WO2025178977A1

WO2025178977A1 - Pet imaging with her2 inhibitor compounds

Info

Publication number: WO2025178977A1
Application number: PCT/US2025/016507
Authority: WO
Inventors: Zhongdong HUANG; Iriny Botrous; Joseph Michael DENNIS; Svitlana KULYK; Laurent Gomez; Shawn Eric WRIGHT; Wallace Deshaughn DERRICOTTE; Thomas Francis MILLER, III; Frederick Roy MANBY; Mary Anderson
Original assignee: Iambic Therapeutics Inc
Current assignee: Iambic Therapeutics Inc
Priority date: 2024-02-20
Filing date: 2025-02-19
Publication date: 2025-08-28
Anticipated expiration: 2026-08-20

Abstract

Provided herein are radiolabeled HER2 inhibitor compounds useful for PET imaging, and methods of selecting a subject for treating HER2 mediated disease, tracking tumor response to a HER2 mediated disease, and obtaining a scanning image.

Description

PET IMAGING WITH HER2 INHIBITOR COMPOUNDS

CROSS-REFERENCE

[0001] This application claims the benefit of U.S. Provisional Application No. 63/555,739, filed February 20, 2024, which is hereby incorporated by reference in its entirety herein.

BACKGROUND

[0002] HER2 (also referred to as Her2) belongs to the epidermal growth factor receptor (EGFR) family. This family is composed of four HER receptors: human epidermal growth factor receptor 1 (Herl) (also termed EGFR), HER2, human epidermal growth factor receptor 3 (HER3), and human epidermal growth factor receptor 4 (Her4). The HER2 receptor is a 185 kDa transmembrane protein that is encoded by the HER2 (also known as erb-b2 receptor tyrosine kinase 2 [ERBB2]) gene. HER2 is normally expressed on cell membranes of epithelial cells of several organs like the lungs, breast and the skin, as well as gastrointestinal, reproductive, and urinary tract . HER2 in normal cells is expressed at low levels, whereas in HER2-positive cancer cells, there is an increase in the number of HER2 gene copies (gene amplification) and HER2 receptors with up to 40-to- 100-fold increase in protein overexpression. The increased amount of cell surface HER2 receptors associated with HER2 overexpression leads to increased receptor-receptor interactions, provoking a sustained tyrosine phosphorylation of the kinase domain and therefore constant activation of the signaling pathways.

[0003] Tumors driven by HER2 mutations or HER2 wild type over expression may benefit from tyrosine kinase inhibitors that target HER2.

[0004] Current irreversible HER2 tyrosine kinase inhibitors in clinical development include Poziotinib and Pyrotinib that both lack selectivity for HER2 mutated tumors vs. EGFR and have adverse event profiles consistent with EGFR-related toxicities. Specifically, patients receiving poziotinib experienced Grade 3 skin rash, among other Grade 3 adverse events, that was difficult to tolerate, leading to significant dose reductions. In addition, patients receiving pyrotinib also experienced various Grade 3 adverse events including an increase of 7 or more stools a day which usually requires hospitalization.

[0005] In many cancer types, tumor cells make extra copies of the gene that produces the HER2 protein, known as gene amplification. The resulting flood of HER2 protein causes cancer cells to grow uncontrollably, and cancer cells may also become dependent on the extra HER2 such that stopping the production of the HER2 protein can cause the cancer cells to stop growing or die. In breast cancer, 30% of tumors overexpress HER2 and HER2 -targeted treatments are commonly used.

[0006] HER2 overexpression has been described in not only breast and gastric/gastroesophageal junction carcinomas, but somatic HER2 mutations have also been described at low frequencies in a variety of human cancers including non-small cell lung cancer , colorectal cancer, and bladder cancer. Breast cancer is a heterogeneous disease comprising various molecular subtypes, with approximately 15-20% of cases characterized by HER2 -positive overexpression. Targeted therapies, such as trastuzumab, have demonstrated substantial clinical benefits for these patients, although challenges persist, including the development of treatment resistance. Given the central role of HER2 expression in driving the disease, combining HER2-directed agents with trastuzumab has gained attention as a strategy to address HER2- related aspects of the disease from multiple angles, offering potential for improved treatment outcomes. Despite recent advances in the treatment of metastatic NSCLC, the absolute number of long-term survivors remains low.

[0007] In metastatic CRC, 3% to 5% of patients present with HER2 alterations, and the prognosis for patients with metastatic colorectal cancer remains poor with 5-year survival rates of 5% or less. The 5-year relative survival rate for patients with metastatic bladder cancer is only 8%. Bladder cancer ranks third among all cancers in terms of HER2 overexpression, carrying as much as 6% to 17% of gene mutations and/or amplification in tumor tissue samples. HER2 overexpression is associated with pathological malignancy and poor prognosis indicators including carcinoma in situ, multifocal tumor, large tumor size, high tumor stage and grade, lymph node metastasis, progression, recurrence, and papillary tumor.

[0008] In recent years, increasing attention has been paid to dual anti-HER2 therapies with the aim of resolving the occurrence of toxic reactions and the development of resistance. Trastuzumab (marketed as Herceptin) is a monoclonal antibody that binds to the extracellular domain of the HER2 receptor. Tucatinib is a specific and reversible inhibitor of the protein tyrosine kinase activity of HER2, and the binding of tucatinib to the intracellular HER2 tyrosine kinase domain occurs intracellularly. Thus, tucatinib and trastuzumab block the activity of HER2 proteins but in different ways. Food and Drug Administration (FDA) granted accelerated approval to the combination of two HER2 targeted drugs, tucatinib (Tukysa) and a trastuzumab (Herceptin) for people with advanced colorectal cancer that produces an excess amount of a protein called HER2. Trastuzumab also been used for the treatment for HER2-postive breast cancer, and tucatinib has also been used in combination with trastuzumab in breast cancer. Combinations of trastuzumab and capecitabine have also been used to treat Her2 positive breast cancer and colorectal cancer.

[0009] Radiolabeled [C¹ ^tucatinib has been synthesized recently and evaluated for use in Her2 PET imaging. No significant tumor accumulation was observed despite its high affinity for HER-2 receptors (IC50 = 6.9 11M). High liver and intestinal uptake indicate that [^{1 !}C] tucatinib was too lipophilic to be used as a tumor targeting PET tracer. The study highlighted the differences between a drug, which needs to be effective, and an imaging agent, which is dependent on contrast.

[0010] However, most existing tyrosine kinase inhibitors, such as neratinib and poziotinib, are dual HER2-EGFR inhibitors and display significant toxicity from inhibition of EGFR. Tucatinib is the only approved HER2-selective tyrosine kinase inhibitor (approved in combination with trastuzumab for the treatment of HER2- positive colorectal cancer), but it lacks potency against exon 20 insertion mutations. There is therefore an urgent unmet need to find and monitor HER2 mediated tumor cells in patients in order to more effectively treat patients with anti-HER2 monotherapies and dual therapies designed to treat patients with HER2-driven cancers. There is also an urgent need for novel anti-HER2 monotherapies and dual therapies designed to treat patients with HER2-driven cancers, which exhibit robust activity against both HER2 and HER2 mutants (such as YVMA HER2 exon20 insertion mutations), while preserving wild-type (WT) EGFR.

SUMMARY

[0011] Disclosed herein are a radiolabeled compounds having the structure of Formula (I): or a pharmaceutically acceptable salt thereof, wherein:

A is selected from the group consisting of CH and N;

B is selected from the group consisting of CH2 ,CHF, and CF2;

E is selected from the group consisting of CH2, CHF, and CF2;

X is selected from the group consisting of CH, CF, C(OH) and N; or X is CH and B and E are both absent;

Q¹ is selected from the group consisting of H, Ci-Cealkyl, F, and Cl;

Q² is selected from the group consisting of halogen, haloalkyl, alkyl, alkene, alkyne, - NR^aR^b, -Ci-C6alkylene-NR^aR^b, -Ci-Cealkylene-OR⁰, cyano, hydroxyalkyl, -Co-Cealkylene- C(O)OH, -Ci-C6alkylene-C(O)O-alkyl, alkoxyalkyl, -Co-C4alkylene-cycloalkyl optionally substituted with 1-3 J⁴ groups, -Co-C4alkylene-cycloalkenyl optionally substituted with 1-3 J⁴ groups, -Co-C4alkylene-7-l l membered spirocyclic cycloalkyl, optionally substituted with 1- 3 J⁴ groups, -Co-C4alkylene-7-l l membered spirocyclic heterocycloalkyl optionally substituted with 1-3 J⁴ groups, -Co-C4alkylene-heterocycloalkyl optionally substituted with 1- 3 J⁴ groups, and -Co-C4alkylene-heterocycloalkenyl optionally substituted with 1-3 J⁴ groups; each J⁴ is independently selected from the group consisting of halogen, alkyl, haloalkyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, oxo, and -Co-C4alkylene-NR^aR^b, provided that J⁴ groups can only include up to two oxo groups and up to one -Co-C4alkylene- NR^aR^b group;

Q³ is selected from H and F;

R^a and R^b each are independently selected from the group consisting of Ci-Cealkyl, Ci-Cehaloalkyl, Ci-Cehydroxyalkyl, and -Ci-Cealkyl-Ci-Cealkoxy;

R^c is selected from the group consisting of H, alkyl, haloalkyl, hydroxyalkyl, alkoxyalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl, wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are each optionally substituted with 1-3 groups selected from the group consisting of halogen, alkyl, alkoxy and alkoxyalkyl;

R¹ is selected from the group consisting of alkyl, haloalkyl and halogen;

R² is selected from the group consisting of is -O-alkyl, -O-aryl, -O-heteroaryl, -O- cycloalkyl, -O-heterocycloalkyl, -O-heteroaryl-alkylene-aryl, -NH-alkyl, -NH-aryl, or-NH- heteroaryl, wherein each of the alkyl, aryl, heteroaryl, cycloalkyl or heterocycloalkyl moieties are optionally substituted with 1-4 J¹ groups; or R¹ and R² join with the carbon atoms to which they are attached to form a saturated or unsaturated carbocyclic or heterocyclic ring, wherein the saturated or unsaturated carbocyclic or heterocyclic ring is optionally substituted with 1-4 J¹ groups; each J¹ is independently selected from the group consisting of halogen, alkyl, haloalkyl, hydroxy, hydroxyalkyl, -Co-C4alkylene-N(H)R^C, alkoxy, and alkoxyalkyl; and wherein at least one carbon atom of Formula (I) is substituted with at least one ¹⁸F, and the at least one ¹⁸F atom can substitute any hydrogen or halogen atom in Formula (I) provided that a C-¹⁸F bond is formed. [0012] Disclosed herein is a method of selecting a subject for treating a HER2 mediated disease, the method comprising administering to the subject a radiolableled compound described herein, or a pharmaceutically acceptable salt thereof, generating a scanning image of the subject, and selecting the subject for treating the HER2 mediated disease when the scanning image shows that the radiolableled compound, or a pharmaceutically acceptable salt thereof, is accumulated in a tumor tissue.

[0013] Disclosed herein is a method of tracking tumor response to a HER2 mediated disease in a subject, the method comprising administering to the subject a radiolableled compound described herein, or a pharmaceutically acceptable salt thereof, and generating a scanning image of the subject.

[0014] Disclosed herein is a method of obtaining a scanning image, comprising administering to a subject the radiolableled compound described herein, or a pharmaceutically acceptable salt thereof, and subsequently generating a scanning image of the subject.

BRIEF DESCRIPTION OF THE DRAWINGS

[0015] FIG. 1 illustrates the concentration of the radiolabeled compound in different tissue samples in relation to time.

DETAILED DESCRIPTION

[0016] Compounds of this disclosure are useful for detecting human epidermal growth factor 2 (HER2) and HER2 mutants, including the exon 20 insertion mutations. Surprisingly, compounds of this disclosure have been found to bind to HER2 that is a new and highly advantageous binding mechanism for HER2 inhibitors. Significantly, this new HER2 binding mechanism unique to the compounds of this disclosure can be useful for probing tumor cells compared to normal cells.

[0017] The compounds described herein can be useful as probes for imaging. For example, in a non-limiting case, the compounds can be useful for monitoring improvements in brain metastasis using tomography imaging, such as PET imaging. PET is a rapidly growing area in medical imaging, and may be useful for a more personalized approach to medical treatment. Use of the novel radioactive probe compounds described herein can be useful for detecting and diagnosing HER2 -mediated diseases such as cancer. Radiolabeled Compounds

[0018] The radiolabeled compounds described herein can be useful as a radioactive probe for imaging. In some embodiments, is a radiolabeled compound having the structure of Formula (I): or a pharmaceutically acceptable salt thereof, wherein:

A is selected from the group consisting of CH and N;

B is selected from the group consisting of CH2 ,CHF, and CF2;

E is selected from the group consisting of CH2, CHF, and CF2;

Q¹ is selected from the group consisting of H, Ci-Cealkyl, F, and Cl;

Q³ is selected from H and F;

R¹ is selected from the group consisting of alkyl, haloalkyl and halogen;

R² is selected from the group consisting of is -O-alkyl, -O-aryl, -O-heteroaryl, -O- cycloalkyl, -O-heterocycloalkyl, -O-heteroaryl-alkylene-aryl, -NH-alkyl, -NH-aryl, or-NH- heteroaryl, wherein each of the alkyl, aryl, heteroaryl, cycloalkyl or heterocycloalkyl moieties are optionally substituted with 1-4 J¹ groups; or R¹ and R² join with the carbon atoms to which they are attached to form a saturated or unsaturated carbocyclic or heterocyclic ring, wherein the saturated or unsaturated carbocyclic or heterocyclic ring is optionally substituted with 1-4 J¹ groups; each J¹ is independently selected from the group consisting of halogen, alkyl, haloalkyl, hydroxy, hydroxyalkyl, -Co-C4alkylene-N(H)R^C, alkoxy, and alkoxyalkyl; and wherein at least one carbon atom of Formula (I) is substituted with at least one ¹⁸F, and the at least one ¹⁸F atom can substitute any hydrogen or halogen atom in Formula (I) provided that a C-¹⁸F bond is formed.

[0019] In some embodiments, the radiolabeled compound of Formula (I) one ¹⁸F atom substitutes a hydrogen atom to form a C-¹⁸F bond. In some embodiments, the radiolabeled compound of Formula (I) one ¹⁸F atom substitutes a halogen atom to form a C-¹⁸F bond. In some embodiments, the radiolabeled compound of Formula (I) one ¹⁸F atom substitutes a flourine atom to form a C-¹⁸F bond.

[0020] In some embodiments, Q³ is selected from H and F. In some embodiments, Q³ is H. In some embodiments, Q³ is F.

[0021] In some embodiments, A is selected from CH and N. In some embodiments, A is CH. In some embodiments, A is N.

[0022] In some embodiments, R¹ is selected from the group consisting of alkyl, haloalkyl and halogen. In some embodiments, R¹ is selected from the group consisting of alkyl and haloalkyl. In some embodiments, R¹ is alkyl. In some embodiments, R¹ is Ci-Ce alkyl. In some embodiments, R¹ is methyl, ethyl, or propyl. In some embodiments, R¹ is methyl or ethyl. In some embodiments, R¹ is methyl.

[0023] In some embodiments, R² is selected from the group consisting of is -O-alkyl, -O-aryl, -O-heteroaryl, -O-cycloalkyl, -O-heterocycloalkyl, -O-heteroaryl-alkylene-aryl, -NH-alkyl, - NH-aryl, or-NH-heteroaryl, wherein each of the alkyl, aryl, heteroaryl, cycloalkyl or heterocycloalkyl moieties are optionally substituted with 1-4 J¹ groups. In some embodiments, R² is selected from the group consisting of is -O-alkyl, -O-aryl, -O-heteroaryl, -O-cycloalkyl, -O-heterocycloalkyl, and -O-heteroaryl-alkylene-aryl, wherein each of the alkyl, aryl, heteroaryl, cycloalkyl or heterocycloalkyl moieties are optionally substituted with 1-4 J¹ groups. In some embodiments, R² is selected from the group consisting of , -O-aryl, - O-heteroaryl, -O-cycloalkyl, and -O-heterocycloalkyl, wherein each of the aryl, heteroaryl, cycloalkyl or heteocycloalkyl moieties are optionally substituted with 1-4 J¹ groups. In some embodiments, R² is selected from the group consisting of , -O-aryl and -O-heteroaryl, wherein each of the aryl or heteroaryl moieties are optionally substituted with 1-4 J¹ groups. In some embodiments, R² is -O-heteroaryl, wherein the heteroaryl moiety is optionally substituted with 1-4 J¹ groups. In some embodiments, R² is -O-heteroaryl, wherein the heteroaryl is a 5- to 9- membered heteroaryl.

[0024] In some embodiments, the radiolabeled compound has a structure of Formula (II): or a pharmaceutically acceptable salt thereof.

[0025] In some embodiments, Q¹ is selected from the group consisting of H, Ci-Cealkyl, F, and Cl. In some embodiments, Q¹ is H or Ci-Ce alkyl. In some embodiments, Q¹ is H. In some embodiments, Q¹ is F. In some embodiments, Q¹ is Cl. In some embodiments, Q¹ is Ci- Ce alkyl. In some embodiments, Q¹ is methyl, ethyl, or propyl. In some embodiments, Q¹ is methyl or ethyl. In some embodiments, Q¹ is methyl.

[0026] In some embodiments, Q² is selected from the group consisting of halogen, haloalkyl, alkyl, alkene, alkyne, -NR^aR^b, -Ci-C6alkylene-NR^aR^b, -Ci-Cealkylene-OR⁰, cyano, hydroxyalkyl, -Co-Cealkylene-C(0)OH, -Ci-C6alkylene-C(O)O-alkyl, alkoxyalkyl, -Co- C4alkylene-cycloalkyl optionally substituted with 1-3 J⁴ groups, -Co-C4alkylene-cycloalkenyl optionally substituted with 1-3 J⁴ groups, -Co-C4alkylene-7-l 1 membered spirocyclic cycloalkyl, optionally substituted with 1-3 J⁴ groups, -Co-C4alkylene-7-l 1 membered spirocyclic heterocycloalkyl optionally substituted with 1-3 J⁴ groups, -Co-C4alkylene- heterocycloalkyl optionally substituted with 1-3 J⁴ groups, and -Co-C4alkylene- heterocycloalkenyl optionally substituted with 1-3 J⁴ groups. In some embodiments, Q² is - Ci-C3alkylene-NR^aR^b, -Ci-Cealkylene-OR⁰, -Co-C4alkylene-cycloalkyl optionally substituted with 1-3 J⁴ groups, -Co-C4alkylene-cycloalkenyl optionally substituted with 1-3 J⁴ groups, or -Co-C4alkylene-heterocycloalkyl optionally substituted with 1-3 J⁴. In some embodiments, Q² is -Ci-C3alkylene-NR^aR^b, -Co-C4alkylene-cycloalkyl optionally substituted with 1-3 J⁴ groups, or -Co-C4alkylene-heterocycloalkyl optionally substituted with 1-3 J⁴. In some embodiments, Q² is -Ci-C3alkylene-NR^aR^b.

[0027] In some embodiments, R^a and R^b each are independently selected from the group consisting of Ci-Cealkyl, Ci-Cehaloalkyl, Ci-Cehydroxyalkyl, and -Ci-Cealkyl-Ci-Cealkoxy. In some embodiments, R^a and R^b each are independently selected from the group consisting of Ci-Cealkyl, Ci-Cehaloalkyl, and Ci-Cehydroxy alkyl. In some embodiments, R^a and R^b each are independently selected from the group consisting of Ci-Cealkyl and Ci-Cehaloalkyl. In some embodiments, R^a and R^b are each independently Ci-Cealkyl. In some embodiments, R^a and R^b are each independently Ci-Cehaloalkyl.

[0028] In some embodiments, Q² is -Co-C4alkylene-heterocycloalkyl optionally substituted with 1-3 J⁴.

[0029] In some embodiments, each J⁴ is independently selected from the group consisting of halogen, alkyl, haloalkyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, oxo, and -Co- C4alkylene-NR^aR^b, provided that J⁴ groups can only include up to two oxo groups and up to one -Co-C4alkylene-NR^aR^b group. In some embodiments, each J⁴ is independently halogen, alkyl, haloalkyl, hydroxy, hydroxyalkyl, alkoxy, or alkoxyalkyl. In some embodiments, each J⁴ is independently halogen, alkyl, haloalkyl, hydroxyalkyl, or alkoxy. In some embodiments, each J⁴ is independently halogen, alkyl, or haloalkyl. In some embodiments, each J⁴ is independently halogen or alkyl. In some embodiments, each J⁴ is independently halogen. In some embodiments, each J⁴ is independently C1-C3 alkyl.

[0030] In some embodiments, X is selected from CH, CF, C(OH) and N. In some embodiments, X is CH, CF, or C(OH). In some embodiments, X is CH or CF. In some embodiments, X is CH. In some embodiments, X is CF.

[0031] In some embodiments, at least one carbon atom of Q¹, Q², B, E, or X that has a single bond to either a hydrogen or halogen atom is substituted with ¹⁸F. In some embodiments, at least one carbon atom of Q¹, Q², B, or E that has a single bond to either a hydrogen or halogen atom is substituted with ¹⁸F. In some embodiments, at least one carbon atom of Q¹ or Q² that has a single bond to either a hydrogen or halogen atom is substituted with ¹⁸F.

[0032] In some embodiments, the radiolabeled compound has the following structure:

or a pharmaceutically acceptable salt thereof.

Methods

[0033] Provided herein are methods which use the radiolabeled compounds described herein. The methods can be useful for treating a disease, tracking response, and scanning an image. [0034] In some embodiments is a method of selecting a subject for treating a HER2 mediated disease, the method comprising administering to the subject a radiolabeled compound described herein, or a pharmaceutically acceptable salt thereof, generating a scanning image of the subject, and selecting the subject for treating the HER2 mediated disease when the scanning image shows that the radiolabeled compound, or a pharmaceutically acceptable salt thereof, is accumulated in a tumor tissue.

[0035] The radiolabeled compound or pharmaceutically acceptable salt thereof, described herein can be administered via systemic administration. In some embodiments, the radiolabeled compound is administered via enteral administration or parental administration. In some embodiments, the radiolabeled compound is administered via enteral administration. In some embodiments, the radiolabeled compound is administered via parental administration.

[0036] In some embodiments, the radiolabeled compound can be accumulated in one area of the subject. In some embodiments, the radiolabeled compound, or a pharmaceutically acceptable salt thereof, is accumulated in the brain.

[0037] In some embodiments, the scanning image is a tomographic image. In some embodiments, the tomographic image is generated with positron emission tomography (PET), X-ray computed tomography (CT), or magnetic resonance imaging (MRI). In some embodiments, the tomographic image is generated with positron emission tomography (PET). [0038] In some embodiments, the HER2 mediated disease is metastatic brain tumor. The metastatic brain tumor can be caused by other types of tumors or cancers. In some embodiments, the metastatic brain tumor is caused by lung cancer, breast cancer, skin cancer, colon cancer, or melanoma. In some embodiments, the metastatic brain tumor is caused by breast cancer. In some embodiments, the metastatic brain tumor is caused by lung cancer. In some embodiments, the metastatic brain tumor is caused by skin cancer. In some embodiments, the metastatic brain tumor is caused by colon cancer. In some embodiments, the metastatic brain tumor is caused by melanoma.

[0039] In some embodiments, the HER2 mediated disease is cancer. In some embodiments, the cancer is brain cancer, colorectal cancer, breast cancer, bladder cancer, biliary cancer, or non-small cell lung cancer. In some embodiments, the cancer is brain cancer. In some embodiments, the cancer is colorectal cancer. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is bladder cancer. In some embodiments, the cancer is non-small cell lung cancer. In some embodiments, the cancer is ovarian cancer.

[0040] In some embodiments is a method of tracking tumor response to a HER2 mediated disease in a subject, the method comprising administering to the subject a radiolabeled compound described herein, or a pharmaceutically acceptable salt thereof, and generating a scanning image of the subject.

[0041] In some embodiments, the scanning image is generated once every two weeks to once every four weeks. In some embodiments, the scanning image is generated once very two weeks. In some embodiments, the scanning image is generated once every four weeks. In some embodiments, the scanning imaging is generated for at least 1 month. In some embodiments, the scanning imaging is generated for at least 2 months. In some embodiments, the scanning image is generated for at least 3 months. In some embodiments, the scanning image is generated for at least 4 months. In some embodiments, the scanning image is generated for at least 5 months. In some embodiments, the scanning image is generated for at least 6 months.

[0042] The radiolabeled compound or pharmaceutically acceptable salt thereof, described herein can be administered via systemic administration. In some embodiments, the radiolabeled compound is administered via enteral administration or parental administration. In some embodiments, the radiolabeled compound is administered via enteral administration. In some embodiments, the radiolabeled compound is administered via parental administration.

[0043] In some embodiments, the radiolabeled compound can be accumulated in one area of the subject. In some embodiments, the radiolabeled compound, or a pharmaceutically acceptable salt thereof, is accumulated in the brain.

[0044] In some embodiments, the scanning image is a tomographic image. In some embodiments, the tomographic image is generated with positron emission tomography (PET), X-ray computed tomography (CT), or magnetic resonance imaging (MRI). In some embodiments, the tomographic image is generated with positron emission tomography (PET). [0045] In some embodiments, the HER2 mediated disease is metastatic brain tumor. The metastatic brain tumor can be caused by other types of tumors or cancers. In some embodiments, the metastatic brain tumor is caused by lung cancer, breast cancer, skin cancer, colon cancer, or melanoma. In some embodiments, the metastatic brain tumor is caused by breast cancer. In some embodiments, the metastatic brain tumor is caused by lung cancer. In some embodiments, the metastatic brain tumor is caused by skin cancer. In some embodiments, the metastatic brain tumor is caused by colon cancer. In some embodiments, the metastatic brain tumor is caused by melanoma.

[0046] In some embodiments is a method of obtaining a scanning image, comprising administering to a subject the radiolabeled compound described herein, or a pharmaceutically acceptable salt thereof, and subsequently generating a scanning image of the subject.

[0047] In some embodiments, the scanning image is a tomographic image. In some embodiments, the tomographic image is generated with positron emission tomography (PET), X-ray computed tomography (CT), or magnetic resonance imaging (MRI). In some embodiments, the tomographic image is generated with positron emission tomography (PET). Definitions

[0048] Unless defined otherwise, all terms of art, notations and other technical and scientific terms or terminology used herein are intended to have the same meaning as is commonly understood by one of ordinary skill in the art to which the claimed subject matter pertains. In some cases, terms with commonly understood meanings are defined herein for clarity and/or for ready reference, and the inclusion of such definitions herein should not necessarily be construed to represent a substantial difference over what is generally understood in the art. [0049] Throughout this application, various embodiments may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the disclosure. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

[0050] As used in the specification and claims, the singular forms “a”, “an” and “the” include plural references unless the context clearly dictates otherwise. For example, the term “a sample” includes a plurality of samples, including mixtures thereof.

[0051] Unless a point of attachment indicates otherwise, the chemical moieties listed in the definitions of the variables of Formula (I) of this disclosure, and all the embodiments thereof, are to be read from left to right, wherein the right-hand side is directly attached to the parent structure as defined. However, if a point of attachment (e.g., a dash is shown on the lefthand side of the chemical moiety (e.g., -Ci-Cealkyl-N(R⁶)2), then the left-hand side of this chemical moiety is attached directly to the parent moiety as defined.

[0052] It is assumed that when considering generic descriptions of compounds described herein for the purpose of constructing a compound, such construction results in the creation of a stable structure. That is, one of ordinary skill in the art would recognize that, theoretically, some constructs would not normally be considered as stable compounds (that is, sterically practical and/or synthetically feasible).

[0053] “Alkyl,” by itself, or as part of another substituent, means, unless otherwise stated, a straight or branched chain hydrocarbon, having the number of carbon atoms designated (i.e. Ci-Ce means one to six carbons). Representative alkyl groups include straight and branched chain alkyl groups having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms. Further representative alkyl groups include straight and branched chain alkyl groups having 1, 2, 3, 4, 5, 6, 7 or 8 carbon atoms. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. For each of the definitions herein (e.g., alkyl, alkoxy, heterocycloalkylalkyl, heteroaryl alkyl, etc.), when a prefix is not included to indicate the number of carbon atoms in an alkyl portion, the alkyl moiety or portion thereof will have 12 or fewer main chain carbon atoms or 8 or fewer main chain carbon atoms or 6 or fewer main chain carbon atoms. For example, Ci-Cealkyl refers to a straight or branched hydrocarbon having 1, 2, 3, 4, 5 or 6 carbon atoms and includes, but is not limited to, -CH3, C2alkyl, Csalkyl, C4alkyl, Csalkyl, C₆alkyl, Ci-C₂alkyl, C₂alkyl, C₃alkyl, Ci-C₃alkyl, Ci-C₄alkyl, Ci-C₅alkyl, Ci-C₆alkyl, C₂. C₃alkyl, C₂.C₄alkyl, C₂.C₅alkyl, C₂.C₆alkyl, C₃.C₄alkyl, C₃.C₅alkyl, C₃.C₆alkyl, C₄.C₅alkyl, C₄.Cealkyl, Cs-Ce alkyl and Cealkyl. It is understood that substitutions are attached at any available atom to produce a stable compound.

[0054] “Alkylene” by itself or as part of another substituent means a linear or branched saturated divalent hydrocarbon moiety derived from an alkane having the number of carbon atoms indicated in the prefix. For example, (i.e., Ci-Ce means one to six carbons; Ci- Cealkylene is meant to include methylene, ethylene, propylene, 2-methylpropylene, pentylene, hexylene and the like). Ci-C₄ alkylene includes methylene -CH₂-, ethylene -CH₂CH₂-, propylene -CH₂CH₂CH₂-, and isopropylene -CH(CH₃)CH₂-, -CH₂CH(CH₃)-, -CH₂-(CH₂)₂CH₂-, -CH₂-CH(CH₃)CH₂-, -CH₂- C(CH₃)₂-CH₂-CH₂CH(CH₃)-. Typically, an alkyl (or alkylene) group will have from 1 to 24 carbon atoms, with those groups having 10 or fewer, 8 or fewer, or 6 or fewer carbon atoms. When a prefix is not included to indicate the number of carbon atoms in an alkylene portion, the alkylene moiety or portion thereof will have 12 or fewer main chain carbon atoms or 8 or fewer main chain carbon atoms, 6 or fewer main chain carbon atoms, or 4 or fewer main chain carbon atoms, or 3 or fewer main chain carbon atoms, or 2 or fewer main chain carbon atoms, or 1 carbon atom.

[0055] “Alkoxy” or “alkoxyl” refers to a -O-alkyl group, where alkyl is as defined herein. By way of example, “Ci-Cealkoxy” refers to a -O-Ci-Cealkyl group, where alkyl is as defined herein. While it is understood that substitutions on alkoxy are attached at any available atom to produce a stable compound, substitution of alkoxy is such that O, S, or N (except where N is a heteroaryl ring atom), are not bound to the alkyl carbon bound to the alkoxy O. Further, where alkoxy is described as a substituent of another moiety, the alkoxy oxygen is not bound to a carbon atom that is bound to an O, S, or N of the other moiety (except where N is a heteroaryl ring atom), or to an alkene or alkyne carbon of the other moiety.

[0056] “Amino” or “amine” denotes the group NH₂.

[0057] “Aryl” by itself, or as part of another substituent, unless otherwise stated, refers to a monocyclic, bicyclic or polycyclic polyunsaturated aromatic hydrocarbon radical containing 6 to 14 ring carbon atoms, which can be a single ring or multiple rings (up to three rings) which are fused together or linked covalently. Aryl, however, does not encompass or overlap in any way with heteroaryl defined below. If one or more aryl rings are fused with a heteroaryl ring, the resulting ring system is heteroaryl. Non-limiting examples of unsubstituted aryl groups include phenyl, 1 -naphthyl and 2-naphthyl. The term “arylene” refers to a divalent aryl, wherein the aryl is as defined herein.

[0058] “Cycloalkyl” or “Carbocycle” or “Carbocyclic” by itself, or as part of another substituent, unless otherwise stated, refers to saturated or partially unsaturated, nonaromatic monocyclic ring, bridged rings, spiro rings, fused rings (e.g., bicyclic or tricyclic carbon ring systems), or cubane, having the number of carbon atoms indicated in the prefix or if unspecified having 3-6, also 4-6, and also 5-6 ring members per ring, such as cyclopropyl, cyclopentyl, cyclohexyl, where one or two ring carbon atoms may optionally be replaced by a carbonyl. Further, the term cycloalkyl is intended to encompass ring systems fused to an aromatic ring (e.g., of an aryl or heteroaryl), regardless of the point of attachment to the remainder of the molecule. Cycloalkyl refers to hydrocarbon rings having the indicated number of ring atoms (e.g., C3-C6 cycloalkyl and 3-6 membered cycloalkyl both mean three to six ring carbon atoms). The term “cycloalkenyl” refers to a cycloalkyl having at least one unit of unsaturation. A substituent of a cycloalkyl or cycloalkenyl may be at the point of attachment of the cycloalkyl or cycloalkenyl group, forming a quaternary center.

[0059] “Halogen” or “halo” refers to all halogens, that is, chloro (Cl), fluoro (F), bromo (Br), or iodo (I).

[0060] “Heteroatom” is meant to include oxygen (O), nitrogen (N), and sulfur (S).

[0061] “Heteroaryl” refers to a monocyclic or bicyclic aromatic ring radical containing 5-9 ring atoms (also referred to in this disclosure as a 5-9 membered heteroaryl, including monocyclic aromatic ring radicals containing 5 or 6 ring atoms (also referred to in this disclosure as a 5-6 membered heteroaryl), containing one or more, 14, 13, or 12, heteroatoms independently selected from the group consisting of O, S, and N. Any aromatic ring or ring system containing at least one heteroatom is a heteroaryl regardless of the point of attachment (i.e., through any one of the fused rings). Heteroaryl is also intended to include oxidized S or N, such as sulfinyl, sulfonyl and N-oxide of a tertiary ring nitrogen. A carbon or nitrogen atom is the point of attachment of the heteroaryl ring structure such that a stable compound is produced. Examples of heteroaryl groups include, but are not limited to, pyridyl, pyridazinyl, pyrazinyl, indolizinyl, benzo[b]thienyl, quinazolinyl, purinyl, indolyl, quinolinyl, pyrimidinyl, pyrrolyl, pyrazolyl, oxazolyl, thiazolyl, thienyl, isoxazolyl, oxathiadi azolyl, isothiazolyl, tetrazolyl, imidazolyl, triazolyl, furanyl, benzofuryl, indolyl, triazinyl, quinoxalinyl, cinnolinyl, phthalazinyl, benzotri azinyl, benzimidazolyl, benzopyrazolyl, benzotri azolyl, benzisoxazolyl, isobenzofuryl, isoindolyl, indolizinyl, benzotriazinyl, thienopyridyl, thienopyrimidinyl, pyrazolopyrimidinyl, imidazopyridines, benzothiaxolyl, benzothienyl, quinolyl, isoquinolyl, indazolyl, pteridinyl and thiadi azolyl. “Nitrogen containing heteroaryl” refers to heteroaryl wherein at least one of the ring heteroatoms is N. [0062] The term “heteroarylalkyl” refers to an alkyl group substituted with a heteroaryl group, where both terms are as defined herein.

[0063] The terms “heterocycle” or “heterocyclic ring” are interchangeable and comprise heterocycloalkyl rings and heteroaryl rings as they are defined herein. A heterocycle may be a saturated, unsaturated, or aromatic ring comprising one or more heteroatoms. Exemplary heteroatoms include N, O, P, and S atoms. Heterocycles include 3- to 10-membered monocyclic rings, 6- to 12-membered bicyclic rings, 5- to 12-membered spiro bicycles, and 5- to 12-membered bridged rings. A bicyclic heterocycle includes any combination of saturated, unsaturated and aromatic bicyclic rings, as valence permits.

[0064] The term “heterocycloalkyl” refers to a saturated or unsaturated non-aromatic cycloalkyl group that contains from one to five heteroatoms selected from N, O, S (including S(O) and S(O)2), or P (including phosphine oxide) wherein the nitrogen, sulfur, and phosphorous atoms are optionally oxidized, and the nitrogen atom(s) are optionally quartemized, the remaining ring atoms being C, where one or two C atoms may optionally be present as a carbonyl. A heterocycloalkyl group can have one or more carbon-carbon double bonds or carbon-heteroatom double bonds in the ring as long as the ring is not rendered aromatic by their presence. Further, the term heterocycloalkyl is intended to encompass any ring or ring system containing at least one heteroatom that is not a heteroaryl, regardless of the point of attachment to the remainder of the molecule. Heterocycloalkyl groups include those having a ring with a formally charge-separated aromatic resonance structure, for example, N-methylpyridonyl. The heterocycloalkyl may be substituted with one or two oxo groups, and can include sulfone and sulfoxide derivatives. The heterocycloalkyl may be a monocyclic, a bridged ring system, a fused bicyclic or a fused polycyclic ring system of 3 to 12, 4 to 10, 5 to 10, or 5 to 6 ring atoms in which one to five ring atoms are heteroatoms selected from -N=, -N-, -O-, -S-, -S(O)-, or -S(O)2- and further wherein one or two ring atoms are optionally replaced by a -C(O)- group. As an example, a 4-9 membered heterocycloalkyl is a heterocycloalkyl with 4-9 ring members having at least one heteroatom. The heterocycloalkyl can also be a heterocyclic alkyl ring fused with a cycloalkyl. Non limiting examples of heterocycloalkyl groups include pyrrolidine, piperidine, morpholine, pyridone, pyrrolidine, azepane, 1,4-diazepane, azetidine, 8-azabicylo[3.2.1]octane, 8- azabicylo[3.2.1]octene, and 3,9-diazabicyclo[4.2.1]nonane and the like. A heterocycloalkyl group can be attached to the remainder of the molecule through a ring carbon or a heteroatom. “Heterocycloalkenyl” refers to a heterocycloalkyl having at least one unit of unsaturation. A substituent of a heterocycloalkyl or heterocycloalkenyl may be at the point of attachment of the heterocycloalkyl or heterocycloalkenyl group, forming a quaternary center. [0065] The term “heterocycloalkylalkyl” refers to an alkyl group substituted with a heterocycloalkyl group. Examples include, but are not limited to, azetidinylmethyl, morpholinomethyl, and the like.

[0066] The term “Ci-Cehaloalkyl” refers to Ci-Ce alkyl as defined herein that is substituted with one or more halogen atoms.

[0067] The term “-Ci-C4alkylene-NR^aR^b” refers to a “-Ci-C4alkylene- that is attached to the parent moiety, and which substituted with NR^aR^b.

[0068] The term “Ci-Cehydroxyalkyl” refers to Ci-Ce alkyl as defined herein that is substituted with one or more hydroxy groups as defined herein.

[0069] The term “-Co-C4alkylene-C3-C7cycloalkyl” refers to -Co-C4alkylene- that is attached to the parent moiety, and which is substituted with a Cs-Cvcycloalkyl group as defined herein. [0070] The term “oxo” refers to C(=O) or (O). In some embodiments, two possible points of attachment on a carbon form an oxo group.

[0071] “Hydroxyl” or “hydroxy” refers to the group OH. The term “hydroxyalkyl” or “hydroxy alkylene” refers to an alkyl group or alkylene group, respectively as defined herein, substituted with 1-5 hydroxy groups.

[0072] The term “substituent” is an atom or group of atoms substituted in place of hydrogen atom(s) of the parent molecule. Non-limiting examples of substituents in this disclosure include J⁴ which can include monovalent or divalent substituents. Monovalent substituents are bonded to the parent moiety by replacing one hydrogen atom of the parent moiety through a single bond. The hydrogen atom that the monovalent substituent replaces may be an available hydrogen atom from a carbon or nitrogen atom of the parent moiety. Divalent substituents are bonded to the parent moiety by replacing two available hydrogen atoms of the parent moiety through a double bond. It is understood that substituents described in this disclosure cannot be attached to a parent moiety in a way that would result in an unstable molecule.

[0073] “Optional substituents” or “optionally substituted” as used throughout the disclosure means that the substitution on a compound may or may not occur, and that the description includes instances where the substitution occurs and instances in which the substitution does not. For example, the phrase “optionally substituted with 1-3 J¹ groups” means that the J¹ group may but need not be present. It is assumed in this disclosure that optional substitution on a compound occurs in a way that would result in a stable compound.

[0074] Unit dosage form” refers to a composition intended for a single administration to treat a subject suffering from a disease or medical condition. Each unit dosage form typically comprises each of the active ingredients of this disclosure plus pharmaceutically acceptable excipients. Examples of unit dosage forms are individual tablets, individual capsules, bulk powders, liquid solutions, ointments, creams, eye drops, suppositories, emulsions or suspensions. Treatment of the disease or condition may require periodic administration of unit dosage forms, for example: one unit dosage form two or more times a day, one with each meal, one every four hours or other interval, or only one per day. The expression “oral unit dosage form” indicates a unit dosage form designed to be taken orally.

[0075] As used herein in connection with compounds of the disclosure, the term “synthesizing” and like terms means chemical synthesis from one or more precursor materials.

[0076] As used herein, the term “composition” refers to a formulation suitable for administration to an intended animal subject for therapeutic purposes that contains at least one pharmaceutically active compound and at least one pharmaceutically acceptable carrier or excipient.

[0077] The term “pharmaceutically acceptable” indicates that the indicated material does not have properties that would cause a reasonably prudent medical practitioner to avoid administration of the material to a patient, taking into consideration the disease or conditions to be treated and the respective route of administration. For example, it is commonly required that such a material be essentially sterile, e.g., for injectables.

[0078] “Pharmaceutically acceptable salt” refers to a salt which is acceptable for administration to a patient, such as a mammal (e.g., salts having acceptable mammalian safety for a given dosage regime). Contemplated pharmaceutically acceptable salt forms include, without limitation, mono, bis, tris, tetrakis, and so on. Pharmaceutically acceptable salts are non-toxic in the amounts and concentrations at which they are administered. The preparation of such salts can facilitate the pharmacological use by altering the physical characteristics of a compound without preventing it from exerting its physiological effect. Useful alterations in physical properties include lowering the melting point to facilitate transmucosal administration and increasing the solubility to facilitate administering higher concentrations of the drug. Such salts can be derived from pharmaceutically acceptable inorganic or organic bases and from pharmaceutically-acceptable inorganic or organic acids, depending on the particular substituents found on the compounds described herein.

[0079] Pharmaceutically acceptable salts can be prepared by standard techniques. For example, the free-base form of a compound can be dissolved in a suitable solvent, such as an aqueous or aqueous-alcohol solution containing the appropriate acid and then isolated by evaporating the solution. In another example, a salt can be prepared by reacting the free base and acid in an organic solvent.

[0080] When compounds of the present disclosure contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base (i.e. a primary, secondary, tertiary, quaternary, or cyclic amine; an alkali metal hydroxide; alkaline earth metal hydroxide; or the like), either neat or in a suitable inert solvent. The desired acid can be, for example, a pyranosidyl acid (such as glucuronic acid or galacturonic acid), an alpha-hydroxy acid (such as citric acid or tartaric acid), an amino acid (such as aspartic acid or glutamic acid), an aromatic acid (such as benzoic acid or cinnamic acid), a sulfonic acid (such as p-toluenesulfonic acid or ethanesulfonic acid), or the like. In some embodiments, salts can be derived from pharmaceutically acceptable acids such as acetic, trifluoroacetic, propionic, ascorbic, benzenesulfonic, benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, glycolic, gluconic, glucoronic, glutamic, hippuric, hydrobromic, hydrochloric, isethionic, lactic, lactobionic, maleic, malic, malonic, mandelic, oxalic, methanesulfonic, mucic, naphthalenesulfonic, nicotinic, nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, sulfamic, hydroiodic, carbonic, tartaric, p-toluenesulfonic, pyruvic, aspartic, benzoic, cinnamic, anthranilic, mesylic, salicylic, p-hydroxybenzoic, phenylacetic, embonic (pamoic), ethanesulfonic, benzenesulfonic, 2-hydroxyethanesulfonic, sulfanilic, stearic, cyclohexylsulfamic, cyclohexylaminosulfonic, quinic, algenic, hydroxybutyric, galactaric and galacturonic acid and the like.

[0081] Also included are salts of amino acids such as arginate and the like, and salts of organic acids like glucuronic or galactunoric acids and the like (see, for example, Berge, S. M. et al., “Pharmaceutical Salts,” J. Pharmaceutical Science, 1977, 66:1-19). Certain specific compounds of the present disclosure contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.

[0082] The neutral forms of the compounds may be regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents, but otherwise the salts are equivalent to the parent form of the compound for the purposes of the present disclosure.

[0083] The pharmaceutically acceptable salt of the different compounds may be present as a complex. Examples of complexes include 8-chlorotheophylline complex (analogous to, e.g., dimenhydrinate: diphenhydramine 8-chlorotheophylline (1 :1) complex; Dramamine) and various cyclodextrin inclusion complexes.

[0084] The term “deuterated” as used herein alone or as part of a group, means substituted deuterium atoms. The term “deuterated analog” as used herein alone or as part of a group, means substituted deuterium atoms in place of hydrogen. The deuterated analog of the disclosure may be a fully or partially deuterium substituted derivative. In some embodiments, the deuterium substituted derivative of the disclosure holds a fully or partially deuterium substituted alkyl, aryl or heteroaryl group.

[0085] The disclosure requires isotopically-labeled compounds in which one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Specifically, the disclosure includes the use of C-¹⁸F moieties, wherein ¹⁸F can be substituted for any hydrogen atom or halogen atom in a carbon-hydrogen or carbon-halogen moiety. Any variable in compound Formula described as, for example, “CH” is understood to include both “CH” and “C-¹⁸F” unless otherwise stated. A variable such as “alkyl” is understood to include an alkyl group with “C-H” moieties and an alkyl group in which at least one “C-H” is replaced with a “C-¹⁸F”. Further, all isotopic variations of the compounds of the present disclosure, whether radioactive or not, are intended to be encompassed within the scope of the present disclosure. Examples of isotopes that can be incorporated into compounds of the disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, and chlorine, such as, but not limited to ²H (deuterium, D), ³H (tritium), ⁿC, ¹³C, ¹⁴C, ¹⁵N, ¹⁸F, ³¹P, ³²P, ³⁵S, ³⁶C1, and ¹²⁵I. Unless otherwise stated, when a position is designated specifically as “H” or “hydrogen,” the position is understood to have hydrogen at its natural abundance isotopic composition or its isotopes, such as deuterium (D) or tritium (³H). Certain isotopically-labeled compounds of the present disclosure (e.g., those labeled with ³H and ¹⁴C) are useful in compound and/or substrate tissue distribution assays. Tritiated (i.e., ³H) and carbon-14 (i.e., ¹⁴C) and fluorine- 18 (¹⁸F) isotopes are useful for their ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium (i.e., ²H) may afford certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and hence may be preferred in some circumstances. Isotopically labeled compounds of the present disclosure can generally be prepared by following procedures analogous to those described in the Schemes and in the Examples herein below, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

[0086] “Prodrugs” means any compound which releases an active parent drug according to Formula (I) in vivo when such prodrug is administered to a subject. Prodrugs of a compound of Formula (I) are prepared by modifying functional groups present in the compound of Formula (I) in such a way, either in routine manipulation or in vivo, that the modifications may be cleaved in vivo to release the parent compound. Prodrugs may proceed from prodrug form to active form in a single step or may have one or more intermediate forms which may themselves have activity or may be inactive. Some prodrugs are activated enzymatically to yield the active compound, or a compound which, upon further chemical reaction, yields the active compound. Prodrugs include compounds of Formula (I) wherein a hydroxy, amino, carboxyl or sulfhydryl group in a compound of Formula (I) is bonded to any group that may be cleaved in vivo to regenerate the free hydroxyl, amino, or sulfhydryl group, respectively. Examples of prodrugs include, but are not limited to esters (e.g., acetate, formate, and benzoate derivatives), amides, guanidines, carbamates (e.g., N,N-dimethylaminocarbonyl) of hydroxy functional groups in compounds of Formula (I), and the like. Other examples of prodrugs include, without limitation, carbonates, ureides, solvates, or hydrates of the active compound. Preparation, selection, and use of prodrugs is discussed in T. Higuchi and V. Stella, “Pro-drugs as Novel Delivery Systems,” Vol. 14 of the A.C.S. Symposium Series; “Design of Prodrugs,” ed. H. Bundgaard, Elsevier, 1985; and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, each of which are hereby incorporated by reference in their entirety.

[0087] As described in The Practice of Medicinal Chemistry, Ch. 31-32 (Ed. Wermuth, Academic Press, San Diego, CA, 2001), prodrugs can be conceptually divided into two nonexclusive categories, bioprecursor prodrugs and carrier prodrugs. Generally, bioprecursor prodrugs are compounds that are inactive or have low activity compared to the corresponding active drug compound, that contain one or more protective groups and are converted to an active form by metabolism or solvolysis. Both the active drug form and any released metabolic products should have acceptably low toxicity. Typically, the formation of active drug compound involves a metabolic process or reaction that is one of the follow types:

(1) Oxidative reactions: Oxidative reactions are exemplified without limitation to reactions such as oxidation of alcohol, carbonyl, and acid functionalities, hydroxylation of aliphatic carbons, hydroxylation of alicyclic carbon atoms, oxidation of aromatic carbon atoms, oxidation of carbon-carbon double bonds, oxidation of nitrogen-containing functional groups, oxidation of silicon, phosphorus, arsenic, and sulfur, oxidative N-dealkylation, oxidative O- and S-dealkylation, oxidative deamination, as well as other oxidative reactions.

(2) Reductive reactions: Reductive reactions are exemplified without limitation to reactions such as reduction of carbonyl functionalities, reduction of alcohol functionalities and carbon-carbon double bonds, reduction of nitrogen-containing functional groups, and other reduction reactions.

(3) Reactions without change in the oxidation state: Reactions without change in the state of oxidation are exemplified without limitation to reactions such as hydrolysis of esters and ethers, hydrolytic cleavage of carbon-nitrogen single bonds, hydrolytic cleavage of non-aromatic heterocycles, hydration and dehydration at multiple bonds, new atomic linkages resulting from dehydration reactions, hydrolytic dehalogenation, removal of hydrogen halide molecule, and other such reactions.

[0088] Carrier prodrugs are drug compounds that contain a transport moiety, e.g., that improves uptake and/or localized delivery to a site(s) of action. Desirably for such a carrier prodrug, the linkage between the drug moiety and the transport moiety is a covalent bond, the prodrug is inactive or less active than the drug compound, the prodrug and any release transport moiety are acceptably non-toxic. For prodrugs where the transport moiety is intended to enhance uptake, typically the release of the transport moiety should be rapid. In other cases, it is desirable to utilize a moiety that provides slow release, e.g., certain polymers or other moieties, such as cyclodextrins. (See, e.g., Cheng et al., U.S. Patent Publ. No.

2004/0077595, incorporated herein by reference.) Such carrier prodrugs are often advantageous for orally administered drugs. Carrier prodrugs can, for example, be used to improve one or more of the following properties: increased lipophilicity, increased duration of pharmacological effects, increased site-specificity, decreased toxicity and adverse reactions, and/or improvement in drug formulation (e.g. stability, water solubility, suppression of an undesirable organoleptic or physiochemical property). For example, lipophilicity can be increased by esterification of hydroxyl groups with lipophilic carboxylic acids, or of carboxylic acid groups with alcohols, e.g., aliphatic alcohols.

[0089] The term “carrier” is also meant to include microspheres, liposomes, micelles, nanoparticles (naturally-equipped nanocarriers, for example, exosomes), and the like. It is known that exosomes can be highly effective drug carriers, and there are various ways in which drugs can be loaded into exosomes, including those techniques described in J Control Release. 2015 December 10; 219: 396-405, the contents of which are incorporated by reference in its entirety.

[0090] Metabolites, e.g., active metabolites, overlap with prodrugs as described above, e.g., bioprecursor prodrugs. Thus, such metabolites are pharmacologically active compounds or compounds that further metabolize to pharmacologically active compounds that are derivatives resulting from metabolic process in the body of a subject. Of these, active metabolites are such pharmacologically active derivative compounds. For prodrugs, the prodrug compound is generally inactive or of lower activity than the metabolic product. For active metabolites, the parent compound may be either an active compound or may be an inactive prodrug.

[0091] Prodrugs and active metabolites may be identified using routine techniques known in the art. See, e.g., Bertolini et al., 1997, J. Med. Chem., 40:2011-2016; Shan et al., 1997, J Pharm Sci 86(7):756-757 ; Bagshawe, 1995, Drug Dev. Res., 34:220-230.

[0092] “ Tautomer” means compounds produced by the phenomenon wherein a proton of one atom of a molecule shifts to another atom. See, Jerry March, Advanced Organic Chemistry: Reactions, Mechanisms and Structures, Fourth Edition, John Wiley & Sons, pages 69-74 (1992). The tautomers also refer to one of two or more structural isomers that exist in equilibrium and are readily converted from one isomeric form to another. Examples of include keto-enol tautomers, such as acetone/propen-2-ol, imine-enamine tautomers and the like, ring-chain tautomers, such as glucose/2,3,4,5,6-pentahydroxy-hexanal and the like, the tautomeric forms of heteroaryl groups containing a -N=C(H)-NH- ring atom arrangement, such as pyrazoles, imidazoles, benzimidazoles, triazoles, and tetrazoles. Where the compound contains, for example, a keto or oxime group or an aromatic moiety, tautomeric isomerism (‘tautomerism’) can occur. The compounds described herein may have one or more tautomers and therefore include various isomers. A person of ordinary skill in the art would recognize that other tautomeric ring atom arrangements are possible. All such isomeric forms of these compounds are expressly included in the present disclosure.

[0093] “ Isomers” mean compounds that have identical molecular Formulae but differ in the nature or sequence of bonding of their atoms or in the arrangement of their atoms in space. Isomers that differ in the arrangement of their atoms in space are termed “stereoisomers.” “Stereoisomer” and “stereoisomers” refer to compounds that exist in different stereoisomeric forms, for example, if they possess one or more asymmetric centers or a double bond with asymmetric substitution and, therefore, can be produced as individual stereoisomers or as mixtures. Stereoisomers include enantiomers and diastereomers. Stereoisomers that are not mirror images of one another are termed “diastereomers” and those that are non- superimposable mirror images of each other are termed “enantiomers.” When a compound has an asymmetric center, for example, an atom such as carbon bonded to four different groups, a pair of enantiomers is possible. An enantiomer can be characterized by the absolute configuration of its asymmetric center and is described by the R- and S-sequencing rules of Cahn and Prelog, or by the manner in which the molecule rotates the plane of polarized light and designated as dextrorotatory or levorotatory (i.e., as (+) or (-)-isomers respectively). A chiral compound can exist as either individual enantiomer or as a mixture thereof. A mixture containing equal proportions of the enantiomers is called a “racemic mixture.” As another example, stereoisomers include geometric isomers, such as cis- or trans- orientation of substituents on adjacent carbons of a double bond. Unless otherwise indicated, the description is intended to include individual stereoisomers as well as mixtures. The methods for the determination of stereochemistry and the separation of stereoisomers are well-known in the art (see discussion in Chapter 4 of ADVANCED ORGANIC CHEMISTRY, 6th edition J. March, John Wiley and Sons, New York, 2007) differ in the chirality of one or more stereocenters.

[0094] In the context of the use, testing, or screening of compounds that are or may be modulators, the term “contacting” means that the compound(s) are caused to be in sufficient proximity to a particular molecule, complex, cell, tissue, organism, or other specified material that potential binding interactions and/or chemical reaction between the compound and other specified material can occur.

[0095] By “assaying” is meant the creation of experimental conditions and the gathering of data regarding a particular result of the exposure to specific experimental conditions. For example, enzymes can be assayed based on their ability to act upon a detectable substrate. A compound can be assayed based on its ability to bind to a particular target molecule or molecules.

[0096] As used herein, the terms “ligand” and “modulator” are used equivalently to refer to a compound that changes (i.e., increases or decreases) the activity of a target biomolecule, e.g., an enzyme such as those described herein. Generally a ligand or modulator will be a small molecule, where “small molecule refers to a compound with a molecular weight of 1500 Daltons or less, 1000 Daltons or less, 800 Daltons or less, or 600 Daltons or less. Thus, an “improved ligand” is one that possesses better pharmacological and/or pharmacokinetic properties than a reference compound, where “better” can be defined by one skilled in the relevant art for a particular biological system or therapeutic use.

[0097] The term “binds” in connection with the interaction between a target and a potential binding compound indicates that the potential binding compound associates with the target to a statistically significant degree as compared to association with proteins generally (i.e., nonspecific binding). Thus, the term “binding compound” refers to a compound that has a statistically significant association with a target molecule. In some embodiments, a binding compound interacts with a specified target with a dissociation constant (KD) of 10 mM or less, 1,000 pM or less, 5000 nM or less, 3000 nM or less, 1500 nM or less, 1,000 nM or less, 500 nM or less, 200 nM or less, 100 nM or less, 50 nM or less, or 25 nM or less.

[0098] The term “selective” indicates that the compound binds more tightly than a reference compound, or than the same compound in a reference condition, i.e., with a lower dissociation constant. Certain compounds of this disclosure selectively inhibit wild-type Her2 and/or mutant Her to over wild-type EGFR thereby reducing EGFR dose-limiting toxicities.

[0099] In some embodiments, the greater affinity of one or more compounds in Table 1 is at least 1.5, 2, 3, 4, 5, 8, 10, 50, 100, 200, 400, 500, 1000, or 10,000-fold greater affinity. Certain compounds of this disclosure selectively inhibit wild-type Her2 and/or mutant Her to over wild-type EGFR thereby reducing EGFR dose-limiting toxicities.

[0100] The terms “modulate,” “modulation,” and the like refer to the ability of a compound to increase or decrease the function and/or expression of a target, such as the interaction between Her2 (including mutated forms thereof), where such function may include transcription regulatory activity and/or binding. Modulation may occur in vitro or in vivo. Modulation, as described herein, includes the inhibition, antagonism, partial antagonism, activation, agonism or partial agonism of a function or characteristic associated with Her2, either directly or indirectly, and/or the upregulation or downregulation of the expression Her2, either directly or indirectly. In another embodiment, the modulation is direct. Inhibitors or antagonists are compounds that, e.g., bind to, partially or totally block stimulation, decrease, prevent, inhibit, delay activation, inactivate, desensitize, or downregulate signal transduction. Activators or agonists are compounds that, e.g., bind to, stimulate, increase, open, activate, facilitate, enhance activation, activate, sensitize or upregulate signal transduction. In another example, compounds that modulate Her2 can do so by inhibiting Her2 by way of irreversible or covalent binding to the Her2 tyrosine kinase. In another example, compounds that modulate Her2 can do so by inhibiting Her2 by way of reversible or non-covalent binding to the Her2 tyrosine kinase.

[0101] As used herein, the terms “treat,” “treating,” “therapy,” “therapies,” and like terms refer to the administration of material, e.g., any one or more compound(s) as described herein in an amount effective to inhibit Her2, including wild-type Her2 and mutant Her2 such as Her2 with YVMA insertion mutations. In other embodiments of this disclosure, these terms apply to the administration of the compounds of this disclosure to subjects that have disease states associated with Her2 overexpression and/or HER2 amplification. In other embodiments, the terms “treat,” “treating,” “therapy,” “therapies,” and like terms refer to the administration of material, e.g., any one or more compound(s) as described herein is an amount effective to prevent, alleviate, or ameliorate one or more symptoms of a disease or condition, i.e., indication, and/or to prolong the survival of the subject being treated. In other embodiments of this disclosure, these terms apply to the administration of the compounds of this disclosure to subjects that have disease states associated with Her2 overexpression and/or HER2 amplification.

[0102] The terms “prevent,” “preventing,” “prevention” and grammatical variations thereof as used herein, refers to a method of partially or completely delaying or precluding the onset or recurrence of a disease, disorder or condition and/or one or more of its attendant symptoms or barring a subject from acquiring or reacquiring a disorder or condition or reducing a subject’s risk of acquiring or requiring a disorder or condition or one or more of its attendant symptoms.

[0103] As used herein, the term “subject,” “animal subject,” and the like refers to a living organism including, but not limited to, human and non-human vertebrates, e.g. any mammal, such as a human, other primates, sports animals and animals of commercial interest such as cattle, horses, ovines, or porcines, rodents, or pets such as dogs and cats.

[0104] The term “administering” refers to oral administration, administration as a suppository, topical contact, intravenous, intraperitoneal, intramuscular, intralesional, intranasal or subcutaneous administration, or the implantation of a slow-release device e.g., a mini-osmotic pump, to a subject. Administration is by any route, including parenteral and transmucosal (e.g., buccal, sublingual, palatal, gingival, nasal, vaginal, rectal, or transdermal). Parenteral administration includes, e.g., intravenous, intramuscular, intraarteriole, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial. Other modes of delivery include, but are not limited to, the use of liposomal formulations, intravenous infusion, transdermal patches, etc.

[0105] In the present context, the term “therapeutically effective” or “effective amount” indicates that a compound or material or amount of the compound or material when administered is sufficient or effective to prevent, alleviate, or ameliorate one or more symptoms of a disease, disorder or medical condition being treated, and/or to prolong the survival of the subject being treated. The therapeutically effective amount will vary depending on the compound, the disease, disorder or condition and its severity and the age, weight, etc., of the mammal to be treated. In general, satisfactory results in subjects are indicated to be obtained at a daily dosage of from about 0.1 to about 10 g/kg subject body weight. In some embodiments, a daily dose ranges from about 0.10 to 10.0 mg/kg of body weight, from about 1.0 to 3.0 mg/kg of body weight, from about 3 to 10 mg/kg of body weight, from about 3 to 150 mg/kg of body weight, from about 3 to 100 mg/kg of body weight, from about 10 to 100 mg/kg of body weight, from about 10 to 150 mg/kg of body weight, or from about 150 to 1000 mg/kg of body weight. The dosage can be conveniently administered, e.g., in divided doses up to four times a day or in sustained-release form.

[0106] As used herein, the term “Her2 mediated disease or condition” (which is also meant to mean “Her2 mediated disease or condition” as well as “wild-type Her2 and/or mutant Her2 mediated disease or condition”) refers to a disease or condition in which the biological function of Her2 affect the development and/or course of the disease or condition, and/or in which modulation of the interaction of Her2 alters the development, course, and/or symptoms. A Her2 mediated disease or condition includes a disease or condition for which the disruption of Her2 interactions (for example, by inhibiting Her2 with YVMA insertion mutations) provides a therapeutic benefit, e.g. wherein treatment with Her2 inhibitors, including compounds described herein, provides a therapeutic benefit to the subject suffering from or at risk of the disease or condition. A Her2 mediated disease or condition is intended to include a cancer or tumor that harbors loss of function mutations in Her2, or a cancer where there is activation of Her2. In another embodiments of this disclosure, Her2 mediated diseases or conditions are associated with Her2 overexpression and/or Her2 gene amplification. A Her2 mediated disease or condition is also intended to include various human carcinomas, including those of the lung, breast, stomach, ovary, colon, bladder, pancreatic cancer, biliary cancer, endometrial cancer, lung, uterine cervix, head and neck, gastric and esophageal cancer as well as uterine serous endometrial carcinoma, as well any associated comorbidities such as pulmonary disorder, hypertension, hypercholesterolemia, cardiovascular disease, renal function disorder, thyroid disorder, obesity, depression anxiety, osteoporosis, liver disorder, autoimmune disease, dementia, Alzheimer’s disease.

[0107] Also in the context of compounds binding to a biomolecular target, the term “greater specificity” indicates that a compound binds to a specified target to a greater extent than to another biomolecule or biomolecules that may be present under relevant binding conditions, where binding to such other biomolecules produces a different biological activity than binding to the specified target. Typically, the specificity is with reference to a limited set of other biomolecules, e.g., in the case of Her2 or Her2+ mutations. In particular embodiments, the greater specificity is at least 2, 3, 4, 5, 8, 10, 20, 50, 100, 200, 400, 500, or 1000-fold greater specificity.

[0108] As used herein in connection with binding compounds or ligands, the terms “specific for Her2,” (which is intended to include either wild-type Her2, mutant Her2, or both wildtype Her2 and mutant Her2) and terms of like import mean that a particular compound binds to Her2 to a statistically greater extent than to other targets that may be present in a particular sample such as wild-type EGFR. Also, where biological activity other than binding is indicated, the terms “specific for Her2” indicates that a particular compound has greater biological effect associated with binding Her2 than to other enzymes, e.g., enzyme activity inhibition.

[0109] In addition, abbreviations as used herein have respective meanings as follows:

[0110] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

Embodiments

[0111] Embodiment 1 of this disclosure relates to a radiolabeled compound having the structure of Formula I: or a pharmaceutically acceptable salt thereof, wherein:

A is selected from CH and N;

B is selected from CH2 ,CHF, and CF2;

E is selected from CH2, CHF, and CF2;

X is selected from CH, CF, C(OH) and N; or X is CH and B and E are both absent;

Q¹ is selected from the group consisting of H, Ci-Cealkyl, F, and Cl;

Q² is selected from the group consisting of halogen, haloalkyl, alkyl, alkene, alkyne, -NR^aR^b, -Ci-C6alkylene-NR^aR^b, -Ci-Cealkylene-OR⁰, cyano, hydroxyalkyl, -Co-Cealkylene-C(0)OH, -Ci-C6alkylene-C(O)O-alkyl, alkoxyalkyl, -Co-C4alkylene-cycloalkyl optionally substituted with 1-3 J⁴ groups, -Co-C4alkylene-cycloalkenyl optionally substituted with 1-3 J⁴ groups, - Co-C4alkylene-7-l l membered spirocyclic cycloalkyl, optionally substituted with 1-3 J⁴ groups, -Co-C4alkylene-7-l l membered spirocyclic heterocycloalkyl optionally substituted with 1-3 J⁴ groups, -Co-C4alkylene-heterocycloalkyl optionally substituted with 1-3 J⁴ groups, and -Co-C4alkylene-heterocycloalkenyl optionally substituted with 1-3 J⁴ groups; each J⁴ is independently selected from the group consisting of halogen, alkyl, haloalkyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, oxo, and -Co-C4alkylene-NR^aR^b, provided that J⁴ groups can only include up to two oxo groups and up to one -Co-C4alkylene-NR^aR^b group; Q³ is selected from H and F; R^a and R^b each are independently selected from the group consisting of Ci-Cealkyl, Ci- Cehaloalkyl, Ci-Cehydroxy alkyl, and -Ci-Cealkyl-Ci-Cealkoxy;

R¹ is selected from the group consisting of alkyl, haloalkyl and halogen;

R² is selected from the group consisting of is -O-alkyl, -O-aryl, -O-heteroaryl, -O-cycloalkyl, -O-heterocycloalkyl, -O-heteroaryl-alkylene-aryl, -NH-alkyl, -NH-aryl, or-NH-heteroaryl, wherein each of the alkyl, aryl, heteroaryl, cycloalkyl or heteocycloalkyl moieties are optionally substituted with 1-4 J¹ groups; or R¹ and R² join with the carbon atoms to which they are attached to form a saturated or unsaturated carbocyclic or heterocyclic ring, wherein the saturated or unsaturated carbocyclic or heterocyclic ring is optionally substituted with 1-4 J¹ groups; each J¹ is independently selected from the group consisting of halogen, alkyl, haloalkyl, hydroxy, hydroxyalkyl, -Co-C4alkylene-N(H)R^C, alkoxy, and alkoxyalkyl; and wherein at least one carbon atom of Formula (I) is substituted with at least one ¹⁸F, and the at least one ¹⁸F atom can substitute any hydrogen or halogen atom in Formula (I) provided that a C-¹⁸F bond is formed.

[0112] Embodiment 2 of this disclosure relates to the radiolabeled compound of Embodiment 1, wherein the radiolableled compound has a structure of Formula (II): or a pharmaceutically acceptable salt thereof.

[0113] Embodiment 3 of this disclosure relates to the radiolabeled compound of Embodiments 1 or 2, wherein Q¹ is H or Ci-Ce alkyl.

[0114] Embodiment 4 of this disclosure relates to the radiolabeled compound of Embodiment 3, wherein Q¹ is H.

[0115] Embodiment 5 of this disclosure relates to the radiolabeled compound of Embodiment 3, wherein Q¹ is methyl, ethyl, or propyl. [0116] Embodiment 6 of this disclosure relates to the radiolabeled compound of Embodiments 1 to 5, wherein Q² is -Ci-C3alkylene-NR^aR^b, -Co-C4alkylene-cycloalkyl optionally substituted with 1-3 J⁴ groups, or -Co-C4alkylene-heterocycloalkyl optionally substituted with 1-3 J⁴.

[0117] Embodiment 7 of this disclosure relates to the radiolabeled compound of Embodiment 6, wherein Q² is -Ci-C3alkylene-NR^aR^b.

[0118] Embodiment 8 of this disclosure relates to the radiolabeled compound of Embodiment 6, wherein Q² is -Co-C4alkylene-heterocycloalkyl optionally substituted with 1-3 J⁴.

[0119] Embodiment 9 of this disclosure relates to the radiolabeled compound of Embodiments 1 to 8, wherein each J⁴ is independently halogen, alkyl, haloalkyl, hydroxy, hydroxy alkyl, alkoxy, or alkoxy alky.

[0120] Embodiment 10 of this disclosure relates to the radiolabeled compound of Embodiment 9, wherein each J⁴ is independently halogen or alkyl.

[0121] Embodiment 11 of this disclosure relates to the radiolabeled compound of Embodiment 10, wherein each J⁴ is independently halogen.

[0122] Embodiment 12 of this disclosure relates to the radiolabeled compound of Embodiment 10, wherein each J⁴ is independently C1-C3 alkyl.

[0123] Embodiment 13 of this disclosure relates to the radiolabeled compound of Embodiments 1 to 12, wherein X is CH, CF, or C(OH).

[0124] Embodiment 14 of this disclosure relates to the radiolabeled compound of Embodiment 13, wherein X is CH or CF.

[0125] Embodiment 15 of this disclosure relates to the radiolabeled compound of Embodiment 14, wherein X is CH.

[0126] Embodiment 16 of this disclosure relates to the radiolabeled compound of Embodiments 1 to 15, wherein at least one carbon atom of Q¹, Q², B, E, or X that has a single bond to either a hydrogen or halogen atom is substituted with ¹⁸F.

[0127] Embodiment 17 of this disclosure relates to the radiolabeled compound of Embodiment 16, wherein at least one carbon atom of Q¹, Q², B, or E that has a single bond to either a hydrogen or halogen atom is substituted with ¹⁸F.

[0128] Embodiment 18 of this disclosure relates to the radiolabeled compound of Embodiment 17, wherein at least one carbon atom of Q¹ or Q² that has a single bond to either a hydrogen or halogen atom is substituted with ¹⁸F.

[0129] Embodiment 19 of this disclosure relates to the radiolabeled compound of Embodiments 1 to 18, wherein the compound has the following structure:

or a pharmaceutically acceptable salt thereof.

[0130] Embodiment 20 of this disclosure relates to a method of selecting a subject for treating a HER2 mediated disease, the method comprising administering to the subject a radiolableled compound of any one of Embodiments 1 to 19, or a pharmaceutically acceptable salt thereof, generating a scanning image of the subject, and selecting the subject for treating the HER2 mediated disease when the scanning image shows that the radiolableled compound, or a pharmaceutically acceptable salt thereof, is accumulated in a tumor tissue. [0131] Embodiment 21 of this disclosure relates to the method of Embodiment 20, wherein the radiolableled compound, or a pharmaceutically acceptable salt thereof, is administered via systemic administration.

[0132] Embodiment 22 of this disclosure relates to the method of Embodiment 20 or 21, wherein the radiolableled compound, or a pharmaceutically acceptable salt thereof, is accumulated in the brain.

[0133] Embodiment 23 of this disclosure relates to the method of Embodiments 20-22, wherein the scanning image is a tomographic image.

[0134] Embodiment 24 of this disclosure relates to the method of Embodiment 23, wherein the tomographic image is generated with positron emission tomography (PET).

[0135] Embodiment 25 of this disclosure relates to the method of Embodiment 20-24, wherein the HER2 mediated disease is metastatic brain tumor.

[0136] Embodiment 26 of this disclosure relates to the method of Embodiments 25, wherein the metastatic brain tumor is caused by lung cancer, breast cancer, skin cancer, colon cancer, or melanoma.

[0137] Embodiment 27 of this disclosure relates to the method of Embodiment 26, wherein the metastatic brain tumor is caused by breast cancer.

[0138] Embodiment 28 of this disclosure relates to the method of Embodiments 20-24, wherein the HER2 mediated disease is cancer.

[0139] Embodiment 29 of this disclosure relates to the method of Embodiment 28, wherein the cancer is brain cancer, colorectal cancer, breast cancer, bladder cancer, biliary cancer, ovarian cancer, or non-small cell lung cancer.

[0140] Embodiment 30 of this disclosure relates to the radiolabeled compound of Embodiment 29, wherein the cancer is brain cancer.

[0141] Embodiment 31 of this disclosure relates to a method of tracking tumor response to a HER2 mediated disease in a subject, the method comprising administering to the subject a radiolableled compound of any one of Embodiments, or a pharmaceutically acceptable salt thereof, and generating a scanning image of the subject.

[0142] Embodiment 32 of this disclosure relates to the method of Embodiment 31, wherein the scanning image is generated once every two weeks.

[0143] Embodiment 33 of this disclosure relates to the method of Embodiment 32, wherein the scanning image is generated for at least 2 months.

[0144] Embodiment 34 of this disclosure relates to the method of Embodiments 31 to 33, wherein the radiolableled compound, or a pharmaceutically acceptable salt thereof, is administered via systemic administration.

[0145] Embodiment 35 of this disclosure relates to the method of Embodiments 31 to 34, wherein the radiolableled compound, or a pharmaceutically acceptable salt thereof, is accumulated in the brain.

[0146] Embodiment 36 of this disclosure relates to the method of Embodiments 31 to 35, wherein the scanning image is a tomographic image.

[0147] Embodiment 37 of this disclosure relates to the method of Embodiment 36, wherein the tomographic image is generated with positron emission tomography (PET).

[0148] Embodiment 38 of this disclosure relates to the method of Embodiments 31 to 37, wherein the HER2 mediated disease is metastatic brain tumor.

[0149] Embodiment 39 of this disclosure relates to the method of Embodiment 38, wherein the metastatic brain tumor is caused by lung cancer, breast cancer, skin cancer, colon cancer, or melanoma.

[0150] Embodiment 40 of this disclosure relates to the method of Embodiment 39, wherein the metastatic brain tumor is caused by breast cancer.

[0151] Embodiment 41 of this disclosure relates to the method of Embodiments 31 to 37, wherein the HER2 mediated disease is cancer.

[0152] Embodiment 42 of this disclosure relates to the method of Embodiments 41, wherein the cancer is brain cancer, colorectal cancer, breast cancer, bladder cancer, biliary cancer, ovarian cancer, or non-small cell lung cancer.

[0153] Embodiment 43 of this disclosure relates to the method of Embodiments 42, wherein the cancer is brain cancer.

[0154] Embodiment 44 of this disclosure relates to a method of obtaining a scanning image, comprising administering to a subject the radiolableled compound of any one of Embodiments 1 to 19, or a pharmaceutically acceptable salt thereof, and subsequently generating a scanning image of the subject.

[0155] Embodiment 45 of this disclosure relates to the method of Embodiment 44, wherein the scanning image is a tomographic image.

[0156] Embodiment 46 of this disclosure relates to the method of Embodiment 45, wherein the tomographic image is generated with positron emission tomography (PET).

EXAMPLES

[0157] The following illustrative examples are representative of embodiments of the stimulation, systems, and methods described herein and are not meant to be limiting in any way.

[0158] Compounds of Formula I and II of the present disclosure may be synthesized in accordance with the schemes and examples described below. The examples may be altered by substitution of the starting materials with other materials having similar structures to result in corresponding products. The structure of the desired product will generally make apparent to a person of skill in the art the required starting materials. The installation of ¹⁸F radiolabels on the compounds of disclosure would be understood by a person of skill in the art (Bioconjugate Chem. 2015, 26, 1, 1-18). General synthetic schemes for compound of the disclosure are provided herein.

General Schemel

[0159] Step 1: Compound (i) can be converted to Compound (ii) through the application of a peptide coupling reagent such as PyBroP (by way of example) in appropriate reaction conditions which may be in the presence of a tertiary amine such as triethylamine. The reaction can take place in an appropriate solvent which may be aprotic solvent such as THF but may vary depending on the starting materials or intermediate compounds. Variables El, E2, A, G, Rl, and R2 in General Scheme 1 are as defined in this disclosure. Variable X in

General Scheme 1 is an appropriate leaving group such as Br or Cl.

[0160] Step 2: Compound (ii) can be converted to Compound I by cross-coupling reactions by way of example such as palladium catalyzed Suzuki coupling with an organoborate such by way of example to arrive at Compound I. Variable G can be further modified one or more times by techniques described in this disclosure or by techniques known in the art. General Scheme 2

[0161] Step 1’: Compound (iv) can be converted to Compound (v) through the application of a peptide coupling reagent such as PyBroP by way of example in the presence of a tertiary amine such as triethylamine. The reaction can take place in an aprotic solvent such as THF. Variables El, E2, A, Rl, and R2 in General Scheme 1 are as defined in this disclosure. Variable G’ in General Scheme 1 can be a BOC-protected G group or another precursor that can be modified one or more times by techniques described in this disclosure or by techniques known in the art. G’ can also the same as variable G as described in this disclosure in which there would be no Step 2’ to modify variable G’.

[0162] Step 2’: Compound (v) can be converted to Compound I by one or more techniques described in this disclosure or known in the art. Such one or more techniques may include by way of example BOC deprotection, peptide coupling reactions with HATU, amide formation with HOBt, or nucleophilic substitution.

Synthesis of Intermediate A Intermediate A

Step 1. Methyl l-amino-3-bromo-lH-pyrrole-2-carboxylate

[0163] A solution of methyl 3-bromo-lH-pyrrole-2-carboxylate (25 g, 122.53 mmol) in DMF (200 mL) and THF (1000 mL) was treated with NaH (60% in mineral oil, 6.37 g, 159.25 mmol) for 1 hour at 0 °C followed by the addition of O-(2,4-dinitrophenyl)hydroxylamine (29.28 g, 147.04 mmol) at 0 °C and the mixture was stirred for 16 hours at room temperature. The reaction was quenched by the addition of saturated ammonium chloride aqueous solution (500 ml) at 0 °C. The resulting mixture was diluted with water (1 L) and extracted with ethyl acetate (1.5 L x 2). The combined organic layers were washed with brine (1.5 L x 2), dried over anhydrous sodium sulfate, concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0 - 20% ethyl acetate in hexanes) to provide methyl l-amino-3-bromo-lH-pyrrole-2-carboxylate (22 g, 81.97%). LCMS (ESI-MS) m/z = 219.0 [M+H]⁺

Step 2. 5-bromopyrrolo[2,l-f][l,2,4]triazin-4(3H)-one H N^NH

[0164] To a stirred solution of methyl l-amino-3-bromo-lH-pyrrole-2-carboxylate (22 g,

100.43 mmol) in iPrOH (150 mL) was added formimidamide acetate (20.91 g, 200.87 mmol). The mixture was stirred at 80 °C overnight. The resulting mixture was diluted with water (300 ml). The precipitated solids were collected by filtration and washed with water (100 ml x 3) and petroleum ether (200 ml) to afford 5-bromopyrrolo[2,l-f][l,2,4]triazin-4(3H)-one (13.2 g crude). LCMS (ESI-MS) m/z = 214.0 [M+H]⁺.

Step 3. N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5- bromopyrrolo[2,l-f][l,2,4]triazin-4-amine

[0165] A solution of 5-bromopyrrolo[2,l-f][l,2,4]triazin-4(3H)-one (13.2 g, 61.97 mmol), 4- ([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylaniline (14.87 g, 61.97 mmol), PyBrop (43.31 g, 92.95 mmol) and EtsN (18.81g, 185.91 mmol) in THF (300 mL) was stirred overnight at 80 °C. The resulting mixture was purified by silica gel column chromatography (0 - 80% ethyl acetate in hexanes) to afford the title compound N-(4-([l,2,4]triazolo[l,5- a]pyridin-7-yloxy)-3-methylphenyl)-5-bromopyrrolo[2,l-f][l,2,4]triazin-4-amine, intermediate A (12 g, 27.33%). LCMS (ESI-MS) m/z = 436.0 [M+H]⁺.

[0166] The following Example numbers correspond to the Example numbers (E#s) in Table 1 of this disclosure.

Example 1

[0167] To a solution of methanesulfonylbenzene (16.7 g, 107 mmol) in anhydrous tetrahydrofuran (160 mL) at -20 °C under nitrogen atmosphere was added to lithium bis(trimethylsilyl)amide (IM solution in THF, 189 mL, 189 mmol) dropwise and the reaction was allowed to stir for 30 minutes at -20 °C. To the reaction mixture was added chlorotimethylsilane (12.6 ml, 99.2 mmol) and slowed to stirred for a further 15 minutes. To the reaction mixture was added a solution of tert-butyl 3 -formylazetidine- 1 -carboxylate (19.8 g, 106.9 mmol) in anhydrous tetrahydrofuran (200 mL) dropwise and allowed to stir at -20 °C for further 3 hours. The operation was repeated twice. The reaction mixture was quenched with saturated aqueous ammonium chloride (I L) and extracted with ethyl acetate (2 x 1 L). The combined organics were dried over sodium sulfate filtered and concentrated under vacuum to afford the crude product. The crude product was purified by column chromatography (silica, petroleum ether / ethyl acetate ,15%) to give /c/7-butyl(/ )-3-(2- (phenylsulfonyl)vinyl)azetidine-l -carboxylate (52 g, 47.6%). LCMS (ES MS) m/z = 324.1 [M+H]⁺.

Step 2. Ethyl 3-(l-(tert-butoxycarbonyl)azetidin-3-yl)-lH-pyrrole-2-carboxylate

[0168] Potassium 2-methylpropan-2-olate (11.0 g, 98.2 mmol) was added to a mixture of ethyl 2-isocy anoacetate (8.4 g, 37.1 mmol) in tetrahydrofuran (100 mL) under nitrogen atmosphere at 0 °C and stirred for 10 minutes. Then tert-butyl (E)-3-(2- (phenylsulfonyl)vinyl)azetidine-l -carboxylate (20 g, 61.8 mmol) in THF (100 mL) was added to mixture and stirred at 25 °C over the time of one hour. The operation was repeated twice. The reaction was quenched by the addition of saturated aqueous ammonium chloride (500 mL) at 0 °C. The resulting mixture was extracted with ethyl acetate (3 x 500 mL). The combined organic layers were washed with brine (3 x 500 mL), dried over anhydrous sodium sulfate. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with petroleum ether / ethyl acetate (10%) to afford ethyl 3-(l-(tert-butoxycarbonyl)azetidin-3-yl)-lH-pyrrole-2-carboxylate (27 g, 42.2%). LCMS (ESI-MS) m/z = 295.2 [M+H]⁺.

Step 3. Ethyl l-amino-3-(l-(tert-butoxycarbonyl)azetidin-3-yl)-lH-pyrrole-2- carboxylate

[0169] To a flask was added MTBE (1 L) and ammonium chloride (30 g, 0.565 mol). The reaction was cooled to -20 °C. Then concentrated aq. ammonium hydroxide (80 mL) was added to the reaction followed by slow addition of commercial-grade sodium hypochlorite solution (750 mL). After addition, the reaction was stirred at -20 °C for additional 30 minutes. The MTBE layer was separated and washed with brine and dried over anhydrous sodium sulfate. In a separate flask under nitrogen was added ethyl 3-(l-(tert- butoxycarbonyl)azetidin-3-yl)-lH-pyrrole-2-carboxylate (27 g, 91.8 mmol) and dry DMF (300 mL). The reaction was cooled to 0 °C and sodium hydroxide (7.3g, 183.6 mmol) was added portionwise to the reaction. The reaction was stirred at 0 °C for additional 1 hour before it was cooled to -20 °C. At this time, the previously prepared MTBE solution of chloramine was added slowly to the reaction and the mixture was stirred at -20 °C for 1 hour. The reaction was quenched with saturated sodium thiosulfate solution. The organic layer of the reaction was separated and washed with water and brine, dried over sodium sulfate, filtered and concentrated to afford ethyl l-amino-3-(l-(tert-butoxycarbonyl)azeti din-3 -yl)- lH-pyrrole-2-carboxylate (21 g, 60.3%). LCMS (ESI-MS) m/z = 310.2 [M+H]⁺.

Step 4. Tert-butyl 3-(4-oxo-3,4-dihydropyrrolo[2,l-f][l,2,4]triazin-5-yl)azetidine-l- carboxylate

[0170] Acetic acid methanimidamide (7.06 g, 67.8 mmol) was added to a mixture of ethyl 1- amino-3-[l-(tert-butoxycarbonyl)azetidin-3-yl]pyrrole-2-carboxylate (4 g, 13.5 mmol) in iPrOH (15 mL).Then the reaction mixture was stirred at 80 °C for 12 hours. The reaction mixture was concentrated under vacuum and purified by silica gel column chromatography, eluted with PE / EA (37%) and concentrated to afford tert-butyl 3-(4-oxo-3,4- dihydropyrrolo[2,l-f][l,2,4]triazin-5-yl)azetidine-l-carboxylate (15 g, 76.1%). LCMS (ESIMS) m/z = 291.1 [M+H]⁺.

Step 5. Tert-butyl 3-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)azetidine-l-carboxylate [0171] Bromotris(pyrrolidin-l-yl)phosphanium; hexafluoro-lambda5-phosphanuide (2.41 g, 5.16 mmol) was added to a mixture of tert-butyl 3-(4-oxo-3,4-dihydropyrrolo[2,l- f][l,2,4]triazin-5-yl)azetidine-l-carboxylate (1 g, 3.44 mmol), 4-([l,2,4]triazolo[l,5- a]pyridin-7-yloxy)-3-methylaniline (830 mg, 3.44 mmol) and triethylamine (1.1g g, 10.33 mmol) in THF (30 mL) .Then the reaction mixture was stirred at 80 °C for overnight. The resulting mixture was cooled down to room temperature, filtered, the filter cake was washed with di chloromethane (3 x 50 mL). The filtrate was concentrated under reduced pressure to afford the crude and then purified by silica gel column chromatography, eluted with petroleum ether / ethyl acetate (1 :6) to afford the crude 1.2 g (contains 40% of SM). Then the crude material was re-purified by reversed-phase flash chromatography with the following conditions: column, Cl 8 silica gel; mobile phase, ACN in Water (10 mmol/L NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm and concentrated to give tert-butyl 3- (4-((4-([l, 2, 4]tri azolof l,5-a]pyridin-7-yloxy)-3-methylphenyl)amino)pyrrolo[2,l- f][l,2,4]triazin-5-yl)azetidine-l-carboxylate (553 mg, 31.25%). LCMS (ESI-MS) m/z = 513.2 [M+H]⁺.

Step 6. N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(azetidin-3- yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine

[0172] A solution of TFA (1 mL) and tert-butyl 3-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7- yloxy)-3-methylphenyl)amino)pyrrolo[2, l-f][l, 2, 4]triazin-5-yl)azetidine-l -carboxylate (300 mg, 0.58 mmol) in DCM (2 mL) was stirred for 1 hour at 25 °C. The resulting mixture was concentrated under vacuum to afford the crude N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)- 3-methylphenyl)-5-(azetidin-3-yl)pyrrolo [2,l-f][l,2,4]triazin-4-amine (250 mg). LCMS (ESI-MS) m/z = 413.2 [M+H]⁺.

Step 7. (E)-l-(3-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)azetidin-l-yl)-4- (dimethylamino)but-2-en-l-one

[0173] A solution of N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5- (azetidin-3-yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine (250 mg, 0.61 mmol), (2E)-4- (dimethylamino)but-2-enoic acid (94 mg, 0.72 mmol), N,N,N,N-Tetramethyl-O-(7- azabenzotriazol-l-yl)uronium hexafluorophospate (277 mg, 0.72 mmol) and N,N- Diisopropylethylamine (237 mg, 1.81 mmol) in DMF (5 mL) was stirred overnight at room temperature. The resulting mixture was purified by reverse phase flash with the following conditions (5 mmol/L NH4HCO3, Flow rate: 50 mL/min, 30%) to afford to afford (E)-l-(3- (4-((4-([l, 2, 4]tri azolof l,5-a]pyridin-7-yloxy)-3-methylphenyl)amino)pyrrolo[2,l- f][l,2,4]triazin-5-yl)azetidin-l-yl)-4-(dimethylamino)but-2-en-l-one, Example 1 (138 mg, 42.9%). LCMS (ESI-MS) m/z = 524.1 [M+H]⁺.

Example 6 step 2 2 step 3 Example 6

Step 1. T ert-butyl-4-(4-((4-( [1 ,2,4] triazolo [1 ,5-a] pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)piperazine-l-carboxylate

[0174] A mixture of N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5- bromopyrrolo[2,l-f][l,2,4]triazin-4-amine (200 mg, 0.45 mmol), tert-butyl piperazine-1- carboxylate (93.92 mg, 0.50 mmol), Pd2(dba)s (41.98 mg, 0.05 mmol), BINAP (57.09 mg, 0.09 mmol) and t-BuONa (88.11 mg, 0.91 mmol) in dioxane (4 mL) was stirred for 72 hours at 100 °C under nitrogen atmosphere. The reaction mixture was filtered and the filtrate was concentrated to afford the crude product. The crude product was purified by column chromatography (silica gel, 25 g, eluted with ethyl acetate in petroleum ether from 0% to 80% with 20 mL/min flow rate), the desired fractions were combined and concentrated under vacuum to afford the desired product tert-butyl 4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7- yloxy)-3-methylphenyl)amino)pyrrolo [2, l-f][l,2,4]triazin -5-yl)piperazine-l-carboxylate (200 mg, 69%). LCMS (ESI-MS) m/z =542.3 [M+H]⁺.

Step 2. N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(piperazin-l- yl)pyrrolo[2,l-f|[l,2,4]triazin-4-amine

[0175] A mixture of tert-butyl 4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)piperazine-l-carboxylate (200 mg, 0.37 mmol) and TFA (3 mL, 39.99 mmol) in DCM (1 mL) was stirred for 1 hour at room temperature. The resulting mixture was concentrated under vacuum to afford the crude product. The crude product was purified by column chromatography (silica-gel, 25 g, eluted with methanol in dichloromethane from 0% to 10% with 20 mL/min flowrate), the desired fractions were combined and concentrated under vacuum to afford N-(4-([l,2,4]triazolo[l,5- a]pyridin-7-yloxy)-3 -methylphenyl)-5-(piperazin- 1 -yl)pyrrolo[2, 1 -f] [ 1 ,2,4]triazin-4-amine (210 mg crude). LCMS (ESI-MS) m/z = 442.2 [M+H]⁺.

Step 3. l-(4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l, 2,4]triazin-5-yl)piperazin-l-yl)prop-2-en-l-one

[0176] A mixture ofN-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5- (piperazin-l-yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine (210 mg, 0.47 mmol), acryloyl chloride (43.05 mg, 0.47 mmol) and EtsN (144.40 mg, 1.41 mmol) in DCM (2 mL) was stirred for 5 minutes at 0°C. The reaction mixture was purified by column chromatography (silica-gel, 25g, eluted with ethyl acetate in petroleum ether from 0% to 80% with 20mL/min). The fractions with desired mass signal were combined and concentrated under vacuum to afford the desired product l-(4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2, 1 -f] [ 1 ,2,4] triazin-5-yl)piperazin- 1 -yl)prop-2-en- 1 -one, Example 6 (27.5 mg, 11.62%). LCMS (ESI-MS) m/z = 496.2 [M+H]⁺.

Example 7 Step 1. tert-butyl 4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)-3,6-dihydropyridine-l(2H)- carboxylate

[0177] A solution of N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5- bromopyrrolo[2,l-f][l,2,4]triazin-4-amine (200 mg, 0.46 mmol), tert-butyl 4-(4,4,5,5- tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-l(2H)-carboxylate (141.75 mg, 0.46 mmol), Pd(dppf)C12 (37.34 mg, 0.05 mmol) and K2CO3 (126.71 mg, 0.92 mmol) in dioxane (4 mL) and H2O (1.2 mL) was stirred for 2 hours at 100 °C under nitrogen atmosphere. The resulting mixture was concentrated under vacuum to afford the crude product. The crude product was purified by Prep-TLC (petroleum ether / ethyl acetate 1 : 10) to afford tert-butyl tert-butyl 4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)-3,6-dihydropyridine-l(2H)- carboxylate (220 mg, purity = 96.8 %). LCMS (ESI-MS) m/z = 539.2 [M+H]⁺.

Step 2. Tert-butyl 4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)piperidine-l-carboxylate [0178] A solution of 4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)-3,6-dihydropyridine-l(2H)- carboxylate (200 mg, 0.37 mmol) and Pd/C (395.16 mg, 3.71 mmol) in MeOH was stirred overnight at room temperature under hydrogen atmosphere. The mixture was filtered off and the filtrate was concentrated under vacuum to afford crude product. The crude product was used in the next step directly without further purification. LCMS (ESI-MS) m/z = 541.3 [M+H]⁺.

Step 3. N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(piperidin-4- yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine

[0179] A solution of tert-butyl 4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)piperidine-l-carboxylate (140 mg, 0.26 mmol) in TFA (2 mL) was stirred for 1 hour at room temperature. The resulting mixture was concentrated under vacuum to afford the crude product. The crude product was used in the next step directly without further purification. LCMS (ESI-MS) m/z = 441.2 [M+H]⁺.

Step 4. l-(4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)piperidin-l-yl)prop-2-en-l-one

[0180] A solution of N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5- (piperidin-4-yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine (70 mg, 0.16 mmol), acryloyl chloride (14.38 mg, 0.16 mmol) and EtsN (32.16 mg, 0.32 mmol) in DCM (2 mL) was stirred for 5 minutes at 0 °C. The resulting mixture was purified by Prep-TLC (ethyl acetate) to afford 1- (4-(4-((4-([ 1 ,2,4]triazolo[ 1 ,5 -a]pyridin-7-yloxy)-3 -methylphenyl) amino)pyrrolo[2, 1 - f][l,2,4]triazin-5-yl)piperidin-l-yl)prop-2-en-l-one, Example 7 (24.6 mg, 30.55%). LCMS (ESI-MS) m/z = 495.2 [M+H]⁺.

Example 10

Step 1. Tert-butyl (E)-3-(3-(dimethylamino)acryloyl)azetidine-l-carboxylate

[0181] A solution of tert-butyl 3 -acetylazetidine- 1 -carboxylate (2 g, 10.03 mmol) in DMF- DMA (15 mL) was stirred overnight at 110 °C. The resulting mixture was concentrated under vacuum to afford tert-butyl 3-[(2E)-3-(dimethylamino)prop-2-enoyl]azetidine-l-carboxylate (2.3 g, crude). .LCMS (ESI-MS) m/z = 255.2 [M+H]⁺

Step 2. Tert-butyl 3-(lH-pyrazol-3-yl)azetidine-l-carboxylate

[0182] A solution of tert-butyl 3-[(2E)-3-(dimethylamino)prop-2-enoyl]azetidine-l- carboxylate (2.1 g, 8.25 mmol) in hydrazine hydrate (20 mL) was stirred overnight at 80 °C. The resulting mixture was concentrated under vacuum. The residue was purified by silica column chromatography (0 - 40% ethyl acetate in hexanes) to provide the title compound tert-butyl 3-(lH-pyrazol-3-yl)azetidine-l-carboxylate (1.5 g, 66.92%). LCMS (ESI-MS) m/z = 447.3 [2M+H]⁺

Step 3. Tert-butyl 3-(l-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)-lH-pyrazol-3-yl)azetidine-l- carboxylate

[0183] To a stirred mixture of tert-butyl 3-(lH-pyrazol-3-yl)azetidine-l-carboxylate (200 mg, 0.89 mmol) and N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5- bromopyrrolo[2,l-f][l,2,4]triazin-4-amine (781.59 mg, 0.89 mmol) in toluene (3 mL) was added (lR,2R)-cyclohexane-l,2-diamine (153.43 mg, 1.34 mmol) and potassium phosphate (380.27 mg, 1.79 mmol) and cuprous iodide (85.30 mg, 0.44 mmol) under nitrogen atmosphere. The resulting mixture was stirred at 100 °C and overnight. The reaction mixture was filtered off and the filtrate was concentrated. The residue was purified by silica column chromatography (0 - 40% ethyl acetate in hexanes) to provide the title compound tert-butyl 3- ( 1 -(4-((4-([ 1 ,2,4]triazolo[ 1 ,5 -a]pyridin-7-yloxy)-3 -methylphenyl)amino)pyrrolo[2, 1 - f][l,2,4]triazin-5-yl)-lH-pyrazol-3-yl)azetidine-l-carboxylate (80 mg, 15.43%). LCMS (ESIMS) m/z = 579.3 [M+H]⁺.

Step 4. N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(3-(azetidin-3-yl)- lH-pyrazol-l-yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine

[0184] To a stirred solution of tert-butyl 3-(l-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-

3-methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)-lH-pyrazol-3-yl)azetidine-l- carboxylate (80 mg, 0.13 mmol) in dichloromethane (10 mL) was added tri fluoroacetic acid (1 mL). The resulting mixture was stirred at room temperature for 0.5 hour and concentrated under vacuum to afford N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(3- (azetidin-3-yl)-lH-pyrazol-l-yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine (80 mg crude). LCMS (ESI-MS) m/z= 479.2 [M+H]⁺ .

Step 5. l-(3-(l-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f|[l,2,4]triazin-5-yl)-lH-pyrazol-3-yl)azetidin-l- yl)prop-2-en-l-one

[0185] To a stirred mixture of N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)- 5-(3-(azetidin-3-yl)-lH-pyrazol-l-yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine (80 mg crude) and EtsN (67.67 mg, 0.66 mmol) in dichloromethane (2 mL) was added acryloyl chloride (15.13 mg, 0.16 mmol) dropwise at 0 °C and stirred for 3 minutes. The reaction mixture was purified by Prep-TLC to afford the crude product. The crude product (60 mg) was purified by Prep- HPLC with the following conditions (Column: XBridge Shield RP18 OBD Column, 30*150 mm, 5pm; Mobile Phase A: Water(10 mmol/L NH4HCO3+0.1%NH3.H2O), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 28% B to 58% B in 7 min, 58% B) to afford l-(3-(l- (4-((4-([l, 2, 4]tri azolof l,5-a]pyridin-7-yloxy)-3-methylphenyl)amino)pyrrolo[2,l- f][l,2,4]triazin-5-yl)-lH-pyrazol-3-yl)azetidin-l-yl)prop-2-en-l-one, Example 10 (11.2 mg, 12.57%). LCMS (ESI-MS) m/z= 533.1 [M+H]⁺ .

Example 21

(£)-l-(4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-/|[l,2,4]triazin-5-yl)piperidin-l-yl)-4-

(dimethylamino)but-2-en-l-one

[0186] To a 20 mL scintillation vial with Teflon-coated stir bar was added A-(4-

([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(piperidin-4-yl)pyrrolo[2,l- /|[l,2,4]triazin-4-amine (described in step 3 of of Example 4) (TFA salt, 50.0 mg, 1 Eq, 90.2 pmol), (E)-4-(dimethylamino)but-2-enoic acid hydrochloride (22.4 mg, 1.5 Eq, 135 pmol), and HATU (51.4 mg, 1.5 Eq, 135 pmol). The vial was then capped with a rubber septum and evacuated and refilled with N2 (3 x). Then DMF (3.0 mL) and Diisopropylethylamine (46.6 mg, 62.3 pL, 4 Eq, 361 pmol) were added via syringe and the reaction was stirred overnight at rt. The resulting mixture was filtered then purified by preparative reverse phase HPLC (acetonitrile/water gradient with 0.1% TFA) to give the title compound, Example 21 (2.0 mg, 3.3% yield. LCMS (ESI) [M+H]⁺ = 552.3.

Example 42

Step 1. (E)-4-(4-methoxypiperidin-l-yl)but-2-enoic acid

[0187] To a stirred mixture of (E)-4-bromobut-2-enoic acid (50 mg, 0.30 mmol) and 4- methoxy-piperidine HC1 (51 mg, 0.33 mmol) in DMF (1 mL), diisopropylethylamine (0.12 g, 0.91 mmol) was added, and the reaction stirred at RT overnight. The reaction mixture was used as is in the following step.

Step 2. 7V-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(azetidin-3- yl)pyrrolo[2,l-/][l,2,4]triazin-4-amine

[0188] TFA (3 mL) was added to tert-butyl 3-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)- 3-methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)azetidine-l-carboxylate (438 mg, 855 pmol) at RT and the mixture was stirred for 10 min. The resulting mixture was concentrated under vacuum to afford the crude product. The crude was diluted with ethyl acetate (2 x 20 mL) and washed with NaHCCf (20 mL). The organic layers were dried over MgSCh, and was concentrated to provide N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5- (azetidin-3-yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine that was used crude in the next step. LCMS (ESI-MS) m/z = 413.2 [M+H]⁺ .

Step 3. (£)-l-(3-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-/|[l,2,4]triazin-5-yl)azetidin-l-yl)-4-(4- methoxypiperidin-l-yl)but-2-en-l-one

[0189] O-(Benzotriazol-l-yl)-N,N,N’,N’-tetramethyluroniumTetrafluoroborate (TBTU) (93 mg, 0.29 mmol) and N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5- (azetidin-3-yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine (40 mg, 97 pmol) were added to the crude mixture of (E)-4-(4-methoxypiperidin-l-yl)but-2-enoic acid (58 mg, 0.29 mmol) in DMF (ImL) and diisopropylethylamine (from step 1). The reaction stirred at room temperature for 30 min.

[0190] The reaction mixture was filtered and purified by prep HPLC, eluted with 10-40%

ACN/ water/0.1% TFA. Fractions were diluted with ethyl acetate (20mL) and washed with saturated NaHCCf (20 mL), the organic layer was filtered over MgSCU, and the solvent was evaporated to provide (E)-l-(3-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2, 1 -f] [ 1 ,2,4]triazin-5-yl)azetidin- 1 -yl)-4-(4-methoxypiperidin- l-yl)but-2-en-l-one, Example 42 (17.3 mg, 29 %). 'HNMR (499 MHz, CHLOROFORM-d) 5 = 8.50 (d, J = 7.4 Hz, 1H), 8.23 (s, 1H), 8.00 (s, 1H), 7.63 (d, J = 2.7 Hz, 1H), 7.60 (dd, J = 2.6, 8.6 Hz, 1H), 7.57 (d, J = 2.5 Hz, 1H), 7.10 (d, J = 8.5 Hz, 2H), 6.94 (td, J = 6.2, 15.3 Hz, 1H), 6.89 (dd, J = 2.7, 7.4 Hz, 1H), 6.85 (d, J = 2.5 Hz, 1H), 6.73 (d, J = 2.7 Hz, 1H), 6.11 (br d, J = 15.3 Hz, 1H), 4.83 - 4.72 (m, 1H), 4.65 (br t, J = 8.5 Hz, 1H), 4.46 - 4.37 (m, 1H), 4.37 - 4.25 (m, 2H), 3.33 (s, 3H), 3.23 (br d, J = 3.3 Hz, 1H), 3.21 - 3.14 (m, 2H), 2.74 (br s, 2H), 2.34 - 2.19 (m, 5H), 1.90 (br d, J = 12.0 Hz, 2H), 1.63 (br d, J = 8.8 Hz, 2H). LCMS (ESIMS) m/z = 594.3 [M+H]⁺.

Example 50

Step 1. tert-butyl 4-(4-chloropyrrolo[2,l-/][l,2,4]triazin-5-yl)-4-hydroxypiperidine-l- carboxylate

[0191] To a reaction tube with a stir bar was added 5-bromo-4-chloropyrrolo[2,l- f][l,2,4]triazine (100 mg, 430 pmol). The tube was capped and evacuated and refilled with N? (3 x). Dry THF (4.0 mL) was added via syringe and the reaction mixture was cooled to - 78 °C for 15 min. Then nBuLi (33.1 mg, 215 pL, 2.4 molar, 516 pmol) was added slowly via syringe and the reaction was stirred for 30 min. Then tert-butyl 4-oxopiperidine-l- carboxylate (103 mg, 516 pmol) in THF (1.0 mL) was added via syringe and the reaction mixture was stirred at -78 °C for 2 h. Then the reaction was quenched with sat NaHCCh and warmed to rt. Water was then added (10 mL) and the reaction mixture was extracted with DCM (3 x 10 mL). The combined organic layers were washed with brine, dried with MgSCh, and concentrated. Purification via column chromatography (10-100% EtOAc in hexanes) provided tert-butyl 4-(4-chloropyrrolo[2, l-f][l,2,4]triazin-5-yl)-4-hydroxypiperidine-l- carboxylate (77 mg, 51 %), used for the next step without further purification. LCMS (ESI) [M+H]⁺ = 352.1.

Step 2. tert-butyl 4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-/|[l,2,4]triazin-5-yl)-4-hydroxypiperidine-l- carboxylate

[0192] To a 40 mL scintillation vial with a stir bar was added tert-butyl 4-(4- chloropyrrolo[2,l-f][l,2,4]triazin-5-yl)-4-hydroxypiperidine-l-carboxylate (656 mg, 1.86 mmol) and 4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylaniline (447 mg, 1.86 mmol). The vial was capped and evacuated and refilled with N2 (3 x), then dry isopropanol (12.0 mL) was added via syringe and the reaction mixture stirred at rt. After 1 h the reaction mixture was washed with sat. NaHCCL (10 mL) and extracted with DCM (3 x 10 mL). Purification via column chromatography 10-100% EtOAcZEtOH (3: 1 mixture) in hexanes provided tertbutyl 4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)amino)pyrrolo[2,l- f][l,2,4]triazin-5-yl)-4-hydroxypiperidine-l-carboxylate (766 mg, 58 %). LCMS (ESI) [M+H]⁺ = 557.3.

Step 3. 4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-/|[l,2,4]triazin-5-yl)piperidin-4-ol [0193] To a 1-dram vial with a stir bar was added tert-butyl 4-(4-((4-([l,2,4]triazolo[l,5- a]pyridin-7-yloxy)-3-methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)-4- hydroxypiperidine-1 -carboxylate (100 mg, 180 pmol) and TFA (246 mg, 166 pL, 2.16 mmol). The reaction mixture was stirred at rt for 1.5 h then concentrated in vacuo. The crude product was used in the next step without any further purification. LCMS (ESI) [M+H]⁺ = 456.2.

Step 4. (£)-l-(4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-/|[l,2,4]triazin-5-yl)-4-hydroxypiperidin-l-yl)-4-

(dimethylamino)but-2-en-l-one

[0194] To a 2-dram vial with a stir bar added the crude product from the previous step, DMF (2.5 mL), diisopropylethylamine (139 mg, 188 pL, 1.08 mmol), (E)-4-(Dimethylamino)but-2- enoic acid hydrochloride (32.7 mg, 198 pmol), and HATU (102 mg, 269 pmol), then the reaction mixture was stirred at rt. After 45 min, the reaction mixture was purified directly via prepHPLC 10-50% ACN in 0.1% TFA water. The fractions containing the desired product mass were combined, neutralized with sat. aq. NaHCCL (lOmL) and extracted with DCM (4 x 15 mL). The combined organic layers were washed with brine, dried over MgSCU, and concentrated to provide (E)-l-(4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2, 1 -f] [ 1 ,2,4]triazin-5-yl)-4-hydroxypiperidin- 1 -yl)-4- (dimethylamino)but-2-en-l-one. Example 50 (59.13 mg, 57 %). 'H NMR (499 MHz, CHLOROFORM-d) 5 ppm 11.22 (s, 1 H) 8.47 (d, J=7.4 Hz, 1 H) 8.06 (s, 1 H) 7.99 (s, 1 H) 7.79 (d, J=2.5 Hz, 1 H) 7.72 (dd, J=8.6, 2.6 Hz, 1 H) 7.48 (d, J=2.7 Hz, 1 H) 7.07 (d, J=8.8 Hz, 1 H) 6.91 (dd, J=7.5, 2.6 Hz, 1 H) 6.67 - 6.81 (m, 2 H) 6.43 - 6.53 (m, 2 H) 5.36 (br s, 1 H) 4.57 (br d, J=11.8 Hz, 1 H) 3.92 (br d, J=12.0 Hz, 1 H) 3.67 (br t, J=12.5 Hz, 1 H) 3.23 (br t, J=12.2 Hz, 1 H) 3.10 (br d, J=6.0 Hz, 2 H) 2.27 (s, 6 H) 2.22 (s, 3 H) 1.89 - 2.19 (m, 4 H). Example 92

Step 1. tert-butyl (Z)-2-(3-ethoxy-2-fluoro-3-oxoprop-l-en-l-yl)pyrrolidine-l- carboxylate

Boc

[0195] To a cold solution (-78 °C) of ethyl 2-(diethoxyphosphoryl)-2-fluoroacetate (5 g, 20.6 mmol) in THF (50 mL) was added n-BuLi (2.5 M in hexane, 9.91 mL, 24.7 mmol) dropwise. The resulting mixture was stirred for 30 min at -78 °C, followed by addition of tert-butyl 2- formylpyrrolidine-1 -carboxylate (4.11 g, 20.6 mmol). The resulting solution was stirred at - 78 °C for another 3 h. The reaction mixture was quenched with sat. aqueous NH4CI at -78 °C and extracted with ethyl acetate (50 mL x 2). The combined organic layers were washed with brine (100 mL), dried over anhydrous Na2SO4 and concentrated to afford crude product tertbutyl (Z)-2-(3-ethoxy-2-fluoro-3-oxoprop-l-en-l-yl)pyrrolidine-l-carboxylate (5 g, crude). The crude product was used for next step without further purification. LCMS (ESI-MS) m/z = 288.1 [M+H]⁺

Step 2. ethyl (Z)-2-fluoro-3-(pyrrolidin-2-yl)acrylate

[0196] To a solution of tert-butyl tert-butyl (Z)-2-(3-ethoxy-2-fluoro-3 -oxoprop- 1-en-l- yl)pyrrolidine-l -carboxylate (5 g, 17.4 mmol) in DCM (50 mL) was added TFA (10 mL). The resulting solution was stirred at room temperature for 3 h and concentrated under vacuum to afford the crude product ethyl (Z)-2-fluoro-3-(pyrrolidin-2-yl)acrylate (4 g). The crude product was used for next step without further purification. LCMS (ESI-MS) m/z = 188.2 [M+H]⁺

Step 3. ethyl (Z)-2-fluoro-3-(l-methylpyrrolidin-2-yl)acrylate

[0197] To a solution of ethyl (Z)-2-fluoro-3-(pyrrolidin-2-yl)acrylate (2 g, 10.7 mmol) in methanol (40 mL) was added formaldehyde (1.28 g, 42.7 mmol) and NaBFFCN (1.01 g, 16.0 mmol). The resulting mixture was stirred at room temperature for 16 h, quenched by addition of sat. aqueous NH4CI (50 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic phase were washed with brine (100 mL), dried with anhydrous Na2SO4, filtered and concentrated to afford ethyl (Z)-2-fluoro-3-(l-methylpyrrolidin-2-yl)acrylate (1 g, 27.7% yield).LCMS (ESI-MS) m/z = 201.9 [M+H]⁺.

Step 4. (Z)-2-fluoro-3-(l-methylpyrrolidin-2-yl)acrylic acid

[0198] Lithium hydroxide (335.6 mg, 14.0 mmol) was added to a stirred mixture of ethyl (2Z)-2-fluoro-3-(l-methylpyrrolidin-2-yl)prop-2-enoate (940 mg, 4.67 mmol) in methanol (10 mL) and water (3 mL). The resulting mixture was stirred at room temperature for 3 h and concentrated to afford (2Z)-2-fluoro-3-(l-methylpyrrolidin-2-yl)prop-2-enoic acid (crude) without neutralization. LCMS (ESI-MS) m/z = 174.1 [M+H]⁺.

Step 5. (Z)-l-(3-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)azetidin-l-yl)-2-fluoro-3-(l- methylpyrrolidin-2-yl)prop-2-en-l-one

[0199] A mixture of (2Z)-2-fluoro-3-(l-methylpyrrolidin-2-yl)prop-2-enoic acid (200 mg, crude, 1.16 mmol), N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(azetidin- 3-yl)py^rr°l°[2,l-f][l,2,4]triazin-4-amine (476.3 mg, 1.16 mmol), EDCI (198.5 mg, 1.39 mmol), HOBT (187.3 mg, 1.39 mmol) and EtsN (233.7 mg, 2.31 mmol) in DMF (5 mL) was stirred at room temperature for 16 h. The reaction mixture was purified by Prep-HPLC, Mobile Phase A: Water (0.1%FA), Mobile Phase B: ACN; Gradient: 11% B to 31% B to afford (Z)-l-(3-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2, 1 -f] [ 1 ,2,4]triazin-5-yl)azetidin- 1 -yl)-2-fluoro-3 -( 1 - methylpyrrolidin-2-yl)prop-2-en-l-one, Example 92 (11.5 mg, 2% yield). LCMS (ESI-MS) m/z= 568.4 [M+H]⁺. 'HNMR (400 MHz, DMSO-t/₆) 8 (ppm) 8.95 (s, 1H), 8.54 (s, 1H), 8.38 (s, 1H), 8.18 (s, 1H), 7.95 (s, 1H), 7.82 (s, 1H), 7.67-7.33 (m, 2H), 7.22-7.12 (m, 1H), 7.10- 6.79 (m, 2H), 6.78-6.56 (m, 1H), 5.70-5.58 (m, 1H), 4.80-4.63 (m, 2H), 4.51-4.37 (m, 2H), 4.08-4.04 (m, 1H), 3.70-3.58 (m, 1H), 3.03-2.95 (m, 1H), 2.19-2.13 (m, 6H), 2.04-1.81 (m, 2H), 1.71 (s, 1H), 1.47 (s, 1H).

Example 93

Step 1. tert-butyl 4-(((trifluoromethyl)sulfonyl)oxy)-2,3,6,7-tetrahydro-lH-azepine-l- carboxylate

[0200] A solution of tert-butyl 4-oxoazepane-l -carboxylate (5 g, 23.47 mmol) in THF (100 mL) was treated with a solution of LiHMDS (1 M in THF, 25.8 mL, 25.78 mmol) for 1 hour at -78 °C under nitrogen atmosphere. A solution of 1,1,1-trifluoro-N-phenyl-N- ((trifluoromethyl)sulfonyl)methanesulfonamide (9.21g, 25.82 mmol) in THF (150 mL) was then added dropwise at -78 °C over a period of 0.5 hour. The resulting mixture was stirred for 3 hours at -78 °C and overnight at room temperature under nitrogen atmosphere. The reaction mixture was quenched with water (1000 mL) at 0 °C and extracted with ethyl acetate (3 x 1000 mL). The combined organic layers were dried over anhydrous ISfeSCU, filtered and concentrated under vacuum to afford the crude product. The crude product was purified by silica gel column chromatography, eluted with petroleum ether / ethyl acetate = 9: 1 to afford the crude product tert-butyl 4-(((trifluoromethyl)sulfonyl)oxy)-2,3,6,7-tetrahydro-lH- azepine-1 -carboxylate (6.8 g crude). LCMS (ESI-MS) m/z =346.0 [M+H]⁺.

Step 2. tert-butyl 4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-2,3,6,7-tetrahydro-lH- azepine-l-carboxylate

[0201] A mixture of tert-butyl 4-(((trifluoromethyl)sulfonyl)oxy)-2,3,6,7-tetrahydro-

IH-azepine-l -carboxylate (6.7 g, 19.40 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(l,3,2- dioxaborolane) (9.85 g, 38.80 mmol), KOAc (3.81 g, 38.80 mmol) and Pd(dppf)C12 (1.42 g, 1.94 mmol) in dioxane (200 mL) was stirred for 2 hours at 80 °C under nitrogen atmosphere. The reaction mixture was allowed to cool down to room temperature, filtered and the filtrate was concentrated under vacuum to afford the crude product. The crude product was purified by silica gel column chromatography, eluted with petroleum ether / ethyl acetate = 2: 1 to afford tert-butyl 4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)- 2,3,6,7-tetrahydro-lH-azepine-l-carboxylate (1.8 g, 23.7% yield). LCMS (ESI-MS) m/z =324.1 [M+H]⁺

Step 3. tert-butyl 4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)-2,3,6,7-tetrahydro-lH-azepine-l- carboxylate

[0202] A mixture of tert-butyl 4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)- 2,3,6,7-tetrahydro-lH-azepine-l-carboxylate (1.7 g, 5.25 mmol), 5-bromo-N-(4-

([1 ,2,4]triazolo[ 1 , 5-a]pyridin-7-yloxy)-3 -methylphenyl)-5-bromopyrrolo[2, 1 -f] [ 1 ,2,4]triazin- 4-amine (2.29 g, 5.25 mmol), K2CO3 (1.45 g, 10.51 mmol) and Pd(dppf)C12 (0.38 g, 0.52 mmol) in 1,4-di oxane (24 mL) and water (6 mL) was stirred for 12 hours at 100 °C under nitrogen atmosphere. The resulting mixture was filtered and the filtrate was concentrated under vacuum to afford the crude product. The crude product was purified by silica gel column chromatography, eluted with petroleum ether / ethyl acetate = 2: 1 to afford tert-butyl 4-(4-((4-([ 1 , 2, 4] tri azolof 1 , 5-a]pyri din-7 -yloxy)-3 -methylphenyl)amino)pyrrolo[2, 1 - f][l,2,4]triazin-5-yl)-2,3,6,7-tetrahydro-lH-azepine-l-carboxylate (1.3 g, 42.2% yield).

LCMS (ESI-MS) m/z =553.2 [M+H]⁺

Step 4. tert-butyl 4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f|[l,2,4]triazin-5-yl)azepane-l-carboxylate

[0203] A mixture of tert-butyl 4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)-2,3,6,7-tetrahydro-lH-azepine-l- carboxylate (1.3 g, 2.35 mmol) and Pd/C (2.5 g, 23.52 mmol) in MeOH (20 mL) was degassed under vacuum and charged with an atmospheric pressure of hydrogen. The resulting mixture was stirred for 24 hours at room temperature. The solids were filtered off and the filtrate was concentrated under vacuum to afford the crude product tert-butyl 4-(4-((4- ([1, 2, 4]tri azolof l,5-a]pyridin-7-yloxy)-3-methylphenyl)amino)pyrrolo[2,l-f][l, 2, 4]tri azin-5- yl)azepane-l -carboxylate (700 mg crude). The crude product was used in the next step directly without further purification. LCMS (ESI-MS) m/z = 555.4 [M+H]⁺.

Step 5. N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(azepan-4- yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine

[0204] A mixture of tert-butyl 4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)azepane-l-carboxylate (300 mg, 0.54 mmol) and TFA (1 mL, 13.46 mmol) in DCM (10 mL) was stirred for 1 hour at room temperature and then concentrated under vacuum to afford the crude product N-(4- ([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(azepan-4-yl)pyrrolo[2,l- f][l,2,4]triazin-4-amine (360 mg crude). The crude product was used in the next step without further purification. LCMS (ES MS) m/z =455.3 [M+H]⁺

Step 6. (E)-l-(4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)azepan-l-yl)-4- (dimethylamino)but-2-en-l-one

[0205] A mixture of N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5- (azepan-4-yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine (360 mg, 0.79 mmol), (E)-4- (dimethylamino)but-2-enoic acid (133 mg, 1.03 mmol), diisopropylethylamine (204.73 mg, 1.58 mmol) and HATU (451.73 mg, 1.18 mmol) in DMF (4 mL) was stirred at room temperature for 2 hours. The reaction mixture was filtered and the filtrate was purified by Prep-HPLC, Mobile Phase A: Water(0.1%FA), Mobile Phase B: ACN; Gradient: 15% B to 30% B to afford the desired product (E)-l-(4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2, 1 -f] [ 1 ,2,4]triazin-5-yl)azepan- 1 -yl)-4-(dimethylamino)but-2- en-l-one, Example 93 (6.7 mg, 2% yield). LCMS (ESI-MS) m/z =566.4 [M+H] .'H NMR (400 MHz, methanol-t/4) 8 (ppm) 8.74 (s, 1H), 8.32 (s, 2H), 7.72 (s, 1H), 7.62 (s, 1H), 7.56- 7.41 (m, 2H), 7.19-7.17 (m, 1H), 7.10-7.06 (m, 1H), 6.95-6.88 (m, 1H), 6.84 (s, 1H), 6.77- 6.70 (m, 1H), 6.64-6.62 (m, 1H), 3.96-3.61 (m, 6H), 3.55-3.39 (m, 1H), 2.82-2.73 (m, 6H), 2.35-2.30 (m, 1H), 2.24 (s, 5H), 1.98-1.73 (m, 3H).

Step 1. tert-butyl 6-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)-2-azaspiro[3.3]heptane-2- carboxylate

[0206] A mixture of 4,4'-di-tert-butyl-2,2'-bipyridine (30.76 mg, 0.11 mmol) and NiCh dme (25.18 mg, 0.11 mmol) in DCE (1 mL) was heated to 60 °C for 10 minutes under nitrogen atmosphere. The solution was allowed to cool to room temperature to produce solution 1.

[0207] Ir[dF(CF3)ppy]2(dtbpy)PFe (128.58 mg, 0.11 mmol) was added to a mixture of of N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-bromopyrrolo[2,l- f][l,2,4]triazin-4-amine (500 mg, 1.14 mmol), tert-butyl 6-iodo-2-azaspiro[3.3]heptane-2- carboxylate (740.76 mg, 2.29 mmol), l,l,l,3,3,3-hexamethyl-2-(trimethylsilyl)trisilane (313.48 mg, 1.26 mmol) and Na2COs (364.41 mg, 3.43 mmol) in DCE (10 mL) under nitrogen atmosphere, followed by addition of the solution 1 via syringe. The resulting mixture was maintained under nitrogen, stirred at room temperature and irradiated by blue LED (450 nm) in Penn Photoreactor m2 for 6 hours. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (3 x 100 mL). The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered and concentrated under vacuum to afford the crude product. The crude product was purified by column chromatography, eluted with ethyl acetate in petroleum ether from 0% to 30% to afford the desired product tert-butyl 6-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)-pyrrolo[2,l-f][l,2,4]triazin-5-yl)-2-azaspiro[3.3]heptane-2-carboxylate (200 mg crude), the crude product was used for next step directly without further purification. LCMS (ESI-MS) m/z = 553.2 [M+H]⁺.

Step 2. N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(2- azaspiro[3.3]heptan-6-yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine

[0208] TFA (6 mL) was added to a stirred mixture of tert-butyl 6-(4-((4-

([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5- yl)-2-azaspiro[3.3]heptane-2-carboxylate (200 mg, 0.36 mmol) in DCM (2 mL). The resulting mixture was stirred for 1 hour at room temperature and concentrated under vacuum to afford the crude product. The crude product was purified by column chromatography, eluted with MeOH in DCM from 0% to 10% to afford the desired product N-(4- ([1, 2, 4]tri azolof l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(2-azaspiro[3.3]heptan-6- yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine (60 mg crude), the crude product was used for next step directly without further purification. LCMS (ESI-MS) m/z = 453.2 [M+H]⁺.

Step 3. (E)-l-(6-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)-2-azaspiro[3.3]heptan-2-yl)-4-

(dimethylamino)but-2-en-l-one

[0209] Diisopropylethylamine (51.4 mg, 0.39 mmol) was added to a stirred mixture of N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(2-azaspiro[3.3]heptan-6- yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine (60 mg, 0.13 mmol), (E)-4-(dimethylamino)but-2- enoic acid hydrochloride (33 mg, 0.20 mmol) and HATU (75.6 mg, 0.20 mmol) in DMF (1 mL). The resulting mixture was stirred for 1 hour at room temperature and purified by Prep- HPLC, Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Gradient: 17% B to 47% B to afford the desired product (E)-l-(6-(4-((4-([l,2,4]triazolo[l,5-a]pyridin- 7-yloxy)-3-methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)-2-azaspiro-[3.3]heptan-2- yl)-4-(dimethylamino)but-2-en-l-one, Example 95 (16.7 mg, 22.1% yield). LCMS (ESI-MS) m/z = 564.4 [M+H]⁺. 'H NMR (400 MHz, DMSO-t/₆) 8 8.57-8.37 (m, 1H), 8.32-8.13 (m, 1H), 8.05-7.88 (m, 1H), 7.71-7.47 (m, 3H), 7.16-7.03 (m, 1H), 7.01-6.78 (m, 4H), 6.68-6.53 (m, 1H), 6.17-5.96 (m, 1H), 4.51-4.36 (m, 1H), 4.31-4.13 (m, 2H), 4.11-4.00 (m, 1H), 3.94- 3.79 (m, 1H), 3.18-3.01 (m, 2H), 2.91-2.73 (m, 2H), 2.70-2.52 (m, 2H), 2.41-2.15 (m, 9H). Example 103

Step 1. tert-butyl (2-(4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazol-l- yl)ethyl)carbamate

[0210] A mixture of 4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazole

(1.00 g, 5.15 mmol), tert-butyl (2-bromoethyl)carbamate (1.15 g, 5.15 mmol) and CS2CO3 (3.36 g, 10.30 mmol) in MeCN (10 mL) was stirred with reflux overnight. The reaction mixture was diluted with water (20 mL), the resulting solution was then extracted with ethyl acetate (3 x 10 mL). The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered and concentrated under vacuum to afford the crude product. The crude product was purified by column chromatography (eluted with ethyl acetate in petroleum ether from 0% to 30%), the desired fractions were combined and concentrated under vacuum to afford the desired product tert-butyl (2-(4-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)-lH-pyrazol-l-yl)ethyl)carbamate (1.25 g, 72% yield). LCMS (ESI-MS) m/z =338.2 [M+H]⁺.

Step 2. tert-butyl (2-(4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f|[l,2,4]triazin-5-yl)-lH-pyrazol-l-yl)ethyl)carbamate

[0211] A mixture of tert-butyl (2-(4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)- lH-pyrazol-l-yl)ethyl)carbamate (400 mg, 1.18 mmol), N-(4-([l,2,4]triazolo[l,5-a]pyridin-7- yloxy)-3-methylphenyl)-5-bromopyrrolo[2,l-f][l,2,4] triazin-4-amine (517.48 mg, 1.18 mmol), Pd(dppf)C12 (86.79 mg, 0.11 mmol), K2CO3 (327.86 mg, 2.37 mmol) in dioxane (4 mL) was stirred overnight at 100 °C under nitrogen atmosphere. The reaction mixture was diluted with water (15 mL), the resulting solution was extracted with ethyl acetate (3 x 5 mL). The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered and concentrated under vacuum to afford the crude product. The crude product was purified by column chromatography (silica gel, 25 g, eluted with ethyl acetate in petroleum ether from 0% to 30%), the desired fractions were combined and concentrated under vacuum to afford the desired product tert-butyl (2-(4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)-lH-pyrazol-l-yl)ethyl)carbamate (480 mg, 71% yield). LCMS (ESI-MS) m/z =567.2 [M+H]⁺.

Step 3. N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(l-(2-aminoethyl)- lH-pyrazol-4-yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine

[0212] A mixture of tert-butyl (2-(4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)-lH-pyrazol-l-yl)ethyl)carbamate (480 mg, 0.84 mmol) and TFA (1 mL, 13.46 mmol) in DCM (3 mL) was stirred for 1 hour at room temperature. The resulting mixture was concentrated under vacuum to afford the crude product N-(4-([l, 2, 4]triazolo[l, 5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(l-(2-aminoethyl)- lH-pyrazol-4-yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine (480 mg), the crude product was used in next step directly without further purification. LCMS (ESI-MS) m/z = 467.2 [M+H]⁺.

Step 4. (E)-N-(2-(4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)-lH-pyrazol-l-yl)ethyl)-4- (dimethylamino)but-2-enamide

[0213] A solution of N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5- (l-(2-aminoethyl)-lH-pyrazol-4-yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine (50 mg, 0.10 mmol), (E)-4-(dimethylamino)but-2-enoic acid (16.61 mg, 0.12 mmol), HATU (81.51 mg, 0.21 mmol) and diisopropylethylamine (41.56 mg, 0.32 mmol) in DMF (2 mL) was stirred for 2 hours at room temperature. The reaction mixture was diluted with water (20 mL), the resulting solution was extracted with ethyl acetate (3 x 20 mL), and the organic layers were concentrated under vacuum. The crude product was purified by Prep-HPLC, Mobile Phase A: Water (0.05% TFA), Mobile Phase B: ACN; Gradient: 14% B to 44% B to afford (E)-N-(2- (4-(4-((4-([ 1 ,2,4]triazolo[ 1 ,5 -a]pyridin-7-yloxy)-3 -methylphenyl)amino)pyrrolo[2, 1 - f][l,2,4]triazin-5-yl)-lH-pyrazol-l-yl)ethyl)-4-(dimethylamino)but-2-enamide, Example 103 (47.2 mg, 76% yield). LCMS (ESI-MS) m/z =578.2 [M+H]⁺. ¹ H NMR (400 MHz, DMSO-t/₆) 5 9.63 (s, 1H), 8.97 (d, J= 7.3 Hz, 1H), 8.45 (d, J= 10.9 Hz, 2H), 8.10 (s, 1H), 8.05 (s, 1H), 7.85 (s, 1H), 7.79 (s, 1H), 7.65 (s, 1H), 7.60 (s, 1H), 7.21 (d, J= 8.7 Hz, 1H), 7.06 (dd, J= 7.5, 2.6 Hz, 1H), 6.81 (d, J= 2.6 Hz, 1H), 6.76 (d, J= 2.7 Hz, 1H), 6.53 (m, J= 14.7, 7.2 Hz, 1H), 6.21 (m, J= 15.4, 1.3 Hz, 1H), 4.30 (t, = 6.1 Hz, 2H), 3.83 (t, J= 5.8 Hz, 2H), 3.64 (q,

J= 6.0 Hz, 2H), 2.72 (d, J= 4.3 Hz, 6H), 2.17 (s, 3H).

Example 105

Step 1. (E)-4-bromo-l-(4-(4-((3-methyl-4-((l-methyl-lH-benzo[d]imidazol-5- yl)oxy)phenyl)amino)-pyrrolo[2,l-f][l,2,4]triazin-5-yl)piperidin-l-yl)but-2-en-l-one

[0214] To a cold (0 °C) solution of (E)-4-bromobut-2-enoic acid (0.73 g, 4.41 mmol) in DMF (0.05 mL) and DCM (10 mL) was added oxalyl chloride (1.12 g, 8.82 mmol) dropwise. The reaction mixture was stirred overnight at room temperature. The resulting crude mixture was concentrated under vacuum. The residue was dissolved in DCM (15mL). The solution was added to a stirred mixture of N-(3-methyl-4-((l-methyl-lH- benzo[d]imidazol-5-yl)oxy)phenyl)-5-(piperidin-4-yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine (2 g, 4.41 mmol) and NaHCCL (0.75 g, 8.92 mmol) in DCM (20 mL). The resulting mixture was stirred for 4 hours at room temperature and concentrated under vacuum to afford the crude product. The crude product was purified by silica gel column chromatography, eluted with petroleum ether / ethyl acetate = 1 :4 to afford the desired product (E)-4-bromo-l-(4-(4-((3- methyl-4-((l-methyl-lH-benzo[d]imidazol-5-yl)oxy)phenyl)amino)pyrrolo[2,l- f][l,2,4]triazin-5-yl)piperidin-l-yl)but-2-en-l-one (800 mg, 27.2% yield). LCMS (ESI-MS) m/z = 600.2 [M+H]⁺

Step 2. (E)-l-(4-(4-((3-methyl-4-((l-methyl-lH-benzo[d]imidazol-5- yl)oxy)phenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)piperidin-l-yl)-4-(tetrahydro-lH- furo[3,4-c]pyrrol-5(3H)-yl)but-2-en-l-one

[0215] Diisopropylethylamine (161 mg, 1.25 mmol) was added to a mixture of (E)-4- bromo-1 -(4-(4-((3-methyl-4-((l -methyl- lH-benzo[d]imidazol-5- yl)oxy)phenyl)amino)pyrrolo[2, 1 -f] [ 1 ,2,4]triazin-5-yl)piperidin- 1 -yl)but-2-en- 1 -one (150 mg, 0.25 mmol), hexahydro-lH-furo[3,4-c]pyrrole hydrochloride (25 mg, 0.22 mmol) in DMF (2 mL). The resulting mixture was stirred overnight at room temperature. The reaction mixture was purified by Prep-HPLC, Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Gradient: 2% B to 25% B to afford the desired product (E)-l-(4-(4-((3-methyl-4-((l-methyl- lH-benzo[d]imidazol-5-yl)oxy)phenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)piperidin-l- yl)-4-(tetrahydro-lH-furo[3,4-c]pyrrol-5(3H)-yl)but-2-en-l-one, Example 105 (15.5 mg, 10% yield). LCMS (ESI-MS) m/z =633.5 [M+H]⁺. 'H NMR (400 MHz, DMSO-t/₆) 5(ppm) 8.43 (s, 1H), 8.17 (s, 2H), 7.85 (s, 1H), 7.68 (d, J = 2.7 Hz, 1H), 7.57 (d, J = 8.7 Hz, 1H), 7.52 (d, J = 2.6 Hz, 1H), 7.40-7.35 (m, 1H), 7.10 (d, J = 2.3 Hz, 1H), 7.00-6.98 (m, 1H), 6.87 (d, J = 8.6 Hz, 1H), 6.70-6.60 (m, 3H), 4.57 (d, J = 12.6 Hz, 1H), 4.12 (s, 1H), 3.71 (s, 1H), 3.39- 3.35 (m, 2H), 3.31 (d, J = 14.0 Hz, 1H), 2.84 (s, 1H), 2.70 (s, 2H), 2.53 (s, 2H), 2.47 (s, 4H), 2.33-2.35 (m, 2H), 2.30 (s, 1H), 2.25 (s, 3H), 1.96 (s, 2H), 1.55 (s, 3H). Example 117

Boc step 4

Step 1. tert-butyl (E)-4-(2-(phenylsulfonyl)vinyl)piperidine-l-carboxylate -20 C to rt, 3.5 h

[0216] A solution of LiHMDS (1 M solution in THF, 384.6 mL, 384.6 mmol) was added dropwise to a solution of (methylsulfonyl)benzene (50 g, 320.5 mmol) in anhydrous THF (2.5 L) at -20 °C under nitrogen atmosphere. The reaction mixture was allowed to stir for 30 minutes at -20 °C followed by addition of TMSCI (48.8 ml, 99.2 mmol). The resulting mixture was slowly allowed to warm up to room temperature and stirred for another 3 hours. The reaction mixture was quenched by addition of saturated aqueous NH4CI (I L) and extracted with EA (2 x 1 L). The combined organic layers were washed with brine (2 L), dried over anhydrous ISfeSCU and concentrated under vacuum to afford the product. The residue was purified by silica gel column chromatography, eluted with ethyl acetate in petroleum ether from 0% to 20% to afford the desired product tert-butyl (E)-4-(2- (phenylsulfonyl)vinyl)piperidine-l -carboxylate (32.0 g, 44.9% yield). LCMS (ESI-MS) m/z = 352.1 [M+H]⁺.

Step 2. tert-butyl 4-(2-(ethoxycarbonyl)-lH-pyrrol-3-yl)piperidine-l-carboxylate

[0217] tBuOK (18.8 g, 168 mmol) was added to a solution of ethyl 2-isocyanoacetate (14.5 g, 128.18 mmol) in THF (320 mL) under nitrogen atmosphere. The resulting mixture was stirred at 0 °C for 30 minutes. Then a solution of tert-butyl 4-(2-(ethoxycarbonyl)-lH-pyrrol- 3 -yl)piperidine-l -carboxylate (32 g, 91.0 mmol) in THF (320 mL) was added to the mixture above. The resulting mixture was warmed to room temperature, stirred for 3 hours and quenched by the addition of saturated aqueous NH4CI (100 mL) at 0 °C. The resulting mixture was extracted with ethyl acetate (3 x 250 mL). The combined organic layers were washed with brine (400 mL), dried over anhydrous ISfeSCU and concentrated under vacuum to afford the crude product. The residue was purified by silica gel column chromatography, eluted with ethyl acetate in petroleum ether from 0% to 20% to afford the desired product tert-butyl 4-(2-(ethoxycarbonyl)-lH-pyrrol-3-yl)piperidine-l-carboxylate (11.2 g, 38.1% yield). LCMS (ESI-MS) m/z = 323.1 [M+H]⁺.

Step 3. tert-butyl 4-(l-amino-2-(ethoxycarbonyl)-lH-pyrrol-3-yl)piperidine-l- carboxylate

[0218] NaH (60% in mineral oil, 5.09 g, 127.17 mmol) in DMF (180 mL) was added to a solution of tert-butyl 4-(2-(ethoxycarbonyl)-lH-pyrrol-3-yl)piperidine-l-carboxylate (20 g, 62.03 mmol) in THF (900 mL) at 0 °C. The resulting mixture was stirred for 1 hour at room temperature, followed by addition of O-(2,4-dinitrophenyl)hydroxylamine (23.47 g, 117.86 mmol) portionwise. The resulting mixture was stirred at room temperature for 8 hours, quenched by the addition of saturated aqueous NH4CI (300 mL) at 0 °C and extracted with ethyl acetate (3 x 500 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous ISfeSCU and concentrated under vacuum. The residue was purified by silica gel column chromatography, eluted with ethyl acetate in petroleum ether from 0% to 20% to afford the desired product tert-butyl 4-(l-amino-2-(ethoxycarbonyl)-lH-pyrrol-3- yl)piperidine-l -carboxylate (14 g, 66.8 % yield). LCMS (ESI-MS) m/z =338.2 [M+H]⁺.

Step 4. tert-butyl 4-(4-oxo-3,4-dihydropyrrolo[2,l-f][l,2,4]triazin-5-yl)piperidine-l- carboxylate

[0219] Formamidine acetate (14.62 g, 332 mmol) was added to a solution of tert-butyl 4-(l-amino-2-(ethoxycarbonyl)-lH-pyrrol-3-yl)piperidine-l-carboxylate (14 g, 41.49 mmol) in iPrOH (14 mL) at 25 °C. The resulting mixture was stirred overnight at 100 °C. After cooled to room temperature, the reaction mixture was quenched by the addition of water (100 mL) and extracted with ethyl acetate (3 x 100 mL). The combined organic layers were washed with brine (200 mL), dried over anhydrous ISfeSCU and concentrated under reduced pressure to afford the crude product. The residue was purified by silica gel column chromatography, eluted with ethyl acetate in petroleum ether from 0% to 50% to afford the desired product tert-butyl 4-(4-oxo-3,4-dihydropyrrolo[2,l-f][l,2,4] triazin-5- yl)piperidine-l -carboxylate (10.4 g, 92.0% yield). LCMS (ESI-MS) m/z = 319.2 [M+H]⁺. ^JH NMR (400 MHz, DMSO-tL) 3 11.47 (d, J = 4.0 Hz, 1H), 7.72 (d, J = 4.0 Hz, 1H), 7.46 (d, J = 2.8 Hz, 1H), 6.45 (d, J = 2.7 Hz, 1H), 4.05 (s, 2H), 3.33-3.34 (m, 1H), 2.79 (s, 2H), 1.83-1.74 (m, 2H), 1.51-1.25 (m, 2H), 1.41 (s, 9H).

Step 5. tert-butyl 4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f] [l,2,4]triazin-5-yl)piperidine-l-carboxylate

[0220] EtsN (3.2 g, 31.5 mmol) was added to a mixture of tert-butyl 4-(4-oxo-3 ,4- dihydropyrrolo[2,l-f][l,2,4]triazin-5-yl)piperidine-l-carboxylate (3.2 g, 10.5 mmo), 4- ([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylaniline (4.1 g, 10.5 mmol) and PyBrop (12 g, 25.8 mmol) in THF (50 mL). The resulting mixture was stirred at 80 °C overnight then cooled to room temperature. The reaction mixture was filtered, the filter cake was washed with DCM (300 mL). The filtrate was concentrated under vacuum to afford the crude product. The residue was purified by silica gel column chromatography, eluted with ethyl acetate in petroleum ether from 0% to 100% to afford the desired product tert-butyl 4-(4-((4- ([1, 2, 4]tri azolof l,5-a]pyridin-7-yloxy)-3-methylphenyl)amino)pyrrolo[2,l-f][l, 2, 4]tri azin-5- yl)piperidine-l -carboxylate (1.7 g, 31.1% yield). LCMS (ESI-MS) m/z = 541.3 [M+H]⁺. ^JH NMR (400 MHz, DMSO-t/₆) d 8.94-8.92 (m, 1H), 8.47 (s, 1H), 8.38 (d, J = 4.4 Hz, 1H), 7.90 (s, 1H), 7.72 (d, J = 2.7 Hz, 1H), 7.66 (d, J = 2.5 Hz, 1H), 7.61-7.60 (m, 1H), 7.22 (d, J = 8.6 Hz, 1H), 7.16-6.99 (m, 1H), 6.79 (d, J = 2.7 Hz, 1H), 4.12-4.03 (m, 4H), 3.56-3.55 (m, 1H), 3.17 (d, J = 3.9 Hz, 1H), 2.19 (s, 3H), 2.12-2.01 (m, 2H), 1.95-1.87 (m, 2H),1.41 (s, 9H). Step 6. N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(piperidin-4- yl)pyrrolo[2,l-f|[l,2,4]triazin-4-amine

[0221] A solution of tert-butyl 4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)piperidine-l-carboxylate (1 g, 1.85 mmol) and TFA (5 mL) in DCM (5mL) was stirred for 1 hour at room temperature. The resulting mixture was concentrated under vacuum to afford the crude product N-(4- ([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(piperidin-4-yl)pyrrolo[2,l- f][l,2,4]triazin-4-amine (1 g crude). LCMS (ESLMS) m/z = 441.2 [M+H]⁺.

Step 7. (E)-l-(4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)piperidin-l-yl)-4-bromobut-2-en- 1-one

[0222] To a cold mixture (0 °C) of (E)-4-bromobut-2-enoic acid (1.46 g, 8.82 mmol) in DMF (0.1 mL) and DCM (15 mL) was added oxalyl chloride (2.42 g, 17.6 mmol) dropwise. The reaction mixture was stirred overnight at room temperature and concentrated under vacuum. The residue was dissolved in DCM (15 mL). The solution was added to a stirred mixture ofN-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(piperidin- 4-yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine (1 g, 2.27 mmol) and NaHCCh (1.14 g, 13.5 mmol) in THF (20 mL). The resulting mixture was stirred for 30 minutes at room temperature and concentrated under vacuum to afford the crude product. The crude product was purified by silica gel column chromatography, eluted with DCM/MeOH (10:1 volume ratio) to afford the desired product (E)-l-(4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2, 1 -f] [ 1 ,2,4]triazin-5-yl)piperidin- 1 -yl)-4-bromobut-2-en- 1 -one (800 mg, 73.6% yield for two steps). LCMS (ESI-MS) m/z = 587.1 [M+H]⁺.

Step 8. (E)-l-(4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)piperidin-l-yl)-4-bromobut-2-en- 1-one

[0223] Diisopropylethylamine (66 mg, 0.51 mmol) was added to a mixture of (E)-l- (4-(4-((4-([ 1 ,2,4]triazolo[ 1 ,5 -a]77midazol-7-yloxy)-3 -methylphenyl)amino)pyrrolo[2, 1 - f][l,2,4]triazin-5-yl)piperidin-l-yl)-4-bromobut-2-en-l-one (100 mg, 0.17 mmol) and 3- methoxyazetidine hydrochloride (19 mg, 0.15 mmol) in DMF (1 mL). The resulting mixture was stirred overnight at room temperature. The reaction mixture was purified by Prep-HPLC; Mobile Phase A: Water (lOmmol/L NH₄HC03+0.05%NH₃H20), Mobile Phase B: ACN; Gradient: 24% B to 54% to afford the desired product (E)-l-(4-(4-((4-([l,2,4]triazolo[l,5- a]77midazol-7-yloxy)-3-methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)piperidin-l- yl)-4-(3-methoxyazetidin-l-yl)but-2-en-l-one, Example 117 (14.4 mg, 13.6% yield). LCMS (ESI-MS) m/z = 594.3 [M+H]⁺. 'H NMR (400 MHz, Chloroforms/) 5 (ppm) 8.50-8.45 (m, 1H), 8.23 (s, 1H), 7.97 (s, 1H), 7.59-7.50 (m, 3H), 7.12 (d, J = 8.6 Hz, 1H), 7.04 (s, 1H), 6.93-6.83 (m, 2H), 6.82-6.74 (m, 1H), 6.57 (d, J = 2.8 Hz, 1H), 4.89 (s, 1H), 4.33 (s, 1H), 4.15 (s, 1H), 3.90 (s, 1H), 3.45 (d, J = 7.1 Hz, 1H), 3.29 (s, 3H), 3.15 (s, 3H), 2.85 (s, 1H), 2.26 (s, 3H), 2.17-2.10 (m, 3H), 1.84-1.80 (m, 3H), 1.25 (s, 2H).

Example 141

Step 1: tert-butyl (lR,4R,5S)-5-{4-[(3-methyl-4-{[l,2,4]triazolo[l,5-a]pyridin-7- yloxy}phenyl)amino]pyrrolo[2,l-f][l,2,4]triazin-5-yl}-2-azabicyclo[2.2.1]heptane-2- carboxylate

[0224] A sealed tube was charged with deoxazole (0.331 g, 0.836 mmol, 1.6 eq) and tert-butyl (1R,4R,5S) 5-hydroxy-2-azabicyclo[2.2.1]heptane-2-carboxylate (0.195 g, 0.915 mmol, 1.75 eq). The mixture was degasses under vacuum and charged with argon. This process was repeated two times. Then dry MTBE (5.23 ml, 0.1 M) was added via syringe to the tube and the mixture was stirred for 5 min at room temperature. A solution of dry pyridine (0.068 ml, 0.836 mmol, 1.6 eq) in dry MTBE (0.99 ml, 15.0 vol) was then added dropwise over 2 min and the resulting solution was stirred for 10 minutes. In a 7 mL vial, 5-bromo-N- (3 -methyl-4-{ [ 1 , 2, 4]tri azolof 1 , 5-a]pyridin-7-yloxy }phenyl)pyrrolo[2, 1 -f] [ 1 ,2,4]triazin-4- amine (0.24 g, 0.523 mmol, 1.0 eq), [Ir(dtbbpy)(ppy)2]PFe (0.007 g, 0.008 mmol, 0.015 eq), [4,4'-bis(tert-butyl)-2,2'-bipyridine]nickel dibromide (0.019 g, 0.039 mmol, 0.075 eq), phthalimide (0.017 g, 0.118 mmol, 0.225 eq) and quinuclidine (0.102 g, 0.915 mmol, 1.75 eq) were added and the mixture was suspended in dry dimethylacetamide (4.8 ml, 20.0 vol) under argon. MTBE solution was transferred to a syringe and filtered prior to addition. The reaction mixture was purged with argon for 15 minutes and sealed with parafilm. The vial was placed in a PennPhD Photoreactor stirring at 1500 rpm under 450 nm LED irradiation at 100% intensity with a fan speed of 2800 rpm. Stirring was carried out for 2 days. Reaction mixture was filtered through celite pad and washed with ethyl acetate. Solvents were evaporated and purification was performed. Purification via flash chromatography (DCM / MeOH 100:0 -> 95:5) was performed to give a crude product. Additional re-purification was performed via preparative TLC (plate was developed 3 times; DCM/MeOH 100:0 -> 96:4). Tert-butyl (lS,4S,5S)-5-{4-[(3-methyl-4-{[l,2,4]triazolo[l,5-a]pyridin-7- yloxy}phenyl)amino]pyrrolo[2,l-f][l,2,4]triazin-5-yl}-2-azabicyclo[2.2.1]heptane-2- carboxylate (0.03 g, 8%) was isolated. 'H NMR (300 MHz, DMSO-d6) 5 8.94 (dd, J = 7.5, 3.1 Hz, 1H), 8.43 (s, 1H), 8.39 (s, 1H), 7.92 (s, 1H), 7.73 - 7.65 (m, 2H), 7.22 (d, J = 8.4 Hz, 1H), 7.03 (dd, J = 7.5, 2.6 Hz, 1H), 6.81 (d, J = 2.0 Hz, 1H), 6.71 (d, J = 2.7 Hz, 1H), 4.19 (d, J = 15.1 Hz, 1H), 3.90 (dd, J = 9.4, 4.1 Hz, 1H), 3.30 (s, 1H), 2.70 (dd, J = 4.9, 2.1 Hz, 1H), 2.19 (s, 4H), 1.88 - 1.78 (m, 1H), 1.66 (d, J = 10.8 Hz, 1H), 1.62 - 1.54 (m, 1H), 1.42 (s, 9H), 1.24 (s, 2H). UPLC (ESI) [M+H]⁺= 553.10

Step 2: 5-[(lS,4S,5S)-2-azabicyclo[2.2.1]heptan-5-yl]-N-(3-methyl-4-{[l,2,4]triazolo[l,5- a]pyridin-7-yloxy}phenyl)pyrrolo[2,l-f][l,2,4]triazin-4-amine

[0225] To a stirring solution of tert-butyl (lS,4S,5S)-5-{4-[(3-methyl-4-

{ [ 1 ,2,4]triazolo[ 1 , 5-a]pyridin-7-yloxy }phenyl)amino]pyrrolo[2, 1 -f] [ 1 ,2,4]triazin-5-yl } -2- azabicyclo[2.2.1]heptane-2-carboxylate (0.03 g, 0.044 mmol, 1.0 eq) in DCM (0.6 ml, 20.0 vol) was added TFA (0.3 ml, 10.0 vol). The reaction was stirred at RT for 2 hours. Solvents were evaporated and the crude was used in the next step without further purification. 5- [(lS,4S,5S)-2-azabicyclo[2.2.1]heptan-5-yl]-N-(3-methyl-4-{[l,2,4]triazolo[l,5-a]pyridin-7- yloxy}phenyl)pyrrolo[2,l-f][l,2,4]triazin-4-amine (0.045 g, crude) was isolated in the form of trifluoroacetic acid salt. ¹ H NMR (300 MHz, Methanol-d4) 5 8.82 (d, J = 7.5 Hz, 1H), 8.42 (s, 1H), 7.77 (s, 1H), 7.67 (d, J = 2.8 Hz, 1H), 7.56 - 7.52 (m, 1H), 7.47 (dd, J = 8.8, 2.8 Hz, 1H), 7.27 (d, J = 8.5 Hz, 1H), 7.17 (dd, J = 7.6, 2.6 Hz, 1H), 6.89 (d, J = 2.5 Hz, 1H), 6.79 (d, J = 2.8 Hz, 1H), 4.25 (s, 1H), 3.91 (dd, J = 8.4, 4.8 Hz, 1H), 3.37 (d, J = 7.8 Hz, 1H), 3.03 (s, 1H), 2.47 (ddd, J = 14.4, 8.9, 2.4 Hz, 1H), 2.30 (s, 1H), 2.27 (s, 3H), 2.18 - 2.11 (m, 2H), 1.87 - 1.78 (m, 1H). UPLC (ESI) [M+H]⁺ = 492.95

Step 3: l-[(lS,4S,5S)-5-{4-[(3-methyl-4-{[l,2,4]triazolo[l,5-a]pyridin-7- yloxy}phenyl)amino]pyrrolo[2,l-f][l,2,4]triazin-5-yl}-2-azabicyclo[2.2.1]heptan-2- y 1] prop-2-en-l-one

[0226] To a stirring solution of 5-[(lS,4S,5S)-2-azabicyclo[2.2.1]heptan-5-yl]-N-(3- methyl-4-{[l,2,4]triazolo[l,5-a]pyridin-7-yloxy}phenyl)pyrrolo[2,l-f][l,2,4]triazin-4-amine (0.045 g, 0.086 mmol, 1.0 eq) in dry DCM (0.9 ml, 20.0 vol) was added tri ethylamine (0.072 ml, 0.513 mmol, 6.0 eq) and acrylic anhydride (0.008 g, 0.064 mmol, 0.75 eq) at room temperature. The reaction was stirred at room temperature for 2h. The solvent was evaporated, followed by purification using preparative TLC (plate was developed 4 times at DCM/MeOH 100:0 -> 98:2 -> 96:4 -> 95:5). l-[(lS,4S,5S)-5-{4-[(3-methyl-4-

{ [ 1 ,2,4]triazolo[ 1 , 5-a]pyridin-7-yloxy }phenyl)amino]pyrrolo[2, 1 -f] [ 1 ,2,4]triazin-5-yl } -2- azabicyclo[2.2.1]heptan-2-yl]prop-2-en-l-one, Example 141 (0.011 g, 25%) was isolated. LCMS (ESI) [M+H]⁺ = 507.33. 'H NMR (300 MHz, DMSO-d6) 5 8.95 (d, J = 7.4 Hz, 1H), 8.41 (d, J = 12.2 Hz, 2H), 7.91 (s, 1H), 7.75 - 7.65 (m, 2H), 7.58 (dt, J = 8.5, 2.1 Hz, 1H), 7.22 (d, J = 8.6 Hz, 1H), 7.03 (dd, J = 7.4, 2.4 Hz, 1H), 6.81 (d, J = 2.4 Hz, 1H), 6.75 (dd, J = 5.7, 2.3 Hz, 1H), 6.16 (ddd, J = 16.7, 5.7, 2.4 Hz, 1H), 5.67 (td, J = 9.7, 2.4 Hz, 1H), 4.60 (d, J = 9.0 Hz, 1H), 3.88 (dd, J = 8.5, 5.2 Hz, 1H), 3.58 (s, 1H), 3.46 (d, J = 11.0 Hz, 1H), 2.79 (d, J = 8.1 Hz, 1H), 2.25 - 2.21 (m, 1H), 2.19 (s, 3H), 1.91 - 1.74 (m, 2H), 1.74 - 1.55 (m, 2H).

Example 151

Step 1. N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(azetidin-3- yl)pyrrolo[2,l-f|[l,2,4]triazin-4-amine

[0227] TFA (2 mL) was added into a stirred mixture of tert-butyl 3-(4-((4-

([1, 2, 4]tri azolof l,5-a]pyridin-7-yloxy)-3-methylphenyl)amino)pyrrolo[2,l-f][l, 2, 4]tri azin-5- yl)azetidine-l -carboxylate (300 mg, 0.58 mmol) in DCM (5 mL). The resulting mixture was stirred for 1 hour at room temperature and then concentrated under high vacuum to afford the crude product N-(4-([l, 2, 4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(azetidin-3- yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine (200 mg crude). The crude product was used in the next step immediately without further purification. LCMS (ESI-MS) m/z =413.2 [M+H]⁺.

Step 2. (E)-l-(3-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)azetidin-l-yl)-4-bromobut-2-en-l- one

[0228] Oxalyl chloride (462 mg, 3.63 mmol) was added to a mixture of (E)-4- bromobut-2-enoic acid (240 mg, 1.45 mmol) in DCM (3 mL) at 0 °C, followed by addition of catalytic amount of DMF (0.01 mL). The resulting mixture was stirred overnight at room temperature and concentrated under vacuum to afford the crude (E)-4-bromobut-2-enoyl chloride. A mixture ofN-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5- (azetidin-3-yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine (200 mg, 0.48 mmol) and NaHCCh (407 mg, 4.85 mmol) in THF (3 mL) was stirred for 1.5 hours at room temperature. Then a solution of (E)-4-bromobut-2-enoyl chloride in 3 mL THF was added to the reaction mixture dropwise and stirred for another hour. The resulting mixture was quenched by addition of water (50 mL) and extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with brine, dried over anhydrous ISfeSCU and concentrated under vacuum to afford the crude product. The residue was purified by Prep-TLC (DCM / MeOHlO: 1) to afford the desired product (E)-l-(3-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)-pyrrolo[2,l-f][l,2,4]triazin-5-yl)azetidin-l-yl)-4-bromobut-2-en-l-one (150 mg, 46.5% yield for two steps). LCMS (ESI-MS) m/z = 559.2 [M+H]⁺.

Step 3. (E)-l-(3-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)azetidin-l-yl)-4- (methylamino)but-2-en-l-one

[0229] A solution of methylamine in EtOH (33% wt, 100 mg, 1.06 mmol) was added to a mixture of (E)-l-(3-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2, 1 -f] [ 1 ,2,4]triazin-5-yl)azetidin- 1 -yl)-4-bromobut-2-en- 1 -one (150 mg, 0.27 mmol) and diisopropylethylamine (104 mg, 0.8 mmol) in DMF (2 mL) at 0 °C. The resulting mixture was stirred for 2 hours at room temperature and concentrated under vacuum to afford the crude product. The residue was purified by reverse phase flash chromatography; Mobile Phase A: Water(0.1%FA), Mobile Phase B: ACN; Gradient: 4% B to 31% B to afford the desired product (E)-l-(3-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7- yloxy)-3 -methylphenyl)amino)pyrrolo[2, 1 -f] [ 1 ,2,4] triazin-5-yl)azetidin- 1 -yl)-4- (methylamino)but-2-en-l-one, Example 151 (9.5 mg, 6.9% yield). LCMS (ESI-MS) m/z = 510.2 [M+H]⁺. 'H NMR (400 MHz, DMSO-t/₆) 8 (ppm) 8.94 (d, J = 7.4 Hz, 1H), 8.56 (s, 1H), 8.38 (s, 1H), 7.95 (s, 1H), 7.81 (s, 1H), 7.68 (s, 2H), 7.20 (d, J = 8.5 Hz, 1H), 7.05-7.04 (m, 1H), 6.95 (s, 1H), 6.80 (s, 1H), 6.69-6.54 (m, 1H), 6.34 (d, J = 15.3 Hz, 1H), 4.70 (s, 3H), 4.42 (s, 1H), 4.26 (s, 1H), 4.05 (s, 1H), 3.61 (d, J = 5.9 Hz, 2H), 2.47 (s, 3H), 2.24-2.13 (s, 3H).

Example 164

Step 1. N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(piperidin-4- yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine

[0230] TFA (3 mL) was added to a stirred solution of tert-butyl 4-(4-((4-([l,2,4]triazolo[l,5- a]pyridin-7-yloxy)-3 -methylphenyl)amino)pyrrolo[2, 1 -f] [ 1 ,2,4]triazin-5-yl)piperidine- 1 - carboxylate (1 g, 1.48 mmol) in DCM (10 mL). The resulting mixture was stirred for 1 hour at room temperature, quenched by the addition of sat. NaHCCh (aq.) (20 mL) and extracted with EA (3 x 15 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na2SO4 and concentrated under vacuum to afford the crude product N-(4- ([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(piperidin-4-yl)pyrrolo[2,l- f][l,2,4]triazin-4-amine (400 mg crude). LCMS (ESI-MS) m/z = 441.2 [M+H]⁺

Step 2. (E)-l-(4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)piperidin-l-yl)-4-bromobut-2-en- 1-one

[0231] To a stirred mixture of (E)-4-bromobut-2-enoic acid (0.73 g, 4.41 mmol) and DMF (0.05 mL) in DCM (10 mL) was added oxalic dichloride (1.12 g, 8.82 mmol) dropwise at 0 °C. The resulting mixture was stirred overnight at 25 °C and concentrated under vacuum. The residue was dissolved in DCM (15mL). The resulting solution was added to a stirred mixture of N-(4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3-methylphenyl)-5-(piperidin-4- yl)pyrrolo[2,l-f][l,2,4]triazin-4-amine (1.92 g, 4.41 mmol) and NaHCCh (0.75 g, 8.92 mmol) in DCM (20 mL). The reaction mixture was stirred for 4 hours at room temperature and concentrated under vacuum to afford the crude product. The crude product was purified by silica gel column chromatography, eluted with PE / EA (1 :4) to afford (E)-l-(4-(4-((4- ([1, 2, 4]tri azolof l,5-a]pyridin-7-yloxy)-3-methylphenyl)amino)pyrrolo[2,l-f][l, 2, 4]tri azin-5- yl)piperidin-l-yl)-4-bromobut-2-en-l-one (800 mg, 27% yield). LCMS (ESI-MS) m/z = 587.2 [M+H]⁺

Step 3. (E)-l-(4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)piperidin-l-yl)-4-(3- fluoropyrrolidin-l-yl)but-2-en-l-one

[0232] DIEA (161 mg, 1.25 mmol) was added to a mixture of (E)-l-(4-(4-((4-

([1, 2, 4]tri azolof l,5-a]pyridin-7-yloxy)-3-methylphenyl)amino)pyrrolo[2,l-f][l, 2, 4]tri azin-5- yl)piperidin-l-yl)-4-bromobut-2-en-l-one (146 mg, 0.25 mmol) and 3 -fluoropyrrolidine hydrochloride (11 mg, 0.25 mmol) in DMF (2 mL). The resulting mixture was stirred overnight at room temperature. The reaction mixture was purified by Prep-HPLC with the following conditions: Column: XB ridge Prep OBD C18 Column, 50*250 mm, 10pm;

Mobile Phase A: Water(10nmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min mL/min; Gradient: 35% B to 65% B in 20 min; Wave Length: 254nm/220nm nm; RTl(min): 17.65, to afford the desired product (E)-l-(4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)piperidin-l-yl)-4-(3-fluoropyrrolidin-l- yl)but-2-en-l-one (15.5 mg, 10.5% yield). LCMS (ESI-MS) m/z =596.2 [M+H]⁺. 'HNMR (400 MHz, DMSO-tfc) 5(ppm) 8.95 (d, J= 7.4 Hz, 1H), 8.51 (s, 1H), 8.39 (s, 1H), 7.90 (s, 1H), 7.72 (d, J= 2.7 Hz, 1H), 7.66 (s, 1H), 7.61 (d, J= 9.3 Hz, 1H), 7.23 (d, J= 8.6 Hz, 1H), 7.04 (dd, J = 7.5, 2.6 Hz, 1H), 6.80 (d, J= 2.6 Hz, 1H), 6.71 (d, J= 2.7 Hz, 1H), 6.67-6.54 (m, 1H), 4.58 (s, 1H), 4.14 (s, 1H), 3.67 (s, 1H), 2.93-2.72 (m, 2H), 2.69 (s, 1H), 2.59-2.55

(m, 1H), 2.36-2.34 (m, 1H), 2.20 (s, 2H), 2.11 (d, J = 19.6 Hz, 1H), 2.04-1.82 (m, 2H), 1.57 (s, 2H).

Alternative Step 3a. (R,E)-l-(4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)piperidin-l-yl)-4-(3- fluoropyrrolidin-l-yl)but-2-en-l-one

[0233] DIEA (132 mg, 1.0 mmol) was added to a mixture of (E)-l-(4-(4-((4-

([1, 2, 4]tri azolof l,5-a]pyridin-7-yloxy)-3-methylphenyl)amino)pyrrolo[2,l-f][l, 2, 4]tri azin-5- yl)piperidin-l-yl)-4-bromobut-2-en-l-one (200 mg, 0.34 mmol) and (R)-3 -fluoropyrrolidine hydrochloride (43 mg, 0.34 mmol) in DMF (2 mL). The mixture was stirred overnight at room temperature. The reaction mixture was purified by Prep-HPLC with the following conditions: Column: X-select CSH Prep OBD C18 Column, 30*150 mm, 5pm; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min mL/min; Gradient: 2% B to 25% B in 9 min; Wave Length: 254nm/220nm nm; RTl(min): 8.53. The fractions with desired mass signals were combined and lyophilized to afford the desired product (R,E)-l-(4- (4-((4-([l, 2, 4]tri azolof l,5-a]pyridin-7-yloxy)-3-methylphenyl)amino)pyrrolo[2,l- f] [ 1 ,2,4]triazin-5-yl)piperidin- 1 -yl)-4-(3 -fluoropyrrolidin- 1 -yl)but-2-en- 1 -one (130.4 mg, 61.7% yield). LCMS (ESI-MS) m/z =596.2 [M+H]⁺. 'HNMR (400 MHz, Methanol-^) 8 8.76 (d, J= 7.4 Hz, 1H), 8.30 (s, 1H), 7.82 (s, 1H), 7.64-7.56 (m, 2H), 7.20 (d, J= 8.7 Hz, 1H), 7.10-7.05 (m, 1H), 6.88 (d, J= 2.6 Hz, 1H), 6.88-6.76 (m, 1H), 6.76-6.68 (m, 2H), 5.28- 5.25 (m, 1H), 4.74 (d, J= 13.1 Hz, 1H), 4.28 (d, J= 13.7 Hz, 1H), 3.61 (s, 1H), 3.37 (d, J= 6.1 Hz, 1H), 2.98-2.95 (m, 3H), 2.74-2.70 (m, 1H), 2.51-2.48 (m, 1H), 2.26 (s, 2H), 2.15 (s, 2H), 1.76-1.72 (m, 1H).

Alternative Step 3b. (S,E)-l-(4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f][l,2,4]triazin-5-yl)piperidin-l-yl)-4-(3- fluoropyrrolidin-l-yl)but-2-en-l-one

[0234] DIEA (132 mg, 1.0 mmol) was added to a mixture of (E)-l-(4-(4-((4-

([1, 2, 4]tri azolof l,5-a]pyridin-7-yloxy)-3-methylphenyl)amino)pyrrolo[2,l-f][l, 2, 4]tri azin-5- yl)piperidin-l-yl)-4-bromobut-2-en-l-one (200 mg, 0.34 mmol) and (S)-3 -fluoropyrrolidine hydrochloride (43 mg, 0.34 mmol) in DMF (2 mL). The mixture was stirred overnight at room temperature. The reaction mixture was purified by Prep-HPLC with the following conditions: Column: X-select CSH Prep OBD C18 Column, 30*150 mm, 5pm; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min mL/min; Gradient: 2% B to 25% B in 9 min; Wave Length: 254nm/220nm nm; RTl(min): 8.53. The fractions with desired mass signals were combined and lyophilized to afford the desired product (S,E)-l-(4- (4-((4-([l, 2, 4]tri azolof l,5-a]pyridin-7-yloxy)-3-methylphenyl)amino)pyrrolo[2,l- f] [ 1 ,2,4]triazin-5-yl)piperidin- 1 -yl)-4-(3 -fluoropyrrolidin- 1 -yl)but-2-en- 1 -one (131.2 mg, 64.7% yield). LCMS (ESI-MS) m/z =596.2 [M+H]⁺. 'HNMR (400 MHz, Methanol-^) 8 8.76 (d, J= 7.4 Hz, 1H), 8.30 (s, 1H), 7.82 (s, 1H), 7.66-7.56 (m, 2H), 7.20 (d, J= 8.7 Hz, 1H), 7.10-7.05 (m, 1H), 6.88 (d, J= 2.6 Hz, 1H), 6.88-6.76 (m, 1H), 6.76-6.68 (m, 2H), 5.28- 5.25 (m, 1H), 4.74 (d, J= 13.1 Hz, 1H), 4.28 (d, J= 13.7 Hz, 1H), 3.61 (s, 1H), 3.37 (d, J= 6.1 Hz, 1H), 2.98-2.95 (m, 3H), 2.73-2.70 (m, 1H), 2.51-2.48 (m, 1H), 2.26 (s, 2H), 2.15-2.13 (m, 2H), 1.76-1.72 (m, 1H).

Radiolabeling Step 3c. (R,E)-l-(4-(4-((4-([l,2,4]triazolo[l,5-a]pyridin-7-yloxy)-3- methylphenyl)amino)pyrrolo[2,l-f|[l,2,4]triazin-5-yl)piperidin-l-yl)-4-(3- fluoropyrrolidin-l-yl)but-2-en-l-one

[0235] To obtain (A)-3-(fluoro-¹⁸F)pyrrolidine in step 1, tert-butyl (S)-3- (tosyloxy)pyrrolidine-l -carboxylate is treated with titanium dioxide followed by ¹⁸F' tetrabutyl ammonium bromide in water upon heating. Then Boc protecting group is removed under treatment with aqueous HC1 followed by basification with NaOH to provide R)-3- (fluoro-¹⁸F)pyrrolidine as a free base. In step 2 DIEA was added to a mixture of (E)-l-(4-(4- ((4-([l, 2, 4]tri azolof l,5-a]pyridin-7-yloxy)-3-methylphenyl)amino)pyrrolo[2,l- f] [ 1 ,2,4]triazin-5-yl)piperidin- 1 -yl)-4-bromobut-2-en- 1 -one and (R)-3 -(fluoro-¹⁸F)pyrrolidine in DMF. The mixture was stirred at room temperature and then purified using HPLC to afford (R,E)~ 1 -(4-(4-((4-([ 1 , 2, 4]tri azolof 1 ,5-a]pyridin-7-yloxy)-3 -methylphenyl)amino)pyrrolo[2, 1 - f] [ 1 ,2,4]triazin-5-yl)piperidin- 1 -yl)-4-(3 -(fluoro-¹⁸F)pyrrolidin- 1 -yl)but-2-en- 1 -one. [0236] All compounds in Table 1 listed below can be made according to the synthetic examples described in this disclosure, and by making any necessary substitutions of starting materials that the skilled artisan would be able to obtain either commercially or otherwise.

TABLE 1

Example 2. Compound Accumulation in Tumor Cells

[0237] BaF3 HER2 YVMA mouse tumor model was used to demonstrate the enrichment of Compound 164 in HER2+ tumor specifically. BaF3 cells overexpressing HER2 YVMA exon 20 insertion mutant were inoculated into SCID mice and grew to tumors with size range of 300-400 mm3. The mice were then dosed with Compound 164 by IV at dosage of 50 mg/kg, and tissue samples (plasma, tumor, liver, kidney, heart, brain) were collected at different timepoints (0.5, 1, 2, 4, 8 and 12 hr post dose) followed by PK analysis for the levels of compound in plasma and tissue. As shown in below figure, Compound 164 exposure level is clearly higher in tumor than any other tissues tested in a time window of 4-12 hr post IV dosing. These data suggest a clinical potential of Compound 164 (and its derivatives with similar tumor enrichment profile) in PET imaging for detecting HER2 altered tumors and metastasis in human patients. FIG. 1 shows the concentration in each cell type in relation to time.

Example 3. Clinical Application for PET Imaging

[0238] Clinical applications based on PET images include, but is not limited ti:

1. Non- invasive full body detection of HER2 altered tumors and metastasis

2. Better signal-noise ratio from all tissues, suggesting a higher sensitivity than current HER2 PET imaging tracers

3. Longitudinally monitor response of a specific HER2 altered tumor to drug treatment, and also monitor appearance of HER2 signaling to non-HER2i drug treatment; and

4. Detecting presence of HER2 overexpression and oncogenic mutants which causing resistance to other HER2 inhibitors, including p95HER2 which is not detectable by current antibody -based HER2 -targeting PET tracers Example 4. Compound Accumulation in Tumor Cells

[0239] A HER2+ mouse tumor model will be used to demonstrate the PET imaging application potential using the radioactive tracer enriched in HER2+ tumor specifically. Human breast cancer model HCC1954 (high HER2 expression) is suitable for this purpose. The radioactive tracer will be dosed by IV at desirable dosage into the mouse with tumor at size of 200-500 mm3. At certain time points (4, 8 and 12 hr), PET imaging will be conducted to image tumor-associated PET signal vs signal from other tissue and organs.

[0240] While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.

Claims

CLAIMS What is claimed is:

1. A radiolabeled compound having the structure of Formula I: or a pharmaceutically acceptable salt thereof, wherein:

A is selected from CH and N;

B is selected from CH2 ,CHF, and CF2;

E is selected from CH2, CHF, and CF2;

X is selected from CH, CF, C(OH) and N; or X is CH and B and E are both absent;

Q¹ is selected from the group consisting of H, Ci-Cealkyl, F, and Cl;

Q³ is selected from H and F;

R¹ is selected from the group consisting of alkyl, haloalkyl and halogen;

R² is selected from the group consisting of is -O-alkyl, -O-aryl, -O-heteroaryl, -O- cycloalkyl, -O-heterocycloalkyl, -O-heteroaryl-alkylene-aryl, -NH-alkyl, -NH-aryl, or-NH- heteroaryl, wherein each of the alkyl, aryl, heteroaryl, cycloalkyl or heteocycloalkyl moieties are optionally substituted with 1-4 J¹ groups; or R¹ and R² join with the carbon atoms to which they are attached to form a saturated or unsaturated carbocyclic or heterocyclic ring, wherein the saturated or unsaturated carbocyclic or heterocyclic ring is optionally substituted with 1-4 J¹ groups; each J¹ is independently selected from the group consisting of halogen, alkyl, haloalkyl, hydroxy, hydroxyalkyl, -Co-C4alkylene-N(H)R^C, alkoxy, and alkoxyalkyl; and wherein at least one carbon atom of Formula (I) is substituted with at least one ¹⁸F, and the at least one ¹⁸F atom can substitute any hydrogen or halogen atom in Formula (I) provided that a C-¹⁸F bond is formed.

2. The radiolabeled compound of claim 1, wherein the radiolableled compound has a structure of Formula (II): or a pharmaceutically acceptable salt thereof.

3. The radiolabeled compound of claim 1 or 2, wherein Q¹ is H or Ci-Ce alkyl.

4. The radiolabeled compound of claim 3, wherein Q¹ is H.

5. The radiolabeled compound of claim 3, wherein Q¹ is methyl, ethyl, or propyl.

6. The radiolabeled compound of any one of claims 1 to 5, wherein Q² is -Ci- C3alkylene-NR^aR^b, -Co-C4alkylene-cycloalkyl optionally substituted with 1-3 J⁴ groups, or - Co-C4alkylene-heterocycloalkyl optionally substituted with 1-3 J⁴.

7. The radiolabeled compound of claim 6, wherein Q² is -Ci-C3alkylene-NR^aR^b.

8. The radiolabeled compound of claim 6, wherein Q² is -Co-C4alkylene- heterocycloalkyl optionally substituted with 1-3 J⁴.

9. The radiolabeled compound of anyone of claims 1 to 8, wherein each J⁴ is independently halogen, alkyl, haloalkyl, hydroxy, hydroxyalkyl, alkoxy, or alkoxyalkyl.

10. The radiolabeled compound of claim 9, wherein each J⁴ is independently halogen or alkyl.

11. The radiolabeled compound of claim 10, wherein each J⁴ is independently halogen.

12. The radiolabeled compound of claim 10, wherein each J⁴ is independently C1-C3 alkyl.

13. The radiolabeled compound of any one of claims 1 to 12, wherein X is CH, CF, or C(OH).

14. The radiolabeled compound of claim 13, wherein X is CH or CF.

15. The radiolabeled compound of claim 14, wherein X is CH.

16. The radiolabeled compound of any one of claims 1 to 15, wherein at least one carbon atom of Q¹, Q², B, E, or X that has a single bond to either a hydrogen or halogen atom is substituted with ¹⁸F.

17. The radiolabeled compound of claim 16, wherein at least one carbon atom of Q¹, Q², B, or E that has a single bond to either a hydrogen or halogen atom is substituted with ¹⁸F.

18. The radiolabeled compound of claim 17, wherein at least one carbon atom of Q¹ or Q² that has a single bond to either a hydrogen or halogen atom is substituted with ¹⁸F.

19. The radiolabeled compound of any one of claims 1 to 18, wherein the compound has the following structure: or a pharmaceutically acceptable salt thereof.

20. A method of selecting a subject for treating a HER2 mediated disease, the method comprising administering to the subject a radiolableled compound of any one of claims 1 to 19, or a pharmaceutically acceptable salt thereof, generating a scanning image of the subject, and selecting the subject for treating the HER2 mediated disease when the scanning image shows that the radiolableled compound, or a pharmaceutically acceptable salt thereof, is accumulated in a tumor tissue.

21. The method of claim 20, wherein the radiolableled compound, or a pharmaceutically acceptable salt thereof, is administered via systemic administration.

22. The method of claim 20 or 21, wherein the radiolableled compound, or a pharmaceutically acceptable salt thereof, is accumulated in the brain.

23. The method of any one of claims 20 to 22, wherein the scanning image is a tomographic image.

24. The method of claim 23, wherein the tomographic image is generated with positron emission tomography (PET).

25. The method of any one of claims 20 to 24, wherein the HER2 mediated disease is metastatic brain tumor.

26. The method of claim 25, wherein the metastatic brain tumor is caused by lung cancer, breast cancer, skin cancer, colon cancer, or melanoma.

27. The method of claim 26, wherein the metastatic brain tumor is caused by breast cancer.

28. The method of any one of claims 20 to 24, wherein the HER2 mediated disease is cancer.

29. The method of claim 28, wherein the cancer is brain cancer, colorectal cancer, breast cancer, bladder cancer, biliary cancer, ovarian cancer, or non-small cell lung cancer.

30. The method of claim 29, wherein the cancer is brain cancer.

31. A method of tracking tumor response to a HER2 mediated disease in a subject, the method comprising administering to the subject a radiolableled compound of any one of claims 1 to 19, or a pharmaceutically acceptable salt thereof, and generating a scanning image of the subject.

32. The method of claim 31, wherein the scanning image is generated once every two weeks.

33. The method of claim 32, wherein the scanning image is generated for at least 2 months.

34. The method of any one of claims 31 to 33, wherein the radiolableled compound, or a pharmaceutically acceptable salt thereof, is administered via systemic administration.

35. The method of any one of claims 31 to 34, wherein the radiolableled compound, or a pharmaceutically acceptable salt thereof, is accumulated in the brain.

36. The method of any one of claims 31 to 35, wherein the scanning image is a tomographic image.

37. The method of claim 36, wherein the tomographic image is generated with positron emission tomography (PET).

38. The method of any one of claims 31 to 37, wherein the HER2 mediated disease is metastatic brain tumor.

39. The method of claim 38, wherein the metastatic brain tumor is caused by lung cancer, breast cancer, skin cancer, colon cancer, or melanoma.

40. The method of claim 39, wherein the metastatic brain tumor is caused by breast cancer.

41. The method of any one of claims 31 to 37, wherein the HER2 mediated disease is cancer.

42. The method of claim 41, wherein the cancer is brain cancer, colorectal cancer, breast cancer, bladder cancer, biliary cancer, ovarian cancer, or non-small cell lung cancer.

43. The method of claim 42, wherein the cancer is brain cancer.

44. A method of obtaining a scanning image, comprising administering to a subject the radiolableled compound of any one of claims 1 to 19, or a pharmaceutically acceptable salt thereof, and subsequently generating a scanning image of the subject.

45. The method of claim 44, wherein the scanning image is a tomographic image.

46. The method of claim 45, wherein the tomographic image is generated with positron emission tomography (PET).