For example, different therapeutic compounds can be administered simultaneously or sequentially with the same preparation or a separate preparation. In certain embodiments, different therapeutic compounds can be administered within 1 hour, 12 hours, 24 hours, 36 hours, 48 ​​hours, 72 hours or a week apart from each other. Therefore, the subject receiving this treatment can benefit from the combined effects of different therapeutic compounds.

[0310] In certain embodiments, provided herein is a compound and one or more other therapeutic agents (e.g., one or more other chemotherapeutic agents) co-administered relative to provided herein provided compound or one or more other therapeutic agents each separately administered, providing improved efficacy. In certain such embodiments, co-administered provides an additive effect, wherein the additive effect refers to the sum of each effect of the compound provided herein and one or more other therapeutic agents administered separately.

[0311] Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

[0312] Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, etc.; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, etc.; (3) metal chelators, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.

[0313] Now that the disclosure is generally described, it will be more readily understood by reference to the following examples, which are intended merely to illustrate certain aspects and embodiments of the disclosure and are not intended to limit the disclosure.

[0314] Example

[0315] Example 1.

[0316] Generation of Ba / F3 stable cell lines. Genes encoding CD74-ROS1 wild type (wt) and CD74-ROS1 G2032R were synthesized (GeneRay), cloned into the retroviral construct pMSCV-puro (Biovector), and packaged into retroviral particles. Ba / F3 cells (RIKEN) were infected with the virus at a multiplicity of infection = 1 for 1 day. The infected cells were revived in a medium (RPMI-1640, containing 10% fetal bovine serum and 1% streptomycin and penicillin) supplemented with mouse IL-3 (10 ng / mL) for 2 days, and stable cell lines were selected by IL-3 withdrawal and puromycin (0.8 μg / mL) for 7 days. Monoclones were selected by single cell dilution in a medium containing puromycin (0.8 μg / mL) without IL-3. The transformation of the desired gene was confirmed by Sanger sequencing and Western blotting using ROS1 antibody (CST#3287).

[0317] Cell proliferation assay. Stable cells were plated at 1,000 cells / well (40 μL) in 384-well plates for 1 day. Test compounds (40 nL) were then added in a 3-fold dilution series using a TECAN EVO200 liquid handler and incubated for 72 hours. The plates were equilibrated at room temperature for 15 minutes, followed by the addition of 40 μL of CellTiter-Glo reagent (Promega). Luminescence was measured on a plate reader. The half-maximal inhibitory concentration (IC) was calculated from the percentage inhibition and inhibitor concentration using a four-parameter logistic regression. 50 ).

[0318] The results for Compound 1 and various ROS1 inhibitors are shown below.

[0319]

[0320] Example 2. In vivo evaluation of compound 1

[0321] Model 2A: Tumor sections (~30 mm) from model LU-01-0414 were implanted subcutaneously into the right flank of female Balb / c nude mice. 3 ), which was derived from a 51-year-old woman with NSCLC and unknown smoking history. The tumor was G3 and the tumor stage was T2N2M0. The tumor carried SDC4-ROS1. In the efficacy study, the tumor grew to a mean tumor volume of 160mm 3 Afterwards, mice (n=5-8 / group) were randomized and administered vehicle or compound 1 by oral gavage twice a day (BID; 12 hours apart) for 28 days. Figure 2A .

[0322] Model 2B: Athymic naked females - Foxn1 nu The mice were implanted subcutaneously in the left flank with tumor fragments from model CTG-0848, which was derived from a 41-year-old Asian male with stage II NSCLC. The tumor carried a CD74-ROS1 fusion. When the tumor grew to approximately 240 mm 3 Afterwards, mice (n=5-5 / group) were randomized and administered vehicle or compound 1 by oral gavage twice a day (BID; 12 hours apart) for 21 days (BID×21 days). The results are shown in Figure 2B .

[0323] Model 2C: Athymic naked females - Foxn1 nu The left flank of mice was subcutaneously implanted with tumor fragments from the model CTG-2532, which was derived from a female Asian non-smoker with stage IV NSCLC whose disease responded to the TKI crizotinib and then progressed. The tumor carried CD74-ROS1 G2032R. After the tumor grew to approximately 215, mice (n=5 / group) were randomized and given vehicle 1, 5 or 15 mg / kg of compound 1 by oral gavage twice a day (BID; 12 hours apart) for 21 days (BID×21 days). The results are shown in Figure 2C .

[0324] Example 3. In vivo evaluation of compound 1

[0325] Model 1 (LU-01-0414PDX SDC4-ROS1): Tumor sections (~30 mm) from model LU-01-0414 were implanted subcutaneously into the right flank of female Balb / c nude mice. 3 ), which was derived from a 51-year-old woman with NSCLC and unknown smoking history. The tumor was G3 and the tumor stage was T2N2M0. The tumor carried SDC4-ROS1. When the tumor grew to an average weight volume of 492mm 3 Afterwards, mice received a single dose or BID x 5 days of vehicle or compound 1, and tumors and blood were collected 1 hour and 12 hours after the last dose (treatment groups only). Tumors were split and snap-frozen and then processed for Western blotting ( Figure 4A and Figure 4B , described below) or fixed in formalin for 24-48 hours, followed by FFPE block preparation and processing for immunohistochemistry ( Figure 5A , described below) or gene expression analysis by NanoString assay ( Fig. 6A , described below). Blood samples were collected into tubes containing EDTA-K2, centrifuged, and the resulting plasma was analyzed by LC / MS / MS to determine the total concentration of Compound 1 in plasma. The free concentration of Compound 1 was calculated by multiplying the unbound fraction of Compound 1 in plasma of female Balb / c mice determined by equilibrium dialysis ( Figure 4A ).

[0326] Model 2 (CTG-0848PDX CD74-ROS1): Female athymic naked-Foxn1 nu Mice were implanted subcutaneously in the left flank with tumor fragments from model CTG-0848, which was derived from a 41-year-old Asian male with stage II NSCLC. The tumor carried a CD74-ROS1 fusion. When the tumor grew to 350 to 500 mm 3 Afterwards, mice received a single dose or BID x 5 days of vehicle or compound 1, and tumors were harvested 1 and 12 hours after the last dose (treatment groups only) and processed as described under Model 1 for Western blot analysis ( Figure 4C ), immunohistochemical analysis ( Figure 5B ) or gene expression analysis ( Figure 6B ).

[0327] Model 3 (CTG-2532PDX CD74-ROS1 G2032R): Athymic female nude-Foxn1 nuMice were implanted subcutaneously in the left flank with tumor fragments from the model CTG-2532, which was derived from a female Asian non-smoker with stage IV NSCLC whose disease responded to the TKI crizotinib and then progressed. The tumor carried CD74-ROS1 G2032R. When the tumor grew to 350 to 500 mm 3 Afterwards, mice received vehicle or compound 1 either once or BID x 5 days (11 doses in total), and tumors were harvested 1 and 12 hours after the last dose (treatment groups only) and processed as described under Model 1 for Western blot analysis ( Figure 4D ), immunohistochemical analysis ( Figure 5C ) or gene expression analysis ( Figure 6C ).

[0328] These three models used the following preparations to evaluate the inhibitory activity of compound 1 on ROS1 signaling in tumors.

[0329] For Western blot analysis, the rapidly frozen tumors were cut, lysed with RIPA buffer (containing a mixture of protease and phosphatase inhibitors), ground and centrifuged. The supernatant was collected, the protein was quantified, and supernatant samples of similar concentrations were separated by electrophoresis and transferred to a nitrocellulose membrane. The membrane was cut, blotted, treated with HRP substrate, and chemiluminescence was detected by Amersham Imager 680. The intensity of each band was quantified using 1D quant (GE) densitometry software. The following antibodies were used: phospho-ROS1 (Tyr2274), Cell Signaling Technology (CST) 3078; ROS1 (D4D6) CST 3287; phospho-AKT (Ser473) (D9E), CST 4060; AKT, CST 9272; phospho-p44 / 42MAPK (Erk1 / 2) (Thr202 / Tyr204), CST 9101; p44 / 42MAPK (Erk1 / 2), CST 9102; β2-microglobulin (β2m), CST 12851; GAPDH (D4C6R), CST 97166; goat anti-rabbit IgG H&L (HRP), Abcam ab6721; and goat anti-mouse IgG H&L (HRP), Abcam ab97023.

[0330] For immunohistochemical analysis, FFPE blocks were sectioned, automatically stained with a Leica Bond RX system, and imaged with a Leica Aperio Versa 8. Tumor areas were manually selected in HALO (Indica Labs), and the percentage of positive cells was automatically recorded. The following antibodies were used: ROS1, CST 63452; rabbit IgG, Abcam 172730; ERK, CST 4695; phospho-ERK, CST 4376; and Ki-67, CST 9027.

[0331] For gene expression analysis by NanoString assay, RNA was extracted from FFPE sections by RNAstorm FFPE RNA extraction kit, quantified by NanoDrop, Qubit and Agilent BioAnalyzer 2100, and analyzed by NanoString nCounter PanCancer Human Pathways Panel according to manufacturer's instructions. NanoString data were analyzed by Excel and data packages from R / Biocondutor, and included quality control of the assay, background correlation and linearity by negative and positive spikes, normalization based on housekeeping genes by the ratio median method, and differential gene expression analysis by the Walt test method (where p values ​​were corrected for multiple testing using the Benjamini-Hochberg method).

[0332] Example 4: Phase 1 / 2 clinical study

[0333] The FIH, Phase 1 / 2, multicenter, open-label, dose-escalation, and expansion studies were conducted to evaluate the safety and tolerability of compound 1, determine the RP2D, and evaluate the antitumor activity in patients with advanced ROS1-positive NSCLC and other advanced ROS1-positive solid tumors.

[0334] Phase 1 Purpose:

[0335] Primary objective: To determine the RP2D and / or maximum tolerated dose (MTD) of Compound 1 in patients with advanced ROS1-positive solid tumors.

[0336] Secondary objectives: To evaluate the overall safety and tolerability of compound 1; to characterize the PK characteristics of compound 1; and to assess the preliminary antitumor activity of compound 1 in patients with advanced ROS1-positive solid tumors.

[0337] Phase 2 Objectives:

[0338] Primary objective: To evaluate the ORR of compound 1 at the RP2D dose in patients with ROS1-positive NSCLC and other solid tumors.

[0339] Secondary objectives: To evaluate additional clinical efficacy indicators in patients with ROS1-positive NSCLC and other solid tumors; To evaluate the intracranial antitumor activity of compound 1 at the RP2D dose in patients with ROS1-positive NSCLC and other solid tumors; To characterize the safety and tolerability of compound 1 at the RP2D dose; To confirm the PK profile of compound 1 at the RP2D dose.

[0340] End point of phase 1:

[0341] Primary endpoint: Incidence of dose-limiting toxicity (DLT) in cycle 1

[0342] Secondary endpoints:

[0343] The incidence and severity of treatment-emergent adverse events (TEAEs) and changes in clinically relevant laboratory parameters

[0344] ●Pharmacokinetic parameters of compound 1: Maximum plasma concentration (C max );C max – Dose-normalized plasma concentration at the end of the dosing interval (C tau ); Time to reach maximum concentration (T max ); the area under the curve at the end of the dosing interval (AUC tau ); AUC tau – Dose-normalized area under the curve (AUC) from time 0 to 24 0-24 ); AUC 0-24 – Dose-normalized area under the curve (AUC) from time 0 to infinity inf ); AUC inf – Dose normalization; oral clearance (CL / F); volume of distribution (V z / F); and half-life (t 1 / 2 )

[0345] Objective response rate (ORR) according to Response Evaluation Criteria in Solid Tumors version 1.1 (RECIST 1.1) - defined as the percentage of patients with a complete response (CR) or partial response (PR) according to RECIST 1.1

[0346] Duration of response (DOR) according to RECIST 1.1 - In responders, this was defined as the time from the first response according to RECIST 1.1 to radiographic disease progression or death

[0347] Intracranial ORR according to RECIST 1.1 (IC-ORR) - In patients with measurable metastatic CNS disease, up to 5 intracranial target lesions were assessed as responders according to RECIST 1.1

[0348] ● Intracranial DOR per RECIST 1.1 (IC-DOR) - in responders with metastatic CNS disease, defined as the time from first IC response per RECIST 1.1 to radiographic IC disease progression or death

[0349] Clinical benefit rate (CBR) according to RECIST 1.1 - defined as the percentage of patients with a confirmed CR or PR according to RECIST 1.1 or stable disease lasting at least 24 weeks

[0350] ●Time to response according to RECIST 1.1 - defined as the time from the first dose to the first confirmed radiographic response according to RECIST 1.1

[0351] Progression-free survival (PFS) according to RECIST 1.1 - defined as the time from first dose to radiographic disease progression or death according to RECIST 1.1

[0352] Exploratory endpoints:

[0353] ● Pharmacodynamic markers associated with response to compound 1

[0354] Extracranial ORR (EC-ORR) according to RECIST 1.1

[0355] ●Among patients with extracranial metastatic disease, defined as the percentage of patients with an extracranial response according to RECIST 1.1 by investigator review

[0356] ●Evaluation of the metabolite profile of compound 1

[0357] Phase 2 endpoints:

[0358] Primary endpoint: ORR according to RECIST 1.1 - defined as the proportion of patients with confirmed CR or PR according to RECIST 1.1 by blinded independent central review (BICR) among patients with measurable disease at baseline.

[0359] Secondary End Points

[0360] ●DOR according to RECIST 1.1 - in responders, defined as the time from the first confirmed response according to RECIST 1.1 assessed by BICR to radiographic disease progression or death

[0361] IC-ORR per RECIST 1.1 - in patients with measurable metastatic CNS disease, defined as up to five intracranial target lesions assessed as responding per RECIST 1.1 by BICR in patients with measurable intracranial disease confirmed by BICR at baseline

[0362] ●IC-DOR according to RECIST 1.1 - In responders with metastatic CNS disease, defined as the time from the first confirmed response according to RECIST 1.1 assessed by BICR to radiographic disease progression or death.

[0363] ● CBR according to RECIST 1.1 - defined as a confirmed CR or PR according to RECIST 1.1 with BICR or SD lasting at least 24 weeks

[0364] ●Time to response according to RECIST 1.1 - defined as the time from the first dose to the first confirmed radiological response according to RECIST 1.1 by BICR

[0365] ●Time to IC response per RECIST 1.1 - In patients with metastatic CNS disease, defined as the time from first dose to first confirmed radiographic IC response per RECIST 1.1 by BICR

[0366] The incidence and severity of TEAEs and changes in clinically relevant laboratory parameters

[0367] ●Pharmacokinetic parameters of compound 1-C max , C max -Dose-normalized, C tau , T max , AUC tau , AUC tau -Dose-normalized, AUC 0-24 , AUC 0-24 – Dose-normalized, AUC inf , AUC inf –Dose-normalized, CL / F, V z / F,t 1 / 2

[0368] ●PFS according to RECIST 1.1 - defined as the time from first dose to radiographic disease progression according to RECIST 1.1 or death based on BICR assessment

[0369] ●Intracranial PFS according to RECIST 1.1 (IC-PFS) - in patients with metastatic CNS disease, defined as the time from first dose to radiographic disease progression according to RECIST 1.1 based on BICR assessment or death

[0370] Overall survival (OS) - defined as the time from first dose to death from any cause

[0371] Study Design: The study was conducted in 2 phases ( Figure 1 ).

[0372] ●Phase 1 with Bayesian Optimal Interval Design (BOIN) and 3+3 lead-in dose escalation: patients with advanced / metastatic ROS1-positive solid tumors treated with at least 1 prior ROS1 TKI therapy.

[0373] Phase 2 will include 5 cohorts:

[0374] ○ Cohort 2a: Patients with advanced / metastatic ROS1-positive NSCLC not treated with TKI therapy. Up to one prior platinum-based chemotherapy with or without immunotherapy was allowed.

[0375] ○ Cohort 2b: Patients with advanced / metastatic ROS1-positive NSCLC treated with 1 prior ROS1 TKI (crizotinib or entrectinib), and no prior platinum-based chemotherapy or immunotherapy was allowed.

[0376] ○ Cohort 2c: Patients with advanced / metastatic ROS1-positive NSCLC treated with 1 prior ROS1 TKI (crizotinib or entrectinib) and 1 prior platinum-based chemotherapy with or without immunotherapy.

[0377] ○ Cohort 2d: Patients with advanced / metastatic ROS1-positive NSCLC treated with ≥2 prior ROS1 TKIs and up to 1 prior platinum-based chemotherapy with or without immunotherapy.

[0378] ○ Cohort 2e (exploratory): Patients with any advanced / metastatic ROS1-positive solid tumor (including patients with ROS1-positive NSCLC who are not otherwise eligible for any other cohort) and who have progressed on any prior therapy (including but not limited to patients who have progressed on a prior ROS1 TKI).

[0379] The Phase 1 portion of the study used a BOIN design with a 3+3 lead-in phase to evaluate dose escalation. The primary objectives of the Phase 1 portion of the study were to determine the RP2D and evaluate the safety of Compound 1 in patients with solid tumors. Safety characteristics were assessed based on: physical examination; ECOGPS; changes from baseline in laboratory parameters, ECG, vital signs, and ophthalmological examinations (baseline only and as clinically indicated); and reporting of AEs. The DLT observation period was defined as the period from the first dose on Day 1 of Cycle 1 (C1D1) to the end of Cycle 1 (28-day cycle time).

[0380] After treating patients at a given dose level and monitoring them for 28 days, all available safety data were reviewed. Whether to start the next dosing group depended on the results of the BOIN analysis.

[0381] Selection of an RP2D will be performed after a sufficient number of patients have been enrolled and completed the first imaging / efficacy assessment and the data have been reviewed.

[0382] Phase 1 Dose Levels: Phase 1 evaluates increasing doses of Compound 1 monotherapy, administered orally once daily (QD), until disease progression, unacceptable toxicity, or voluntary withdrawal of consent. Each treatment cycle is 28 days. Under the BOIN design, up to approximately 53 patients will be enrolled and treated at the dose levels listed in the table below.

[0383] Dosage Levels Compound 1 Dosage frequency* DL1 25mg QD DL2 50mg QD DL3 75mg QD DL4 100mg QD DL5 125mg QD DL6** 150mg QD

[0384] *BID dosing can be explored at any DL.

[0385] **Higher DL can be explored.

[0386] Intermediate dose levels can be explored during dose escalation; lower dose levels (eg, 10 mg QD and 5 mg QD) can be explored as additional dose levels based on the results of BOIN.

[0387] Recommended Phase 2 Dose: The recommended Phase 2 dose (RP2D) is expected to be the optimal biological dose at or below the MTD, or the maximum feasible dose, and is the dose level considered appropriate for testing in the Phase 2 portion of the study. Based on PK modeling simulations, the RP2D is expected to achieve adequate target coverage of the G2032R ROS1 mutation in the periphery and CNS, and the RP2D also demonstrates a tolerable safety profile as well as preliminary antitumor activity. The selection of the RP2D is made using all data obtained from Phase 1 and includes evaluations of clinical PK, pharmacodynamics, safety, and antitumor activity.

[0388] Duration of treatment: In both phases, patients received study drug continuously from the first dose until independent radiologically confirmed disease progression, unacceptable toxicity, patient withdrawal, study termination, or commercial availability. Patients could continue to receive Compound 1 after progression in the body and brain amenable to local ablation.

[0389] Study End Definition: Study end was defined as the date of the last visit of the last patient in the study or study termination.

[0390] Inclusion Criteria: Patients must meet all of the following criteria to be eligible for inclusion in the study:

[0391] 1. Age ≥ 18 years. (Phase 2 Cohort 2e only: age ≥ 12 years and weight > 40 kg.)

[0392] 2. Disease criteria

[0393] a. Stage 1: Histologically or cytologically confirmed metastatic solid tumors with documented ROS1 rearrangement as determined by testing using local diagnostic tests or commercial tests or by regulatory approved fluorescence in situ hybridization (FISH) or next generation sequencing (NGS) or reverse transcription polymerase chain reaction (RT-PCR) in a Clinical Laboratory Improvement Amendments (CLIA) laboratory in the United States or an equivalent certified diagnostic laboratory outside the United States.

[0394] b. Cohorts 2a, 2b, 2c, and 2d: Histologically or cytologically confirmed metastatic NSCLC with a ROS1 rearrangement as determined by testing in a CLIA or equivalent certified diagnostic laboratory using a local diagnostic test or commercial test or by regulatory approved FISH or NGS or RT-PCR.

[0395] c. Cohort 2e: Histologically or cytologically confirmed metastatic solid tumors (except NSCLC) with ROS1 rearrangement as determined by testing in a CLIA or equivalent certified diagnostic laboratory using a local diagnostic test or commercial test or by regulatory approved FISH or NGS or RT-PCR.

[0396] 3. Previous anticancer treatment

[0397] a. Stage 1: Must have received at least 1 prior ROS1 TKI therapy; any number of prior platinum-based chemotherapy with or without immunotherapy is allowed.

[0398] b. Cohort 2a: Must be naive to TKI therapy and treated with at most one prior platinum-based chemotherapy with or without immunotherapy.

[0399] c. Cohort 2b: Must have received 1 prior ROS1 TKI therapy (crizotinib or entrectinib) and no prior platinum-based chemotherapy or immunotherapy.

[0400] d. Cohort 2c: Must have received 1 prior ROS1 TKI therapy (crizotinib or entrectinib) and 1 prior platinum-based chemotherapy with or without immunotherapy

[0401] e. Cohort 2d: Must have received at least 2 prior ROS1 TKI therapies and up to 1 prior platinum-based chemotherapy with or without immunotherapy.

[0402] f. Cohort 2e: Must have progressed on any prior therapy (including but not limited to patients who have progressed on prior ROS1 TKI).

[0403] Phase 4.1: Must have evaluable disease (target or non-target) according to RECIST 1.1. Phase 2: Must have measurable disease, defined as ≥1 radiologically measurable target lesion according to RECIST 1.1 (Oken et al., Am. J. Clin. Oncol. 1982, 5(6): 649-655). In all study parts, patients with asymptomatic CNS-only measurable disease of ≥10 mm as defined by RECIST 1.1 are eligible.

[0404] 5. The patient agrees to provide tumor tissue (archived (if available) or fresh biopsy) for ROS1 status confirmation and is willing to consider tumor biopsy during treatment if it is considered safe and medically optional.

[0405] 6. Eastern Cooperative Oncology Group (ECOG) performance status (PS) of 0, 1 or 2 (Eisenhauer et al., Eur. J. Cancer 2009, 45(2):228-247).

[0406] 7. Adequate organ function and bone marrow reserve, as demonstrated by the following laboratory assessments performed prior to the first dose of study drug:

[0407] a. Bone marrow function: absolute neutrophil count (ANC) ≥1500 / μL; platelet count >75,000 / μL; hemoglobin ≥8 g / dL.

[0408] b. Renal function: estimated creatinine clearance calculated according to the Cockroft-Gault formula ≥ 60 mL / min.

[0409] c. Liver function: bilirubin <1.5×ULN, unless there is evidence of Gilbert's syndrome, in which case the patient's total bilirubin must be <3.0 mg / dL; aspartate aminotransferase and alanine aminotransferase ≤3.0×ULN (≤5.0×ULN if liver metastases are involved).

[0410] 8. All clinically relevant toxicities related to previous anticancer therapy must have recovered to Grade ≤1 or baseline (except alopecia or ototoxicity).

[0411] 9. Women of childbearing potential (WOCBP) and male patients must be willing to abstain from sexual activity or use effective contraceptive methods or surgical sterilization from the time of signing the informed consent form (ICF) to the duration of the study and 30 days (females) and 90 days (males) after the last administration of study drug. Effective contraception for WOCBP includes 1 "highly effective method" or 2 "effective" methods based on World Health Organization (WHO) standards.

[0412] 10. Provided written informed consent and willing to comply with the requirements of the study protocol. Patients <18 years of age (Phase 2 Cohort 2e only) must Assent, and parents / guardians must provide written consent.

[0413] Exclusion criteria: Patients meeting any of the following criteria were excluded from the study:

[0414] 1. The patient's cancer has a known primary driver alteration other than ROS1. For example, NSCLC with targetable mutations of EGFR, ALK, MET, RET, or BRAF; colorectal cancer with oncogenic KRAS, NRAS, or BRAF mutations.

[0415] 2. Known allergic / hypersensitivity reaction to the excipients of Compound 1.

[0416] 3. Major surgical procedure within 4 weeks prior to study entry. Minor surgical procedures (eg, port insertion) are permitted but with adequate wound healing time deemed clinically appropriate.

[0417] 4. Continuous or recent anticancer therapy within the following timeframes prior to first dose of study drug (Compound 1 may be started within the limits of prior TKI or chemotherapy if deemed safe and in the best interest of the patient):

[0418] a. TKI or other non-chemotherapy / non-immunotherapy anticancer agent < 5 half-lives or < 7 days, whichever is longer

[0419] b. Chemotherapy < 21 days

[0420] c. Immunotherapy or cell therapy < 28 days

[0421] 5. Continuous or recent radiotherapy within the following time frame before the first dose of study drug:

[0422] a. Radiation therapy (except palliative radiation to relieve bone pain) < 14 days

[0423] b. Palliative radiation to relieve bone pain < 48 hours

[0424] c. Stereotactic or small-field brain radiation < 7 days

[0425] d. Whole brain radiation < 14 days

[0426] 6. Previous high-dose chemotherapy, requiring stem cell rescue.

[0427] 7. Poorly controlled clinically relevant bacterial or fungal infection requiring systemic therapy.

[0428] 8. Have known active tuberculosis or active hepatitis B or C. Active hepatitis B is defined by a known positive HBsAg result. Active hepatitis C is defined by a known hepatitis C Ab result and a known quantitative HCV RNA result greater than the detection limit of the assay.

[0429] 9. The patient's QTcF is always >450 milliseconds. The patient has a history of long QT syndrome or torsades de pointes.

[0430] 10. Patients with clinically significant cardiovascular disease are as follows:

[0431] a. Within 3 months before enrollment: cerebrovascular accident / stroke; myocardial infarction; unstable angina; poorly controlled atrial fibrillation of any grade

[0432] b. Congestive heart failure (New York Heart Association classification ≥ II); 2nd or 3rd degree AV block (unless rhythmic) or any AV block with a PR consistently > 220 msec; or sustained arrhythmia of NCI-CTCAE grade ≥ 2.

[0433] 11. Patients with central nervous system (CNS) metastases or primary CNS tumors associated with progressive neurological symptoms or requiring escalating doses of steroids to control CNS disease. If patients require steroids to control CNS disease, the dose must be stable for at least 2 weeks prior to C1D1.

[0434] 12. Symptomatic spinal cord compression.

[0435] 13. Patients with moderate to severe cognitive impairment or psychiatric disorders that compromise their ability to comply with study requirements.

[0436] 14. Evidence of active malignancy (except current ROS1-positive solid malignancy) requiring systemic therapy within the previous 2 years.

[0437] a. Exceptions: non-melanoma skin cancer, melanoma in situ, cervical cancer in situ, papillary thyroid cancer, ductal carcinoma in situ of the breast, or localized and presumed cured prostate cancer. Patients receiving long-term antihormonal therapy for a prior malignancy are permitted as long as the malignancy has not been active within the previous 2 years.

[0438] 15. Concurrent use (within 12 days before enrollment) of strong CYP3A4 inducers or strong CYP3A4 inhibitors.

[0439] 16. Patients with manifestations of malabsorption due to previous gastrointestinal surgery, disease, or other diseases that may affect the oral absorption, distribution, metabolism, or excretion of the study drug.

[0440] 17. Patient is pregnant or breastfeeding. WOCBP must have a negative serum pregnancy test at screening and a negative urine test before the first dose of study drug.

[0441] 18. Actively receiving systemic therapy or direct medical intervention in another therapeutic clinical study.

[0442] 19. Any medical condition or laboratory abnormality that would pose a risk to the study patients or confound the ability to interpret the study results.

[0443] Study Drug, Dosage and Route of Administration: Compound 1 is supplied as oral tablets in 2 strengths: 5 mg and 25 mg in 32-tablet high-density polyethylene bottles with induction seal / child-resistant caps for oral administration. Compound 1 should be taken on an empty stomach, at least 1 hour before and no less than 2 hours after ingestion of food and / or beverages other than water.

[0444] Prior and concurrent medications and therapies:

[0445] Compound 1 is not considered a substrate for CYP2B6, CYP2C9, CYP2C19, or CYP2D6. Compound 1 is a substrate for CYP3A4, with possible contributions from CYP1A2 and CYP2C8.

[0446] Compound 1 is not considered to be an inhibitor of CYP1A2, CYP2B6, or CYP2D6 (IC 50 >100 μM), and it is CYP2C8 (IC 50 =70 μM), CYP2C9 (IC 50 =70.3μM), CYP3A4 (testosterone IC 50 =62.7 μM), CYP2C19 (IC 50=38.4 μM) and CYP3A4 (midazolam IC 50 =30 μM). Compound 1 is a weak inhibitor of CYP3A4 (testosterone) and CYP3A4 (midazolam). Compound 1 is a potential inducer of CYP3A4.

[0447] In certain embodiments, the concomitant use (within 12 days prior to enrollment) of drugs that are known strong CYP3A4 inducers (such as phenobarbital, rifampicin, phenytoin, carbamazepine, rifabutin, rifapentine, clevidipine, St. John's wort) is prohibited. In certain embodiments, the concomitant use (within 12 days prior to enrollment) of drugs that are known strong CYP3A4 inducers (such as phenobarbital, rifampicin, phenytoin, carbamazepine, rifabutin, rifapentine, clevidipine, St. John's wort) is allowed. In certain embodiments, the concomitant use (within 12 days prior to enrollment) of drugs that are known strong CYP3A4 inducers (such as phenobarbital, rifampicin, phenytoin, carbamazepine, rifabutin, rifapentine, clevidipine, St. John's wort) should be used with caution.

[0448] In certain embodiments, the concomitant use (within 12 days prior to enrollment) of the following drugs should be used with caution: substrates or inducers of CYP3A4, inhibitors of CYP3A4 or CYP2C8, substrates of P-glycoprotein (P-gp), breast cancer resistance protein (BCRP) or MATE1 transporters, substrates of CYP2C19 with narrow therapeutic indicators. In certain embodiments, the concomitant use (within 12 days prior to enrollment) or co-administration of Compound 1 and any one or more of the following drugs is permitted: substrates or inducers of CYP3A4, inhibitors of CYP3A4 or CYP2C8, substrates of P-glycoprotein (P-gp), breast cancer resistance protein (BCRP) or MATE1 transporters, substrates of CYP2C19 with narrow therapeutic indicators. In certain embodiments, concomitant use (within 12 days prior to enrollment) or co-administration of the following drugs is prohibited: substrates or inducers of CYP3A4, inhibitors of CYP3A4 or CYP2C8, substrates of P-glycoprotein (P-gp), breast cancer resistance protein (BCRP), or MATE1 transporters, substrates of CYP2C19 with a narrow therapeutic indication.

[0449] In certain embodiments, simultaneous use (within 12 days prior to enrollment) or co-administration of gastric acid inhibitors and Compound 1 should be avoided. In certain embodiments, simultaneous use (within 12 days prior to enrollment) or co-administration of gastric acid inhibitors and Compound 1 should be used with caution. In certain embodiments, simultaneous use (within 12 days prior to enrollment) or co-administration of gastric acid inhibitors and Compound 1 is permitted.

[0450] The agents provided herein include, but are not limited to:

[0451] CYP3A4 inducers: Phenobarbital, rifampin, phenytoin, carbamazepine, rifabutin, rifapentine, clevidipine, St. John's wort

[0452] CYP3A4 inhibitors: Clarithromycin, erythromycin, diltiazem, itraconazole, ketoconazole, ritonavir, verapamil, goldenseal, grapefruit (including juice)

[0453] CYP3A4 substrates (sensitivity): Buspirone, everolimus, lovastatin, midazolam, simvastatin, triazolam, maraviroc, conivaptan, darifenacin

[0454] P-gp substrates: digoxin, fexofenadine, loperamide, quinidine, talinolol, vinblastine

[0455] BCRP substrates: daidzein, dantrolene, estrone-3-sulfate, prazosin, sulfasalazine

[0456] MATE1 substrates: Metformin, tetraethylammonium (TEA), cimetidine, procainamide

[0457] Example 5: Phase 1 / 2 Clinical Study

[0458] The FIH, Phase 1 / 2, multicenter, open-label, dose-escalation, and expansion studies were conducted to evaluate the safety and tolerability of Compound 1, determine the RP2D and (if applicable) MTD, and evaluate antitumor activity in patients with advanced ROS1-positive NSCLC and other advanced ROS1-positive solid tumors.

[0459] Phase 1 Purpose:

[0460] Primary objective: To determine the RP2D and / or maximum tolerated dose (MTD) of Compound 1 in patients with advanced ROS1-positive solid tumors.

[0461] Secondary objectives: To evaluate the overall safety and tolerability of compound 1; to characterize the PK characteristics of compound 1; and to assess the preliminary antitumor activity of compound 1 in patients with advanced ROS1-positive solid tumors.

[0462] Phase 2 Objectives:

[0463] Primary objective: To evaluate the ORR of compound 1 at the RP2D dose in patients with ROS1-positive NSCLC and other solid tumors.

[0464] Secondary objectives: To evaluate additional clinical efficacy indicators in patients with ROS1-positive NSCLC and other solid tumors; To evaluate the intracranial antitumor activity of compound 1 at the RP2D dose in patients with ROS1-positive NSCLC and other solid tumors; To characterize the safety and tolerability of compound 1 at the RP2D dose; To confirm the PK profile of compound 1 at the RP2D dose.

[0465] End point of phase 1:

[0466] Primary endpoints: RP2D and, if applicable, MTD as determined by DLT incidence in Cycle 1, overall safety profile, PK, PD, and preliminary efficacy.

[0467] Secondary endpoints:

[0468] The incidence and severity of treatment-emergent adverse events (TEAEs) and changes in clinically relevant laboratory parameters

[0469] ●Pharmacokinetic parameters of compound 1: Maximum plasma concentration (C max );C max_DN (C max – dose-normalized), plasma concentration at the end of the dosing interval (C tau ); Time to reach maximum concentration (T max ); the area under the curve at the end of the dosing interval (AUC tau ); AUC tau_DN (AUC tau -dose-normalized), area under the curve (AUC) from time 0 to 24 0-24 ); AUC 0-24_DN (AUC 0-24 – dose-normalized), area under the curve (AUC) from time 0 to infinity inf ); AUC inf_DN (AUC inf – dose-normalized); oral clearance (CL / F); volume of distribution (V z / F); and half-life (t 1 / 2 )

[0470] Objective response rate (ORR) according to Response Evaluation Criteria in Solid Tumors version 1.1 (RECIST 1.1) - defined as the percentage of patients with a complete response (CR) or partial response (PR) according to RECIST 1.1

[0471] Duration of response (DOR) according to RECIST 1.1 - In responders, this was defined as the time from the first response according to RECIST 1.1 to radiographic disease progression or death

[0472] Intracranial ORR per RECIST 1.1 (IC-ORR) - defined as the proportion of patients with a confirmed intracranial response (IC-CR or IC-PR) based on an assessment of up to five intracranial target lesions (as assessed by the investigator per RECIST 1.1 criteria) among patients with measurable metastatic disease at baseline

[0473] ●Intracranial DOR according to RECIST 1.1 (IC-DOR) - defined as the time from the first investigator-assessed IC response according to RECIST 1.1 criteria to radiographic IC disease progression or death in patients with an intracranial response

[0474] Clinical benefit rate (CBR) according to RECIST 1.1 - defined as the percentage of patients with a confirmed CR or PR according to RECIST 1.1 or stable disease lasting at least 24 weeks

[0475] ●Time to response according to RECIST 1.1 - defined as the time from the first dose to the first confirmed radiographic response according to RECIST 1.1

[0476] Progression-free survival (PFS) according to RECIST 1.1 - defined as the time from first dose to radiographic disease progression or death according to RECIST 1.1

[0477] Exploratory endpoints:

[0478] ● Pharmacodynamic markers associated with response to compound 1

[0479] Extracranial ORR (EC-ORR) according to RECIST 1.1

[0480] ●Among patients with extracranial metastatic disease, defined as the percentage of patients with an extracranial response according to RECIST 1.1 by investigator review

[0481] ●Evaluation of the metabolite profile of compound 1

[0482] Phase 2 endpoints:

[0483] Primary endpoint: ORR according to RECIST 1.1 - defined as the proportion of patients with confirmed CR or PR according to RECIST 1.1 by blinded independent central review (BICR) among patients with measurable disease at baseline.

[0484] Secondary End Points

[0485] ●DOR according to RECIST 1.1 - in responders, defined as the time from the first confirmed response according to RECIST 1.1 assessed by BICR to radiographic disease progression or death

[0486] IC-ORR according to RECIST 1.1 - among patients with measurable metastatic disease at baseline, defined as the proportion of patients with a confirmed intracranial response (IC-CR or IC-PR) based on evaluation of up to five intracranial target lesions (by BICR according to RECIST 1.1 criteria)

[0487] IC-DOR according to RECIST 1.1 - in patients with an intracranial response, defined as the time from the first confirmed IC response according to RECIST 1.1 criteria assessed by BICR to radiographic IC disease progression or death

[0488] ● CBR according to RECIST 1.1 - defined as a confirmed CR or PR according to RECIST 1.1 with BICR or SD lasting at least 24 weeks

[0489] ●Time to response according to RECIST 1.1 - defined as the time from first dose to first confirmed radiographic IC response according to RECIST 1.1 criteria by BICR in patients with measurable metastatic CNS disease

[0490] ●Time to IC response per RECIST 1.1 - In patients with metastatic CNS disease, defined as the time from first dose to first confirmed radiographic IC response per RECIST 1.1 by BICR

[0491] The incidence and severity of TEAEs and changes in clinically relevant laboratory parameters

[0492] ●Pharmacokinetic parameters of compound 1-C max , C max_DN (Dose-normalized C max ), C tau , T max , AUC tau , AUC tau_DN (AUC tau -dose-normalized), AUC 0-24 , AUC 0-24_CN (Dose-normalized AUC 0-24 ), AUC inf , AUC inf_DN (AUC inf -dose-normalized), CL / F, V z / F,t 1 / 2

[0493] ●PFS according to RECIST 1.1 - defined as the time from first dose to radiographic disease progression according to RECIST 1.1 or death based on BICR assessment

[0494] Intracranial PFS according to RECIST 1.1 (IC-PFS) - defined as the time from first dose to radiographic disease progression according to RECIST 1.1 criteria or death based on BICR assessment in patients with measurable metastatic CNS disease

[0495] Overall survival (OS) - defined as the time from first dose to death from any cause

[0496] Study Design: The study was conducted in 2 phases ( Figure 1 , except for period 1 where n=54).

[0497] ● Phase 1 with Bayesian Optimal Interval Design (BOIN) and 3+3 lead-in dose escalation: patients with advanced / metastatic ROS1-positive NSCLC treated with at least 1 prior ROS1 TKI therapy, or patients with other ROS1-positive solid tumors who have progressed on any prior therapy (including but not limited to patients who have progressed on prior ROS1 TKIs).

[0498] Phase 2 will include 5 cohorts:

[0499] ○ Cohort 2a: Patients with advanced / metastatic ROS1-positive NSCLC not treated with TKI therapy. Up to one prior platinum-based chemotherapy with or without immunotherapy was allowed.

[0500] ○ Cohort 2b: Patients with advanced / metastatic ROS1-positive NSCLC treated with 1 prior ROS1 TKI (crizotinib or entrectinib), and no prior platinum-based chemotherapy or immunotherapy was allowed.

[0501] ○ Cohort 2c: Patients with advanced / metastatic ROS1-positive NSCLC treated with 1 prior ROS1 TKI (crizotinib or entrectinib) and 1 prior platinum-based chemotherapy with or without immunotherapy.

[0502] ○ Cohort 2d: Patients with advanced / metastatic ROS1-positive NSCLC treated with ≥2 prior ROS1 TKIs and up to 1 prior platinum-based chemotherapy with or without immunotherapy.

[0503] ○ Cohort 2e (exploratory): Patients with any advanced / metastatic ROS1-positive solid tumor (including patients with ROS1-positive NSCLC who are not otherwise eligible for any other cohort) and who have progressed on any prior therapy (including but not limited to patients who have progressed on a prior ROS1 TKI).

[0504] The Phase 1 portion of the study used a BOIN design with a 3+3 lead-in phase to evaluate dose escalation. The primary objectives of the Phase 1 portion of the study were to determine the RP2D and, if applicable, the MTD, and to evaluate the safety of Compound 1 in patients with solid tumors. Safety characteristics were assessed based on: physical examination; ECOGPS; changes from baseline in laboratory parameters, ECG, vital signs, and ophthalmological examinations (baseline only and as clinically indicated); and reporting of AEs. The DLT observation period was defined as the period from the first dose on Day 1 of Cycle 1 (C1D1) to the end of Cycle 1 (28-day cycle time).

[0505] After patients were treated and monitored for 28 days at a given dose level, available safety data (including but not limited to the incidence of DLT / other AEs and preliminary PK) were reviewed. Whether to start the next dosing group depends on the incidence of DLT and the results of the BOIN analysis and takes into account the overall safety profile. During dose escalation, additional patients (up to a total of 12 patients, including patients already included in the BOIN dose escalation) may be enrolled at that dose level when available data support the safety, PK, and clinical activity of a given dose level.

[0506] BOIN dose escalation is declared complete when the number of evaluable patients treated at the current dose reaches 9 and the decision to stay at the current dose is made, or when the maximum sample size is reached. However, if the optimal biological dose or maximum feasible dose is reached before the completion of BOIN dose escalation, an RP2D can be declared. Selection of an RP2D is made after a sufficient number of patients have been enrolled and completed the first imaging / efficacy assessments and the data have been reviewed.

[0507] To further evaluate safety and efficacy and to confirm the RP2D, up to 20 additional patients may be treated at the RP2D or at one or more candidate RP2Ds prior to initiation of the Phase 2 portion of the study.

[0508] Phase 1 Dose Levels: Phase 1 evaluates increasing doses of Compound 1 monotherapy, administered orally once daily (QD), until disease progression, unacceptable toxicity, or voluntary withdrawal of consent. Each treatment cycle is 28 days. Under the BOIN design, up to approximately 53 patients will be enrolled and treated at the dose levels listed in the table below.

[0509] Dosage Levels Compound 1 Dosage frequency* DL1 25mg QD DL2 50mg QD DL3 75mg QD DL4 100mg QD DL5 125mg QD DL6** 150mg QD

[0510] *BID dosing can be explored at any DL.

[0511] **Higher DL can be explored.

[0512] In the process of dose escalation, intermediate dose levels can be explored; lower dose levels (e.g., 10 mg Qd and 5 mg QD) can be explored as additional dose levels based on the results of BOIN. For example, the following dose reduction levels are provided herein.

[0513] Initial dose level Dose Level-1 Dose Level - 2** 25mg QD 10mg QD 5mg QD 50mg QD 25mg QD 10mg QD 75mg QD 50mg QD 25mg QD 100mg QD 75mg QD 50mg QD 125mg QD 100mg QD 75mg QD 150 mg QD* 125mg QD 100mg QD

[0514] *Higher DLs may be explored. If a dose reduction is necessary, the dose should be reduced to a lower dose level.

[0515] **If treatment is not tolerated after 2 dose reductions, a third dose may be administered.

[0516] Recommended Phase 2 Dose: The recommended Phase 2 dose (RP2D) is expected to be the optimal biological dose at or below the MTD, or the maximum feasible dose, and is the dose level considered appropriate for testing in the Phase 2 portion of the study. Based on PK modeling simulations, the RP2D is expected to achieve adequate target coverage of the G2032R ROS1 mutation in the periphery and CNS, and the RP2D also demonstrates a tolerable safety profile as well as preliminary antitumor activity. The selection of the RP2D is made using all data obtained from Phase 1 and includes evaluations of clinical PK, pharmacodynamics, safety, and antitumor activity.

[0517] Duration of treatment: In both phases, patients received study drug continuously from the first dose until disease progression, unacceptable toxicity, patient withdrawal, study termination, or commercial availability. Patients could continue to receive Compound 1 after progression in the body and brain amenable to local ablation.

[0518] Study End Definition: Study end was defined as the date of the last visit of the last patient in the study or study termination.

[0519] Inclusion Criteria: Patients must meet all of the following criteria to be eligible for inclusion in the study:

[0520] 1. Age ≥ 18 years old.

[0521] a. Phase 2 Cohort 2e only: Age ≥12 years and weight >40 kg. (Patients aged 12 to 17 years were enrolled only in countries and centers where regulatory approval was granted.)

[0522] 2. Disease criteria

[0523] a. Stage 1: Histologically or cytologically confirmed locally advanced or metastatic solid tumors with documented ROS1 rearrangement as determined by testing in a Clinical Laboratory Improvement Amendments (CLIA) laboratory in the United States or an equivalent certified diagnostic laboratory outside the United States using a local diagnostic test or commercial test or by a regulatory agency approved test such as fluorescence in situ hybridization (FISH) or next generation sequencing (NGS) or reverse transcription polymerase chain reaction (RT-PCR).

[0524] b. Cohorts 2a, 2b, 2c, and 2d: Histologically or cytologically confirmed locally advanced or metastatic NSCLC with ROS1 rearrangement as determined by testing in a CLIA or equivalent certified diagnostic laboratory using a local diagnostic test or commercial test or by a regulatory agency approved test such as FISH or NGS or RT-PCR.

[0525] c. Cohort 2e: Histologically or cytologically confirmed locally advanced or metastatic solid tumors (including NSCLC not eligible for Cohorts 2a-2d) with ROS1 rearrangement as determined by testing in a CLIA or equivalent certified diagnostic laboratory using a local diagnostic test or commercial test or by a regulatory agency approved test such as FISH or NGS or RT-PCR.

[0526] 3. Previous anticancer treatment

[0527] a. Phase 1: Patients with ROS1-positive NSCLC must have previously received at least 1 prior ROS1 TKI, and patients with other ROS1-positive solid tumors must have progressed on any prior therapy (including but not limited to patients who have progressed on a prior ROS1 TKI); any number of prior platinum-based chemotherapy with or without immunotherapy is allowed.

[0528] b. Cohort 2a: Must be naive to TKI therapy and treated with at most one prior platinum-based chemotherapy with or without immunotherapy.

[0529] c. Cohort 2b: Must have received 1 prior ROS1 TKI therapy (crizotinib or entrectinib) and no prior platinum-based chemotherapy or immunotherapy.

[0530] d. Cohort 2c: Must have received 1 prior ROS1 TKI therapy (crizotinib or entrectinib) and 1 prior platinum-based chemotherapy with or without immunotherapy

[0531] e. Cohort 2d: Must have received at least 2 prior ROS1 TKI therapies and up to 1 prior platinum-based chemotherapy with or without immunotherapy.

[0532] f. Cohort 2e: Must have progressed on any prior therapy (including but not limited to patients who have progressed on prior ROS1 TKI).

[0533] Phase 4.1: Must have evaluable disease (target or non-target) according to RECIST 1.1. Phase 2: Must have measurable disease, defined as ≥1 radiologically measurable target lesion according to RECIST 1.1 (Eisenhauer et al., Eur. J. Cancer 2009, 45(2): 228-247). Patients with CNS-only disease are eligible if the disease is evaluable (Phase 1) or measurable (Phase 2) and do not meet Exclusion Criterion #11.

[0534] 5. The patient agrees to provide tumor tissue (archived (if available) or fresh biopsy) for central analysis. Preferably, the submitted tumor tissue is obtained during or after the latest disease progression. If suitable tissue cannot be obtained and if biopsy is not considered safe and medically feasible, the patient may be approved for enrollment in consultation with the medical monitor.

[0535] 6. Eastern Cooperative Oncology Group (ECOG) performance status (PS) of 0, 1 or 2 (Oken et al., Am. J. Clin. Oncol. 1982, 5(6): 649-655).

[0536] 7. Adequate organ function and bone marrow reserve, as demonstrated by the following laboratory values ​​last assessed before the first dose of study drug:

[0537] a. Bone marrow function: absolute neutrophil count (ANC) ≥1500 / μL; platelet count >75,000 / μL; hemoglobin ≥8g / dL (without blood transfusion).

[0538] b. Renal function: estimated creatinine clearance calculated by the Cockroft-Gault formula, modified Cockroft-Gault formula, or 24-hour creatinine clearance ≥ 60 mL / min.

[0539] c. Liver function: bilirubin <1.5×ULN, unless there is evidence of Gilbert's syndrome, in which case the patient's total bilirubin must be <3.0 mg / dL; aspartate aminotransferase and alanine aminotransferase ≤3.0×ULN (≤5.0×ULN if liver metastases are involved).

[0540] 8. All clinically relevant toxicities related to previous anticancer therapy must have recovered to Grade ≤1 or baseline (except alopecia or ototoxicity).

[0541] 9. Women of childbearing potential (WOCBP) and male patients must be surgically sterilized or willing to abstain from sexual activity or use effective abstention methods from the time of signing the informed consent form (ICF) to at least 30 days (females) and at least 90 days (males) after the last administration of study drug (or longer, as required by laws or regulations). Effective contraception for WOCBP includes 1 "highly effective method" or 2 "effective" methods based on the World Health Organization (WHO) standards.

[0542] 10. Provided written informed consent and willing to comply with the requirements of the study protocol. Assent must be obtained from patients <18 years of age (or minors under the age of consent as defined by local regulations), and parents / guardians must provide written consent.

[0543] Exclusion criteria: Patients meeting any of the following criteria were excluded from the study:

[0544] 1. The patient's cancer has a known oncogenic driver alteration other than ROS1. For example, NSCLC with a targetable mutation of EGFR, ALK, MET, RET, or BRAF; colorectal cancer with an oncogenic KRAS, NRAS, or BRAF mutation.

[0545] 2. Known allergic / hypersensitivity reaction to the excipients of Compound 1.

[0546] 3. Major surgery within 4 weeks prior to first dose of study drug. Minor surgery (e.g., port insertion) is permitted but with adequate wound healing time deemed clinically appropriate.

[0547] 4. Continuous or recent anticancer therapy within the following timeframes prior to first dose of study drug (Compound 1 may be started within the limits of prior TKI or chemotherapy if deemed safe and in the best interest of the patient):

[0548] a. TKI or other anticancer drugs (excluding chemotherapy, immunotherapy or cell therapy for which guidelines are given below) < 5 half-lives or < 7 days, whichever is longer

[0549] b. Chemotherapy < 21 days

[0550] c. Immunotherapy or cell therapy < 28 days

[0551] 5. Continuous or recent radiotherapy within the following time frame before the first dose of study drug:

[0552] a. Radiation therapy (except palliative radiation to relieve bone pain) < 14 days

[0553] b. Palliative radiation to relieve bone pain < 48 hours

[0554] c. Stereotactic or small-field brain radiation < 7 days

[0555] d. Whole brain radiation < 14 days

[0556] 6. Previous high-dose chemotherapy, requiring stem cell rescue.

[0557] 7. Poorly controlled clinically relevant bacterial or fungal infection requiring systemic therapy.

[0558] 8. Have known active tuberculosis or active hepatitis B or C. Active hepatitis B is defined by a known positive HBsAg result and a known quantitative HBV DNA result greater than the detection limit of the assay. Active hepatitis C is defined by a known hepatitis C Ab result and a known quantitative HCV RNA result greater than the detection limit of the assay.

[0559] 9. The patient has a QTcF > 450 msec (repeatedly demonstrated during multiple evaluations). The patient has a history of long QT syndrome or torsades de pointes.

[0560] 10. Patients with clinically significant cardiovascular disease are as follows:

[0561] a. Within 3 months before enrollment: cerebrovascular accident / stroke; myocardial infarction; unstable angina; poorly controlled atrial fibrillation of any grade

[0562] b. Congestive heart failure (New York Heart Association classification ≥ II); 2nd or 3rd degree AV block (unless rhythmic) or any AV block with a PR consistently > 220 msec; or sustained arrhythmia of NCI-CTCAE grade ≥ 2.

[0563] 11. Patients have central nervous system (CNS) metastases or primary CNS tumors associated with progressive neurological symptoms or requiring escalating doses of steroids to control CNS disease. If patients require steroids to control CNS disease, the dose must be stable for at least 2 weeks prior to C1D1. Asymptomatic leptomeningeal carcinomatosis is permitted.

[0564] 12. Symptomatic spinal cord compression.

[0565] 13. Patients with moderate to severe cognitive impairment or psychiatric disorders that compromise their ability to comply with study requirements.

[0566] 14. Evidence of active malignancy (except current ROS1-positive solid malignancy) requiring systemic therapy within the previous 2 years.

[0567] a. Exceptions: non-melanoma skin cancer, melanoma in situ, cervical cancer in situ, papillary thyroid cancer, ductal carcinoma in situ of the breast, or localized and presumed cured prostate cancer. Patients receiving long-term antihormonal therapy for a prior malignancy are permitted as long as the malignancy has not been active within the previous 2 years.

[0568] 15. Concomitant use (within 12 days before the first dose of study drug) of strong CYP3A4 inducers or strong CYP3A4 inhibitors.

[0569] 16. Patients with manifestations of malabsorption due to previous gastrointestinal surgery, disease, or other diseases that may affect the oral absorption, distribution, metabolism, or excretion of the study drug.

[0570] 17. Patient is pregnant or breastfeeding. WOCBP must have a negative serum pregnancy test at screening and a negative serum or urine test before the first dose of study drug.

[0571] 18. Actively receiving systemic therapy or direct medical intervention in another therapeutic clinical study.

[0572] 19. Any medical condition or laboratory abnormality that would pose a risk to the study patients or confound the ability to interpret the study results.

[0573] Study Drug, Dosage and Administration Route: Compound 1 is supplied as film-coated tablets (for oral administration) in 2 strengths: 5 mg and 25 mg in 32-tablet high-density polyethylene bottles with induction seal / child-resistant caps. Compound 1 should be taken on an empty stomach, at least 1 hour before and no less than 2 hours after ingestion of food and / or beverages other than water.

[0574] Prior and concurrent medications and therapies:

[0575] Compound 1 is not considered a substrate for CYP2B6, CYP2C9, CYP2C19, or CYP2D6. Compound 1 is a substrate for CYP3A4, with possible contributions from CYP1A2 and CYP2C8.

[0576] Compound 1 is not considered to be an inhibitor of CYP1A2, CYP2B6, or CYP2D6 (IC 50 >100 μM), and it is CYP2C8 (IC 50 =70 μM), CYP2C9 (IC 50 =70.3μM), CYP3A4 (testosterone IC 50 =62.7 μM), CYP2C19 (IC 50 =38.4 μM) and CYP3A4 (midazolam IC 50=30 μM). Compound 1 is a weak inhibitor of CYP3A4 (testosterone) and CYP3A4 (midazolam). Compound 1 is a potential inducer of CYP3A4.

[0577] In certain embodiments, the concomitant use (within 12 days prior to enrollment) of drugs that are known strong CYP3A4 inducers (such as phenobarbital, rifampicin, phenytoin, carbamazepine, rifabutin, rifapentine, clevidipine, St. John's wort) is prohibited. In certain embodiments, the concomitant use (within 12 days prior to enrollment) of drugs that are known strong CYP3A4 inducers (such as phenobarbital, rifampicin, phenytoin, carbamazepine, rifabutin, rifapentine, clevidipine, St. John's wort) is allowed. In certain embodiments, the concomitant use (within 12 days prior to enrollment) of drugs that are known strong CYP3A4 inducers (such as phenobarbital, rifampicin, phenytoin, carbamazepine, rifabutin, rifapentine, clevidipine, St. John's wort) should be used with caution.

[0578] In certain embodiments, the concomitant use (within 12 days prior to enrollment) of the following drugs should be used with caution: substrates or inducers of CYP3A4, inhibitors of CYP3A4 or CYP2C8, substrates of P-glycoprotein (P-gp), breast cancer resistance protein (BCRP) or MATE1 transporters, substrates of CYP2C19 with narrow therapeutic indicators. In certain embodiments, the concomitant use (within 12 days prior to enrollment) or co-administration of Compound 1 and any one or more of the following drugs is permitted: substrates or inducers of CYP3A4, inhibitors of CYP3A4 or CYP2C8, substrates of P-glycoprotein (P-gp), breast cancer resistance protein (BCRP) or MATE1 transporters, substrates of CYP2C19 with narrow therapeutic indicators. In certain embodiments, concomitant use (within 12 days prior to enrollment) or co-administration of the following drugs is prohibited: substrates or inducers of CYP3A4, inhibitors of CYP3A4 or CYP2C8, substrates of P-glycoprotein (P-gp), breast cancer resistance protein (BCRP), or MATE1 transporters, substrates of CYP2C19 with a narrow therapeutic indication.

[0579] In certain embodiments, simultaneous use (within 12 days prior to enrollment) or co-administration of gastric acid inhibitors and Compound 1 should be avoided. In certain embodiments, simultaneous use (within 12 days prior to enrollment) or co-administration of gastric acid inhibitors and Compound 1 should be used with caution. In certain embodiments, simultaneous use (within 12 days prior to enrollment) or co-administration of gastric acid inhibitors and Compound 1 is permitted.

[0580] The agents provided herein include, but are not limited to:

[0581] CYP3A4 inducers: Phenobarbital, rifampin, phenytoin, carbamazepine, rifabutin, rifapentine, clevidipine, St. John's wort

[0582] CYP3A4 inhibitors: Clarithromycin, erythromycin, diltiazem, itraconazole, ketoconazole, ritonavir, verapamil, goldenseal, grapefruit (including juice)

[0583] CYP3A4 substrates (sensitivity): Buspirone, everolimus, lovastatin, midazolam, simvastatin, triazolam, maraviroc, conivaptan, darifenacin

[0584] P-gp substrates: digoxin, fexofenadine, loperamide, quinidine, talinolol, vinblastine

[0585] BCRP substrates: daidzein, dantrolene, estrone-3-sulfate, prazosin, sulfasalazine

[0586] MATE1 substrates: Metformin, tetraethylammonium (TEA), cimetidine, procainamide

[0587] Results. Twenty patients (19 NSCLC, 1 pancreatic cancer) have received compound 1 orally at dose levels of 25-100 mg once daily. Patients have received a median of 3 (range: 1-9) prior anticancer therapies, including any ROS1 TKI (100%); investigational ROS1 TKI (85%, including 55% lorlatinib, 40% lopatinib); ≥2 ROS1 TKIs (75%); any chemotherapy (80%); ≥2 lines of chemotherapy (50%). At baseline, 55% had CNS metastases and 45% had ROS1 G2032R. No DLTs, dose reductions, or drug-related treatment discontinuations were reported. All treatment-related adverse events (TRAEs) were grade 1. The only TRAE in >1 patient was nausea (n=2). Compound 1 PK analysis showed that exposure was dose-dependent. In 12 efficacy-evaluable ROS1+NSCLC patients treated with 25-75mg QD, 6 confirmed partial responses (PRs) were achieved. Shrinkage or remission of intracranial metastases was observed; no patient experienced intracranial progression. PRs were achieved in the majority (n=5 / 7) of ROS1G2032R mutant cancers, including tumors pretreated with lorlatinib or lopatinib. Circulating tumor DNA analysis showed a reduction in ROS1 variant allele frequency. RP2D has not yet been determined, and dose escalation continues.

[0588] Based on data from September 13, 2022, for patients treated through September 1, 2022, the Phase 1 patient population had heavily pretreated ROS1-positive solid tumors ( Fig. 20 The safety profile was favorable and consistent with the highly ROS1-selective, TRK-sparing Compound 1. No DLTs were observed, no treatment-related SAEs, no AEs leading to dose reduction or discontinuation, and no treatment-related dizziness ( Fig.21 ).

[0589] Based on data through September 13, 2022, for patients treated through September 1, 2022, exposure to Compound 1 in clinical trials exceeded target levels that provided resolution in preclinical models ( Fig.19 Compound 1 exhibited favorable pharmacokinetics, including low intra-cohort patient PK variability, increasing exposure with increasing dose levels, and a half-life of approximately 20 hours supporting QD dosing.

[0590] For example, a 54-year-old patient with metastatic lung adenocarcinoma who responded to two cycles of carboplatin, paclitaxel, bevacizumab, and atezolizumab tested positive for ROS1 fusion on tumor tissue by fluorescence in situ hybridization. The patient received crizotinib for two months, complicated by pneumonia, and subsequently received lorlatinib with an initial partial response (PR). Intrathoracic disease progression occurred 32 months later. She then received two cycles of carboplatin and pemetrexed and continued on lorlatinib with stable disease. Next-generation sequencing of lorlatinib-resistant lung nodules showed CD74-ROS1 fusion and ROS1 G2032R. The patient started treatment with compound 1 at the lowest dose level of 25 mg per day. A clinical response was achieved within 2 weeks of treatment, and the patient's dyspnea improved significantly. Imaging 4 weeks later showed a PR with a reduction in multiple dual lung metastases (31% reduction by RECIST 1.1). As permitted by the protocol, her Compound 1 dose was escalated to 75 mg daily, and she continued treatment for more than 5 months with ongoing confirmed PRs (reduction of 60 by RECIST 1.1; Fig. 8A ) without neurotoxicity. The unbound plasma concentration of compound 1 within 24 hours on day 15 of cycle 1 is shown in Fig.18A .

[0591] In addition to extracranial antitumor activity, intracranial activity was observed in another patient with metastatic NSCLC harboring EZR-ROS1 G2032R. A 65-year-old patient with stage IV lung adenocarcinoma and multiple brain metastases had EZR-ROS1 identified by ctDNA testing. Entrectinib treatment was initiated with an initial response, but the patient’s disease progressed 9 months later. Biopsy of growing liver lesions confirmed the known EZR-ROS1 fusion and identified the ROS1 G2032R mutation. She was switched to a clinical trial of entrectinib. The patient rapidly requested a switch in systemic therapy due to disease progression and was started on a combination of carboplatin, pemetrexed, bevacizumab, and entrectinib. Unfortunately, mixed responses, growing brain metastases, and new liver metastases were observed. Repeat ctDNA testing again detected ROS1 G2032R. Compound 1 treatment was initiated at 50 mg daily. At week 4, regression of the right occipital lobe metastasis was observed, in addition to reduction of several metastases in the liver and lung (38% reduction by RECIST 1.1). She continued treatment for approximately 5 months with a confirmed PR and further disease regression in the brain and liver (58% reduction by RECIST 1.1; Figure 8B ). No neurological toxicity was reported. The corresponding ctDNA analysis is shown in Fig.17 , and the unbound plasma concentration of compound 1 over a 24-hour period on day 15 of cycle 1 is shown in Fig.18B .

[0592] The antitumor activity of compound 1 extends to the absence of known ROS1 resistance mutations. A 75-year-old patient with metastatic EZR-ROS1 fusion-positive lung adenocarcinoma was initially treated with alectinib by the referring physician. As expected, major progression was observed, and treatment was switched to crizotinib. Clinical and radiographic responses were maintained until 39 months when bilateral growing lung nodules were noted on imaging. ctDNA analysis did not reveal any ROS1 kinase domain mutations. Compound 1 treatment was initiated at 25 mg daily. Treatment was well tolerated with no evidence of dizziness, orthostatic syncope, or paresthesias. A PR was observed at week 4 (33% reduction by RECIST 1.1); this PR was confirmed and maintained with continued treatment for more than 7 months (48% reduction by RECIST 1.1; Figure 8C The unbound plasma concentration of compound 1 during the 24-hour period on day 15 of cycle 1 is shown in Fig.18C .

[0593] Compound 1 was well tolerated and had favorable pharmacokinetics up to 125 mg daily. Activity was demonstrated in heavily pretreated patients (70% of whom had received ≥2 ROS1 TKIs plus chemotherapy), including patients with brain metastases and the G2032R mutation.

[0594] Whole blood samples were collected at the clinical trial research center to STRECK cell-free DNA The tubes were shipped at ambient temperature to a central laboratory for plasma generation and stored at -80° C. The plasma samples were then analyzed to determine genomic alternations in circulating tumor DNA (ctDNA) via a hybridization capture-based next-generation sequencing approach.

[0595] ctDNA analysis showed that compound 1 potently suppressed ROS1 variant allele frequency (VAF) at all dose levels tested compared to baseline (Cycle 1 Day 1 "C1D1"). At 25 mg, compound 1 treatment resulted in complete clearance of EZR-ROS1 and ROS1 G2032R variants in ctDNA. At higher dose levels, complete clearance of ROS1 fusions and G2032R was also observed. In summary, ctDNA data indicate that compound 1 is active against ROS1 fusions and ROS1 G2032R solvent front mutations at all dose levels tested, supporting the on-target activity of this compound in patients ( Fig.11 ).like Fig.11 As used in, C1D15 represents cycle 1 day 15, and C3D1 represents cycle 3 day 1.

[0596] As of the data cutoff point of September 19, 2022, preliminary clinical pharmacokinetic analyses are summarized in the table below:

[0597]

[0598]

[0599] N = number of patients in treatment group, n = number of patients for whom PK parameters were determined.

[0600] When n=1, a single value is reported, and when n≥2, a range is reported.

[0601] Except t 1 / 2 , R ac and T max In addition, the geometric means (geometric %CV) of all parameters are shown.

[0602] t 1 / 2 and R ac = Arithmetic mean ± SD, T max =Median (range)

[0603] The human free fraction of Compound 1 was calculated to be equal to 20.6% of the total plasma concentration.

[0604] As used herein, C maxrepresents the maximum plasma concentration; C max_DN represents the dose-normalized C max ; AUC 最后 AUC is the area under the plasma concentration-time curve from time zero to the last measurable concentration. 最后_DN represents the dose-normalized AUC 最后 ; AUC 0-24 AUC represents the area under the curve from time 0 to 24. tau AUC represents the area under the curve during the dosing interval; tau_DN represents the dose-normalized AUC tau ; AUC inf Represents the area under the curve from time 0 to infinity; AUC inf_DN represents the dose-normalized AUC inf ; CL / F represents oral clearance; V z / F represents the volume of distribution (V z / F); t 1 / 2 represents half-life; C min It represents the minimum plasma concentration of the drug achieved during the dosing interval.

[0605] Additionally, based on a data cutoff date of September 8, 2022 for patients treated until September 1, 2022, a preliminary efficacy analysis based on radiographic assessments indicated that Compound 1 induced tumor responses in an overly pretreated patient population (see table below). As used herein: CNS means central nervous system; PD means progressive disease; PR means confirmed partial response; SD means stable disease; TKI means tyrosine kinase inhibitor.

[0606]

[0607] a Includes 2 ongoing partial reactions awaiting confirmation.

[0608] b Includes 1 ongoing partial reaction awaiting confirmation.

[0609] c Patients discontinued treatment due to symptomatic worsening without post-baseline radiographic assessment

[0610] Based on a data cutoff of September 13, 2022, for patients treated through September 1, 2022, the primary efficacy analysis demonstrated radiographic tumor regressions across all dose levels of Compound 1 (see Fig.12 Two patients (25 mg QD and 125 mg QD dose cohorts, both with prior therapy consisting of crizotinib, lorlatinib, and chemotherapy) are not shown due to incomplete or missing post-baseline tumor assessments in the setting of PD and symptomatic progression.

[0611] Based on data as of September 13, 2022, for NSCLC patients treated to September 1 for response evaluable, Compound 1 provided sustained duration of treatment. The median (range) duration of treatment was approximately 3.6 (1.0, 8.3+) months. No treatment was discontinued due to toxicity. 76% (16 / 21) of response evaluable patients continued treatment. The median time to response was approximately 3.6 (range 3.1, 4.6) weeks (see Fig.13 ).

[0612] Based on data as of September 13, 2022, for NSCLC patients who were evaluable for response as of September 1, 2022, Compound 1 was found to induce rapid responses in TKI-resistant patients (see Fig.14 ). For the subgroup of patients with known ROS1 G2032R resistance mutations, the ORR was 78% (7 / 9), and 100% (9 / 9) were found to have tumor shrinkage. In central ctDNA analysis, complete clearance of G2032R was detected in all 7 patients with G2032R. One patient with ROS1 D2033N with an ongoing PR (-40%) is pending confirmation.

[0613] Based on data from patients treated until September 1, 2022 as of September 13, 2022, compound 1 was found to induce a response in intracranial lesions. Intracranial PR in 3 / 3 patients with measurable (>10mm) CNS metastases (one patient with an ongoing intracranial PR is pending confirmation). An ORR of 73% (8 / 11) was observed in patients with any CNS disease whose response could be evaluated. No CNS progression was observed in any of the 35 treated patients.

[0614] An intracranial response was observed in a 65-year-old female with CD74-ROS1 fusion NSCLC who had previously been treated with chemotherapy, crizotinib, and lorlatinib and had CNS progression and no known ROS1 resistance mutations (see Fig.15 ). The patient continued Compound 1 treatment (100 mg QD) at 3.2 months with an ongoing response.

[0615] Another case study of compound 1 treatment provided intracranial and extracranial activity in TKI-refractory ROS1 G2032R+NSCLC in a patient diagnosed with EZR-ROS1 fusion NSCLC. The patient had previously been treated with entrectinib (disease progression and ROS1G2032R identified) and lopatinib (rapid disease progression observed). The patient had also previously been treated with platinum-based chemotherapy + bevacizumab + entrectinib (disease progression observed). Based on data as of September 13, 2022, compound 1 (50 mg) was observed to produce a PR after 4 weeks (38% reduction per RECIST 1.1), including shrinkage of right occipital lobe metastases and reduction of several liver / lung metastases. Further disease regression was observed at 16 weeks (-58%), including near-complete regression of brain lesions. The treatment was well tolerated with no dizziness, orthostatic syncope, and paresthesia. The patient continues to receive compound 1 and has an ongoing response at 5.3 months. Radiological images are shown in Fig.16 .

[0616] Example 6: Phase 1 Clinical Study

[0617] A Phase 1 study was conducted to investigate the food effect, potential impact of pH shifts, and drug-drug interaction potential of Compound 1 in healthy subjects. 28 healthy subjects were planned (14 subjects in Part A and 14 subjects in Part B).

[0618] Primary objectives: Part A: To determine the effect of food on the single oral PK of Compound 1 in healthy subjects; To determine the effect of multiple oral administrations of the PPI lansoprazole (mutagenic drug molecule) on the single oral PK of Compound 1 (mutagenic drug molecule) in healthy subjects. Part B: To determine the effect of multiple oral administrations of Compound 1 (mutagenic drug molecule) on the single oral PK of midazolam (mutagenic drug molecule) in healthy subjects.

[0619] Secondary objectives: Part A and Part B: Further evaluate the safety and tolerability of compound 1 in healthy subjects.

[0620] Primary endpoint: Part A: Pharmacokinetic parameters of compound 1 determined using non-compartmental analysis (C max , T max , AUC 0-24 , AUC 最后 , AUC inf ,t 1 / 2 Part B: Pharmacokinetic parameters of midazolam determined using non-compartmental analysis (C max , AUC 0-24 , AUC inf ).

[0621] Secondary endpoints: Part A and Part B: incidence and severity of TEAEs, changes in ECG parameters, and changes in clinically relevant laboratory parameters.

[0622] Study Design: The study was conducted in two parts ( Figure 7 ).

[0623] This is a phase 1, open-label study. Parts A and B are conducted in parallel:

[0624] Part A - Food Effects, Proton Pump Inhibitor (PPI) Drug Interactions: Part A used a randomized, balanced, crossover study design. Fourteen (14) healthy subjects were studied in a single cohort as follows:

[0625] ● Subjects stayed in the CRU from Day -1 to Day 19.

[0626] In the food effect portion of the study, each subject received a single oral dose of Compound 1 (25 mg) in the fed (high-fat breakfast) or fasted state on Days 1 and 6. Half of the subjects (n=7) were randomized on Day 1 to receive Compound 1 under fed conditions and then crossed over on Day 6 to receive Compound 1 under fasted conditions, and half of the subjects (n=7) were randomized on Day 1 to receive Compound 1 under fasted conditions and then crossed over on Day 6 to receive Compound 1 under fed conditions.

[0627] In the PPI drug interaction portion of the study, subjects received lansoprazole (30 mg Qd) from Days 11 to 17, followed by a third dose of Compound 1 (25 mg) under fasting conditions on Day 17. Post-dose PK sampling continued to 48 hours (morning of Day 19).

[0628] • All subjects had a follow-up visit (clinic visit or phone call) 5-7 days after the last dose of Compound 1 for safety review.

[0629] Part B - Compound 1 Mutagenic Drug-Drug Interactions: Part B Sampling Fixed Sequence, Multiple Oral Dosing Study Design for Compound 1. Fourteen (14) healthy subjects were studied in a single group:

[0630] • Subjects reside in the CRU from Day -1 to the morning of Day 10.

[0631] • Midazolam (2 mg) was administered and PK time points were collected over 24 hours on Days 1 and 9. Compound 1 (25 mg QD) was administered under fed conditions on Days 2 to 9.

[0632] • All subjects had a follow-up visit (clinic visit or phone call) 5-7 days after the last dose of Compound 1 for safety review.

[0633] Duration of treatment: Subjects in Part A received treatment for 17 days: three single daily doses of Compound 1 (on study days 1, 6, and 17), and lansoprazole from day 11 to day 17. Subjects in Part B received treatment for 9 days: once daily dosing of Compound 1 (study days 2-9) and midazolam on days 1 and 9.

[0634] Inclusion Criteria: Subjects must meet all of the following criteria to be eligible for inclusion in the study:

[0635] 1. Healthy men and women.

[0636] 2. Age 18-55 (inclusive).

[0637] 3. BMI 18 to 32 kg / m2 (inclusive).

[0638] 4. Any ethnic origin.

[0639] 5. Non-smoker.

[0640] 6. Female and male subjects of childbearing potential must be surgically sterilized or willing to refrain from sexual activity or use effective contraceptive methods from the time of signing the ICF until 30 days (female) and 90 days (male) after the last administration of study drug.

[0641] 7. Male subjects must agree not to donate sperm throughout the study period and within 90 days after the last dose of Compound 1.

[0642] 8. Provide written informed consent and be willing and able to comply with the requirements of the study protocol, including but not limited to outlining study restrictions regarding food consumption, alcohol, caffeine, tobacco / nicotine, recreational drug use, exercise, and donation of sperm (for male subjects) and / or blood, plasma, or platelets.

[0643] Exclusion criteria: Subjects meeting any of the following criteria were excluded from the study:

[0644] 1. History of significant hypersensitivity, intolerance, or allergy to any drug compound, food, or other substance, or any contraindication to lansoprazole (Part A) or midazolam (Part B), unless approved by the Investigator (or designee).

[0645] 2. Hospitalization within 2 months before admission to CRU and any type of major surgical operation within 6 months.

[0646] 3. Have a history of the following: chronic or recurrent infection considered a risk by the investigator; severe or life-threatening infection within 6 months before admission to the CRU; or any suspected current active bacterial or viral infection that requires antibiotics, antivirals or other active treatment or that the investigator considers to be a risk to study participation.

[0647] 4. Have known active tuberculosis, positive HIV test, positive hepatitis B combination test, and / or antibodies to hepatitis C. Subjects with hepatitis B results consistent with prior immunization may be included.

[0648] 5. The subject's QTcF is consistently >450 ms (male) or >470 ms (female). If the value is out of range, the ECG can be repeated twice and the average of the 3 results used to determine whether to exclude the subject. The subject has a history of long QT syndrome or torsades de pointes.

[0649] 6. Subjects with moderate to severe cognitive impairment or psychiatric disorders that compromise the subject's ability to comply with study requirements.

[0650] 7. Subjects who have received the following:

[0651] a. Received medication (such as St. John's wort) or vaccines (including COVID vaccine and / or booster vaccine) within 30 days prior to admission to the CRU.

[0652] b. Received OTC medications, multivitamins, or homeopathic preparations within 7 days prior to admission to the CRU.

[0653] c. Received any other medication (including prescription medications, oral contraceptives or other hormonal contraceptive treatments) within 14 days prior to admission to the CRU.

[0654] d. Note: Permitted exceptions include occasional use of acetaminophen up to 2 g per day for no more than 3 consecutive days and other medications when ≥ 5 half-lives have passed since the last dose (upon approval by the investigator).

[0655] 8. Self-reported drug or alcohol abuse and / or dependence within 1 year before CRU admission, including subjects who were in a drug or alcohol rehabilitation program within 1 year before CRU admission.

[0656] 9. Active alcohol consumption exceeds 14 units per week or drinking within 48 hours before admission to CRU.

[0657] 10. Use of any tobacco or nicotine-containing substances at least 3 months before admission to the CRU until the last discharge from the CRU.

[0658] 11. Positive test result for drugs of abuse, alcohol, or cotinine during screening or admission to the CRU.

[0659] 12. Consumption of caffeine within 48 hours prior to admission to the CRU.

[0660] 13. Consumption of food and beverages containing grapefruit within 7 days prior to admission to the CRU.

[0661] 14. Donated or lost >500mL of whole blood within 30 days before admission to CRU; donated plasma within 2 weeks before admission to CRU; or donated platelets within 6 weeks before admission to CRU.

[0662] 15. Participated in strenuous exercise within 72 hours before admission to the CRU, and agreed not to start a new exercise program or participate in any unusually strenuous physical activity during the entire study period.

[0663] 16. Subject is pregnant or breastfeeding. Female subjects must have a negative serum pregnancy test at screening and at the time of CRU check-in. A positive BHcG test is exclusionary unless consistent with postmenopausal status at the time of quantitative testing.

[0664] 17. Actively enroll in another clinical study involving investigational drugs.

[0665] 18. Any of the following laboratory abnormalities at screening or at CRU admission examination (repeat testing may be performed to confirm results):

[0666] a. Total bilirubin >1.5×ULN (Note: Patients with known or suspected Gilbert's syndrome are excluded.)

[0667] b. AST, ALT or alkaline phosphatase>2×ULN

[0668] 19. Previous gastrointestinal surgery (excluding uncomplicated appendectomy, uncomplicated cholecystectomy or hernia repair or cosmetic surgery such as abdominoplasty), disease, or other illness that may affect the oral absorption, distribution, metabolism, or excretion of the study drug as judged by the investigator.

[0669] 20. Any other medical illness or laboratory abnormality or other cause that, in the opinion of the investigator, may pose a risk to the study subjects or confound the ability to judge the study results.

[0670] Study Drug, Dosage and Route of Administration: Compound 1 is supplied as 1 strength of tablets for oral administration: 25 mg (oval) supplied in 32-tablet high-density polyethylene bottles with induction seal / child-resistant caps.

[0671] Study Medications, Doses, and Routes of Administration: Subjects should refrain from taking any medications during the study. The Investigator may, at his / her discretion, administer any medication deemed necessary for the safety and well-being of the subject, taking into account the following guidelines. Administration of all medications (including investigational products) must be listed on the appropriate CRF page.

[0672] Medications to be avoided or taken with caution: The following medications, supplements, and foods must be avoided or taken with caution (illustrative agents are provided in Example 4):

[0673] ●Drugs that are strong inhibitors of CYP3A4.

[0674] ● Herbal supplements and foods that are strong inhibitors of CYP3A4, including but not limited to grapefruit and grapefruit juice.

[0675] ●Drugs that are known substrates of CYP3A4.

[0676] ●Drugs that are CYP2C19 substrates with a narrow therapeutic index.

[0677] • Drugs that are known substrates of P-gp / MDR1 and BCRP / ABCG2.

[0678] ●Drugs that are known substrates of the MATE1 transporter (including but not limited to metformin, cimetidine, procainamide, etc.).

[0679] ● Gastric acid suppressants, such as PPIs. Co-administration of gastric acid suppressants with Compound 1 should be avoided due to the potential for drug-drug interactions between Compound 1 and gastric acid suppressants.

[0680] ●Warfarin and low molecular weight heparin.

[0681] Prohibited drugs and therapies: The following drugs and therapies must be excluded during the study:

[0682] Any other investigational therapy

[0683] ●Drugs that are strong inducers of CYP3A4

[0684] Statistical methods:

[0685] Analysis Set: The PK analysis set was defined as all subjects who received study treatment and had at least one evaluable plasma PK sample. PK was examined in the following subject subsets: Part A: PK of Compound 1 in fed subjects (n=14); PK of Compound 1 in fasted subjects (n=14); PK of Compound 1 in subjects receiving lansoprazole (PPI) to steady state (n=14). Part B: PK of midazolam (n=14); PK of midazolam in subjects receiving Compound 1 to steady state (n=14). The safety analysis set consisted of subjects who received at least 1 dose of Compound 1.

[0686] Pharmacokinetic Analysis: PK parameters were determined using Phoenix 64 (Certara, version 8.2) and non-compartmental methods. All PK concentration and parameter analyses were performed on the PK analysis set. max The terminal elimination half-life was estimated using at least three time points after the time point of . For graphs and derivations of PK parameters, values ​​below the limit of quantitation occurring before the first measurable concentration were set to zero. Missing values ​​occurring after the first measurable concentration were set to missing. Actual sampling times were used. Actual post-dose sampling times were expressed in hours and rounded to three significant figures.

[0687] Derived PK parameter: C max : Maximum observed plasma concentration. T max : Reach C max Time. AUC 0-24 : The area under the plasma concentration-time curve from the time of administration to 24 hours after administration. AUC 最后 : The area under the plasma concentration-time curve from the time of administration to the last measurable non-zero concentration. AUC inf : The area under the plasma concentration-time curve from the time of administration to infinity. 1 / 2 : Terminal elimination half-life. CL / F: Apparent total clearance of drug from plasma after oral administration.

[0688] Summary of PK parameters: The derived PK parameters were summarized using the number of observations (n) and the following statistics: mean, median, standard deviation, coefficient of variation, minimum, maximum, geometric mean.

[0689] Comparison of PK parameters:

[0690] Part A: Food Effect: The C of compound 1 was calculated in subjects in the fed and fasted states. max , AUC 最后 and AUC inf Data are presented as the geometric mean ratio of the fed to the fasted state.

[0691] Part A: PPI drug interactions: The C of compound 1 was calculated in fasting subjects and in fasting subjects receiving lansoprazole (PPI) (at steady state). max , AUC 最后 and AUC inf The data are presented as the geometric mean ratio of compound 1 alone to compound 1 + lansoprazole.

[0692] Part B: Compound 1 mutator DDIL: The C of midazolam was calculated for subjects before receiving Compound 1 and for subjects after administration of Compound 1 (at steady state). max , AUC 0-24 , AUC inf Data are presented as the geometric mean ratio of midazolam alone to midazolam + compound 1.

[0693] Other derived PK parameters may be evaluated and included in the data set as needed or deemed useful in the analysis.

[0694] Preliminary analysis of Part A study:

[0695] A preliminary analysis of data through September 21, 2022 found that AUC 0-inf The geometric mean ratio of is about 1.12; AUC 0-48 The geometric mean ratio of C is about 1.08; max The geometric mean ratio of the subjects in the fed state and the subjects in the fasted state was calculated as the value (e.g., C max , AUC 0-48 and AUC inf ). Preliminary results indicate that compound 1 can be taken with or without food.

[0696] A preliminary analysis of data through September 21, 2022 found that AUC 0-inf The geometric mean ratio of 0-48 The geometric mean ratio of C is about 0.92; max The geometric mean ratio of the subjects receiving Compound 1 and lansoprazole and the subjects receiving Compound 1 alone was calculated as the value (e.g., C max , AUC 0-48 and AUC inf ). Preliminary results indicate that compound 1 can be taken simultaneously with PPIs.

[0697] Preliminary analysis of Part B study:

[0698] A preliminary analysis of data through September 21, 2022 found that AUC 0-inf The geometric mean ratio of is about 1.12; AUC 0-24 The geometric mean ratio of C is about 1.11; max The geometric mean ratio of C was approximately 1.20. For subjects receiving midazolam alone and subjects receiving both midazolam and Compound 1, the geometric mean ratio was calculated as the value (e.g., C max , AUC 0-48 and AUC inf ). Preliminary results indicate that compound 1 should not be taken simultaneously with sensitive CYP3A4 substrates.

[0699] Example 7: Cell viability assay

[0700] Cell culture: All cells were maintained at 37°C with 5% CO2. Ba / F3 cells were provided by RIKEN BRC through the National Bio-Resource Project of MEXT, Japan. Ba / F3 cells were maintained in RPMI-1640 + 10% FBS. Genes encoding human CD74-ROS1 (containing wild-type kinase domain or containing G2032R, D2033N, L2026M or S1986F mutations), TPM3-TRKA, ETV6-TRKB, ETV6-TRKC or TRKB (full length) were synthesized, cloned into a retroviral vector with a puromycin resistance marker, and packaged into retroviral particles. Ba / F3 cells were infected with the virus. Stable cell lines were selected by interleukin (IL)-3 withdrawal and puromycin for at least 7 days. Polyclonal cultures were used directly in the assay, or monoclonal cultures were established by limiting dilution before use in the assay. Successful transformants were confirmed by Sanger sequencing and Western blotting. All cells were confirmed to express the desired protein in full. Ba / F3CD74-ROS1 contained a small C-terminal truncation and Ba / F3 ETV6-TRKC contained a small C-terminal frameshift. Both were confirmed to have no discernible effect on inhibitor activity by orthogonal assays. HCC78 and A549 were obtained from Pharmaron and cultured in 1640 medium + 10% FBS or F12K medium + 10% FBS, respectively.

[0701] Alternatively, Ba / F3 cells expressing the following genes were prepared respectively. ROS1 fusion mutants were prepared using site-directed mutagenesis (Agilent, New England Biolabs). Biotool DNA transfection reagent was used to transfect platinum-E cells (Cell Biolabs, Inc) with pBABE CD74-ROS1, pMIG CEP85L-ROS1, pCX4 EZR-ROS1, pBABE GOPC (S)-ROS1, pCX4 GOPC (S)-ROS1, pBABE GOPC (L)-ROS1, pCX4 GOPC (L)-ROS1 or pMIG SLC34A2-ROS1 wild-type or mutant constructs to generate a non-replicative ecotropic retrovirus. Ba / F3 parental cells were infected with retrovirus. Cells were treated with puromycin to select cells stably expressing each fusion. Cells surviving in IL-3 withdrawal were used for in vitro assays. All transformed cell lines were sequenced to verify the presence of the desired mutation. Cells were harvested, pelleted, and purified using QuickExtract TM DNA was extracted using DNA extraction solution (Lucigen). ROS1 kinase and C-terminal domains were PCR amplified. Chromatograms were aligned using Benchling software to confirm the presence of the desired mutations and to verify that no unwanted mutations were introduced during viral transduction.

[0702] Cell viability assay: A549 or stable Ba / F3 cells were seeded into 384-well plates and test compounds were added in the form of a 3-fold dilution series in complete medium containing 10% FBS. Cell viability was measured using CellTiter-Glo reagent (Promega) after 72 h of incubation with inhibitors. Untreated cells served as negative controls (no inhibition of proliferation), while cells treated with high concentrations of the nonspecific kinase inhibitor staurosporine served as positive controls (complete inhibition of proliferation). IC was calculated from the percentage inhibition and log(inhibitor concentration) using 4-parameter logistic regression 50 .

[0703] Alternatively, all inhibitors were prepared as 1 mM stock solutions in DMSO. 25 μL complete medium was pre-seeded for each well of the plate using a Multidrop Combi reagent dispenser (Thermo Scientific). Inhibitors were distributed at 2 times the indicated concentration in 25 μL complete medium in each well of a 384-well plate using a D300 digital dispenser (Hewlett-Packard). Ba / F3 cell lines expressing wild-type or mutant ROS1 fusions were inoculated at 1,000 cells per well in a 25 μL volume using a Multidrop Combi reagent dispenser (Thermo Scientific). The plates were incubated for 72 hours. Viability was measured using an assay (Bimake) based on WST-8 [2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolyl monosodium salt] and read on a Biotek Synergy 2 plate reader. Each condition was measured in triplicate. Data were normalized using Microsoft Excel, and IC was calculated using nonlinear regression analysis in GraphPad Prism. 50 value.

[0704] Results: Six ROS1 TKIs (crizotinib, entrectinib, lorlatinib, taretinib, lopatinib, and compound 1) were analyzed for inhibition in a cell viability assay against one human cancer cell line and 18 engineered Ba / F3 cell lines covering six ROS1 fusion partners (SLC34A2, EZR, CD74, GOPC(L), GOPC(S), and CEP85L) and eight ROS1 mutant variants (wild type, G2032R, S1986F, F2004C, F2004V, L2026M, D2033N, and G2101A).

[0705] In seven cell lines expressing wild-type ROS1 fusions, all six TKIs had significant growth inhibitory activity despite varying levels of potency. Compound 1 and lorlatinib were the most potent (mean IC 50 =0.4 nM and 0.5 nM), which is 7-fold more potent than lopretinib (IC 50 =3.3 nM), which is >19-fold higher than crizotinib, entrectinib or tadalafil (IC 50 =9.7–30 nM).

[0706] Compound 1 was the only TKI to achieve single-digit nanomolar potency in six cell lines harboring ROS1 fusions and G2032R mutations (mean IC 50 =1.6 nM), which is >10-fold higher than lopretinib and tacrolimus (IC 50=18–44 nM), >60-fold higher than lorlatinib (IC 50 =98 nM), and >500-fold more potent than crizotinib and entrectinib (IC 50 =846nM–916nM).

[0707] The IC values ​​of compound 1 and several ROS1 inhibitors are shown below: 50 (nM) Results.

[0708]

[0709]

[0710] To measure the effect of the G2032R substitution on TKI sensitivity, the IC values ​​of six TKIs from wild-type ROS1 to ROS1G2032R were analyzed in three matched fusion pairs (CD74-ROS1, EZR-ROS1, and GOPC(L)-ROS1 cell lines, wild-type and G2032R positive). 50 The analysis showed two different groups of ROS1 TKIs based on their resistance to G2032R. For the first group, which included crizotinib, entrectinib, and lorlatinib, G2032R was deleterious, resulting in a mean IC of 1.34% compared with wild-type ROS1. 50 The second group, which included tarotik, lopretinib, and compound 1, was tolerable to G2032R, showing only modest IC compared to wild-type ROS1. 50 However, in contrast to lopretinib and taretinib, compound 1 provided a 4-fold change in ROS1 G2032R and maintained single-digit nanomolar potency due to its subnanomolar potency in inhibiting wild-type ROS1.

[0711] In addition to G2032R, other substitutions observed after disease progression on crizotinib and / or entrectinib treatment included S1986F, F2004C, F2004V, L2026M, D2033N, and G2101A. CD74-ROS1 or EZR-ROS1 fusion proteins carrying these mutations were expressed in Ba / F3 cells and tested against six TKIs in a cell viability assay. Compound 1 potently inhibited non-G2032R ROS1 mutants with IC 50 ≤1.5nM. IC 50 Mutation analysis showed that most non-G2032R mutations made ROS1 more resistant to crizotinib and entrectinib, but the degree of resistance was similar to that of G2032R (IC 50 Change = 24–110-fold) compared to a moderate (IC50 In contrast, non-G2032R mutations did not confer resistance to lorlatinib, taloratatinib, lopretinib, and compound 1 (IC 50 change = 0.01–1.3 fold).

[0712] The IC values ​​of compound 1 and several ROS1 inhibitors are shown below: 50 (nM) Results.

[0713]

[0714] Given the diversity of upstream ROS1 fusion partners, ROS1 TKIs were studied in Ba / F3 cells expressing CD74-ROS1, EZR-ROS1, GOPC(L)-ROS1, GOPC(S)-ROS1, CEP85L-ROS1, or SLC34A2-ROS1 fusions with either a wild-type kinase domain or with the G2032R mutation. Compound 1 showed potent activity against all ROS1 fusions evaluated (IC 50 <10nM).

[0715] Example 8: Anchorage-independent growth

[0716] Summary: Anoikis avoidance is a hallmark of cancer and potentially indicative of migratory or metastatic propensity. To investigate the effect of ROS1 inhibition on anoikis, an anchorage-independent colony formation assay was performed in NIH3T3 cells expressing CD74-ROS1 or EZR-ROS1 fusions.

[0717] Colony formation assay: Plates were pre-seeded with 0.8% agarose in complete medium containing DMSO or inhibitors (10, 100, or 1000 nM of crizotinib, entrectinib, lorlatinib, or compound 1). Each inhibitor was paired with its own DMSO condition as an accurate control. NIH3T3 cells expressing CD74-ROS1 or EZR-ROS1 wild-type or mutant fusions were seeded in 0.4% agarose in complete medium at a density of 2,000 cells per 0.5 mL of agarose containing the same concentration of DMSO or inhibitor as the bottom layer. The plates were incubated for 4 weeks, and 75 μL of complete medium with or without inhibitors was added to each well 3 times per week to prevent the agarose from drying out. The cells were then plated using a GelCount TM (Oxford Optronix) to read the plates. The colony counts were averaged by condition and normalized to the colony counts from the paired DMSO condition. Data analysis and visualization were performed using Microsoft Excel and GraphPad Prism.

[0718] Results: NIH3T3 cells transformed with ROS1 lost contact inhibition and formed colonies on soft agar. For NIH3T3 cells expressing wild-type ROS1 fusions, all ROS1 TKIs evaluated inhibited colony formation by >80% at <100 nM, with compound 1 and lorlatinib having the highest potency, followed by crizotinib and then entrectinib. This potency trend correlated with the degree of target engagement and signaling modulation measured by western blotting. In contrast, for NIH3T3 cells expressing ROS1 fusions containing G2032R, only compound 1 was able to inhibit colony formation by ≥90% at 100 nM, while other TKIs (crizotinib, entrectinib, and lorlatinib) had weaker effects at the same concentrations. This potency trend was confirmed by pathway analysis. Although tarazolitinib was not evaluated in colony formation assays, Western blot analysis indicated that Compound 1 was more potent than tarazolitinib in ROS1-driven NIH3T3 cells, consistent with the observations in Example 7.

[0719] Example 9: Cellular phosphorylation assay

[0720] Summary: The relative potency of compound 1 and other ROS1 inhibitors between TRK and ROS1 was evaluated.

[0721] Methods: For Ba / F3 TRKB cell phosphorylation assay, cells were seeded into 384-well plates and test compounds were added in the form of a 3-fold dilution series in complete medium + 10% FBS. Cells were stimulated with 100 ng / mL BDNF for 20 minutes. TRK phosphorylation was measured using phospho-TRKA (Tyr674 / 675) / phospho-TRKB (Tyr706 / Tyr707) AlphaLISA reagents (PerkinElmer #ALSU-PTRKAB). Untreated wells served as negative controls (no inhibition), while wells treated with high concentrations of the nonspecific kinase inhibitor staurosporine served as positive controls (complete inhibition). IC was calculated from the percentage inhibition and inhibitor concentration using 4-parameter logistic regression 50 .

[0722] NIH3T3 cells expressing EZR-ROS1 wild-type or mutant fusions were treated with the indicated concentrations of inhibitors for 3 h before harvesting. Cells were washed with PBS and harvested with cell lysis buffer supplemented with 0.25% deoxycholate, 0.05% SDS, and protease and phosphatase inhibitors. TMProtein concentration was determined by BCA protein assay (ThermoFisher Scientific).Lysates were extracted at 75°C for 10 min using Laemelli sample buffer supplemented with β-mercaptoethanol and run on 4-20% precast gradient Bis-tris gels (Invitrogen; ThermoFisher Scientific). Proteins were transferred to nitrocellulose membranes (Prometheus) and stained with phospho-ROS1 Y2274 (3078; 1:1000; Cell Signaling Technology), DYKDDDDK [Tag] (8H8L17; 1:1000; Invitrogen), phospho-SHP2 (A5278; 1:1000; Bimake), phospho-ERK1 / 2 (9101; 1:1000; Cell Signaling Technology), ERK2 (sc-1647; 1:1000; Santa Cruz), phospho-S6 (4858; 1:1000; Cell Signaling Technology), S6 (2216; 1:1000, Cell Signaling Technology), phospho-Akt (4060; 1:1000, Cell Signaling Technology), Akt (9272; 1:1000, Cell Signaling Technology). Signaling Technology) or actin (JLA-20; 1:5000; Developmental Studies Hybridoma Bank) were used for detection. Signals were detected using HRP-conjugated or IR-dye secondary antibodies on a BioRad ChemiDoc imaging station or LI-COR Odyssey imaging station, respectively.

[0723] Results: TRK-related neurotoxicity is most consistent with the known functions of the TRKB signaling pathway. Therefore, an assay was developed that used Ba / F3 cells stably expressing full-length TRKB. Stimulation of this cell line with brain-derived neuronal factor (BDNF) promoted autophosphorylation (pTRKB). Consistent with our biochemical and cell viability assays, compound 1 only weakly inhibited cellular TRKB phosphorylation (IC 50 =850 nM), providing wide selectivity windows (670-fold and 240-fold) for wild-type ROS1 and ROS1G2032R over pTRKB that were not achieved with any other ROS1 TKI tested.

[0724] The IC values ​​of compound 1 and several ROS1 inhibitors are shown below: 50 (nM) measurements.

[0725]

[0726] Example 10: Preclinical intracranial activity

[0727] Methods: All procedures related to animal handling, management, and treatment in these studies were performed according to guidelines approved by Pharmaron's IACUC in accordance with AAALAC guidelines.

[0728] Compound 1 was formulated as a 1 mg / mL suspension in 20% hydroxypropyl-β-cyclodextrin (HP-β-CD) in deionized water. The compound was orally administered to male Wistar Han rats (n=3 each). After 1 hour, brain samples and plasma were collected and brain samples were homogenized in phosphate buffered saline (PBS). Brain and plasma samples were precipitated by acetonitrile and centrifuged (4700 rpm, 15 minutes). The drug concentration in the supernatant was quantified by LC / MS / MS. The unbound fraction was determined using rapid equilibrium dialysis. K p,uu It was calculated as the ratio between the unbound drug concentration in brain and the unbound drug concentration in plasma.

[0729] Ba / F3 CD74-ROS1 G2032R cells were transduced with viral particles containing the firefly luciferase gene and a neomycin resistance marker. Infected cells were selected with neomycin, and monoclonal cultures were established by limiting dilution. Successful transformants were confirmed by Sanger sequencing and bioluminescence. For in vivo studies, 1 × 10 5 Ba / F3 CD74-ROS1 G2032R luciferase cells. After 5 days, mice were randomized into 3 groups based on the average bioluminescent signal, with n=7-10 mice per group, and received vehicle or compound 1, 2 mg / kg, PO, BID. Bioluminescence and body weight were measured at regular intervals until the end of the study (61 days after the start of treatment) or until the animals met the criteria for euthanasia.

[0730] Results: After a single oral dose of 10 mg / kg in Wistar-Han rats, Compound 1 showed a favorable K of 0.16 measured at 1 hour. p,uu(Unbound brain-plasma distribution). To evaluate intracranial antitumor activity, a Ba / F3CD74-ROS1 G2032R luciferase model was generated, which allows monitoring of brain tumor burden by bioluminescent imaging of living animals. Ba / F3CD74-ROS1 G2032R luciferase cells were injected intracranially into mice, and rapid tumor growth was observed. All mice treated with vehicle lost weight and died of disease at 19 days, with a median overall survival (mOS) of 16.5 days. In contrast, 2 mg / kg of compound 1 inhibited intracranial tumor growth, with no significant changes in body weight, and all mice survived until the end of the study (mOS>61 days), equivalent to a >4-fold extension of survival compared to vehicle treatment. These results indicate that compound 1 has strong intracranial antitumor activity.

[0731] Example 11: In vivo pharmacology

[0732] Methods: Mice bearing PDX tumors with SDC4-ROS1 were treated with a single dose (PO) of vehicle (20% HP-β-CD) or compound 1, followed by tumor collection and Western blot analysis at the indicated time points (n=3). Anti-β2-microglobulin (β2m) is a loading control that does not recognize mouse β2-microglobulin. pERK, pAKT, and cleaved PARP are markers of MAPK signaling, PI3K signaling, and apoptosis, respectively.

[0733] Results: The results of this study are shown in Fig. 9 Direct in vivo inhibition of ROS1 fusion by compound 1 was supported by downstream inhibition of signaling pathways MAPK and PI3K, as measured by reduced levels of phospho-p44 / 42 MAPK ERK1 / 2 (pERK) and phospho-AKT (pAKT), respectively. These results indicate that compound 1 inhibits signaling through the MAPK and PI3K / AKT pathways and induces apoptosis in PDX tumors bearing SDC4-ROS1.

[0734] Example 12: Cell viability assay of LTK cell line

[0735] Generation of Ba / F3 stable cell lines: The gene encoding CLIP1-LTK was synthesized, cloned into the retroviral construct pMSCV-puro (Biovector), and packaged onto retroviral particles. Ba / F3 cells (RIKEN) were infected with the virus at a multiplicity of infection = 1 or 10 for 1 day. The infected cells were revived in medium (RPMI-1640, containing 10% fetal bovine serum and 1% streptomycin and penicillin) supplemented with mouse IL-3 (10 ng / mL) for 2 days, and stable cell lines were selected by IL-3 withdrawal and puromycin (0.8 μg / mL) for 7 days. Transformation of the desired gene was confirmed by Sanger sequencing and Western blotting.

[0736] Cell proliferation assay: Stable cells were plated at 1,000 cells / well (40 μL) in 384-well plates for 1 day. Test compounds (40 nL) were then added in a 3-fold dilution series using a TECAN EVO200 liquid handler and incubated for 72 hours. The plates were equilibrated at room temperature for 15 minutes, followed by the addition of 40 μL of CellTiter-Glo reagent (Promega). Luminescence was measured on a plate reader. The half-maximal inhibitory concentration (IC) was calculated from the percent inhibition and inhibitor concentration using a four-parameter logistic regression. 50 ).

[0737] Results: The IC of compound 1 for inhibiting Ba / F3 CLIP1-LTK cell line was determined 50 It is 9.57nM.

[0738] Example 13: Ba / F3 stable cell line with L2086F mutation

[0739] Biochemical kinase assay

[0740] ROS1 L2086F (amino acids 1881–2347) was custom made at SignalChem (Catalog # NP72-092G / Y4232-2). The kinase reaction was initiated by mixing ATP (1 mM) with the fluorescent phosphorylation substrate AQT0101 (15 μM, AssayQuant) and ROS1 L2086F (0.5 nM, SignalChem) in buffer (54 mM HEPES pH 7.5, 0.012% Brij-35, 0.52 mM EGTA, 1.2 mM DTT, 1% glycerol, 0.2 mg / mL BSA, 10 mM MgCl2). The plate was sealed and read at λ = 485 nm by SpectraMax Paradigm at 30°C, every 2 minutes for 120 minutes. The initial reaction rate (v) was calculated from the change in fluorescence intensity over time during the initial linear portion of the reaction. Finally, the apparent inhibition constant (K) was determined from regression ...

Claims

1. A method for treating a patient suffering from a solid tumor, comprising administering to the patient a therapeutically effective amount of Compound 1: or a stereoisomer thereof, or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof.

2. The method of claim 1, wherein the solid tumor is an advanced solid tumor.

3. The method of claim 2, wherein the advanced solid tumor is relapsed, refractory or resistant following the prior treatment with a tyrosine kinase inhibitor (TKI).

4. The method according to any one of claims 1 to 3, wherein the solid tumor is non-small cell lung cancer (NSCLC).

5. The method according to any one of claims 1 to 4, wherein the solid tumor is metastatic.

6. The method of claim 5, wherein the solid tumor is CNS metastatic.

7. The method according to any one of claims 1 to 6, wherein the solid tumor is ROS1 positive.

8. The method according to any one of claims 1 to 7, wherein the solid tumor has a ROS1 mutation.

9. The method of claim 8, wherein the ROS1 mutation is G2032R.

10. The method of any one of claims 1 to 9, wherein the solid tumor has a ROS1 fusion.

11. The method of any one of claims 1 to 6, wherein the solid tumor is ALK positive (eg, ALK fusion).

12. The method according to any one of claims 1 to 6, wherein the solid tumor is LTK positive.

13. The method of any one of claims 1 to 12, wherein the patient is not treated with tyrosine kinase inhibitor (TKI) therapy.

14. The method of any one of claims 1 to 12, wherein the patient has been treated with one prior TKI therapy.

15. The method of any one of claims 1 to 12, wherein the patient has been treated with at least one prior TKI therapy.

16. The method of any one of claims 1 to 12, wherein the patient has been treated with at least two prior TKI therapies.

17. The method according to any one of claims 13 to 16, wherein the TKI is a ROS1 TKI.

18. The method of any one of claims 14 to 17, wherein the prior TKI therapy is one or more selected from the group consisting of crizotinib, ceritinib, alectinib, brigatinib, lorlatinib, entrectinib, lopatinib, cabozantinib, foreitinib, mesartinib, taretinib, masitinib or ensartinib.

19. The method of any one of claims 1 to 18, wherein the patient has not been treated with prior platinum-based chemotherapy.

20. The method of any one of claims 1 to 18, wherein the patient has been treated with at most one prior platinum-based chemotherapy.

21. The method of any one of claims 1 to 18, wherein the patient has been treated with a prior platinum-based chemotherapy.

22. The method of any one of claims 1 to 21, wherein the patient has not been treated with immunotherapy.

23. The method of any one of claims 1 to 21, wherein the patient has been treated with immunotherapy.

24. The method of any one of claims 1 to 23, wherein the patient has not been treated with chemotherapy.

25. The method of any one of claims 1 to 23, wherein the patient has been treated with at least one prior line of chemotherapy.

26. The method of any one of claims 1 to 23, wherein the patient has been treated with at least two prior lines of chemotherapy.

27. The method of any one of claims 1 to 26, wherein the patient has been treated with at least three prior lines of anti-cancer therapy.

28. The method of any one of claims 1 to 27, wherein the patient has been treated with at least two prior lines of anticancer therapy selected from the group consisting of ROS1 TKI (e.g., investigational ROS1 TKI, crizotinib, lorlatinib, entrectinib, lopatinib, and taretinib) and chemotherapy.

29. The method of any one of claims 1 to 28, wherein the patient has been treated with at least one line of ROS1 TKI and one line of chemotherapy.

30. The method of any one of claims 1 to 29, wherein the patient has been treated with at least two lines of ROS1 TKI and one line of chemotherapy.

31. The method of any one of claims 1 to 30, wherein the patient has been treated with at least three lines of ROS1 TKI and one line of chemotherapy.

32. The method of any one of claims 1 to 31, wherein the patient has been treated with at least two lines of chemotherapy.

33. The method of any one of claims 1 to 32, wherein the patient has been treated with at least one line of ROS1 TKI and two lines of chemotherapy.

34. The method of any one of claims 1 to 33, wherein the patient has been treated with at least two lines of ROS1 TKI and two lines of chemotherapy.

35. The method of any one of claims 1 to 34, wherein the patient has been treated with at least three lines of ROS1 TKI and two lines of chemotherapy.

36. The method of any one of claims 17 to 35, wherein the ROS1 TKI is crizotinib.

37. The method of any one of claims 17 to 35, wherein the ROS1 TKI is entrectinib.

38. The method of any one of claims 17 to 35, wherein the ROS1 TKI is lorlatinib.

39. The method of any one of claims 17 to 35, wherein the ROS1 TKI is lopatinib.

40. The method of any one of claims 1 to 39, wherein the patient has been treated with lorlatinib and lopatinib.

41. The method of any one of claims 1 to 40, wherein the solid tumor is an advanced or metastatic ROS1 -positive solid tumor and the patient has been treated with at least one prior ROS1 TKI therapy.

42. The method of any one of claims 1 to 40, wherein the solid tumor is advanced or metastatic ROS1 -positive NSCLC and the patient has not been treated with TKI therapy and has been treated with at most one prior platinum-based chemotherapy with or without immunotherapy.

43. The method of any one of claims 1 to 40, wherein the solid tumor is advanced or metastatic ROS1 -positive NSCLC and the patient has been treated with one prior ROS1 TKI therapy and has not been treated with a prior platinum-based chemotherapy or immunotherapy.

44. The method of any one of claims 1 to 40, wherein the solid tumor is advanced or metastatic ROS1 -positive NSCLC and the patient has been treated with one prior ROS1 TKI therapy and one prior platinum-based chemotherapy with or without immunotherapy.

45. The method of any one of claims 1 to 40, wherein the solid tumor is advanced or metastatic ROS1 -positive NSCLC and the patient has been treated with at least two prior ROS1 TKI therapies and at most one prior platinum-based chemotherapy with or without immunotherapy.

46. ​​The method of any one of claims 1 to 45, wherein the solid tumor is an advanced or metastatic ROS1 -positive solid tumor and the patient has progressed on prior therapy.

47. The method of any one of claims 1 to 46, wherein Compound 1 is administered to the patient for one or more days.

48. The method of any one of claims 1 to 47, wherein Compound 1 is administered to the patient for at least one treatment cycle.

49. The method of claim 48, wherein a treatment cycle is at least 7 days.

50. The method of claim 48, wherein a treatment cycle is at least 14 days.

51. The method of claim 48, wherein a treatment cycle is at least 21 days.

52. The method of claim 48, wherein a treatment cycle is at least 28 days.

53. The method of any one of claims 1 to 52, wherein the patient does not experience a Grade 4 adverse event (e.g., TRAE) following administration of Compound 1.

54. The method of any one of claims 1 to 53, wherein the patient does not experience a Grade 3 adverse event (e.g., TRAE) following administration of Compound 1.

55. The method of any one of claims 1 to 54, wherein the patient does not experience a Grade 2 adverse event (e.g., TRAE) following administration of Compound 1.

56. The method of any one of claims 1 to 55, wherein the patient does not experience a Grade 1 adverse event (e.g., TRAE) following administration of Compound 1.

57. The method of any one of claims 1 to 55, wherein the patient experiences no more than a Grade 1 adverse event (e.g., TRAE) following administration of Compound 1.

58. The method of claim 57, wherein the Grade 1 adverse event is fatigue, myalgia, or nausea.

59. The method of any one of claims 1 to 58, wherein the patient experiences only nausea following administration of Compound 1.

60. The method of any one of claims 1 to 59, wherein the patient does not experience CNS adverse events following administration of Compound 1.

61. The method of claim 60, wherein the CNS adverse event is one or more selected from the group consisting of dizziness, ataxia, gait disturbance, paresthesia, weight gain, increased appetite, paresthesia, movement abnormalities, cognitive changes, speech effects (e.g., dysarthria, bradyspeech, or speech disorders), mood disorders (e.g., irritability, anxiety, depression, mood swings, personality changes, mood swings, affective disorders, aggression, restlessness, mood changes, depressed mood, euphoria, or mania), and cognitive disorders (e.g., memory impairment, cognitive disorders, amnesia, confusion, attention disorders, delirium, mental impairment, attention-deficit / hyperactivity disorder, dementia, sleep disorders, or reading disorders).

62. The method of any one of claims 1 to 61, wherein the patient does not experience adverse events of weight gain and / or glucose metabolism disorders.

63. The method of any one of claims 1 to 62, wherein the patient is a patient population.

64. The method of claim 63, wherein less than 30% of the patient population experiences a Grade 1 TRAE following administration of Compound 1.

65. The method of claim 63, wherein less than 20% of the patient population experiences a Grade 2 TRAE following administration of Compound 1.

66. The method of claim 63, wherein the patient population does not experience Grade 3 or 4 TRAEs following administration of Compound 1.

67. The method of any one of claims 1 to 66, wherein the patient has a complete response after one or more treatment cycles.

68. The method of any one of claims 1 to 66, wherein the patient has a partial response after one or more treatment cycles.

69. The method of any one of claims 1 to 68, wherein the patient achieves stable disease after one or more treatment cycles.

70. The method of any one of claims 1 to 69, wherein the patient has brain metastases.

71. The method of any one of claims 1 to 70, wherein the patient has brain metastases and does not experience intracranial progression after at least one treatment cycle.

72. The method of any one of claims 1 to 71, wherein the patient has at least about 5% to about 100% reduction in ROS1 allelic variants in circulating tumor DNA after at least one treatment cycle.

73. The method of any one of claims 1 to 72, wherein the patient has at least about a 35% reduction in ROS1 allelic variants in circulating tumor DNA after at least one treatment cycle.

74. The method of any one of claims 1 to 73, wherein the patient has at least about a 55% reduction in ROS1 allelic variants in circulating tumor DNA after at least one treatment cycle.

75. The method of any one of claims 1 to 74, wherein the patient has at least about a 65% reduction in ROS1 allelic variants in circulating tumor DNA after at least one treatment cycle.

76. The method of any one of claims 1 to 75, wherein the patient has at least about a 100% reduction in ROS1 allelic variants in circulating tumor DNA after at least one treatment cycle.

77. The method of any one of claims 1 to 76, wherein the patient has undetectable ROS1 allelic variant in circulating tumor DNA after at least one treatment cycle.

78. The method of claim 77, wherein the ROS1 allelic variant is G2032R.

79. The method of any one of claims 46 to 78, wherein the prior therapy is prior ROS1 TKI therapy.

80. The method of claim 79, wherein the ROS1 TKI is crizotinib, entrectinib, taloratatinib, lorlatinib, or lopatinib.

81. The method of any one of claims 1 to 80, wherein administration of Compound 1 provides an area under the curve (AUC) of the compound from 0 to 24 hours. 0-24 ) ranged from (80% to 125% of 500ng*h / mL) to (80% to 125% of 30000ng*h / mL).

82. The method of any one of claims 1 to 81, wherein administration of Compound 1 provides an area under the curve (AUC) of the compound from 0 to 24 hours. 0-24 ) is in the range of about 500 to about 30000 ng*h / mL.

83. The method of any one of claims 1 to 82, wherein the area under the curve (AUC) of the compound from 0 to 24 hours after about half a treatment cycle. 0-24 ) is in the range of about 1000 to 13000 ng*h / mL.

84. The method of any one of claims 1 to 83, wherein for each mg of Compound 1 administered, such administration provides an area under the curve (AUC 100) from 0 to 24 hours after administration of the compound. 0-24 ) ranged from (80% to 125% of 20 ng*h / mL) to (80% to 125% of 500 ng*h / mL).

85. The method of any one of claims 1 to 84, wherein for each mg of Compound 1 administered, such administration provides an area under the curve (AUC 100) from 0 to 24 hours after administration of the compound. 0-24 ) is in the range of from about 20 to about 500 ng*h / mL.

86. The method of any one of claims 1 to 85, wherein after about half a treatment cycle, such administration provides an area under the curve (AUC) from 0 to 24 hours after administration of the compound. 0-24 ) is in the range of from about 50 to about 200 ng*h / mL.

87. The method of any one of claims 1 to 86, wherein such administration provides an AUC of the compound of 最后,未结合 In the range of about 400 to about 7000 ng*h / mL.

88. The method of any one of claims 1 to 87, wherein for each mg of Compound 1 administered, such administration provides an AUC of the compound of 最后,未结合 In the range of from about 10 to about 80 ng*h / mL.

89. The method of any one of claims 1 to 88, wherein such administration provides an AUC of the compound of tau In the range of about 2000 to about 8000 ng*h / mL.

90. The method of any one of claims 1 to 89, wherein for each mg of Compound 1 administered, such administration provides an AUC of said compound of tau In the range of about 50 to about 150 ng*h / mL.

91. The method of any one of claims 1 to 90, wherein such administration provides an AUC of the compound of inf In the range of about 4000 to about 15000 ng*h / mL.

92. The method of any one of claims 1 to 91, wherein for each mg of Compound 1 administered, such administration provides an AUC of the compound of inf In the range of about 50 to about 250 ng*h / mL.

93. The method of any one of claims 1 to 92, wherein such administration provides a maximum plasma concentration (C max ) ranged from (80% to 125% of 100 ng / mL) to (80% to 125% of 1500 ng / mL).

94. according to the method described in any one of claims 1 to 93, wherein such administration provides the maximum plasma concentration (C max ) is in the range of about 100 to about 1500 ng / mL.

95. The method of any one of claims 1 to 94, wherein for each mg of Compound 1 administered, such administration provides a maximum plasma concentration (C max ) is in the range of about 2 to about 50 ng / mL.

96. The method of any one of claims 1 to 95, wherein such administration provides a minimum plasma concentration (C min ) is in the range of about 50 to about 350 ng / mL.

97. The method of any one of claims 1 to 96, wherein such administration provides an unbound maximum plasma concentration (C max,未结合 ) is in the range of about 30 ng / mL to about 400 ng / mL.

98. The method of any one of claims 1 to 97, wherein such administration provides a T of the compound after administration of max In the range of about 0.25h to about 5h or about 0.7h to about 1.3h.

99. The method of any one of claims 1 to 98, wherein such administration provides a t of the compound after administration of 1 / 2 In the range of about 2h to about 50h or about 10h to about 24h.

100. The method of any one of claims 1 to 99, wherein said administration provides a dose plasma concentration of said compound that is at least 70%, 80%, 90%, 95%, 97%, 98% or 99% higher than the IC of said compound against the ROS1 G2032R mutant during about 24 hours immediately following said administration. 50 At least 10%, 20%, 30%, 40% or 50% higher.

101. A method for reducing lesions in a subject having a ROS1-positive solid tumor, comprising: (i) obtaining a first radiological measurement of the size of the lesion; (ii) administering a pharmaceutically effective amount of Compound 1 once a day for one or more days; and (iii) obtaining a second radiological measurement of the size of the lesion; Wherein the second measurement is at most 100% of the first measurement.

102. The method of claim 101, wherein the second measurement is at most about 90% of the first measurement.

103. The method of claim 101, wherein the second measurement is at most about 80% of the first measurement.

104. The method of claim 101, wherein the second measurement is at most about 70% of the first measurement.

105. The method of claim 101, wherein the second measurement is at most about 60% of the first measurement.

106. The method of claim 101, wherein the second measurement is at most about 50% of the first measurement.

107. The method of claim 101, wherein the second measurement is about 0.01% to about 90% of the first measurement.

108. The method of claim 101, wherein the second measurement shows no detectable lesions.

109. The method of any one of claims 1 to 108, wherein the compound is administered once daily in an amount from about 5 mg to about 500 mg (by weight of Compound 1 free base).

110. The method of claim 109, wherein the compound is administered once daily in an amount from about 25 mg to about 250 mg (by weight of free base Compound 1).

111. The method of claim 109, wherein the compound is administered once daily in an amount from about 25 mg to about 200 mg (by weight of free base Compound 1).

112. The method of claim 109, wherein the compound is administered once daily in an amount of about 50 mg (by weight of free base Compound 1).

113. The method of claim 109, wherein the compound is administered once daily in an amount of about 75 mg (by weight of free base Compound 1).

114. The method of claim 109, wherein the compound is administered once daily in an amount of about 100 mg (by weight of free base Compound 1).

115. The method of claim 109, wherein the compound is administered once daily in an amount of about 125 mg (by weight of free base Compound 1).

116. The method of claim 109, wherein the compound is administered once daily in an amount of about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 75 mg, about 100 mg, about 125 mg, or about 150 mg (based on the weight of free base Compound 1).

117. The method of any one of claims 1 to 108, wherein the compound is administered twice daily (BID) in an amount from about 5 mg to about 500 mg (by weight of Compound 1 free base).

118. The method of claim 117, wherein the compound is administered twice daily (BID) in an amount from about 25 mg to about 250 mg (by weight of free base Compound 1).

119. The method of claim 117, wherein the compound is administered twice daily (BID) in an amount from about 25 mg to about 200 mg (by weight of free base Compound 1).

120. The method of claim 117, wherein the compound is administered twice daily (BID) in an amount from about 25 mg to about 150 mg (by weight of free base Compound 1).

121. The method of claim 117, wherein the compound is administered twice daily (BID) in an amount from about 25 mg to about 100 mg (by weight of free base Compound 1).

122. The method of claim 117, wherein the compound is administered twice a day (BID) in an amount of about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 75 mg, about 100 mg, about 125 mg, or about 150 mg (by weight of free base Compound 1).

123. The method of any one of claims 1 to 122, wherein the compound is administered orally.

124. The method of claim 123, wherein the compound is administered in the form of one or more tablets.

125. The method of claim 124, wherein the tablet has a unit dosage strength of about 5 mg to about 25 mg by weight of free base Compound 1.

126. The method of any one of claims 1 to 125, wherein the compound is administered to a patient in a fasting state.

127. The method of any one of claims 1 to 125, wherein the compound is administered to a patient in a fed state.

128. The method of any one of claims 1 to 127, wherein the patient is not taking any of a strong inhibitor of CYP3A4, a strong inducer of CYP3A4, a sensitive substrate of CYP3A4, a substrate of P-gp / multidrug resistance protein (MDR1), a substrate of BCRP / breast cancer resistance protein (ABCG2), a substrate of MATE1, or a gastric acid reducing agent.

129. The method of any one of claims 1 to 127, wherein the patient is taking any of a strong inhibitor of CYP3A4, a strong inducer of CYP3A4, a sensitive substrate of CYP3A4, a substrate of P-gp / multidrug resistance protein (MDR1), a substrate of BCRP / breast cancer resistance protein (ABCG2), a substrate of MATE1, or a gastric acid reducing agent.

130. The method of any one of claims 1 to 127, wherein the compound is administered in the absence of a sensitive substrate of CYP3A4.

131. The method of claim 130, wherein the CYP3A4 sensitive substrate comprises one or more of buspirone, everolimus, lovastatin, midazolam, simvastatin, triazolam, maraviroc, conivaptan, and darifenacin.

132. The method of any one of claims 1 to 131, wherein the patient does not experience one or more symptoms selected from the group consisting of cognitive impairment, mood disorders, sleep disturbances, dizziness, ataxia, and weight gain following administration of the compound.

133. The method of any one of claims 1 to 132, wherein the patient experiences a decrease in the level of one or more of pROS1, ROS1, pAKT, and pERK following administration of the compound.

134. The method of any one of claims 1 to 133, wherein the patient experiences a decrease in expression levels of one or more MAP kinase pathway genes in the tumor following administration of the compound.

135. The method of any one of claims 1 to 134, wherein the patient experiences a decrease in expression levels of one or more MAP kinase pathway genes in a solid tumor following administration of the compound.

136. The method of claim 134 or 135, wherein the one or more MAP kinase pathway genes are selected from the group consisting of DUSP6, FOS, IL1R1, and SPRY4.

137. The method of any one of claims 1 to 136, wherein the compound is Compound 1 free base.

138. The method of any one of claims 1 to 137, wherein the compound administered is a solid form of Compound 1, characterized in that An XRPD pattern comprising peaks at approximately 10.7, 15.0 and 21.2° 2Θ (± 0.2°).

139. The method of any one of claims 1 to 138, wherein the compound is administered in the form of a pharmaceutical composition comprising Compound 1, or a stereoisomer thereof, or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, and a diluent, a binder, a disintegrant, and a lubricant.

140. The method of claim 139, wherein the diluent is mannitol, the binder is a mixture of HPC and SMCC, the disintegrant is sodium starch glycolate (SSG), and the lubricant is magnesium stearate.

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