Methods and compounds for restoring mutant p53 function

Abstract

Mutations in oncogenes and tumor suppressor genes contribute to the development and progression of cancer. The present disclosure describes compounds and methods for restoring wild-type function to p53 mutants. The compounds of the invention can bind to mutant p53 and restore the ability of p53 mutants to bind DNA and activate downstream effectors involved in tumor suppression. The disclosed compounds can be used to reduce the progression of cancers containing p53 mutations.

Description

[0001] Cross-referencing

[0002] This application claims the benefit of U.S. Provisional Application No. 62 / 904,369, filed September 23, 2019; and U.S. Provisional Application No. 63 / 038,388, filed June 12, 2020, the entire contents of which are incorporated herein by reference.

[0003] sequence list

[0004] This application contains a sequence list, which has been electronically submitted in ASCII format, the entire contents of which are hereby incorporated by reference. The ASCII copy was created on September 18, 2020, and is named 44727-705.601 SequenceListing.txt, with a size of 2,578 bytes.

[0005] background

[0006] Cancer, an uncontrolled proliferation of cells, is a multifactorial disease characterized by tumor formation, growth, and in some cases, metastasis. The replication of cells carrying activated oncogenes, damaged genomes, or other cancer-promoting alterations can be prevented by a complex tumor suppressor network. A core component of this network is p53, one of the most potent tumor suppressor genes in cells. Both wild-type and mutant conformations of p53 are associated with cancer progression.

[0007] Incorporation

[0008] Every patent, publication and non-patent document cited in this application is hereby incorporated in its entirety by reference, as if each patent, publication and non-patent document were individually incorporated by reference. Summary of the Invention

[0009] In some embodiments, this document describes a compound comprising a heterocyclic group containing a halogenated substituent, wherein the compound binds to mutant p53 protein and increases the wild-type p53 activity of mutant p53 protein.

[0010] In some embodiments, compounds of the following formula are described herein:

[0011]

[0012] in:

[0013] -each It can be a single bond or a double bond independently;

[0014] -X 1 For CR5 CR 5 R 6 , N, NR 5 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0015] -X 2 For CR 7 CR 7 R 8 , N, NR 7 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0016] -X 3 For CR 9 CR 9 R 10 , N, NR 9 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0017] -X 4 For CR 11 CR 11 R 12 , N, NR 11 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0018] -X 5 For CR 13 , N or NR 13 ;

[0019] Where X 1 X 2 X 3 and X 4 At least one of them is connected to Q 1 carbon atoms;

[0020] -Q 1 For C=O, C=S, C=CR 14 R 15 C=NR 14 Alkylene, alkenylene, or ynylene groups, each of which is independently substituted or unsubstituted; or they may be bonds;

[0021] -m is 1, 2, 3 or 4;

[0022] -Y is N, O, or does not exist;

[0023] -R 1 -C(O)R 16 -C(O)OR 16-C(O)NR 16 R 17 -OR 16 -SR 16 -NR 16 R 17 -NR 16 C(O)R 16 -OC(O)R 16 C=O, C=S, -CN, -SiR 16 R 17 R 18 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen;

[0024] - Each R 3 and R 4 Independently for -C(O)R 19 -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 -SO2R 19 Alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen, or R 3 and R 4 Together with R 3 and R 4 The combined Y atoms form a ring, wherein the ring is substituted or unsubstituted, or R 3 It does not exist;

[0025] - Each R 2 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 R 13 R 14 R 15 R 16 R 17 and R 18 Independently for -C(O)R 21 -C(O)OR 21 -C(O)NR 21 R 22 -OR 21 -SR 21 -NR 21 R 22 -NR 21C(O)R 22 -OC(O)R 21 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0026] - Each R 19 and R 20 For C(O)R 23 -C(O)OR 23 -C(O)NR 23 R 24 -OR 23 -SR 23 -NR 23 R 24 -NR 23 C(O)R 24 -OC(O)R 23 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0027] - Each R 21 and R 22 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic, each of which is independently substituted or unsubstituted; or hydrogen; and

[0028] - Each R 23 and R 24 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each independently either substituted or unsubstituted; or hydrogen.

[0029] Or a pharmaceutically acceptable salt thereof, wherein the compound is not one of the compounds listed in Table 1.

[0030] In some embodiments, compounds of the following formula are described herein:

[0031]

[0032] in:

[0033] -each It can be a single bond or a double bond independently;

[0034] -X 1 For CR 5 CR 5 R 6 , N, NR 5 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0035] -X 2 For CR7 CR 7 R 8 , N, NR 7 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0036] -X 3 For CR 9 CR 9 R 10 , N, NR 9 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0037] -X 4 For CR 11 CR 11 R 12 , N, NR 11 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0038] -X 5 For CR 13 , N or NR 13 ;

[0039] Where X 1 X 2 X 3 and X 4 At least one of them is connected to Q 1 carbon atoms;

[0040] -Q 1 For C=O, C=S, C=CR 14 R 15 C=NR 14 Alkylene, alkenylene, or ynylene groups, each of which is independently substituted or unsubstituted; or they may be bonds;

[0041] -m is 1, 2, 3 or 4;

[0042] -Y is N, O, or does not exist;

[0043] -R 1 -C(O)R 16 -C(O)OR 16 -C(O)NR 16 R 17 -OR 16 -SR 16 -NR 16 R 17 -NR 16 C(O)R 16 -OC(O)R16 C=O, C=S, -CN, -SiR 16 R 17 R 18 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen;

[0044] - Each R 3 and R 4 Independently for -C(O)R 19 -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 -SO2R 19 Alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen, or R 3 and R 4 Together with R 3 and R 4 The combined nitrogen atoms form a ring, wherein the ring is substituted or unsubstituted, or R 3 It does not exist, where R 3 and R 4 At least one of them is an alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclic group, each of which is at least halogenated;

[0045] - Each R 2 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 R 13 R 14 R 15 R 16 R 17 and R 18 Independently for -C(O)R 21 -C(O)OR 21 -C(O)NR 21 R 22 -OR 21 -SR 21 -NR 21 R 22 -NR 21 C(O)R 22 -OC(O)R 21Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0046] - Each R 19 and R 20 For C(O)R 23 -C(O)OR 23 -C(O)NR 23 R 24 -OR 23 -SR 23 -NR 23 R 24 -NR 23 C(O)R 24 -OC(O)R 23 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0047] - Each R 21 and R 22 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic, each of which is independently substituted or unsubstituted; or hydrogen; and

[0048] - Each R 23 and R 24 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each independently either substituted or unsubstituted; or hydrogen.

[0049] Or its pharmaceutically acceptable salt.

[0050] In some embodiments, this document describes a compound comprising a heterocyclic group containing a halogenated substituent, wherein the compound binds to the mutant p53 protein and increases the wild-type p53 activity of the mutant p53.

[0051] In some implementations, this document describes a method for treating cancer, which includes administering a compound of formula (I) to a subject in need:

[0052]

[0053] in:

[0054] -each It can be a single bond or a double bond independently;

[0055] -X 1 For CR 5 CR 5 R 6 , N, NR 5, O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0056] -X 2 For CR 7 CR 7 R 8 , N, NR 7 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0057] -X 3 For CR 9 CR 9 R 10 , N, NR 9 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0058] -X 4 For CR 11 CR 11 R 12 , N, NR 11 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0059] -X 5 For CR 13 , N or NR 13 ;

[0060] Where X 1 X 2 X 3 and X 4 At least one of them is connected to Q 1 carbon atoms;

[0061] -Q 1 For C=O, C=S, C=CR 14 R 15 C=NR 14 Alkylene, alkenylene, or ynylene groups, each of which is independently substituted or unsubstituted; or they may be bonds;

[0062] -m is 1, 2, 3 or 4;

[0063] -Y is N, O, or does not exist;

[0064] -R 1 -C(O)R 16 -C(O)OR 16 -C(O)NR 16 R 17 -OR 16 -SR16 -NR 16 R 17 -NR 16 C(O)R 16 -OC(O)R 16 C=O, C=S, -CN, -SiR 16 R 17 R 18 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen;

[0065] - Each R 3 and R 4 Independently for -C(O)R 19 -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 -SO2R 19 Alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen, or R 3 and R 4 Together with R 3 and R 4 The combined Y atoms form a ring, wherein the ring is substituted or unsubstituted, or R 3 It does not exist;

[0066] - Each R 2 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 R 13 R 14 R 15 R 16 R 17 and R 18 Independently for -C(O)R 21 -C(O)OR 21 -C(O)NR 21 R 22 -OR 21 -SR 21 -NR 21 R 22 -NR 21 C(O)R 22 -OC(O)R 21Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0067] - Each R 19 and R 20 For C(O)R 23 -C(O)OR 23 -C(O)NR 23 R 24 -OR 23 -SR 23 -NR 23 R 24 -NR 23 C(O)R 24 -OC(O)R 23 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0068] - Each R 21 and R 22 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic, each of which is independently substituted or unsubstituted; or hydrogen; and

[0069] - Each R 23 and R 24 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each independently either substituted or unsubstituted; or hydrogen.

[0070] Or its pharmaceutically acceptable salt;

[0071] Among them, the SC of this compound against p53 Y220C, as measured by homogeneous time-resolved fluorescence (HTRF) assay, was... 150 The value is less than 1 μM.

[0072] In some embodiments, compounds of the following formula are described herein:

[0073]

[0074] in:

[0075] -each It can be a single bond or a double bond independently;

[0076] -X 1 For CR 5 CR 5 R 6 , N, NR 5 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0077] -X 2 For CR 7 CR 7 R 8 , N, NR 7 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0078] -X 3 For CR 9 CR 9 R 10 , N, NR 9 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0079] -X 4 For CR 11 CR 11 R 12 , N, NR 11 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0080] -X 5 For CR 13 , N or NR 13 ;

[0081] Where X 1 X 2 X 3 and X 4 At least one of them is connected to Q 1 carbon atoms;

[0082] -A is a linking group;

[0083] -Q 1 For C=O, C=S, C=CR 14 R 15 C=NR 14 Alkylene, alkenylene, or ynylene groups, each of which is independently substituted or unsubstituted; or they may be bonds;

[0084] -m is 1, 2, 3 or 4;

[0085] -Y is N, O, or does not exist;

[0086] -R 1 -C(O)R 16 -C(O)OR 16 -C(O)NR 16 R 17 -OR 16 -SR 16 -NR16 R 17 -NR 16 C(O)R 16 -OC(O)R 16 -SiR 16 R 17 R 18 Alkyl, alkenyl, alkoxy, aryl, heteroaryl, heterocyclic, or halogenated, each of which is independently substituted or unsubstituted; or hydrogen;

[0087] - Each R 3 and R 4 Independently for -C(O)R 19 -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 -SO2R 19 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen, or R 3 and R 4 Together with R 3 and R 4 The combined nitrogen atoms form a ring, wherein the ring is substituted or unsubstituted, or R 3 It does not exist; where R 3 and R 4 At least one of them is an alkyl, alkylene, alkenyl, alkenylene, alkynyl, alkynylene, aryl, heteroaryl, or heterocyclic group, each of which is at least halogenated;

[0088] - Each R 2 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 R 13 R 14 R 15 R 16 R 17 and R 18 Independently for -C(O)R 21 -C(O)OR 21 -C(O)NR 21 R 22 -OR 21 -SR 21 -NR 21 R 22 -NR 21C(O)R 22 -OC(O)R 21 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0089] - Each R 19 and R 20 Independently for -C(O)R 23 -C(O)OR 23 -C(O)NR 23 R 24 -OR 23 -SR 23 -NR 23 R 24 -NR 23 C(O)R 24 -OC(O)R 23 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0090] - Each R 21 and R 22 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic, each of which is independently substituted or unsubstituted; or hydrogen; and

[0091] - Each R 23 and R 24 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each independently either substituted or unsubstituted; or hydrogen.

[0092] Or its pharmaceutically acceptable salt.

[0093] In some embodiments, compounds of the following formula are described herein:

[0094]

[0095] in:

[0096] -each It can be a single bond or a double bond independently;

[0097] -X 1 For CR 5 CR 5 R 6 , N, NR 5 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0098] -X 2 For CR 7 CR7 R 8 , N, NR 7 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0099] -X 3 For CR 9 CR 9 R 10 , N, NR 9 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0100] -X 4 For CR 11 CR 11 R 12 , N, NR 11 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0101] -X 5 For CR 13 , N or NR 13 ;

[0102] Where X 1 X 2 X 3 and X 4 At least one of them is connected to Q 1 carbon atoms;

[0103] -A is a linking group;

[0104] -Q 1 For C=O, C=S, C=CR 14 R 15 C=NR 14 Alkylene, alkenylene, or ynylene groups, each of which is independently substituted or unsubstituted; or they may be bonds;

[0105] -m is 1, 2, 3 or 4;

[0106] -Y is N, O, or does not exist;

[0107] -R 1 -C(O)R 16 -C(O)OR 16 -C(O)NR 16 R 17 -OR 16 -SR 16 -NR 16 R 17 -NR 16 C(O)R16 -OC(O)R 16 -SiR 16 R 17 R 18 Alkyl, alkenyl, alkoxy, aryl, heteroaryl, heterocyclic, or halogenated, each of which is independently substituted or unsubstituted; or hydrogen;

[0108] - Each R 3 and R 4 Independently for -C(O)R 19 -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 -SO2R 19 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen, or R 3 and R 4 Together with R 3 and R 4 The combined nitrogen atoms form a ring, wherein the ring is substituted or unsubstituted, or R 3 It does not exist;

[0109] - Each R 2 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 R 13 R 14 R 15 R 16 R 17 and R 18 Independently for -C(O)R 21 -C(O)OR 21 -C(O)NR 21 R 22 -OR 21 -SR 21 -NR 21 R 22 -NR 21 C(O)R 22 -OC(O)R 21 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0110] - Each R 19 and R20 Independently for -C(O)R 23 -C(O)OR 23 -C(O)NR 23 R 24 -OR 23 -SR 23 -NR 23 R 24 -NR 23 C(O)R 24 -OC(O)R 23 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0111] - Each R 21 and R 22 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic, each of which is independently substituted or unsubstituted; or hydrogen; and

[0112] - Each R 23 and R 24 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each independently either substituted or unsubstituted; or hydrogen.

[0113] Or a pharmaceutically acceptable salt thereof, wherein the compound is not one of the compounds listed in Table 1.

[0114] In some embodiments, this document describes a method for inducing apoptosis of cells, the method comprising contacting cells with a therapeutically effective amount of a compound of the present disclosure that binds to a p53 mutant, wherein the compound increases the ability of the p53 mutant to bind DNA, wherein the cells express the p53 mutant.

[0115] In some embodiments, this document describes a method for treating cancer, which includes administering a therapeutically effective amount of the disclosed compound to a subject in need. Detailed Implementation

[0116] This invention provides compounds and methods for restoring the wild-type function of mutant p53. The compounds of this invention can bind to mutant p53 and restore the ability of the p53 mutant to bind DNA. Restoration of p53 mutant activity allows for the activation of downstream effectors of p53, leading to inhibition of cancer progression. This invention also provides methods for treating cancerous lesions or tumors carrying p53 mutations.

[0117] Cancer is a group of related diseases characterized by the uncontrolled proliferation of cells with the potential to metastasize throughout the body. Cancers can be classified into five main categories, including, for example: carcinomas, which can arise from cells both inside and outside the body (such as the lungs, breast, and colon); sarcomas, which can arise from cells located in bone, cartilage, fat, connective tissue, muscle, and other supporting tissues; lymphomas, which can occur in lymph nodes and immune system tissues; leukemias, which can occur in the bone marrow and accumulate in the bloodstream; and adenomas, which can occur in the thyroid gland, pituitary gland, adrenal glands, and other glandular tissues.

[0118] Although different cancers can develop in almost any tissue of the body and have unique characteristics, the basic processes that lead to cancer are likely similar in all forms of the disease. Cancer begins when a cell breaks free from the normal limits of cell division and begins to grow and divide uncontrollably. Genetic mutations in a cell can prevent it from repairing damaged DNA or initiating apoptosis, potentially leading to uncontrolled cell growth and division.

[0119] The proliferative capacity of tumor cell populations depends not only on the cell proliferation rate but also on the cell consumption rate. Programmed cell death, or apoptosis, represents the main mechanism of cell consumption. Cancer cells can evade apoptosis through various strategies, such as inhibiting p53 function, thereby suppressing the expression of pro-apoptotic proteins.

[0120] Oncogenes and tumor suppressor genes regulate cell proliferation. Gene mutations can affect both oncogenes and tumor suppressor genes, potentially causing abnormal activation or inhibition of their activity, further promoting uncontrolled cell division. Oncogenes contribute to cell growth, while tumor suppressor genes slow cell division by repairing damaged DNA and activating apoptosis. Oncogenes that can mutate in cancer include, for example, Cdk1, Cdk2, Cdk3, Cdk4, Cdk6, EGFR, PDGFR, VEGF, HER2, Raf kinase, K-Ras, and myc. Tumor suppressor genes that can mutate in cancer include, for example, BRCA1, BRCA2, cyclin-dependent kinase inhibitor 1C, retinoblastoma protein (pRb), PTEN, p16, p27, p53, and p73.

[0121] Tumor suppressor gene p53.

[0122] The tumor suppressor gene p53 is a 393-amino acid transcription factor that regulates cell growth in response to cellular stresses, including, for example, UV radiation, hypoxia, oncogene activation, and DNA damage. p53 possesses multiple mechanisms for inhibiting cancer progression, including, for example, initiation of apoptosis, maintenance of genomic stability, cell cycle arrest, senescence induction, and inhibition of angiogenesis. Due to its crucial role in tumor suppression, p53 is inactivated in almost all cancers either through direct mutations or by perturbation of relevant signaling pathways involved in tumor suppression. Homozygous loss of the p53 gene occurs in almost all types of cancer, including breast, colon, and lung cancer. In several types of human cancers, the presence of certain p53 mutations may be associated with a less favorable patient prognosis.

[0123] In the absence of stress signals, p53 levels are maintained at a low level through the interaction between p53 and Mdm2 (an E3 ubiquitin ligase). In unstressed cells, Mdm2 can target p53 for degradation by the proteasome. Under stress conditions, the interaction between Mdm2 and p53 is disrupted, leading to p53 accumulation. A key event leading to p53 activation is the phosphorylation of the N-terminal domain of p53 by a protein kinase, thereby transducing upstream stress signals. Phosphorylation of p53 results in a conformational change, which can promote p53 binding to DNA and allow transcription of downstream effectors. Activation of p53 can induce, for example, intrinsic and extrinsic apoptosis pathways, cell cycle arrest, senescence, and DNA repair. p53 can activate proteins involved in the aforementioned pathways, including, for example, Fas / Apo1, Killer / DR5, Bax, Puma, Noxa, Bid, caspase-3, caspase-6, caspase-7, caspase-8, caspase-9, and p21(WAF1). Furthermore, p53 can inhibit the transcription of various genes, such as c-MYC, cyclin B, VEGF, RAD51, and hTERT.

[0124] Each strand of the p53 tetramer consists of several functional domains, including a transactivation domain (amino acids 1-100), a DNA-binding domain (amino acids 101-306), and a tetramerizing domain (amino acids 307-355). These functional domains are highly mobile and primarily unstructured. Most p53 oncogenes are located in the protein's core DNA-binding domain, which contains a central β-sandwich of antiparallel β-sheets that acts as the basic scaffold of the DNA-binding surface. The DNA-binding surface consists of two β-turn loops, L2 and L3, which are stabilized by zinc ions at, for example, Arg175 and Arg248, as well as loop-sheet-helical motifs. In summary, these structural elements form an extended DNA-binding surface enriched with positively charged amino acids that specifically contact various p53 response elements.

[0125] Because p53 mutations are prevalent in almost all types of cancer, reactivating wild-type p53 function in cancer cells may be an effective therapy. Mutations in p53 located in the DNA-binding domains or peripheries of the DNA-binding surface of proteins result in abnormal protein folding required for DNA recognition and binding. For example, mutations in p53 can occur at the amino acids Val143, His168, Arg175, Tyr220, Gly245, Arg248, Arg249, Phe270, Arg273, and Arg282. p53 mutations that can invalidate p53 activity include, for example, R175H, Y220C, G245S, R248Q, R248W, R273H, and R282H. These p53 mutations may distort the structure of the DNA-binding site or cause thermodynamic instability of the folded protein at body temperature. Wild-type function of p53 mutants can be restored by combining p53 mutants with a compound that can shift the fold-unfold balance to the folded state, thereby reducing the rate of unfolding and instability.

[0126] Non-limiting examples of amino acids include: alanine (A, Ala); arginine (R, Arg); asparagine (N, Asn); aspartic acid (D, Asp); cysteine ​​(C, Cys); glutamic acid (E, Glu); glutamine (Q, Gln); glycine (G, Gly); histidine (H, His); isoleucine (I, Ile); leucine (L, Leu); lysine (K, Lys); methionine (M, Met); phenylalanine (F, Phe); proline (P, Pro); serine (S, Ser); threonine (T, Thr); tryptophan (W, Trp); tyrosine (Y, Tyr); and valine (V, Val).

[0127] The mechanism of action of the compounds of this invention.

[0128] The compounds of this invention selectively bind to p53 mutants and restore wild-type activity of the p53 mutants, including, for example, DNA binding function and activation of downstream targets involved in tumor suppression. In some embodiments, the compounds of this invention selectively bind to the p53 Y220C mutant. The Y220C mutant is a temperature-sensitive mutant that binds to DNA at lower temperatures and denatures at body temperature. The compounds of this invention stabilize the Y220C mutant to reduce the likelihood of protein denaturation at body temperature.

[0129] The aromatic ring of Y220 is located at the periphery of the p53 β-sandwich connecting β chains S7 and S8, and is a component of the hydrophobic core of the β-sandwich. Due to the formation of an inner surface cavity, the Y220C mutation can be highly unstable. The compounds of this invention can bind to and occupy this surface cleft to stabilize the β-sandwich, thereby restoring wild-type p53 DNA binding activity.

[0130] To determine the ability of the compounds of the present invention to bind to and stabilize mutant p53, assays can be used to detect, for example, conformational changes in the p53 mutant or activation of the wild-type p53 target. Conformational changes in p53 can be measured, for example, by differential scanning fluorescence (DSF), isothermal titration calorimetry (ITC), nuclear magnetic resonance spectroscopy (NMR), or X-ray crystallography. Furthermore, wild-type specific antibodies against the p53 mutant conformation can be used to detect conformational changes by, for example, immunoprecipitation (IP), immunofluorescence (IF), or Western blotting.

[0131] Methods for detecting the ability of p53 mutants to bind DNA may include, for example, DNA affinity immunoblotting, modified enzyme-linked immunosorbent assay (ELISA), electrophoretic mobility shift assay (EMSA), fluorescence resonance energy transfer (FRET), homogeneous time-resolved fluorescence (HTRF), and chromatin immunoprecipitation (ChIP) assays.

[0132] To determine whether the compounds described herein can reactivate the transcriptional activity of p53, the activation of downstream targets in the p53 signaling cascade can be measured. Activation of p53 effector proteins can be detected, for example, by immunohistochemistry (IHC-P), reverse transcription polymerase chain reaction (RT-PCR), and Western blotting. Measurement of p53 activation can also be performed via apoptosis induction through the caspase cascade system, using methods including, for example, annexin V staining, TUNEL assay, caspaseogen and caspase levels, and cytochrome c levels. Another consequence of p53 activation is senescence, which can be measured using methods such as β-galactosidase staining.

[0133] The p53 mutant that can be used to determine the effectiveness of the compounds of the present invention in enhancing the DNA-binding ability of p53 mutants is a p53 truncated mutant containing only amino acids 94-312, covering the DNA-binding domain of p53. For example, the sequence of the p53 Y220C mutant used to test the efficacy of the compounds may be:

[0134] SSSVPSQ KTYQGSYGFR LGFLHSGTAK SVTCTYSPAL NKMFCQLAKT CPVQLWVDSTPPPGTRVRAM AIYKQSQHMT EVVRRCPHHE RCSDSDGLAP PQHLIRVEGN LRVEYLDDRN TFRHSVVVPCEPPEVGSDCT TIHYNYMCNS SCMGGMNRRP ILTIITLEDS SGNLLGRNSF EVHVCACPGR DRRTEEENLRKKGEPHHELP PGSTKRALSN NT(SEQ ID NO.1)

[0135] Compared to the ability of p53 mutants to bind DNA in the absence of the compounds of the present invention, the compounds of the present invention can increase the ability of p53 mutants to bind DNA by at least or at most about 0.1%, at least or at most about 0.2%, at least or at most about 0.3%, at least or at most about 0.4%, at least or at most about 0.5%, at least or at most about 0.6%, at least or at most about 0.7%, at least or at most about 0.8%, at least or at most about 0.9%, at least or at most about 1%, at least or at most about 2%, at least or at most about 3%, at least or at most about 4%, at least or at most about 5%, at least or at most about 6%, at least or at most about 7%, at least or at most about 8%, at least or at most about 9%, at least or at most about 10%, at least or At most about 11%, at least or at most about 12%, at least or at most about 13%, at least or at most about 14%, at least or at most about 15%, at least or at most about 16%, at least or at most about 17%, at least or at most about 18%, at least or at most about 19%, at least or at most about 20%, at least or at most about 21%, at least or at most about 22%, at least or at most about 23%, at least or at most about 24%, at least or at most about 25%, at least or at most about 26%, at least or at most about 27%, at least or at most about 28%, at least or at most about 29%, at least or at most about 30%, at least or at most about 31%, at least or at most about 32%, at least or at most about 33%, at least or at most about 34%, at least or at most about 35%, at least Or at most about 36%, at least or at most about 37%, at least or at most about 38%, at least or at most about 39%, at least or at most about 40%, at least or at most about 41%, at least or at most about 42%, at least or at most about 43%, at least or at most about 44%, at least or at most about 45%, at least or at most about 46%, at least or at most about 47%, at least or at most about 48%, at least or at most about 49%, at least or at most about 50%, at least or at most about 51%, at least or at most about 52%, at least or at most about 53%, at least or at most about 54%, at least or at most about 55%, at least or at most about 56%, at least or at most about 57%, at least or at most about 58%, at least or at most about 59%, at least or at most about 60%, to At least or at most about 61%, at least or at most about 62%, at least or at most about 63%, at least or at most about 64%, at least or at most about 65%, at least or at most about 66%, at least or at most about 67%, at least or at most about 68%, at least or at most about 69%, at least or at most about 70%, at least or at most about 71%, at least or at most about 72%, at least or at most about 73%, at least or at most about 74%, at least or at most about 75%, at least or at most about 76%, at least or at most about 77%, at least or at most about 78%, at least or at most about 79%, at least or at most about 80%, at least or at most about 81%, at least or at most about 82%, at least or at most about 83%, at least or at most about 84%, at least or at most about 85%.At least or at most about 86%, at least or at most about 87%, at least or at most about 88%, at least or at most about 89%, at least or at most about 90%, at least or at most about 91%, at least or at most about 92%, at least or at most about 93%, at least or at most about 94%, at least or at most about 95%, at least or at most about 96%, at least or at most about 97%, at least or at most about 98%, at least or at most about 99%, at least or at most about 100%, at least or at most about 125%, at least or at most about 150%, at least or at most about 175%, at least or at most about 200%, at least or at most about 225%, or at least or at most about 250%.

[0136] The compounds described herein can increase the activity of p53 mutants by, for example, at least or at most about 2 times, at least or at most about 3 times, at least or at most about 4 times, at least or at most about 5 times, at least or at most about 6 times, at least or at most about 7 times, at least or at most about 8 times, at least or at most about 9 times, at least or at most about 10 times, at least or at most about 11 times, at least or at most about 12 times, at least or at most about 13 times, at least or at most about 14 times, at least or at most about 15 times, at least or at most about 16 times, at least or at most about 17 times, or at least... Or at most about 18 times, at least or at most about 19 times, at least or at most about 20 times, at least or at most about 25 times, at least or at most about 30 times, at least or at most about 35 times, at least or at most about 40 times, at least or at most about 45 times, at least or at most about 50 times, at least or at most about 55 times, at least or at most about 60 times, at least or at most about 65 times, at least or at most about 70 times, at least or at most about 75 times, at least or at most about 80 times, at least or at most about 85 times, at least or at most about 90 times, at least or at most about 95 times, at least or at most about 100 times, at least or at most about 110 times, at least or At most approximately 120 times, at least or at most approximately 130 times, at least or at most approximately 140 times, at least or at most approximately 150 times, at least or at most approximately 160 times, at least or at most approximately 170 times, at least or at most approximately 180 times, at least or at most approximately 190 times, at least or at most approximately 200 times, at least or at most approximately 250 times, at least or at most approximately 300 times, at least or at most approximately 350 times, at least or at most approximately 400 times, at least or at most approximately 450 times, at least or at most approximately 500 times, at least or at most approximately 550 times, at least or at most approximately 600 times, at least or at most approximately 650 times, at least or at most approximately 7 00 times, at least or at most about 750 times, at least or at most about 800 times, at least or at most about 850 times, at least or at most about 900 times, at least or at most about 950 times, at least or at most about 1,000 times, at least or at most about 1,500 times, at least or at most about 2,000 times, at least or at most about 3,000 times, at least or at most about 4,000 times, at least or at most about 5,000 times, at least or at most about 6,000 times, at least or at most about 7,000 times, at least or at most about 8,000 times, at least or at most about 9,000 times, or at least or at most about 10,000 times.

[0137] For example, the compounds of the present invention can be used to induce apoptosis, cell cycle arrest, or senescence in cells. In some embodiments, the cells are cancer cells. In some embodiments, the cells carry a mutation in p53.

[0138] The compounds of this invention.

[0139] In some embodiments, the compounds of this disclosure comprise a heterocyclic group containing a halogenated substituent, wherein the compound binds to the mutant p53 protein and increases the wild-type p53 activity of the mutant protein. In some embodiments, the compound further comprises an indole group. In some embodiments, the indole group has a 1,1,1-trifluoroethyl substituent at the 1-position of the indole group.

[0140] In some embodiments, the indole group has a propargyl substituent at the 2-position of the indole group. In some embodiments, the propargyl substituent is attached to the indole group via the sp carbon atom of the propargyl substituent. In some embodiments, the propargyl substituent is attached to the nitrogen atom of the aniline group via the methylene group of the propargyl substituent. In some embodiments, the indole group includes an amino substituent at the 4-position of the indole group. In some embodiments, the amino substituent is attached to a heterocyclic group. In some embodiments, the heterocyclic group is a piperidine group. In some embodiments, the halogenated substituent is a fluorinated group. In some embodiments, the halogenated substituent is a chlorinated group. In some embodiments, the oral bioavailability of the compound is at least about 50% higher than that of similar compounds lacking a halogenated substituent on the heterocyclic group.

[0141] Non-limiting examples of compounds of the present invention include compounds of any of the following formulas:

[0142]

[0143]

[0144]

[0145]

[0146]

[0147] In some embodiments, this disclosure provides compounds of the following formula:

[0148]

[0149] in:

[0150] -each It can be a single bond or a double bond independently;

[0151] -X 1 For CR 5 CR 5 R 6 , N, NR 5 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0152] -X 2 For CR 7 CR 7 R 8 , N, NR 7 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0153] -X 3 For CR 9 CR 9 R 10 , N, NR 9 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0154] -X 4 For CR 11 CR 11 R 12 , N, NR 11 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0155] -X 5 For CR 13 , N or NR 13 ;

[0156] Where X 1 X 2 X 3 and X 4 At least one of them is connected to Q 1 carbon atoms;

[0157] -Q 1 For C=O, C=S, C=CR 14 R 15 C=NR 14 Alkylene, alkenylene, or ynylene groups, each of which is independently substituted or unsubstituted; or they may be bonds;

[0158] -m is 1, 2, 3 or 4;

[0159] -Y is N, O, or does not exist;

[0160] -R 1 -C(O)R 16 -C(O)OR 16 -C(O)NR 16 R 17 -OR 16 -SR 16 -NR 16 R 17-NR 16 C(O)R 16 -OC(O)R 16 C=O, C=S, -CN, -SiR 16 R 17 R 18 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen;

[0161] - Each R 3 and R 4 Independently for -C(O)R 19 -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 -SO2R 19 Alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen, or R 3 and R 4 Together with R 3 and R 4 The combined Y atoms form a ring, wherein the ring is substituted or unsubstituted, or R 3 It does not exist;

[0162] - Each R 2 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 R 13 R 14 R 15 R 16 R 17 and R 18 Independently for -C(O)R 21 -C(O)OR 21 -C(O)NR 21 R 22 -OR 21 -SR 21 -NR 21 R 22 -NR 21 C(O)R 22 -OC(O)R 21 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0163] - Each R 19 and R 20 For C(O)R 23 -C(O)OR 23 -C(O)NR 23 R 24 -OR 23 -SR 23 -NR 23 R 24 -NR 23 C(O)R 24 -OC(O)R 23 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0164] - Each R 21 and R 22 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic, each of which is independently substituted or unsubstituted; or hydrogen; and

[0165] - Each R 23 and R 24 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each independently either substituted or unsubstituted; or hydrogen.

[0166] Or a pharmaceutically acceptable salt thereof, wherein the compound is not one of the compounds listed in Table 1.

[0167] In some embodiments, compounds of the following formula are disclosed herein:

[0168]

[0169] in:

[0170] -each It can be a single bond or a double bond independently;

[0171] -X 1 For CR 5 CR 5 R 6 , N, NR 5 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0172] -X 2 For CR 7 CR 7 R 8 , N, NR 7 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0173] -X 3 For CR 9 CR 9 R 10 , N, NR 9 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0174] -X 4 For CR 11 CR 11 R 12 , N, NR 11 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0175] -X 5 For CR 13 , N or NR 13 ;

[0176] Where X 1 X 2 X 3 and X 4 At least one of them is connected to Q 1 carbon atoms;

[0177] -Q 1 For C=O, C=S, C=CR 14 R 15 C=NR 14 Alkylene, alkenylene, or ynylene groups, each of which is independently substituted or unsubstituted; or they may be bonds;

[0178] -m is 1, 2, 3 or 4;

[0179] -Y is N, O, or does not exist;

[0180] -R 1 -C(O)R 16 -C(O)OR 16 -C(O)NR 16 R 17 -OR 16 -SR 16 -NR 16 R 17 -NR 16 C(O)R 16 -OC(O)R 16 C=O, C=S, -CN, -SiR 16 R 17 R 18Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen;

[0181] - Each R 3 and R 4 Independently for -C(O)R 19 -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 -SO2R 19 Alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen, or R 3 and R 4 Together with R 3 and R 4 The combined nitrogen atoms form a ring, wherein the ring is substituted or unsubstituted, or R 3 It does not exist, where R 3 and R 4 At least one of them is an alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclic group, each of which is at least halogenated;

[0182] - Each R 2 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 R 13 R 14 R 15 R 16 R 17 and R 18 Independently for -C(O)R 21 -C(O)OR 21 -C(O)NR 21 R 22 -OR 21 -SR 21 -NR 21 R 22 -NR 21 C(O)R 22 -OC(O)R 21 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0183] - Each R 19 and R20 For C(O)R 23 -C(O)OR 23 -C(O)NR 23 R 24 -OR 23 -SR 23 -NR 23 R 24 -NR 23 C(O)R 24 -OC(O)R 23 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0184] - Each R 21 and R 22 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic, each of which is independently substituted or unsubstituted; or hydrogen; and

[0185] - Each R 23 and R 24 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each independently either substituted or unsubstituted; or hydrogen.

[0186] Or its pharmaceutically acceptable salt.

[0187] In some embodiments, the compound has the formula:

[0188]

[0189] in:

[0190] -each It can be a single bond or a double bond independently;

[0191] -X 1 For CR 5 CR 5 R 6 , N, NR 5 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0192] -X 2 For CR 7 CR 7 R 8 , N, NR 7 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0193] -X 3 For CR 9 CR9 R 10 , N, NR 9 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0194] -X 4 For CR 11 CR 11 R 12 , N, NR 11 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0195] -X 5 For CR 13 , N or NR 13 ;

[0196] Where X 1 X 2 X 3 and X 4 At least one of them is connected to Q 1 carbon atoms;

[0197] -A is a linking group;

[0198] -Q 1 For C=O, C=S, C=CR 14 R 15 C=NR 14 Alkylene, alkenylene, or ynylene groups, each of which is independently substituted or unsubstituted; or they may be bonds;

[0199] -m is 1, 2, 3 or 4;

[0200] -Y is N, O, or does not exist;

[0201] -R 1 -C(O)R 16 -C(O)OR 16 -C(O)NR 16 R 17 -OR 16 -SR 16 -NR 16 R 17 -NR 16 C(O)R 16 -OC(O)R 16 -SiR 16 R 17 R 18 Alkyl, alkenyl, alkoxy, aryl, heteroaryl, heterocyclic, or halogenated, each of which is independently substituted or unsubstituted; or hydrogen;

[0202] - Each R 3 and R 4 Independently for -C(O)R 19 -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 -SO2R 19 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen, or R 3 and R 4 Together with R 3 and R 4 The combined nitrogen atoms form a ring, wherein the ring is substituted or unsubstituted, or R 3 It does not exist; where R 3 and R 4 At least one of them is an alkyl, alkylene, alkenyl, alkenylene, alkynyl, alkynylene, aryl, heteroaryl, or heterocyclic group, each of which is at least halogenated;

[0203] - Each R 2 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 R 13 R 14 R 15 R 16 R 17 and R 18 Independently for -C(O)R 21 -C(O)OR 21 -C(O)NR 21 R 22 -OR 21 -SR 21 -NR 21 R 22 -NR 21 C(O)R 22 -OC(O)R 21 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0204] - Each R 19 and R 20 Independently for -C(O)R 23 -C(O)OR 23 -C(O)NR23 R 24 -OR 23 -SR 23 -NR 23 R 24 -NR 23 C(O)R 24 -OC(O)R 23 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0205] - Each R 21 and R 22 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic, each of which is independently substituted or unsubstituted; or hydrogen; and

[0206] - Each R 23 and R 24 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each independently either substituted or unsubstituted; or hydrogen.

[0207] Or its pharmaceutically acceptable salt.

[0208] In some embodiments, the compound has the formula:

[0209]

[0210] in:

[0211] -each It can be a single bond or a double bond independently;

[0212] -X 1 For CR 5 CR 5 R 6 , N, NR 5 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0213] -X 2 For CR 7 CR 7 R 8 , N, NR 7 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0214] -X 3 For CR 9 CR 9 R 10 , N, NR 9 , O, S, C=O, C=S or connected to Q1 carbon atoms;

[0215] -X 4 For CR 11 CR 11 R 12 , N, NR 11 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0216] -X 5 For CR 13 , N or NR 13 ;

[0217] Where X 1 X 2 X 3 and X 4 At least one of them is connected to Q 1 carbon atoms;

[0218] -A is a linking group;

[0219] -Q 1 For C=O, C=S, C=CR 14 R 15 C=NR 14 Alkylene, alkenylene, or ynylene groups, each of which is independently substituted or unsubstituted; or they may be bonds;

[0220] -m is 1, 2, 3 or 4;

[0221] -Y is N, O, or does not exist;

[0222] -R 1 -C(O)R 16 -C(O)OR 16 -C(O)NR 16 R 17 -OR 16 -SR 16 -NR 16 R 17 -NR 16 C(O)R 16 -OC(O)R 16 -SiR 16 R 17 R 18 Alkyl, alkenyl, alkoxy, aryl, heteroaryl, heterocyclic, or halogenated, each of which is independently substituted or unsubstituted; or hydrogen;

[0223] - Each R 3 and R 4 Independently for -C(O)R 19-C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 -SO2R 19 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen, or R 3 and R 4 Together with R 3 and R 4 The combined nitrogen atoms form a ring, wherein the ring is substituted or unsubstituted, or R 3 It does not exist;

[0224] - Each R 2 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 R 13 R 14 R 15 R 16 R 17 and R 18 Independently for -C(O)R 21 -C(O)OR 21 -C(O)NR 21 R 22 -OR 21 -SR 21 -NR 21 R 22 -NR 21 C(O)R 22 -OC(O)R 21 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0225] - Each R 19 and R 20 Independently for -C(O)R 23 -C(O)OR 23 -C(O)NR 23 R 24 -OR 23 -SR 23 -NR 23 R 24 -NR 23 C(O)R 24 -OC(O)R 23Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0226] - Each R 21 and R 22 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic, each of which is independently substituted or unsubstituted; or hydrogen; and

[0227] - Each R 23 and R 24 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each independently either substituted or unsubstituted; or hydrogen.

[0228] Or a pharmaceutically acceptable salt thereof, wherein the compound is not one of the compounds listed in Table 1.

[0229] In some embodiments, A is an alkylene, alkenylene, or ynylene group, each of which may be substituted or unsubstituted. In some embodiments, A is an alkylene group. In some embodiments, A is an alkenylene group. In some embodiments, A is an ynylene group.

[0230] In some embodiments, the compound of this formula is:

[0231]

[0232] in:

[0233] -each It can be a single bond or a double bond independently;

[0234] -X 1 For CR 5 CR 5 R 6 , N, NR 5 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0235] -X 2 For CR 7 CR 7 R 8 , N, NR 7 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0236] -X 3 For CR 9 CR 9 R 10 , N, NR 9 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0237] -X 4 For CR 11 CR 11 R 12 , N, NR 11 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0238] -X 5 For CR 13 , N or NR 13 ;

[0239] Where X 1 X 2 X 3 and X 4 At least one of them is connected to Q 1 carbon atoms;

[0240] -Q 1 For C=O, C=S, C=CR 14 R 15 C=NR 14 Alkylene, alkenylene, or ynylene groups, each of which is independently substituted or unsubstituted; or they may be bonds;

[0241] -m is 1, 2, 3 or 4;

[0242] -Y is N, O, or does not exist;

[0243] -A is a cyclic group that has been at least halogenated;

[0244] -R 1 -C(O)R 16 -C(O)OR 16 -C(O)NR 16 R 17 -OR 16 -SR 16 -NR 16 R 17 -NR 16 C(O)R 16 -OC(O)R 16 C=O, C=S, -CN, -SiR 16 R 17 R 18 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen;

[0245] -R 3 -C(O)R 19 -C(O)OR 19-C(O)NR 19 R 20 -SOR 19 -SO2R 19 Alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen, or R 3 and A together with R 3 The nitrogen atom that combines with A forms a ring, wherein the ring is substituted or unsubstituted, or R 3 It does not exist.

[0246] - Each R 2 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 R 13 R 14 R 15 R 16 R 17 and R 18 Independently for -C(O)R 21 -C(O)OR 21 -C(O)NR 21 R 22 -OR 21 -SR 21 -NR 21 R 22 -NR 21 C(O)R 22 -OC(O)R 21 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0247] - Each R 19 and R 20 For C(O)R 23 -C(O)OR 23 -C(O)NR 23 R 24 -OR 23 -SR 23 -NR 23 R 24 -NR 23 C(O)R 24 -OC(O)R 23Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0248] - Each R 21 and R 22 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic, each of which is independently substituted or unsubstituted; or hydrogen; and

[0249] - Each R 23 and R 24 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each independently either substituted or unsubstituted; or hydrogen.

[0250] Or its pharmaceutically acceptable salt.

[0251] In some embodiments, the compound has the formula:

[0252]

[0253] in:

[0254] -each It can be a single bond or a double bond independently;

[0255] -X 1 For CR 5 CR 5 R 6 , N, NR 5 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0256] -X 2 For CR 7 CR 7 R 8 , N, NR 7 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0257] -X 3 For CR 9 CR 9 R 10 , N, NR 9 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0258] -X 4 For CR 11 CR 11 R 12 , N, NR 11 , O, S, C=O, C=S or connected to Q1 carbon atoms;

[0259] -X 5 For CR 13 , N or NR 13 ;

[0260] Where X 1 X 2 X 3 and X 4 At least one of them is connected to Q 1 carbon atoms;

[0261] -Q 1 For C=O, C=S, C=CR 14 R 15 C=NR 14 Alkylene, alkenylene, or ynylene groups, each of which is independently substituted or unsubstituted; or they may be bonds;

[0262] -m is 1, 2, 3 or 4;

[0263] -Y is N, O, or does not exist;

[0264] -A is a cyclic group that has been at least halogenated;

[0265] -R 1 -C(O)R 16 -C(O)OR 16 -C(O)NR 16 R 17 -OR 16 -SR 16 -NR 16 R 17 -NR 16 C(O)R 16 -OC(O)R 16 C=O, C=S, -CN, -SiR 16 R 17 R 18 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen;

[0266] -R 3 -C(O)R 19 -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 -SO2R 19Alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen, or R 3 and A together with R 3 The nitrogen atom that combines with A forms a ring, wherein the ring is substituted or unsubstituted, or R 3 It does not exist.

[0267] - Each R 2 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 R 13 R 14 R 15 R 16 R 17 and R 18 Independently for -C(O)R 21 -C(O)OR 21 -C(O)NR 21 R 22 -OR 21 -SR 21 -NR 21 R 22 -NR 21 C(O)R 22 -OC(O)R 21 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0268] - Each R 19 and R 20 For C(O)R 23 -C(O)OR 23 -C(O)NR 23 R 24 -OR 23 -SR 23 -NR 23 R 24 -NR 23 C(O)R 24 -OC(O)R 23 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0269] - Each R 21 and R 22Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic, each of which is independently substituted or unsubstituted; or hydrogen; and

[0270] - Each R 23 and R 24 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each independently either substituted or unsubstituted; or hydrogen.

[0271] Or its pharmaceutically acceptable salt.

[0272] In some implementations, the pattern of dashed bonds is chosen to provide an aromatic system, such as indole, indole-indole, pyrrolopyridine, pyrrolopyrimidine, or pyrrolopyrazine.

[0273] In some implementation schemes, X 1 For CR 5 CR 5 R 6 Or connect to Q 1 The carbon atom. In some implementations, X 2 For CR 7 CR 7 R 8 Or connect to Q 1 The carbon atom. In some implementations, X 3 For CR 9 CR 9 R 10 Or connect to Q 1 The carbon atom. In some implementations, X 4 For CR 11 CR 11 R 12 Or connect to Q 1 The carbon atom. In some implementations, X 5 For CR 13 , N or NR 13 In some implementations, X 1 To connect to Q 1 The carbon atom. In some implementations, X 2 To connect to Q 1 The carbon atom. In some implementations, X 3 To connect to Q 1 The carbon atom. In some implementations, X 4 To connect to Q 1 The carbon atom. In some implementations, X 5 Let N be the number of elements in the array.

[0274] In some implementation schemes, Q 1 As a key. In some implementations, Q 1It is a C1-alkylene group.

[0275] In some implementations, m is 1. In some implementations, m is 2. In some implementations, m is 3. In some implementations, m is 4.

[0276] In some embodiments, ring A is an aryl, heteroaryl, or heterocyclic group, each of which may be substituted or unsubstituted. In some embodiments, ring A is a substituted aryl group. In some embodiments, ring A is a fluorinated aryl group. In some embodiments, ring A is a chlorinated aryl group. In some embodiments, ring A is a substituted heterocyclic group. In some embodiments, ring A is a fluorinated heterocyclic group. In some embodiments, ring A is a chlorinated heterocyclic group.

[0277] In some implementation schemes, R 1 Alkyl, alkenyl, -C(O)R 16 -C(O)OR 16 or -C(O)NR 16 R 17 Each of them is either unsubstituted or substituted. In some implementations, R 1 The substituted alkyl group. In some embodiments, R... 1 For being NR 16 R 17 Substituted alkyl groups.

[0278] In some implementations, each R 16 and R 17 Independently alkyl, alkenyl, aryl, heteroaryl, or heterocyclic, each of which is independently substituted or unsubstituted; or hydrogen. In some embodiments, R 16 It is hydrogen or alkyl. In some embodiments, R 17 It is an aryl, heteroaryl, or heterocyclic group, each of which is independently substituted or unsubstituted. In some embodiments, R 17 For the substituted aryl group. In some embodiments, R 17 For substituted phenyl groups. In some embodiments, R 17 The phenyl group is substituted with a sulfoxide group, carboxyl group, amide group, amino group, alkyl group, alkoxy group, hydroxyl group, halogenated group, cyano group, or heterocyclic group, each of which is independently substituted or unsubstituted. In some embodiments, R 17 It is a phenyl group substituted with a methoxy group. In some embodiments, R 17 It is a phenyl group substituted with a substituted sulfoxide group. In some embodiments, R 17It is a phenyl group substituted with a carboxyl group. In some embodiments, R 17 It is a phenyl group that has been substituted with an amide group.

[0279] In some embodiments, the compound has the formula:

[0280]

[0281] In some implementation schemes, Q 1 For C=O, C=S, C=CR 14 R 15 C=NR 14 alkylene, alkenylene, or ynylene groups, each independently substituted or unsubstituted; or bonded. In some embodiments, Q 1 It is alkylene, alkenylene, or ynylene. In some embodiments, Q 1 It is a C1-alkylene group or a bond. In some embodiments, Q 1 It is a C1-alkylene group. In some embodiments, Q 1 For key.

[0282] In some implementations, Y is N. In some implementations, Y is 0. In some implementations, Y does not exist.

[0283] In some implementation schemes, R 2 It is hydrogen or alkyl. In some embodiments, R 2 It is an alkyl group. In some embodiments, R... 2 For substituted C1-C5 alkyl groups. In some embodiments, R 2 It is trifluoroethyl. In some embodiments, R 2 It is a cycloalkyl group. In some embodiments, R 2 It is cyclopropyl.

[0284] In some implementation schemes, R 13 It is alkyl, alkenyl, hydrogen, or halogen. In some embodiments, R 13 It is hydrogen.

[0285] In some implementation schemes, R 2 It is a C1-C5-alkyl group, and R 13 It is a C1-C5 alkyl group. In some embodiments, R 2 It is a C1-C5-alkyl group, and R 13 It is hydrogen. In some implementations, R 2 For substituted C1-C5 alkylene groups. In some embodiments, R 2 The monomer is methyl, ethyl, propyl, isopropyl, butyl, or tert-butyl, each of which may be substituted or unsubstituted. In some embodiments, R...13 It is methyl, ethyl, propyl, isopropyl, butyl, or tert-butyl. In some embodiments, R 2 It is hydrogen, and R 13 It is hydrogen. In some implementations, R 2 It is trifluoroethyl, and R 13 It is hydrogen.

[0286] In some embodiments, the compound has the formula:

[0287]

[0288] In some embodiments, the compound has the formula:

[0289]

[0290] In some implementation schemes, R 3 Let H be the number of 'R', and R be the number of 'R'. 4 -C(O)R 19 -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 -SO2R 19 Alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted. In some embodiments, each R 3 and R 4 Independently, it is a substituted or unsubstituted C1-C6 alkylene group. In some embodiments, R 3 For H; and R 4 It is a substituted or unsubstituted C1-C4 alkylene group. In some embodiments, R 3 Let H be the number of 'R', and R be the number of 'R'. 4 It can be a substituted or unsubstituted heterocyclic group. In some embodiments, R 3 Let H be the number of 'R', and R be the number of 'R'. 4 It may be a substituted or unsubstituted piperidinyl group. In some embodiments, R 3 Let H be the number of 'R', and R be the number of 'R'. 4 It can be a substituted or unsubstituted cycloalkyl group. In some embodiments, R 3 Let H be the number of 'R', and R be the number of 'R'. 4 It is a cycloalkyl group substituted with an amino group. In some embodiments, R 3 Let H be the number of 'R', and R be the number of 'R'. 4 The cyclobutyl group may be substituted or unsubstituted. In some embodiments, R 3 Let H be the number of 'R', and R be the number of 'R'. 4 It is a cyclobutyl group substituted with an amino group. In some embodiments, R 3 Let H be the number of 'R', and R be the number of 'R'. 4For substituted or unsubstituted cyclohexyl groups. In some embodiments, R... 3 Let H be the number of 'R', and R be the number of 'R'. 4 It is a cyclohexyl group substituted with an amino group.

[0291] In some embodiments, the compound has the formula:

[0292]

[0293] In some embodiments, the compound has the formula:

[0294]

[0295] R 1 It can be a group substituted by one or more substituents, said substituents being selected from hydroxyl groups, mercapto groups, halogens, amino groups, nitro groups, nitroso groups, cyano groups, azide groups, sulfoxide groups, sulfone groups, sulfonamide groups, carboxyl groups, formaldehyde groups, imine groups, alkyl groups, haloalkyl groups, cycloalkyl groups, alkenyl groups, haloalkenyl groups, alkynyl groups, haloalkynyl groups, alkoxy groups, aryl groups, aryloxy groups, arylalkyl groups, arylalkoxy groups, heterocyclic groups, acyl groups, acyloxy groups, carbamate groups, amide groups, ethyl carbamate groups, and ester groups. In some embodiments, R 1 Alkyl, alkenyl, -C(O)R 16 -C(O)OR 16 or -C(O)NR 16 R 17 In some implementations, R 1 It is a substituted or unsubstituted C1-C3 alkyl group. In some embodiments, R 1 It is a C1-C3-alkyl group substituted with an amine group. In some embodiments, R 1 For being NR 16 R 17 Substituted C1-alkyl groups. In some embodiments, each R 16 and R 17 It is independently aryl, heteroaryl, or heterocyclic, each independently substituted or unsubstituted; or hydrogen. In some embodiments, R 16 Let H be the number of 'R', and R be the number of 'R'. 17 For the substituted aryl group. In some embodiments, R 16 Let H be the number of 'R', and R be the number of 'R'. 17 For substituted phenyl groups. In some embodiments, R 16 Let H be the number of 'R', and R be the number of 'R'. 17 It is a phenyl group substituted with an alkyl, alkoxy, halogenated, sulfonamide, sulfone, or carboxyl group. In some embodiments, R 16 Let H be the number of 'R', and R be the number of 'R'.17 For substituted heteroaryl groups. In some embodiments, R 16 Let H be the number of 'R', and R be the number of 'R'. 17 For substituted heterocyclic groups.

[0296] In some implementation schemes, R 3 -C(O)R 19 -C(O)OR 19 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen, and R 4 -C(O)R 19 -C(O)OR 19 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen. In some embodiments, R 3 It is an alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic group, each of which is independently substituted or unsubstituted; or it is hydrogen. In some embodiments, R 3 The substituted alkyl group. In some embodiments, R... 3 For H.

[0297] In some implementation schemes, R 3 It is hydrogen, and R 4 For ring A. In some implementations, R 4 Alternatively, ring A may be cycloalkyl, aryl, heteroaryl, or heterocyclic, each independently being substituted or unsubstituted. In some embodiments, R... 4 Or ring A may be a substituted or unsubstituted aryl group. In some embodiments, R 4 Or ring A may be a substituted or unsubstituted phenyl group. In some embodiments, R... 4 Alternatively, ring A may be a substituted or unsubstituted cycloalkyl group. In some embodiments, R... 4 Alternatively, ring A may be a substituted or unsubstituted cyclopropyl group. In some embodiments, R... 4 Or ring A is a substituted cyclopropyl group. In some embodiments, R 4 Or ring A is a substituted cyclohexyl group. In some embodiments, R 4 Or ring A is a cyclohexyl group substituted with an amino group.

[0298] In some implementation schemes, R 3 Let H be the number of 'R', and R be the number of 'R'. 4 Alternatively, ring A may be an unsubstituted or substituted heterocyclic group. In some embodiments, R... 4 Or ring A is a heterocyclic group. In some embodiments, R 4 Alternatively, ring A may be piperidinyl, piperazine, tetrahydropyranyl, morpholinyl, or pyrrolidinyl, each independently either substituted or unsubstituted. In some embodiments, R...3 Let H be the number of 'R', and R be the number of 'R'. 4 Or ring A is a substituted piperidinyl group. In some embodiments, R 3 Let H be the number of 'R', and R be the number of 'R'. 4 Alternatively, ring A may be piperidine substituted with an alkyl, carboxyl, heterocyclic, or amide group. In some embodiments, R... 3 Let H be the number of 'R', and R be the number of 'R'. 4 Alternatively, ring A may be an unsubstituted or substituted methylpiperidinyl group. In some embodiments, R 3 Let H be the number of 'R', and R be the number of 'R'. 4 Alternatively, ring A may be 3-fluoro-1-methylpiperidinyl. In some embodiments, R 3 Let H be the number of 'R', and R be the number of 'R'. 4 Alternatively, ring A may be a piperidinyl group substituted with methoxypropanol. In some embodiments, R... 3 Let H be the number of 'R', and R be the number of 'R'. 4 Alternatively, ring A may be 3-fluoro-1-(2-hydroxy-3-methoxypropyl)piperidinyl. In some embodiments, R 3 Let H be the number of 'R', and R be the number of 'R'. 4 Alternatively, ring A may be an unsubstituted or substituted tetrahydropyranyl group. In some embodiments, R... 3 Let H be the number of 'R', and R be the number of 'R'. 4 Alternatively, ring A may be an unsubstituted tetrahydropyranyl group. In some embodiments, R... 3 Let H be the number of 'R', and R be the number of 'R'. 4 Alternatively, ring A may be a tetrahydropyranyl group substituted with an alkyl group. In some embodiments, R... 3 Let H be the number of 'R', and R be the number of 'R'. 4 Or ring A may be tetrahydrothiaran-1,1-dioxide.

[0299] In some implementation schemes, R 4 Alternatively, ring A may be cycloalkyl, aryl, heteroaryl, or heterocyclic, each of which is at least halogenated. In some embodiments, R... 4 Alternatively, ring A may be a C4-C6-cycloalkyl group that has been at least halogenated-substituted. In some embodiments, R... 4 Or ring A is a cyclohexyl group that has been at least halogenated. In some embodiments, R 4 Or ring A is an aryl group that has been at least halogenated. In some embodiments, R 4 Or ring A is a phenyl group that has been at least halogenated. In some embodiments, R 4 Alternatively, ring A may be a fluorinated aryl group. In some embodiments, R 4 Alternatively, ring A may be a fluorinated phenyl group. In some embodiments, R... 4 Or ring A is an aryl group substituted with chlorine. In some embodiments, R 4 Or ring A is a phenyl group substituted with chlorine. In some embodiments, R 4Or ring A is a heteroaryl group that has been at least halogenated-substituted. In some embodiments, R 4 Alternatively, ring A may be a fluorinated heteroaryl group. In some embodiments, R... 4 Or ring A is a heteroaryl group substituted with chlorine. In some embodiments, R 4 Or ring A is a C4-C6 heterocyclic group that has been at least halogenated. In some embodiments, R 4 Alternatively, ring A may be a fluorine-substituted heterocyclic group. In some embodiments, R... 4 Or ring A is a heterocyclic group substituted with chlorine.

[0300] In some implementation schemes, R 4 Alternatively, ring A may be piperidinyl, piperazine, tetrahydropyranyl, morpholinyl, or pyrrolidinyl, each independently halogenated. In some embodiments, R... 4 Alternatively, ring A may be a halogenated piperidinyl group. In some embodiments, R... 4 Alternatively, ring A may be a halogenated-substituted methylpiperidinyl group. In some embodiments, R 4 Alternatively, ring A may be 3-fluoro-1-methylpiperidinyl. In some embodiments, R 4 Alternatively, ring A may be 3-fluoro-1-(2-hydroxy-3-methoxypropyl)piperidinyl. In some embodiments, R 4 Or ring A is a tetrahydropyranyl group that is at least halogenated.

[0301] In some implementation schemes, R 4 Or ring A is a ring, and the ring is:

[0302]

[0303] The ring may be substituted or unsubstituted. In some embodiments, the ring is halogenated. In some embodiments, the ring is fluorinated. In some embodiments, R... 3 Let H be the number of 'R', and R be the number of 'R'. 4 It is a ring, and the ring is The ring may be substituted or unsubstituted. In some embodiments, the ring is halogenated. In some embodiments, the ring is fluorinated. In some embodiments, R... 3 Let H be the number of 'R', and R be the number of 'R'. 4 It is a ring, and the ring is The ring may be substituted or unsubstituted. In some embodiments, R a It is an alkylene group. In some embodiments, R a The methyl group is used. In some embodiments, the ring is halogenated. In some embodiments, the ring is fluorinated. In some embodiments, R... 3 Let H be the number of 'R', and R be the number of 'R'. 4 It is a ring, and the ring is The ring may be substituted or unsubstituted. In some embodiments, the ring is halogenated. In some embodiments, the ring is fluorinated. In some embodiments, R... 3 Let H be the number of 'R', and R be the number of 'R'. 4 It is a ring, and the ring is The rings can be substituted or unsubstituted.

[0304] In some implementation schemes, R 4 Alternatively, ring A may be substituted by one or more substituents selected from hydroxyl groups, mercapto groups, halogens, amino groups, nitro groups, nitroso groups, cyano groups, azide groups, sulfoxide groups, sulfone groups, sulfonamide groups, carboxyl groups, formaldehyde groups, imine groups, alkyl groups, haloalkyl groups, cycloalkyl groups, alkenyl groups, haloalkenyl groups, alkynyl groups, haloalkynyl groups, alkoxy groups, aryl groups, aryloxy groups, aralkyl groups, arylalkoxy groups, heterocyclic groups, acyl groups, acyloxy groups, carbamate groups, amide groups, ethyl carbamate groups, and ester groups.

[0305] In some implementation schemes, R 3 and R 4 Together with R 3 and R 4 The combined nitrogen atoms form a ring, which may be substituted or unsubstituted. In some embodiments, R 3 and R 4 Together with R 3 and R 4 The combined nitrogen atoms together form a substituted heterocycle. In some embodiments, R 3 and R 4 Together with R 3 and R 4 The combined nitrogen atoms together form a heterocycle substituted with a hydroxyl group, halogen group, amino group, or alkyl group. In some embodiments, R 3 and R 4 Together with R 3 and R 4 The combined nitrogen atoms form a heterocycle, in which the heterocycle is replaced by a substituted or unsubstituted heterocycle.

[0306] In some embodiments, the compound has the formula:

[0307]

[0308] in:

[0309] -R 1 -C(O)R 16 -C(O)OR 16-C(O)NR 16 R 17 -OR 16 -SR 16 -NR 16 R 17 -NR 16 C(O)R 16 -OC(O)R 16 C=O, C=S, -CN, -SiR 16 R 17 R 18 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen;

[0310] - Each R Q Independently for -C(O)R 21 -C(O)OR 21 -C(O)NR 21 R 22 -OR 21 -SR 21 -NR 21 R 22 -NR 21 C(O)R 22 -OC(O)R 21 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0311] -y can be 0, 1, 2, 3 or 4;

[0312] - Each R 16 R 17 and R 18 Independently for -C(O)R 21 -C(O)OR 21 -C(O)NR 21 R 22 -OR 21 -SR 21 -NR 21 R 22 -NR 21 C(O)R 22 -OC(O)R 21 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0313] - Each R 19 and R 20 For C(O)R 23 -C(O)OR 23-C(O)NR 23 R 24 -OR 23 -SR 23 -NR 23 R 24 -NR 23 C(O)R 24 -OC(O)R 23 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0314] - Each R 21 and R 22 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic, each of which is independently substituted or unsubstituted; or hydrogen; and

[0315] - Each R 23 and R 24 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each independently either substituted or unsubstituted; or hydrogen.

[0316] Or its pharmaceutically acceptable salt.

[0317] In some implementation schemes, R 1 -C(O)R 16 -C(O)OR 16 -C(O)NR 16 R 17 -OR 16 -SR 16 -NR 16 R 17 -NR 16 C(O)R 16 -OC(O)R 16 -SiR 16 R 17 R 18 Alkyl, alkenyl, alkynyl, alkoxy, aryl, heteroaryl, or heterocyclic groups, each independently substituted or unsubstituted; or hydrogen. In some embodiments, R 1 Alkyl, alkylene, alkoxy, -NR 21 R 22 Or aryl groups, each of which is independently substituted or unsubstituted; halogens or hydrogen.

[0318] In some implementation schemes, R 1 For substituted C1-C3-alkyl groups. In some embodiments, R 1 For being NR 16 R 17Substituted C1-C3-alkyl groups. In some embodiments, R 1 For being NR 16 R 17 Substituted methyl groups, wherein each R 16 and R 17 Independently, it is an alkyl, cycloalkyl, aryl, heteroaryl, heterocyclic, alkoxy, carboxyl, amino, acyl, acyloxy, or amide group, any one of which is unsubstituted or substituted; or it is hydrogen. In some embodiments, R 1 For being NR 16 R 17 Substituted methyl group, wherein R 16 It is hydrogen, and R 17 The carboxyl group is substituted. In some embodiments, R 1 For being NR 16 R 17 Substituted methyl group, wherein R 16 It is hydrogen, and R 17 For the substituted aryl group. In some embodiments, R 1 For being NR 16 R 17 Substituted methyl group, wherein R 16 It is hydrogen, and R 17 For substituted phenyl groups. In some embodiments, R 1 For being NR 16 R 17 Substituted methyl group, wherein R 16 It is hydrogen, and R 17 The phenyl group is substituted with a sulfoxide group, carboxyl group, amide group, amino group, alkyl group, alkoxy group, hydroxyl group, halogenated group, cyano group, or heterocyclic group, each of which is independently substituted or unsubstituted. In some embodiments, R 17 It is a phenyl group substituted with a methoxy group. In some embodiments, R 17 It is a phenyl group substituted with a substituted sulfoxide group. In some embodiments, R 17 It is a phenyl group substituted with a carboxyl group. In some embodiments, R 17 The substituted amide group. In some embodiments, R 17 It is replaced by methoxy and sulfonamide.

[0319] In some implementation schemes, R 2 It is hydrogen or alkyl. In some embodiments, R 2 For substituted C1-C5 alkylene groups. In some embodiments, R 2 It is trifluoroethyl.

[0320] In some embodiments, the compound has the formula:

[0321]

[0322] Or its pharmaceutically acceptable salt, wherein the variables are as defined above.

[0323] In some implementations, each R Q Independently for -C(O)R 21 -C(O)OR 21 -C(O)NR 21 R 22 -OR 21 -SR 21 -NR 21 R 22 -NR 21 C(O)R 22 -OC(O)R 21 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen. In some embodiments, each R Q for

[0324] In some implementations, y is 1. In some implementations, y is 2. In some implementations, y is 3. In some implementations, y is 4.

[0325] In some implementation schemes, R 1 -C(O)R 16 -C(O)OR 16 -C(O)NR 16 R 17 -OR 16 -SR 16 -NR 16 R 17 -NR 16 C(O)R 16 -OC(O)R 16 -SiR 16 R 17 R 18 Alkyl, alkenyl, alkynyl, alkoxy, aryl, heteroaryl, or heterocyclic groups, each independently substituted or unsubstituted; or hydrogen. In some embodiments, R 1 Alkyl, alkylene, alkoxy, -NR 21 R 22 Or aryl groups, each of which is independently substituted or unsubstituted; halogens or hydrogen.

[0326] In some implementation schemes, R 1 The substituted alkyl group. In some embodiments, R... 1 For substituted C1-C3-alkyl groups. In some embodiments, R1 For being NR 16 R 17 Substituted alkyl groups. In some embodiments, R 1 For being NR 16 R 17 Substituted C1-C3-alkyl groups. In some embodiments, R 1 For being NR 16 R 17 Substituted methyl groups, wherein each R 16 and R 17 Independently, it is an alkyl, cycloalkyl, aryl, heteroaryl, heterocyclic, alkoxy, carboxyl, amino, acyl, acyloxy, or amide group, any one of which is unsubstituted or substituted; or it is hydrogen. In some embodiments, R 1 For being NR 16 R 17 Substituted methyl group, wherein R 16 It is hydrogen, and R 17 The substituted carboxyl group.

[0327] In some implementation schemes, R 16 It is alkyl, alkenyl, aryl, heteroaryl, heterocyclic, or hydrogen, and R 17 It is aryl, heteroaryl, or heterocyclic. In some embodiments, R 16 It is hydrogen, and R 17 It can be phenyl, indolyl, piperidinyl, imidazolyl, thiazolyl, morpholinyl, pyrroleyl, or pyridinyl, each of which may be substituted or unsubstituted.

[0328] In some embodiments, the compound has the formula:

[0329]

[0330] In some implementations, each R 16 and R 17 Independently, it is alkyl, alkenyl, aryl, heteroaryl, heterocyclic, or hydrogen. In some embodiments, R 16 It is aryl, and R 17 It is an alkyl group. In some embodiments, R... 16 It is aryl, and R 17 It is hydrogen. In some implementations, R 16 It is a heteroaryl group, and R 17 It is an alkyl group. In some embodiments, R... 16 It is a heteroaryl group, and R 17 It is hydrogen. In some implementations, R 16 For substituted heteroaryl groups, and R 17 It is hydrogen. In some implementations, R 16 For substituted alkyl groups, and R17 It is hydrogen. In some implementations, R 17 It is an aryl, heteroaryl, or heterocyclic group, each independently substituted or unsubstituted by a halogen, alkyl, or hydroxyl group. In some embodiments, R 16 It is hydrogen, and R 17 It is an aryl or heteroaryl group that is substituted with or unsubstituted with halogens or alkyl groups. In some embodiments, R 16 It is an alkyl group, and R 17 It is a heteroaryl group substituted with halogen or alkyl. In some embodiments, R 16 It is hydrogen. In some implementations, R 17 It is an aryl, heteroaryl, or heterocyclic group, each independently substituted or unsubstituted by an alkyl group. In some embodiments, R 17 It is aryl or heteroaryl, each independently substituted with an alkyl group, wherein the alkyl group is optionally substituted with fluorine, chlorine, bromine, iodine, or cyano. In some embodiments, R 16 It is alkyl, alkenyl, aryl, heteroaryl, heterocyclic, or hydrogen, and R 17 It is aryl, heteroaryl, or heterocyclic. In some embodiments, R 16 It is hydrogen, and R 17 It is phenyl, indolyl, piperidinyl, imidazolyl, thiazolyl, morpholinyl, pyrroleyl, or pyridinyl, each of which may be substituted or unsubstituted. In some embodiments, R 16 It is hydrogen, and R 17 For substituted phenyl groups. In some embodiments, R 16 It is hydrogen, and R 17 The phenyl group is substituted with a sulfoxide group, carboxyl group, amide group, amino group, alkyl group, alkoxy group, hydroxyl group, halogenated group, cyano group, or heterocyclic group, each of which is independently substituted or unsubstituted. In some embodiments, R 17 It is a phenyl group substituted with a methoxy group. In some embodiments, R 17 It is a phenyl group substituted with a substituted sulfoxide group. In some embodiments, R 17 It is a phenyl group substituted with a carboxyl group. In some embodiments, R 17 The substituted amide group. In some embodiments, R 17 It is replaced by methoxy and sulfonamide.

[0331] In some implementations, each R 3 and R 4 Independently, it is an unsubstituted or substituted alkyl group. In some embodiments, R 3 It is hydrogen, and R 4 -C(O)R 19 -C(O)OR 19Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which may be substituted or unsubstituted independently. In some embodiments, R 3 It is hydrogen, and R 4 It is an alkyl, aryl, heteroaryl, or heterocyclic group, each of which is independently substituted or unsubstituted. In some embodiments, R 3 Let H be the number of 'R', and R be the number of 'R'. 4 For the substitution of heterocyclic groups. In some embodiments, R 3 Let H be the number of 'R', and R be the number of 'R'. 4 It is a substituted or unsubstituted C4-C6 heterocyclic group. In some embodiments, R 3 Let H be the number of 'R', and R be the number of 'R'. 4 The substituted alkyl group. In some embodiments, R... 3 Let H be the number of 'R', and R be the number of 'R'. 4 For substituted C1-C6-alkyl groups. In some embodiments, R 3 Let H be the number of 'R', and R be the number of 'R'. 4 It can be a substituted or unsubstituted cycloalkyl group. In some embodiments, R 3 Let H be the number of 'R', and R be the number of 'R'. 4 It is a substituted or unsubstituted C4-C6-cycloalkyl group. In some embodiments, R 3 Let H be the number of 'R', and R be the number of 'R'. 4 It is a C4-C6-cycloalkyl group substituted with an amino group.

[0332] In some embodiments, the compound has the formula:

[0333]

[0334] in:

[0335] -Q 1 For C=O, C=S, C=CR 14 R 15 C=NR 14 Alkylene, alkenylene, or ynylene groups, each of which is independently substituted or unsubstituted; or they may be bonds;

[0336] -R 1 -C(O)R 16 -C(O)OR 16 -C(O)NR 16 R 17 -OR 16 -SR 16 -NR 16 R 17 -NR 16 C(O)R 16 -OC(O)R 16 C=O, C=S, -CN, -SiR16 R 17 R 18 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen;

[0337] - Each R 3 and R 4 Independently for -C(O)R 19 -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 -SO2R 19 Alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen, or R 3 and R 4 Together with R 3 and R 4 The combined nitrogen atoms form a ring, wherein the ring is substituted or unsubstituted, or R 3 It does not exist, where R 3 and R 4 At least one of them is an alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclic group, each of which is at least halogenated;

[0338] - Each Z 1 and Z 2 Independent for CR 28 CR 29 Or N;

[0339] - Each R 2 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 R 13 R 14 R 15 R 16 R 17 and R 18 Independently for -C(O)R 21 -C(O)OR 21 -C(O)NR 21 R 22 -OR 21 -SR 21 -NR 21 R 22 -NR 21C(O)R 22 -OC(O)R 21 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0340] - Each R 19 and R 20 For C(O)R 23 -C(O)OR 23 -C(O)NR 23 R 24 -OR 23 -SR 23 -NR 23 R 24 -NR 23 C(O)R 24 -OC(O)R 23 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0341] - Each R 21 and R 22 It is independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic, each of which is independently substituted or unsubstituted; or it is hydrogen;

[0342] - Each R 23 and R 24 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic, each of which is independently substituted or unsubstituted; or hydrogen; and

[0343] - Each R 25 R 26 R 27 R 28 and R 29 Independently, it is hydrogen or a substituent, said substituent being selected from hydroxyl groups, mercapto groups, halogens, amino groups, nitro groups, nitroso groups, cyano groups, azide groups, sulfoxide groups, sulfone groups, sulfonamide groups, carboxyl groups, formaldehyde groups, imine groups, alkyl groups, halo-alkyl groups, alkenyl groups, halo-alkenyl groups, alkynyl groups, halo-alkynyl groups, alkoxy groups, aryl groups, aryloxy groups, aralkyl groups, arylalkoxy groups, heterocyclic groups, acyl groups, acyloxy groups, carbamate groups, amide groups, ureoyl groups, epoxy groups, and ester groups.

[0344] Or its pharmaceutically acceptable salt.

[0345] In some implementation schemes, Z 1 For N. In some implementations, Z1 and Z 2 For N. In some implementations, each R 25 and R 26 Independently halogenated. In some implementations, R 25 for In some implementation schemes, R 25 The substituted sulfone group. In some embodiments, R 25 The sulfone group is substituted with an alkyl group. In some embodiments, R 25 It is a methanesulfonyl group. In some embodiments, R 25 It is a sulfone group substituted with an amino group. In some embodiments, R 25 It is a sulfonamide. In some embodiments, R 25 It is a carboxyl group. In some embodiments, R 25 It is a methoxycarbonyl group.

[0346] In some embodiments, the compound has the formula:

[0347]

[0348] in:

[0349] -R 2 -C(O)R 21 -C(O)OR 21 -C(O)NR 21 R 22 -OR 21 -SR 21 -NR 21 R 22 -NR 21 C(O)R 22 -OC(O)R 21 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0350] - Each R Q Independently for -C(O)R 21 -C(O)OR 21 -C(O)NR 21 R 22 -OR 21 -SR 21 -NR 21 R 22 -NR 21 C(O)R 22 -OC(O)R 21Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted;

[0351] -y can be 0, 1, 2, 3 or 4;

[0352] - Each R 21 and R 22 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic, each of which is independently substituted or unsubstituted; or hydrogen; and

[0353] - Each R 25 R 26 R 27 R 28 and R 29 Independently, it is hydrogen or a substituent, said substituent being selected from hydroxyl groups, mercapto groups, halogens, amino groups, nitro groups, nitroso groups, cyano groups, azide groups, sulfoxide groups, sulfone groups, sulfonamide groups, carboxyl groups, formaldehyde groups, imine groups, alkyl groups, halo-alkyl groups, alkenyl groups, halo-alkenyl groups, alkynyl groups, halo-alkynyl groups, alkoxy groups, aryl groups, aryloxy groups, aralkyl groups, arylalkoxy groups, heterocyclic groups, acyl groups, acyloxy groups, carbamate groups, amide groups, ureoyl groups, epoxy groups, and ester groups.

[0354] Or its pharmaceutically acceptable salt.

[0355] In some embodiments, the compound has the formula:

[0356]

[0357] In some implementation schemes, R 25 The substituted sulfone group. In some embodiments, R 25 The sulfone group is substituted with an alkyl group. In some embodiments, R 25 It is a methanesulfonyl group. In some embodiments, R 25 It is a sulfone group substituted with an amino group. In some embodiments, R 25 It is a sulfonamide. In some embodiments, R 25 It is a carboxyl group. In some embodiments, R 25 It is a methoxycarbonyl group.

[0358] In some embodiments, the compound has the formula:

[0359]

[0360] in:

[0361] - Each RQ Independently for -C(O)R 21 -C(O)OR 21 -C(O)NR 21 R 22 -OR 21 -SR 21 -NR 21 R 22 -NR 21 C(O)R 22 -OC(O)R 21 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted;

[0362] -y can be 0, 1, 2, 3 or 4;

[0363] - Each R 21 and R 22 It is independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic, each of which is independently substituted or unsubstituted; or it is hydrogen;

[0364] - Each R 26 R 27 R 28 and R 29 Independently, it is hydrogen or a substituent, said substituent being selected from hydroxyl groups, mercapto groups, halogens, amino groups, nitro groups, nitroso groups, cyano groups, azide groups, sulfoxide groups, sulfone groups, sulfonamide groups, carboxyl groups, formaldehyde groups, imine groups, alkyl groups, halo-alkyl groups, alkenyl groups, halo-alkenyl groups, alkynyl groups, halo-alkynyl groups, alkoxy groups, aryl groups, aryloxy groups, aralkyl groups, arylalkoxy groups, heterocyclic groups, acyl groups, acyloxy groups, carbamate groups, amide groups, ureyl groups, epoxy groups, and ester groups; and

[0365] -R 30 It is an alkyl or amino group, which may be substituted or unsubstituted, or a pharmaceutically acceptable salt thereof.

[0366] In some implementation schemes, R 30 It is methyl. In some embodiments, R 30 For NH2. In some implementations, R 30 For NHMe. In some implementations, R 30 It is NMe2.

[0367] In some embodiments, the compound has the formula:

[0368]

[0369] Where R 30 The group is an alkyl or amino group, which may be unsubstituted or substituted. In some embodiments, R 30 It is a methyl group.

[0370] Non-limiting examples of compounds disclosed herein include the following:

[0371]

[0372] Or its pharmaceutically acceptable salt.

[0373] Non-limiting examples of compounds disclosed herein include the following:

[0374]

[0375] Or its pharmaceutically acceptable salt.

[0376] Non-limiting examples of compounds disclosed herein include the following:

[0377]

[0378] Or its pharmaceutically acceptable salt.

[0379] Non-limiting examples of compounds disclosed herein include the following:

[0380]

[0381] Or its pharmaceutically acceptable salt.

[0382] Non-limiting examples of compounds disclosed herein include the following:

[0383]

[0384] Or its pharmaceutically acceptable salt.

[0385] Non-limiting examples of compounds disclosed herein include the following:

[0386]

[0387] Or its pharmaceutically acceptable salt.

[0388] Non-limiting examples of compounds disclosed herein include the following:

[0389]

[0390] Or its pharmaceutically acceptable salt.

[0391] The compounds described herein may include all their stereoisomers, enantiomers, diastereomers, mixtures, racemates, blocked isomers, and tautomers.

[0392] Non-limiting examples of optional substituents include hydroxyl groups, mercapto groups, halogens, amino groups, nitro groups, nitroso groups, cyano groups, azide groups, sulfoxide groups, sulfone groups, sulfonamide groups, carboxyl groups, formaldehyde groups, imine groups, alkyl groups, halo-alkyl groups, alkenyl groups, halo-alkenyl groups, alkynyl groups, halo-alkynyl groups, alkoxy groups, aryl groups, aryloxy groups, aralkyl groups, arylalkoxy groups, heterocyclic groups, acyl groups, acyloxy groups, carbamate groups, amide groups, urea groups, epoxy groups, and ester groups.

[0393] Non-limiting examples of alkyl and alkylene groups include straight-chain, branched, and cyclic alkyl and alkylene groups. Alkyl or alkylene groups can be, for example, C1, C2, C3, C4, C5, C6, C7, C8, C9 ... 10 C 11 C 12 C 13 C 14 C 15 C 16 C 17 C 18 C 19 C 20 C 21 C 22 C 23 C 24 C 25 C 26 C 27 C 28 C 29 C 30 C 31 C 32 C 33 C 34 C 35 C 36 C 37 C 38 C 39 C 40 C 41 C 42 C 43 C 44 C 45 C 46 C 47 C 48 C 49 Or C 50 Groups, which may be substituted or unsubstituted.

[0394] Non-limiting examples of straight-chain alkyl groups include methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, and decyl.

[0395] Branched alkyl groups include any straight-chain alkyl group substituted with any number of alkyl groups. Non-limiting examples of branched alkyl groups include isopropyl, isobutyl, sec-butyl, and tert-butyl.

[0396] Non-limiting examples of substituted alkyl groups include hydroxymethyl, chloromethyl, trifluoromethyl, aminomethyl, 1-chloroethyl, 2-hydroxyethyl, 1,2-difluoroethyl and 3-carboxypropyl.

[0397] Non-limiting examples of cyclic alkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups. Cyclic alkyl groups also include fused-, bridged-, and spiro-bicyclic groups, as well as higher fused-ring, bridged-ring, and spirocyclic systems. Cyclic alkyl groups can be substituted with any number of straight-chain, branched, or cyclic alkyl groups. Non-limiting examples of cyclic alkyl groups include cyclopropyl, 2-methyl-cyclopropyl-1-yl, cyclopropyl-2-en-1-yl, cyclobutyl, 2,3-dihydroxycyclobut-1-yl, cyclobut-2-en-1-yl, cyclopentyl, cyclopent-2-en-1-yl, cyclopent-2,4-dien-1-yl, cyclohexyl, cyclohex-2-en-1-yl, cycloheptyl, cyclooctyl, 2,5-dimethylcyclopenten-1-yl, 3,5-dichlorocyclohexyl-1-yl, 4-hydroxy Cyclohexyl-1-yl, 3,3,5-trimethylcyclohexyl-1-yl, octahydrocyclopentadienyl, octahydro-1H-indenyl, 3a,4,5,6,7,7a-hexahydro-3H-inden-4-yl, decahydroazyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[3.1.1]heptyl, 1,3-dimethyl[2.2.1]hept-2-yl, bicyclo[2.2.2]octyl and bicyclo[3.3.3]undecyl.

[0398] Non-limiting examples of alkenyl and alkenyl groups include straight-chain, branched, and cyclic alkenyl groups. One or more alkenes of the alkenyl group may be, for example, E-, Z-, cis-, trans-, terminal-, or exo-methylene. The alkenyl or alkenyl group may be, for example, C2, C3, C4, C5, C6, C7, C8, C9 ... 10 C 11 C 12 C 13 C 14 C 15 C 16 C 17 C 18 C 19 C 20 C 21 C22 C 23 C 24 C 25 C 26 C 27 C 28 C 29 C 30 C 31 C 32 C 33 C 34 C 35 C 36 C 37 C 38 C 39 C 40 C 41 C 42 C 43 C 44 C 45 C 46 C 47 C 48 C 49 Or C 50 Groups, which may be substituted or unsubstituted. Non-limiting examples of alkenyl and alkenyl groups include vinyl, propen-1-yl, isopropenyl, buten-4-yl; 2-chlorovinyl, 4-hydroxybuten-1-yl, 7-hydroxy-7-methyloct-4-en-2-yl, and 7-hydroxy-7-methyloct-3,5-dien-2-yl.

[0399] Non-limiting examples of alkynyl or ynylene groups include straight-chain alkynyl, branched-chain alkynyl, and cyclic alkynyl groups. The triple bond of the alkynyl or ynylene group can be internal or terminal. The alkynyl or ynylene group can be, for example, C2, C3, C4, C5, C6, C7, C8, C9, C1, C2, C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C1 ... 10 C 11 C 12 C 13 C 14 C 15 C 16 C 17 C 18 C 19 C 20 C 21 C 22 C 23 C 24 C 25 C 26 C 27 C 28 C 29 C 30 C31 C 32 C 33 C 34 C 35 C 36 C 37 C 38 C 39 C 40 C 41 C 42 C 43 C 44 C 45 C 46 C 47 C 48 C 49 Or C 50 Groups, which may be substituted or unsubstituted. Non-limiting examples of ynyl or ynylene groups include ethynyl, prop-2-yn-1-yl, prop-1-yn-1-yl, and 2-methyl-hex-4-yn-1-yl; 5-hydroxy-5-methylhex-3-yn-1-yl, 6-hydroxy-6-methylhept-3-yn-2-yl, and 5-hydroxy-5-ethylhept-3-yn-1-yl.

[0400] The halo-alkyl group can be any alkyl group substituted with any number of halogen atoms, such as fluorine, chlorine, bromine, and iodine atoms. The halo-alkenyl group can be any alkenyl group substituted with any number of halogen atoms. The halo-alkynyl group can be any alkynyl group substituted with any number of halogen atoms.

[0401] The alkoxy group can be, for example, an oxygen atom substituted with any alkyl, alkenyl, or alkynyl group. Ethers or ether groups contain alkoxy groups. Non-limiting examples of alkoxy groups include methoxy, ethoxy, propoxy, isopropoxy, and isobutoxy.

[0402] The aryl group can be heterocyclic or non-heterocyclic. The aryl group can be monocyclic or polycyclic. The aryl group can be substituted by any number of substituents described herein, such as hydrocarbon groups, alkyl groups, alkoxy groups, and halogen atoms. Non-limiting examples of aryl groups include phenyl, tolueneyl, naphthyl, pyrrolyl, pyridyl, imidazolyl, thiophene, and furanyl. Non-limiting examples of substituted aryl groups include 3,4-dimethylphenyl, 4-tert-butylphenyl, 4-cyclopropylphenyl, 4-diethylaminophenyl, 4-(trifluoromethyl)phenyl, 4-(difluoromethoxy)phenyl, 4-(trifluoromethoxy)phenyl, 3-chlorophenyl, 4-chlorophenyl, 3,4-dichlorophenyl, 2-fluorophenyl, 2-chlorophenyl, 2-iodophenyl, 3-iodophenyl, 4-iodophenyl, 2-methylphenyl, 3-fluorophenyl, 3-methylphenyl, 3-methoxyphenyl, 4-fluorophenyl, 4-methylphenyl, 4-methoxyphenyl, 2- 3-Difluorophenyl, 3,4-Difluorophenyl, 3,5-Difluorophenyl, 2,3-Dichlorophenyl, 3,4-Dichlorophenyl, 3,5-Dichlorophenyl, 2-Hydroxyphenyl, 3-Hydroxyphenyl, 4-Hydroxyphenyl, 2-Methoxyphenyl, 3-Methoxyphenyl, 4-Methoxyphenyl, 2,3-Dimethoxyphenyl, 3,4-Dimethoxyphenyl, 3,5-Dimethoxyphenyl, 2,4-Difluorophenyl, 2,5-Difluorophenyl, 2,6-Difluorophenyl, 2,3,4-Trifluorophenyl, 2,3,5-Trifluorophenyl, 2,3,6-Trifluorophenyl 2,4,5-Trifluorophenyl, 2,4,6-Trifluorophenyl, 2,4-Dichlorophenyl, 2,5-Dichlorophenyl, 2,6-Dichlorophenyl, 3,4-Dichlorophenyl, 2,3,4-Trichlorophenyl, 2,3,5-Trichlorophenyl, 2,3,6-Trichlorophenyl, 2,4,5-Trichlorophenyl, 3,4,5-Trichlorophenyl, 2,4,6-Trichlorophenyl, 2,3-Dimethylphenyl, 2,4-Dimethylphenyl, 2,5-Dimethylphenyl, 2,6-Dimethylphenyl, 2,3,4-Trimethylphenyl, 2,3,5-Trimethylphenyl, 2 3,6-Trimethylphenyl, 2,4,5-Trimethylphenyl, 2,4,6-Trimethylphenyl, 2-Ethylphenyl, 3-Ethylphenyl, 4-Ethylphenyl, 2,3-Diethylphenyl, 2,4-Diethylphenyl, 2,5-Diethylphenyl, 2,6-Diethylphenyl, 3,4-Diethylphenyl, 2,3,4-Triethylphenyl, 2,3,5-Triethylphenyl, 2,3,6-Triethylphenyl, 2,4,5-Triethylphenyl, 2,4,6-Triethylphenyl, 2-Isopropylphenyl, 3-Isopropylphenyl, and 4-Isopropylphenyl.

[0403] Non-limiting examples of substituted aryl groups include 2-aminophenyl, 2-(N-methylamino)phenyl, 2-(N,N-dimethylamino)phenyl, 2-(N-ethylamino)phenyl, 2-(N,N-diethylamino)phenyl, 3-aminophenyl, 3-(N-methylamino)phenyl, 3-(N,N-dimethylamino)phenyl, 3-(N-ethylamino)phenyl, 3-(N,N-diethylamino)phenyl, 4-aminophenyl, 4-(N-methylamino)phenyl, 4-(N,N-dimethylamino)phenyl, 4-(N-ethylamino)phenyl and 4-(N,N-diethylamino)phenyl.

[0404] Heterocycles can be any ring containing non-carbon ring atoms, such as N, O, S, P, Si, B, or any other heteroatom. Heterocycles can be substituted with any number of substituents, such as alkyl groups and halogen atoms. Heterocycles can be aromatic (heteroaryl) or non-aromatic. Non-limiting examples of heterocycles include pyrrole, pyrrolidine, pyridine, piperidine, succinamide, maleimide, morpholine, imidazole, thiophene, furan, tetrahydrofuran, pyran, and tetrahydropyran.

[0405] Non-limiting examples of heterocycles include: heterocyclic units having a monocyclic ring containing one or more heteroatoms, including bis-acridinyl, acridine, azacyclic butyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolinyl, thiazolyl, isothiazolinyl, oxazolyl ketone, hydantoin, tetrahydrofuranyl, pyrrolidinyl, morpholinyl, piperazinyl, piperidinyl, dihydropyranyl, tetrahydropyranyl, piperidin-2-one, 2,3,4,5-tetrahydro-1H - Aza-heptatrienyl, 2,3-dihydro-1H-indole, and 1,2,3,4-tetrahydroquinoline; and ii) a heterocyclic unit having two or more rings, one of which is a heterocycle, non-limiting examples of which include hexahydro-1H-pyrrolazinyl, 3a,4,5,6,7,7a-hexahydro-1H-benzo[d]imidazolyl, 3a,4,5,6,7,7a-hexahydro-1H-indoleyl, 1,2,3,4-tetrahydroquinolineyl, and decahydro-1H-cyclooctatetraen[b]pyrroleyl.

[0406] Non-limiting examples of heteroaryl groups include: i) heteroaryl rings containing a monocyclic ring, including 1,2,3,4-tetrazolyl, [1,2,3]triazolyl, [1,2,4]triazolyl, triazinyl, thiazolyl, 1H-imidazolyl, oxazolyl, isoxazolyl, isothiazolyl, furanyl, thiophene, pyrimidinyl, 2-phenylpyrimidinyl, pyridinyl, 3-methylpyridinyl, and 4-dimethylaminopyridinyl; and ii) heteroaryl rings containing two or more monocyclic rings. Multi-fused-ring heteroaryl rings, wherein one of the fused rings is a heteroaryl ring, include, without limitation, 7H-purinyl, 9H-purinyl, 6-amino-9H-purinyl, 5H-pyrrolo[3,2-d]pyrimidinyl, 7H-pyrrolo[2,3-d]pyrimidinyl, pyridolo[2,3-d]pyrimidinyl, 4,5,6,7-tetrahydro-1-H-indolyl, quinoxalinyl, quinazolinyl, quinolinyl, 8-hydroxy-quinolinyl, and isoquinolinyl.

[0407] Any compound described herein may be purified. The compounds described herein may have a purity of at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21%, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, and at least 28%. % purity, at least 29% purity, at least 30% purity, at least 31% purity, at least 32% purity, at least 33% purity, at least 34% purity, at least 35% purity, at least 36% purity, at least 37% purity, at least 38% purity, at least 39% purity, at least 40% purity, at least 41% purity, at least 42% purity, at least 43% purity, at least 44% purity, at least 45% purity, at least 46% purity, at least 47% purity, at least 48% purity, at least 49% purity, at least 50% purity, at least 51% purity, at least 52% purity, at least 53% purity, at least 54% purity, at least 55% purity, to At least 56% purity, at least 57% purity, at least 58% purity, at least 59% purity, at least 60% purity, at least 61% purity, at least 62% purity, at least 63% purity, at least 64% purity, at least 65% purity, at least 66% purity, at least 67% purity, at least 68% purity, at least 69% purity, at least 70% purity, at least 71% purity, at least 72% purity, at least 73% purity, at least 74% purity, at least 75% purity, at least 76% purity, at least 77% purity, at least 78% purity, at least 79% purity, at least 80% purity, at least 81% purity, at least 82% purity, at least 83% purity. Purity: at least 84% purity, at least 85% purity, at least 86% purity, at least 87% purity, at least 88% purity, at least 89% purity, at least 90% purity, at least 91% purity, at least 92% purity, at least 93% purity, at least 94% purity, at least 95% purity, at least 96% purity, at least 97% purity, at least 98% purity, at least 99% purity, at least 99% purity, at least 99.1% purity, at least 99.2% purity, at least 99.3% purity, at least 99.4% purity, at least 99.5% purity, at least 99.6% purity, at least 99.7% purity, at least 99.8% purity, or at least 99.9% purity.

[0408] In some embodiments, the compounds disclosed herein do not include the compounds in Table 1 or their pharmaceutically acceptable salts.

[0409] Table 1. List of compounds

[0410]

[0411]

[0412]

[0413]

[0414]

[0415]

[0416]

[0417]

[0418]

[0419]

[0420]

[0421]

[0422]

[0423]

[0424]

[0425]

[0426]

[0427]

[0428]

[0429]

[0430]

[0431]

[0432]

[0433]

[0434]

[0435]

[0436]

[0437]

[0438]

[0439]

[0440]

[0441]

[0442]

[0443]

[0444]

[0445]

[0446]

[0447]

[0448]

[0449]

[0450]

[0451]

[0452]

[0453]

[0454]

[0455]

[0456]

[0457]

[0458]

[0459]

[0460]

[0461]

[0462]

[0463]

[0464]

[0465]

[0466]

[0467]

[0468]

[0469]

[0470]

[0471]

[0472]

[0473]

[0474]

[0475]

[0476]

[0477]

[0478]

[0479]

[0480]

[0481]

[0482]

[0483]

[0484]

[0485]

[0486]

[0487]

[0488]

[0489]

[0490]

[0491]

[0492]

[0493]

[0494]

[0495]

[0496]

[0497]

[0498]

[0499]

[0500]

[0501]

[0502]

[0503]

[0504]

[0505]

[0506]

[0507]

[0508]

[0509]

[0510]

[0511]

[0512]

[0513]

[0514]

[0515]

[0516]

[0517]

[0518]

[0519]

[0520]

[0521]

[0522]

[0523]

[0524]

[0525] In some embodiments, this document discloses a method for treating cancer, which includes administering a therapeutically effective amount of the disclosed compound to a subject in need. For example, the compounds of the present invention can slow the proliferation of cancer cell lines or kill cancer cells.Non-limiting examples of cancers that can be treated by the compounds of the present invention include: acute lymphoblastic leukemia, acute myeloid leukemia, adrenocortical carcinoma, AIDS-related cancer, AIDS-related lymphoma, anal cancer, appendiceal cancer, astrocytoma, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain tumors (such as cerebellar astrocytoma, cerebral astrocytoma / malignant glioma, ependymoma, medulloblastoma, supratentorial primitive neuroectodermal tumor, visual pathway and hypothalamic glioma), breast cancer, bronchial adenoma, Burkitt lymphoma. Lymphoma, cancer of unknown origin, central nervous system lymphoma, cerebellar astrocytoma, cervical cancer, childhood cancer, chronic lymphocytic leukemia, chronic myeloid leukemia, chronic myeloproliferative disorders, colon cancer, cutaneous T-cell lymphoma, desmoplastic small round cell tumor, endometrial cancer, ependymoma, esophageal cancer, Ewing's sarcoma Sarcoma), germ cell tumors, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors, gliomas, hairy cell leukemia, head and neck cancer, heart cancer, hepatocellular carcinoma, Hodgkin's lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell carcinoma, Kaposi's sarcoma, kidney cancer, laryngeal cancer, lip and oral cancer, liposarcoma, liver cancer, lung cancer (e.g., non-small cell and small cell lung cancer), lymphoma, leukemia, macroglobulinemia, malignant fibrous histiocytoma / osteosarcoma of bone, medulloblastoma, melanoma, mesothelioma, latent primary metastatic squamous neck cancer, oral cancer, multiple endocrine neoplasia syndrome, myelodysplastic syndrome, myeloid leukemia, nasal and paranasal sinus cancer, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer, oral cancer Cancer, oropharyngeal cancer, osteosarcoma / malignant fibrous histiocytoma of bone, ovarian cancer, ovarian epithelial cancer, ovarian germ cell tumor, pancreatic cancer, islet cell pancreatic cancer, paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, pineal astrocytoma, pineal germ cell tumor, pituitary adenoma, pleural pulmonary blastoma, plasmacytoma, primary central nervous system lymphoma, prostate cancer, rectal cancer, renal cell carcinoma, transitional cell carcinoma of the renal pelvis and ureter, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma, skin cancer, Merkel cell skin carcinoma. Cell), small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, gastric cancer, T-cell lymphoma, laryngeal cancer, thymoma, thymic carcinoma, thyroid cancer, trophoblastic tumor (pregnancy), cancer of unknown primary site, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer, Waldenström macroglobulinemia. Macroglobulinemia and Wilms tumor.

[0526] In some embodiments, the compounds of the present invention exhibit non-lethal toxicity.

[0527] In some implementations, this document discloses a method for treating cancer, which includes administering a compound of formula (I) to a subject in need:

[0528]

[0529] in:

[0530] -each It can be a single bond or a double bond independently;

[0531] -X 1 For CR 5 CR 5 R 6 , N, NR 5 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0532] -X 2 For CR 7 CR 7 R 8 , N, NR 7 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0533] -X 3 For CR 9 CR 9 R 10 , N, NR 9 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0534] -X 4 For CR 11 CR 11 R 12 , N, NR 11 , O, S, C=O, C=S or connected to Q 1 carbon atoms;

[0535] -X 5 For CR 13 , N or NR 13 ;

[0536] Where X 1 X 2 X 3 and X4 At least one of them is connected to Q 1 carbon atoms;

[0537] -Q 1 For C=O, C=S, C=CR 14 R 15 C=NR 14 Alkylene, alkenylene, or ynylene groups, each of which is independently substituted or unsubstituted; or they may be bonds;

[0538] -m is 1, 2, 3 or 4;

[0539] -Y is N, O, or does not exist;

[0540] -R 1 -C(O)R 16 -C(O)OR 16 -C(O)NR 16 R 17 -OR 16 -SR 16 -NR 16 R 17 ,

[0541] -NR 16 C(O)R 16 -OC(O)R 16 C=O, C=S, -CN, -SiR 16 R 17 R 18 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen;

[0542] - Each R 3 and R 4 Independently for -C(O)R 19 -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 ,

[0543] -SO2R 19 Alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen, or R 3 and R 4 Together with R 3 and R 4 The combined Y atoms form a ring, wherein the ring is substituted or unsubstituted, or R 3 It does not exist;

[0544] - Each R2 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 R 13 R 14 R 15 R 16 R 17 and R 18 Independently for -C(O)R 21 -C(O)OR 21 -C(O)NR 21 R 22 -OR 21 -SR 21 ,

[0545] -NR 21 R 22 -NR 21 C(O)R 22 -OC(O)R 21 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0546] - Each R 19 and R 20 For C(O)R 23 -C(O)OR 23 -C(O)NR 23 R 24 -OR 23 -SR 23 ,

[0547] -NR 23 R 24 -NR 23 C(O)R 24 -OC(O)R 23 Alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each of which is independently substituted or unsubstituted; or hydrogen or halogen;

[0548] - Each R 21 and R 22 Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic, each of which is independently substituted or unsubstituted; or hydrogen; and

[0549] - Each R 23 and R 24Independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclic groups, each independently either substituted or unsubstituted; or hydrogen.

[0550] Or its pharmaceutically acceptable salt;

[0551] Among them, the SC of this compound against p53 Y220C, as measured by homogeneous time-resolved fluorescence (HTRF) assay, was... 150 The value is less than 1 μM.

[0552] In some embodiments, a method for inducing apoptosis of cells is disclosed herein, the method comprising contacting cells with a therapeutically effective amount of a compound that binds to a p53 mutant, wherein the compound is a compound disclosed herein. In some embodiments, the compound increases the ability of the p53 mutant to bind DNA. In some embodiments, the cells express p53. In some embodiments, the p53 mutant has a mutation at amino acid 220. In some embodiments, the p53 mutant is p53Y220C. In some embodiments, the compound induces a conformational change in the p53 mutant. In some embodiments, the compound selectively binds to the p53 mutant compared to wild-type p53. In some embodiments, the therapeutically effective amount is from about 50 mg to about 3000 mg. In some embodiments, the compound increases the stability of the bioactive conformation of the p53 mutant relative to the stability of the bioactive conformation of the p53 mutant in the absence of the compound.

[0553] Pharmaceutically acceptable salt.

[0554] This invention provides the use of pharmaceutically acceptable salts of any of the therapeutic compounds described herein. Pharmaceutically acceptable salts include, for example, acid addition salts and base addition salts. The acid added to the compound to form an acid addition salt can be an organic or inorganic acid. The base added to the compound to form a base addition salt can be an organic or inorganic base. In some embodiments, the pharmaceutically acceptable salt is a metal salt. In some embodiments, the pharmaceutically acceptable salt is an ammonium salt.

[0555] Metal salts can be produced by adding an inorganic base to the compounds of the present invention. The inorganic base consists of a metal cation paired with a basic counterion, such as a hydroxide, carbonate, bicarbonate, or phosphate ion. The metal can be an alkali metal, alkaline earth metal, transition metal, or main group metal. In some embodiments, the metal is lithium, sodium, potassium, cesium, cerium, magnesium, manganese, iron, calcium, strontium, cobalt, titanium, aluminum, copper, cadmium, or zinc.

[0556] In some implementations, the metal salt is a lithium salt, sodium salt, potassium salt, cesium salt, cerium salt, magnesium salt, manganese salt, iron salt, calcium salt, strontium salt, cobalt salt, titanium salt, aluminum salt, copper salt, cadmium salt, or zinc salt.

[0557] Ammonium salts can be produced by adding ammonia or an organic amine to the compounds of the present invention. In some embodiments, the organic amine is triethylamine, diisopropylamine, ethanolamine, diethanolamine, triethanolamine, morpholine, N-methylmorpholine, piperidine, N-methylpiperidine, N-ethylpiperidine, dibenzylamine, piperazine, pyridine, pyrazole, piperazole, imidazole, pyrazine, or piperazine.

[0558] In some embodiments, the ammonium salt is a triethylamine salt, diisopropylamine salt, ethanolamine salt, diethanolamine salt, triethanolamine salt, morpholine salt, N-methylmorpholine salt, piperidine salt, N-methylpiperidine salt, N-ethylpiperidine salt, dibenzylamine salt, piperazine salt, pyridine salt, pyrazole salt, piperpyrazole salt, imidazole salt, pyrazine salt, or piperpyridine salt.

[0559] Acid addition salts can be produced by adding an acid to the compounds of the present invention. In some embodiments, the acid is organic. In some embodiments, the acid is inorganic. In some embodiments, the acid is hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, nitrous acid, sulfuric acid, sulfurous acid, phosphoric acid, isonicotinic acid, lactic acid, salicylic acid, tartaric acid, ascorbic acid, gentian acid, gluconic acid, glucuronic acid, saccaric acid, formic acid, benzoic acid, glutamic acid, pantothenic acid, acetic acid, propionic acid, butyric acid, fumaric acid, succinic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, oxalic acid, or maleic acid.

[0560] In some embodiments, the salt is a hydrochloride, hydrobromide, hydroiodide, nitrate, nitrite, sulfate, sulfite, phosphate, isonicotinate, lactate, salicylate, tartrate, ascorbate, gentianate, gluconate, glucuronide, glycosylate, formate, benzoate, glutamate, pantothenate, acetate, propionate, butyrate, fumarate, succinate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, citrate, oxalate, or maleate.

[0561] The pharmaceutical composition of the present invention.

[0562] The pharmaceutical compositions of the present invention may be used, for example, before, during, or after treatment of a subject with another pharmaceutical agent.

[0563] For example, the subjects can be the elderly, adults, adolescents, pre-adolescent children, children, toddlers, infants, newborns, and non-human animals. In some implementations, the subjects are patients.

[0564] The pharmaceutical compositions of the present invention can be any pharmaceutical compound described herein combined with other chemical components, such as carriers, stabilizers, diluents, dispersants, suspending agents, thickeners, and / or excipients. The pharmaceutical compositions facilitate the administration of the compound to a living organism. The pharmaceutical compositions can be administered in various forms and routes, including, for example, intravenous, subcutaneous, intramuscular, oral, parenteral, ophthalmic, subcutaneous, transdermal, nasal, vaginal, and topical administration to a therapeutically effective amount.

[0565] The pharmaceutical composition can be administered topically, for example, by injecting the compound directly into an organ, optionally as a stock or sustained-release formulation or implant. The pharmaceutical composition can be provided as a rapid-release formulation, a prolonged-release formulation, or an intermediate-release formulation. Rapid-release formulations provide immediate release. Prolonged-release formulations provide controlled release or sustained delayed release.

[0566] For oral administration, pharmaceutical compositions can be formulated by combining the active compound with a pharmaceutically acceptable carrier or excipient. Such carriers can be used to formulate liquids, gels, syrups, elixirs, slurries, or suspensions for oral ingestion by the recipient. Non-limiting examples of solvents used in oral soluble formulations include water, ethanol, isopropanol, saline, physiological saline, DMSO, dimethylformamide, potassium phosphate buffer, phosphate-buffered saline (PBS), sodium phosphate buffer, 4-2-hydroxyethyl-1-piperazine ethanesulfonic acid buffer (HEPES), 3-(N-morpholino)propanesulfonic acid buffer (MOPS), piperazine-N,N′-bis(2-ethanesulfonic acid) buffer (PIPES), and saline sodium citrate buffer (SSC). Non-limiting examples of co-solvents used in oral soluble formulations include sucrose, urea, cremaphore, DMSO, and potassium phosphate buffer.

[0567] Pharmaceutical formulations can be formulated for intravenous administration. Pharmaceutical compositions can be in forms suitable for parenteral injection, as sterile suspensions, solutions, or emulsions in oily or aqueous media, and may contain formulations such as suspending agents, stabilizers, and / or dispersants. Pharmaceutical formulations for parenteral administration include aqueous solutions of the active compound in a water-soluble form. Suspensions of the active compound can be prepared as oily injectable suspensions. Suitable lipophilic solvents or media include fatty oils such as sesame oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. Suspensions may also contain suitable stabilizers or reagents that increase the solubility of the compound to allow for the preparation of high-concentration solutions. Alternatively, the active ingredient may be in powder form, prepared prior to use with a suitable media such as sterile, pyrogen-free water.

[0568] The active compound can be applied topically and can be formulated into a variety of topically applicable compositions, such as solutions, suspensions, lotions, gels, pastes, sticks, balms, creams, and ointments. Such pharmaceutical compositions may contain solubilizers, stabilizers, tonic agents, buffers, and preservatives.

[0569] The compounds of the present invention can be applied topically to the skin or body cavities of an object, such as the mouth, vagina, bladder, skull, spine, thoracic cavity, or pelvic cavity. The compounds of the present invention can be applied to accessible body cavities.

[0570] The compounds can also be formulated into rectal compositions, such as enemas, rectal gels, rectal foams, rectal aerosols, suppositories, jelly suppositories, or retention enemas, comprising a conventional suppository base, such as cocoa butter or other glycerides, and a synthetic polymer, such as polyvinylpyrrolidone and PEG. In the suppository form of the composition, a mixture of low-melting-point waxes, such as fatty acid glycerides optionally combined with cocoa butter, can be melted.

[0571] When performing the treatment or methods of use provided herein, a therapeutically effective amount of the compounds described herein is administered to a subject suffering from the disease or condition to be treated, in the form of a pharmaceutical composition. In some embodiments, the subject is a mammal, such as a human. The therapeutically effective amount can vary considerably depending on the severity of the disease, the subject's age and relative health status, the potency of the compound used, and other factors. These compounds may be used alone or as components of a mixture in combination with one or more therapeutic agents.

[0572] Pharmaceutical compositions can be formulated using one or more physiologically acceptable carriers, including excipients and adjuvants, which facilitate the processing of the active compound into a pharmaceutically usable formulation. The formulation may be modified depending on the chosen route of administration. Pharmaceutical compositions comprising the compounds described herein can be manufactured, for example, by mixing, dissolving, emulsifying, encapsulating, embedding, or compressing processes.

[0573] The pharmaceutical composition may comprise at least one pharmaceutically acceptable carrier, diluent, or excipient, and a compound described herein as a free base or a pharmaceutically acceptable salt. The pharmaceutical composition may comprise a solubilizer, stabilizer, tonicant, buffer, and preservative.

[0574] Methods for preparing compositions comprising the compounds described herein include formulating the compounds with one or more inert, pharmaceutically acceptable excipients or carriers to form solid, semi-solid, or liquid compositions. Solid compositions include, for example, powders, tablets, dispersible granules, capsules, and granules. Liquid compositions include, for example, solutions in which the compounds are dissolved, emulsions containing the compounds, or solutions containing liposomes, micelles, or nanoparticles containing the compounds disclosed herein. Semi-solid compositions include, for example, gels, suspensions, and creams. Compositions may be liquid solutions or suspensions, solid forms suitable for dissolving or suspending in a liquid prior to use, or as emulsions. These compositions may also contain small amounts of non-toxic excipients, such as wetting agents or emulsifiers, pH buffers, and other pharmaceutically acceptable additives.

[0575] Non-limiting examples of dosage forms suitable for use in this invention include liquids, powders, gels, nanosuspensions, nanoparticles, microgels, aqueous or oily suspensions, emulsions, and any combination thereof.

[0576] Non-limiting examples of pharmaceutically acceptable excipients suitable for use in this invention include binders, disintegrants, anti-adhesion agents, antistatic agents, surfactants, antioxidants, coating agents, colorants, plasticizers, preservatives, suspending agents, emulsifiers, antimicrobial agents, spheroidizing agents, and any combination thereof.

[0577] The compositions of the present invention can be, for example, immediate-release or controlled-release formulations. Immediate-release formulations can be formulated to allow the compounds to act rapidly. Non-limiting examples of immediate-release formulations include readily soluble formulations. Controlled-release formulations can be pharmaceutical formulations that have been adapted such that the release rate and release profile of the active agent can be matched to physiological and time-dependent therapeutic requirements, or alternatively, have been formulated to achieve the release of the active agent at a planned rate. Non-limiting examples of controlled-release formulations include particles, delayed-release particles, hydrogels (e.g., synthetic or naturally derived), other gelling agents (e.g., gel-forming dietary fiber), matrix-based formulations (e.g., formulations comprising a polymeric material dispersed with at least one active ingredient), intramatrix particles, polymer mixtures, and particulate matter.

[0578] In some cases, controlled-release formulations are in a delayed-release form. Delayed-release forms can be formulated to prolong the time period by which the action of a compound is delayed. Delayed-release forms can be formulated to delay the release of an effective dose of one or more compounds, for example, by about 4, about 8, about 12, about 16, or about 24 hours.

[0579] Controlled-release formulations can be in a sustained-release form. For example, a sustained-release form can be formulated to maintain the effect of the compound over an extended period of time. Sustained-release forms can be formulated to provide an effective dose (e.g., to provide physiologically effective blood distribution) of any of the compounds described herein over approximately 4 hours, 8 hours, 12 hours, 16 hours, or 24 hours.

[0580] Non-limiting examples of pharmaceutically acceptable excipients can be found, for example, in Remington: The Science and Practice of Pharmacy, 19th edition (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, HA and Lachman, L., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th edition (Lippincott Williams & Wilkins 1999), the entire contents of which are incorporated herein by reference.

[0581] Multiple therapeutic agents can be administered in any order or simultaneously. In some embodiments, the compounds of the present invention are administered in combination with another therapeutic agent or before or after treatment with another therapeutic agent. If administered simultaneously, multiple therapeutic agents can be provided in a single, uniform, or multiple forms, for example, as multiple individual pills. These agents can be packaged together or separately, in a single package or in multiple packages. One or all of the therapeutic agents can be given in multiple doses. If not administered simultaneously, the timing between multiple doses may vary up to approximately one month.

[0582] The therapeutic agents described herein can be administered before, during, or after the onset of a disease or condition, and the timing of administration of the composition containing the therapeutic agent can vary. For example, the composition can be used as a preventative agent and can be continuously administered to subjects predisposed to a disease or condition to reduce the likelihood of its occurrence. The composition can be administered to the subject as soon as possible during or after the onset of symptoms. The therapeutic agent can be initiated within the first 48 hours, the first 24 hours, the first 6 hours, or the first 3 hours of symptom onset. Initial administration can be via any practical route, such as using any of the formulations described herein via any of the routes described herein.

[0583] Once a disease or condition is detected or suspected to have begun, the compound may be administered as soon as practicable and for the duration required to treat the disease, such as, for example, about 1 month to about 3 months. In some embodiments, the duration for which the compound may be administered may be about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 1 month, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 2 months, about 9 weeks, about 10 weeks, about 11 weeks, about 12 weeks, about 3 months, about 13 weeks, about 14 weeks, about 15 weeks, about 16 weeks, about 4 months, about 17 weeks, about 18 weeks, about 19 weeks, about 20 weeks, about 5 months, about 21 weeks, about 2 Treatment duration can vary depending on the individual patient, ranging from approximately 2 weeks, 23 weeks, 24 weeks, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, 2 years, 2.5 years, 3 years, 3.5 years, 4 years, 4.5 years, 5 years, 6 years, 7 years, 8 years, 9 years, or 10 years.

[0584] The pharmaceutical compositions described herein can be unit dosage forms suitable for precise single-dose administration. In a unit dosage form, the formulation is divided into unit doses containing appropriate amounts of one or more compounds. A unit dose can be a packaging form containing discrete amounts of the formulation. Non-limiting examples are packaged injections, vials, or ampoules. Aqueous suspension compositions can be packaged in single-dose, non-resealable containers. For example, multi-dose, resealable containers can be used with or without preservatives. Injectable formulations can be presented in unit dosage forms, for example, in ampoules or in multi-dose containers containing preservatives.

[0585] The pharmaceutical compositions described herein can be administered in combination with other therapies, such as chemotherapy, radiation therapy, surgery, anti-inflammatory agents, and selected vitamins. Other agents may be administered before, after, or concurrently with the pharmaceutical compositions.

[0586] Depending on the intended method of administration, the pharmaceutical composition may be in the form of a solid, semi-solid, or liquid dosage form, such as, for example, tablets, suppositories, pills, capsules, powders, liquids, suspensions, lotions, creams, or gels, for example, in a unit dosage form suitable for a precise single-dose administration.

[0587] For solid compositions, non-toxic solid carriers include, for example, pharmaceutical-grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, talc, cellulose, glucose, sucrose, and magnesium carbonate.

[0588] Non-limiting examples of pharmaceutically active agents suitable for combination with the compositions of this disclosure include anti-infective agents, i.e., aminoglycosides, antiviral agents, antimicrobial agents, anticholinergic drugs / antispasmodics, antidiabetic agents, antihypertensive agents, antitumor agents, cardiovascular drugs, central nervous system agents, coagulation modulators, hormones, immunomodulators, immunosuppressants, and ophthalmic preparations.

[0589] Compounds can be delivered via liposome technology. Using liposomes as drug carriers can improve the therapeutic index of compounds. Liposomes consist of natural phospholipids and can contain mixed lipid chains with surfactant properties (e.g., lecithin ethanolamine). Liposomes can be designed to attach to unhealthy tissues using surface ligands. Non-limiting examples of liposomes include multilayered vesicles (MLVs), small unlayered vesicles (SUVs), and large unlayered vesicles (LUVs). The physicochemical properties of liposomes can be tuned to optimize penetration across biological barriers and retention at the application site, and to reduce the likelihood of premature degradation and toxicity to non-target tissues. Optimal liposome properties depend on the route of administration: large-size liposomes exhibit good retention upon local injection, while small-size liposomes are better suited for passive targeting. Polyethylene glycolization reduces liposome uptake by the liver and spleen and increases circulation time, resulting in increased localization to inflamed sites due to enhanced permeability and retention (EPR) effects. Furthermore, the liposome surface can be modified to achieve selective delivery of encapsulated drugs to specific target cells. Non-limiting examples of targeted ligands include monoclonal antibodies, vitamins, peptides, and polysaccharides that are specific to receptors concentrated on the cell surface associated with disease.

[0590] Non-limiting examples of dosage forms applicable to this disclosure include liquids, elixirs, nanosuspensions, aqueous or oily suspensions, drops, syrups, and any combination thereof. Non-limiting examples of pharmaceutically acceptable excipients applicable to this disclosure include granulators, binders, lubricants, disintegrants, sweeteners, gliding agents, anti-adhesion agents, antistatic agents, surfactants, antioxidants, gums, coating agents, colorants, flavoring agents, plasticizers, preservatives, suspending agents, emulsifiers, plant cellulose materials, and spheronizing agents, and any combination thereof.

[0591] The compositions of the present invention can be packaged as kits. In some embodiments, the kit includes written instructions for the administration / use of the composition. For example, the written material may be a label. The written material may suggest the conditions and methods of administration. The instructions provide best guidance to the subject and attending physician to achieve the best clinical outcomes through the administration of the therapy. The written material may be a label. In some embodiments, the label may be approved by a regulatory agency, such as the U.S. Food and Drug Administration (FDA), the European Medicines Agency (EMA), or other regulatory agencies.

[0592] dose.

[0593] The pharmaceutical compositions described herein can be unit dosage forms suitable for precise single-dose administration. In a unit dosage form, the formulation is divided into unit doses containing appropriate amounts of one or more compounds. A unit dose can be a packaging form containing discrete amounts of the formulation. A non-limiting example is a liquid in a vial or ampoule. Aqueous suspension compositions can be packaged in single-dose, non-resealable containers. For example, multi-dose, resealable containers can be used in conjunction with preservatives. Formulations for parenteral injection can be presented in unit dosage forms, for example, in ampoules or in multi-dose containers containing preservatives.

[0594] Dosage can be expressed as the amount of drug divided by the mass of the subject, for example, milligrams of drug per kilogram of subject body weight. The compounds described herein can be in quantities of about 1 mg to about 2000 mg; about 100 mg to about 2000 mg; about 10 mg to about 2000 mg; about 5 mg to about 1000 mg; about 10 mg to about 500 mg; about 50 mg to about 250 mg; about 100 mg to about 200 mg; about 1 mg to about 50 mg; about 50 mg to about 100 mg; about 100 mg to about 150 mg; about 150 mg to about 200 mg; about 200 mg to about 250 mg; about 250 mg to about 300 mg; about 300 mg to about 350 mg. The range of 0 mg, about 350 mg to about 400 mg, about 400 mg to about 450 mg, about 450 mg to about 500 mg, about 500 mg to about 550 mg, about 550 mg to about 600 mg, about 600 mg to about 650 mg, about 650 mg to about 700 mg, about 700 mg to about 750 mg, about 750 mg to about 800 mg, about 800 mg to about 850 mg, about 850 mg to about 900 mg, about 900 mg to about 950 mg, or about 950 mg to about 1000 mg is present in the composition.

[0595] In some embodiments, the compound is applied in amounts ranging from about 5 mg / kg to about 50 mg / kg, 250 mg / kg to about 2000 mg / kg, about 10 mg / kg to about 800 mg / kg, about 50 mg / kg to about 400 mg / kg, about 100 mg / kg to about 300 mg / kg, or about 150 mg / kg to about 200 mg / kg. In some embodiments, the compound described herein may be present in the composition in the range of about 20 mg / kg to about 400 mg / kg. In some embodiments, the compound described herein may be present in the composition in the range of about 20 mg / kg to about 240 mg / kg. In some embodiments, the compound described herein may be present in the composition in the range of about 75 mg / kg to about 150 mg / kg. In some embodiments, the compound described herein may be present in the composition in the range of about 75 mg / kg to about 150 mg / kg. In some embodiments, the compound described herein may be present in the composition in the range of about 100 mg / kg to about 150 mg / kg.

[0596] In some embodiments, the compound described herein may be present in the composition at an amount of about 75 mg / kg. In some embodiments, the compound described herein may be present in the composition at an amount of about 100 mg / kg. In some embodiments, the compound described herein may be present in the composition at an amount of about 150 mg / kg. In some embodiments, the compound described herein may be present in the composition at an amount of about 200 mg / kg. In some embodiments, the compound described herein may be present in the composition at an amount of about 250 mg / kg. In some embodiments, the compound described herein may be present in the composition at an amount of about 400 mg / kg.

[0597] The compounds described herein may be present in doses of approximately 1 mg, approximately 2 mg, approximately 3 mg, approximately 4 mg, approximately 5 mg, approximately 10 mg, approximately 15 mg, approximately 20 mg, approximately 25 mg, approximately 30 mg, approximately 35 mg, approximately 40 mg, approximately 45 mg, approximately 50 mg, approximately 55 mg, approximately 60 mg, approximately 65 mg, approximately 70 mg, approximately 75 mg, approximately 80 mg, approximately 85 mg, approximately 90 mg, approximately 95 mg, approximately 100 mg, approximately 125 mg, approximately 150 mg, approximately 175 mg, approximately 200 mg, approximately 250 mg, approximately 300 mg, approximately 350 mg, approximately 400 mg, approximately 450 mg, approximately 500 mg, approximately 550 mg, approximately 6 mg, approximately 6 mg, approximately 1 mg, approximately 2 mg, approximately 3 mg, approximately 4 mg, approximately 450 mg, approximately 500 mg, approximately 550 mg, approximately 6 mg, approximately 1 mg, approximately 2 mg, approximately 3 mg, approximately 4 mg, approximately 5 ...5 mg, approximately 5 mg, approximately 6 mg, approximately 1 mg, approximately 1 mg, approximately 2 mg, approximately 3 mg, approximately 4 mg, approximately 5 mg, approximately 4 mg, approximately The composition is present in amounts of 00 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, about 900 mg, about 950 mg, about 1000 mg, about 1050 mg, about 1100 mg, about 1150 mg, about 1200 mg, about 1250 mg, about 1300 mg, about 1350 mg, about 1400 mg, about 1450 mg, about 1500 mg, about 1550 mg, about 1600 mg, about 1650 mg, about 1700 mg, about 1750 mg, about 1800 mg, about 1850 mg, about 1900 mg, about 1950 mg, or about 2000 mg.

[0598] In some embodiments, the compound described herein may be present in the composition in amounts of about 100 mg, about 120 mg, about 140 mg, about 160 mg, about 180 mg, about 200 mg, about 220 mg, about 240 mg, about 260 mg, about 280 mg, or about 300 mg. In some embodiments, the compound described herein may be present in the composition in an amount of about 150 mg. In some embodiments, the compound described herein may be present in the composition in an amount of about 170 mg. In some embodiments, the compound described herein may be present in the composition in an amount of about 280 mg. In some embodiments, the compound described herein may be present in the composition in an amount of about 300 mg.

[0599] Example

[0600] A. Synthesis of alkynyl reagents.

[0601] Example A1: Synthesis of 3-(fluoromethoxy)-N-methyl-4-(prop-2-yn-1-ylamino)benzamide.

[0602]

[0603] Step 1. At -10°C, BBr3 (3.62 g, 14.43 mmol, 1.39 mL, 3.5 equivalents) was added dropwise to a solution of 3-methoxy-N-methyl-4-(prop-2-yn-1-ylamino)benzamide (947.37 mg, 4.12 mmol, 1 equivalent) in DCM (50 mL). The mixture was stirred at 0°C for 2 hours. TLC analysis (DCM:MeOH = 20:1, R f =0.4) indicates that approximately 10% of the starting material remains, and a new spot with a lower polarity than the starting material was detected. Saturated NaOH solution was added until the pH of the mixture was greater than 11. The mixture was extracted with DCM (50 mL x 3), and the organic layer was discarded. 12 M HCl was added to the aqueous phase until the pH reached 8. The aqueous phase was extracted with EtOAc (150 mL x 3), the combined organic layers were washed with brine (150 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was stirred in PE (50 mL) at 25 °C for 12 hours. The mixture was then filtered to give 3-hydroxy-N-methyl-4-(prop-2-yn-1-ylamino)benzamide (0.7 g, 3.08 mmol, 74.81% yield) as a yellow solid.

[0604] Step 2. K₂CO₃ (365.45 mg, 2.64 mmol, 2 equivalents) was added to a solution of 3-hydroxy-N-methyl-4-(prop-2-yn-1-ylamino)benzamide (0.3 g, 1.32 mmol, 1 equivalent) and bromofluoromethane (298.60 mg, 2.64 mmol, 251.45 μL, 2 equivalents) in DMF (10 mL). The mixture was stirred at 50 °C for 1 hour, followed by TLC analysis (DCM:MeOH = 20:1, R0). f =0.5) indicates complete consumption of the starting phenol, and a new spot was observed. The mixture was poured into water (20 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated. The mixture was then analyzed by preparative TLC (DCM:MeOH = 20:1, R f =0.5) The residue obtained after purification was used to give 3-(fluoromethoxy)-N-methyl-4-(prop-2-yn-1-ylamino)benzamide (0.3 g, 1.08 mmol, 81.64% yield), as a yellow solid.

[0605] Example A2: Synthesis of 3-(2-cyanoethoxy)-N-methyl-4-(prop-2-yn-1-ylamino)benzamide.

[0606]

[0607] Under N2, a mixture of 3-hydroxy-N-methyl-4-(prop-2-yn-1-ylamino)benzamide (200 mg, 685.52 μmol, 1 equivalent) and benzyl(trimethyl)ammonium hydroxide (3.82 mg, 6.86 μmol, 4.15 μL, 30% purity, 0.01 equivalent) was stirred in acrylonitrile (1.09 g, 20.57 mmol, 1.36 mL, 30 equivalent) at 85 °C for 16 h. TLC analysis showed that approximately 50% of the starting phenol remained, and a new spot with a lower polarity than the starting material was observed. The mixture was concentrated under reduced pressure to provide the residue, which was then analyzed by preparative TLC (DCM:MeOH = 20:1, R... f Purification with 0.5 g yielded 3-(2-cyanoethoxy)-N-methyl-4-(prop-2-yn-1-ylamino)benzamide (100 mg, 349.80 μmol, 39.69% yield), a yellow solid.

[0608] Example A3: Synthesis of 3-(cyanomethoxy)-4-(prop-2-yn-1-ylamino)benzenesulfonamide.

[0609]

[0610] Step 1. At -10°C, BBr3 (1.24 g, 4.95 mmol, 477.20 μL, 3.5 equivalents) was added dropwise to a solution of 3-methoxy-4-(prop-2-yn-1-ylamino)benzenesulfonamide (0.4 g, 1.42 mmol, 1 equivalent) in DCM (10 mL). The mixture was stirred at 0°C for 2 hours, followed by TLC analysis (DCM:MeOH = 10:1, R0). f =0.4) indicates approximately 10% of the starting methyl ether remains, and two new spots with a polarity greater than that of the starting material were observed. 1N NaOH was added until the pH of the mixture was greater than 11. The mixture was extracted with DCM (50 mL x 3), and the organic layer was discarded. 12M HCl was added to the aqueous phase until the pH equaled 8, and the aqueous phase was extracted with EtOAc (150 mL x 3). The combined organic layers were washed with brine (150 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The mixture was then analyzed by preparative TLC (DCM:MeOH = 10:1, R f =0.4) The residue obtained after purification was used to give 3-hydroxy-4-(prop-2-yn-1-ylamino)benzenesulfonamide (0.25 g, 983.42 μmol, 69.50% yield), which was a yellow solid.

[0611] Step 2. K₂CO₃ (271.84 mg, 1.97 mmol, 2 equivalents) was added to a solution of 3-hydroxy-4-(prop-2-yn-1-ylamino)benzenesulfonamide (0.25 g, 983.42 μmol, 1 equivalent) and bromoacrylonitrile (235.92 mg, 1.97 mmol, 131.07 μL, 2 equivalents) in DMF (10 mL). The mixture was stirred at 50 °C for 2 hours. TLC analysis (DCM:MeOH = 10:1, R f =0.5) indicates complete consumption of the starting phenol, and a new spot was observed. The mixture was poured into water (30 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with brine (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The solution was then analyzed by preparative TLC (DCM:MeOH = 10:1, R f =0.5) The residue obtained after purification was used to give 3-(cyanomethoxy)-4-(prop-2-yn-1-ylamino)benzenesulfonamide (0.2 g, 716.20 μmol, 72.83% yield), which was a yellow solid.

[0612] Example A4: General procedure for the preparation of 2-(fluoromethoxy)-4-(methanesulfonyl)-N-(prop-2-yn-1-yl)aniline and 2-(5-(methanesulfonyl)-2-(prop-2-yn-1-ylamino)phenoxy)acetonitrile.

[0613]

[0614] Synthesis of 5-(methanesulfonyl)-2-(prop-2-yn-1-ylamino)phenol: BBr3 (12.43 g, 49.63 mmol, 4.78 mL, 2.5 equivalents) was added dropwise to a solution of 2-methoxy-4-(methanesulfonyl)-N-(prop-2-yn-1-yl)aniline (5 g, 19.85 mmol, 1 equivalent) in DCM (50 mL) at -10 °C. The mixture was stirred at 0 °C for 2 hours. TLC analysis (PE:EtOAc = 1:1, Rm) was performed. f=0.4) indicates approximately 10% of the starting methyl ether remains, and a major new spot with a polarity greater than that of the starting material is observed. 1N NaOH is added until the pH of the mixture is greater than 11. The mixture is extracted with DCM (50 mL x 3), and the organic layer is discarded. 12M HCl is added to the aqueous phase until the pH equals 8, and the aqueous phase is extracted with EtOAc (150 mL x 3). The combined organic layers are washed with brine (150 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue is stirred in PE (50 mL) at 25 °C for 12 hours. The mixture is then filtered and dried to give 5-(methanesulfonyl)-2-(prop-2-yn-1-ylamino)phenol (8 g, 30.19 mmol, 76.04% yield) as a yellow solid.

[0615] Synthesis of 2-(fluoromethoxy)-4-(methanesulfonyl)-N-(prop-2-yn-1-yl)aniline and 2-(5-(methanesulfonyl)-2-(prop-2-yn-1-ylamino)phenoxy)acetonitrile: K₂CO₃ (521.5 mg, 3.77 mmol, 2 equivalents) was added to a solution of 5-(methanesulfonyl)-2-(prop-2-yn-1-ylamino)phenol (0.5 g, 2.22 mmol, 1 equivalent) and 2-bromoacetonitrile (450 mg, 3.77 mmol, 2 equivalents) or bromofluoromethane (422 mg, 3.77 mmol, 2 equivalents) in DMF (10 mL). The mixture was stirred at 50 °C for 2 hours and then poured into water (50 mL). The mixture was extracted with EtOAc (30 mL x 3), the combined organic layers were washed with brine (40 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by column chromatography to give 2-(fluoromethoxy)-4-(methanesulfonyl)-N-(prop-2-yn-1-yl)aniline (700 mg, crude) or 2-(5-(methanesulfonyl)-2-(prop-2-yn-1-ylamino)phenoxy)acetonitrile as a yellow gel.

[0616] Example A5: General procedure for the preparation of N-((3-methoxy-4-(prop-2-yn-1-ylamino)phenyl)sulfonyl)acetamide and N-((3-methoxy-4-(prop-2-yn-1-ylamino)phenyl)sulfonyl)propionamide.

[0617]

[0618] At 25°C and under N2, a mixture of 3-methoxy-4-(prop-2-yn-1-ylamino)benzenesulfonamide (1 equivalent), DMAP (0.1 equivalent), and TEA (1 equivalent) in THF (4 mL) was added (R 12O (2 equivalents). The mixture was stirred at 25°C for 2 hours, and TLC analysis indicated that the reaction was complete. The reaction mixture was diluted with water (20 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC to give the desired product as a yellow oil.

[0619] Example A6: Preparation of N-((3-methoxy-4-(prop-2-yn-1-ylamino)phenyl)sulfonyl)-N-methylpropionamide.

[0620]

[0621] DMAP (2.88 mg, 23.59 μmol, 0.1 equivalent), TEA (23.87 mg, 235.94 μmol, 1 equivalent), and propionic anhydride (61.41 mg, 471.87 μmol, 60.80 μL, 2 equivalent) were added to a solution of 3-methoxy-N-methyl-4-(prop-2-yn-1-ylamino)benzenesulfonamide (0.06 g, 235.94 μmol, 32.84 μL, 1 equivalent) in THF (2 mL). The reaction mixture was stirred at 25 °C for 10 h. The desired mass was detected by LC-MS analysis. The reaction mixture was concentrated under reduced pressure and analyzed by preparative TLC (SiO2, PE:EtOAc = 1:1, R f =0.43) The residue obtained after purification was used to give N-((3-methoxy-4-(prop-2-yn-1-ylamino)phenyl)sulfonyl)-N-methylpropionamide (0.04 g, 90.22 μmol, 38.24% yield), which was a yellow solid.

[0622] Example A7: Preparation of N-methyl-5-(methanesulfonyl)-2-(prop-2-yn-1-yloxy)aniline, N,N-dimethyl-5-(methanesulfonyl)-2-(prop-2-yn-1-yloxy)aniline and N-(5-(methanesulfonyl)-2-(prop-2-yn-1-yloxy)phenyl)acetamide.

[0623]

[0624] Synthesis of 4-(methanesulfonyl)-2-nitro-1-(prop-2-yn-1-yloxy)benzene: K₂CO₃ (1.91 g, 13.80 mmol, 3 equivalents) was added to a mixture of propargyl bromide (2.74 g, 23 mmol, 1.98 mL, 5 equivalents) and 4-(methanesulfonyl)-2-nitrophenol (1 g, 4.60 mmol, 1 equivalent) in DMF (10 mL). The mixture was stirred at 50 °C for 2 h, and subsequent TLC (EtOAc, Rf = 0.43) indicated that the reaction was complete. The reaction mixture was quenched with water (150 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give crude 4-(methanesulfonyl)-2-nitro-1-(prop-2-yn-1-yloxy)benzene as a pale yellow solid.

[0625] Synthesis of 5-(methanesulfonyl)-2-(prop-2-yn-1-yloxy)aniline: Fe (1.31 g, 23.51 mmol, 5 equivalents) was added to a solution of 4-(methanesulfonyl)-2-nitro-1-(prop-2-yn-1-yloxy)benzene (1.2 g, 4.70 mmol, 1 equivalent) in AcOH (10 mL). The mixture was stirred at 70 °C for 2 hours, followed by TLC analysis (PE:EtOAc = 1:1, R f =0.43) indicates that the reaction was complete. The reaction mixture was concentrated under reduced pressure to remove the solvent and diluted with EtOAc (50 mL). The reaction mixture was quenched at 25 °C by adding a saturated NaHCO3 solution (200 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, PE:EtOAc = 1:1) to give 5-(methanesulfonyl)-2-(prop-2-yn-1-yloxy)aniline (0.86 g, 3.44 mmol, 73.09% yield) as a pale yellow solid.

[0626] Synthesis of N-(5-(methanesulfonyl)-2-(prop-2-yn-1-yloxy)phenyl)acetamide: A mixture of 5-(methanesulfonyl)-2-(prop-2-yn-1-yloxy)aniline (100 mg, 399.53 μmol, 1 equivalent), acetic anhydride (203.94 mg, 2 mmol, 187.10 μL, 5 equivalent) and TEA (80.86 mg, 799.06 μmol, 111.22 μL, 2 equivalent) in DCM (3 mL) was stirred at 50 °C for 2 h. TLC analysis (EtOAc, R f=0.24) indicates that the reaction is complete. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was purified by preparative TLC to give N-(5-(methylsulfonyl)-2-(prop-2-yn-1-yloxy)phenyl)acetamide (100 mg, 374.11 μmol, 93.64% yield) as a pale yellow solid.

[0627] Synthesis of N-methyl-5-(methanesulfonyl)-2-(prop-2-yn-1-yloxy)aniline: AcOH (53.3 mg, 887.8 μmol, 50.8 μL, 0.4 equivalents) and NaBH3CN (418.5 mg, 6.66 mmol, 3 equivalents) were added to a solution of 5-(methanesulfonyl)-2-(prop-2-yn-1-yloxy)aniline (0.5 g, 2.2 mmol, 1 equivalent) in MeOH (5 mL). The mixture was stirred at 25 °C for 0.5 h, and then formaldehyde (234.2 mg, 2.9 mmol, 214.8 μL, 1.3 equivalents) was added. The mixture was stirred at 25 °C for a further 9.5 h, and subsequent LC-MS analysis indicated that the reaction was complete. The reaction mixture was separated into layers by adding saturated NaHCO3 solution (30 mL) and EtOAc (10 mL), and extracted with EtOAc (5 mL x 3). The combined organic layers were washed with brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was first subjected to preparative TLC (PE:EtOAc = 2:1, R...). f =0.6) was purified and further purified by preparative HPLC to obtain N-methyl-5-(methanesulfonyl)-2-(prop-2-yn-1-yloxy)aniline (100 mg, 376.11 μmol, 16.94% yield), which was a colorless oil.

[0628] Synthesis of N,N-dimethyl-5-(methanesulfonyl)-2-(prop-2-yn-1-yloxy)aniline: AcOH (21.3 mg, 355.1 μmol, 20.3 μL, 0.4 equivalents) and NaBH3CN (167.4 mg, 2.66 mmol, 3 equivalents) were added to a solution of 5-(methanesulfonyl)-2-(prop-2-yn-1-yloxy)aniline (0.2 g, 887.8 μmol, 1 equivalent) in MeOH (5 mL). The mixture was stirred at 25 °C for 0.5 h, and then formaldehyde (216.2 mg, 2.7 mmol, 198.3 μL, 3 equivalents) was added. The mixture was further stirred at 25 °C for 9.5 h, and subsequent LC-MS analysis indicated that the reaction was complete. The reaction mixture was separated into layers by adding saturated NaHCO3 solution (30 mL) and EtOAc (10 mL), and extracted with EtOAc (5 mL x 3). The combined organic layers were washed with brine (10 mL x 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was first analyzed by preparative TLC (PE:EtOAc = 2:1, R0). f =0.6) Purification yielded N,N-dimethyl-5-(methanesulfonyl)-2-(prop-2-yn-1-yloxy)aniline (0.1 g, 355.29 μmol, 40% yield), as a colorless oil.

[0629] Example A8: Preparation of 4-(methylsulfonyl)-N-(prop-2-yn-1-yl)-2-(trifluoromethyl)aniline.

[0630]

[0631] Synthesis of methyl(4-nitro-3-(trifluoromethyl)phenyl)thion: Under N2 at 0 °C, a solution of 4-fluoro-1-nitro-2-(trifluoromethyl)benzene (10 g, 47.82 mmol, 1 equivalent) in DMF (100 mL) was added once to NaSMe (33.52 g, 95.65 mmol, 30.47 mL, 20% purity, 2 equivalents). The mixture was stirred at 25 °C for 60 min, and subsequent TLC analysis indicated that the reaction was complete. The reaction mixture was diluted with water (100 mL) and extracted with EtOAc (100 mL x 3). The organic layer was then washed with semi-saturated brine (100 mL x 5), dried over anhydrous sodium sulfate, filtered, and concentrated to give 4-methylthioalkyl-1-nitro-2-(trifluoromethyl)benzene (11.4 g, crude), a brown liquid, which was used unpurified for the next step.

[0632] Synthesis of 4-(methylsulfonyl)-1-nitro-2-(trifluoromethyl)benzene: Under N2 and at 0 °C, potassium persulfate (51.84 g, 84.32 mmol, 2 equivalents) was added in a single addition to a solution of 4-methylthioalkyl-1-nitro-2-(trifluoromethyl)benzene (10 g, 42.16 mmol, 1 equivalent) in acetone (100 mL), water (100 mL), and MeOH (10 mL). The mixture was stirred at 25 °C for 60 min, and subsequent TLC and LC-MS analysis indicated that the reaction was complete. The reaction was quenched by adding a saturated Na2S2O3 solution. The reaction mixture was slowly added to a saturated NaHCO3 solution (15 mL), followed by a saturated Na2S2O3 solution (20 mL). The completion of the reaction was monitored using KI starch paper. The mixture was then extracted with EtOAc (50 mL x 3) and the organic phase was washed with brine (40 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give crude 4-methylsulfonyl-1-nitro-2-(trifluoromethyl)benzene (9.8 g, 36.40 mmol, 86.35% yield), which was a white solid.

[0633] Synthesis of 4-(methylsulfonyl)-2-(trifluoromethyl)aniline: A solution of 4-methylsulfonyl-1-nitro-2-(trifluoromethyl)benzene (9.5 g, 35.29 mmol, 1 equivalent) in EtOH (200 mL) and NH4Cl (aq) (50 mL) was heated to 90 °C, and Fe (9.85 g, 176.45 mmol, 5 equivalents) was added in one step at 90 °C. The reaction mixture was stirred at 90 °C for 1 hour, and subsequent TLC analysis indicated that the reaction was complete. The reaction mixture was filtered while still hot. The filtrate was diluted with water (100 mL) and extracted with EtOAc (200 mL x 4). The combined organic layers were washed with brine (200 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated to give crude 4-methylsulfonyl-2-(trifluoromethyl)aniline (7.2 g) as a yellow solid. The crude product was purified by column chromatography (SiO2, PE:EtOAc = 2:1 to 1:1) to give 4-methylsulfonyl-2-(trifluoromethyl)aniline (6.8 g, 28.43 mmol, 97.14% yield) as a yellow solid.

[0634] Synthesis of tert-butyl (4-(methylsulfonyl)-2-(trifluoromethyl)phenyl)carbamate and (tert-butoxycarbonyl)(4-(methylsulfonyl)-2-(trifluoromethyl)phenyl)carbamate: Boc₂O (4.38 g, 20.07 mmol, 4.61 mL, 1.2 equivalents) and DMAP (2.45 g, 20.07 mmol, 1.2 equivalents) were added to a solution of 4-methylsulfonyl-2-(trifluoromethyl)aniline (4 g, 16.72 mmol, 1 equivalent) in THF (25 mL). The reaction mixture was stirred at 70 °C for 1 hour, and subsequent TLC analysis indicated completion. The reaction mixture was diluted with water (30 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with water (30 mL x 3) and brine (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by column chromatography (SiO2, PE:EtOAc = 6:1 to 4:1) to obtain a mixture of N-[4-methylsulfonyl-2-(trifluoromethyl)phenyl]carbamate tert-butyl ester and N-tert-butoxycarbonyl-N-[4-methylsulfonyl-2-(trifluoromethyl)phenyl]carbamate tert-butyl ester, which was a yellow gel.

[0635] Synthesis of tert-butyl N-[4-(methylsulfonyl)-2-(trifluoromethyl)phenyl]carbamate: K₂CO₃ (2.36 g, 17.07 mmol, 3 equivalents) was added in a single step to the mixture from the previous step (2.5 g, 5.69 mmol, 1 equivalent) dissolved in MeOH (40 mL). The mixture was stirred at 25 °C for 6 hours, and subsequent LC-MS analysis indicated that the reaction was complete. The reaction mixture was filtered and concentrated to give tert-butyl N-[4-methylsulfonyl-2-(trifluoromethyl)phenyl]carbamate (2.0 g, crude product) as a red solid.

[0636] Synthesis of tert-butyl (4-(methylsulfonyl)-2-(trifluoromethyl)phenyl)(prop-2-yn-1-yl)carbamate: Cs₂CO₃ (5.76 g, 17.68 mmol, 3 equivalents) and propargyl bromide (2.10 g, 17.68 mmol, 1.52 mL, 3 equivalents) were added to a solution of N-[4-methylsulfonyl-2-(trifluoromethyl)phenyl]carbamate (2 g, 5.89 mmol, 1 equivalent) in DMF (12 mL). The reaction mixture was stirred at 25 °C for 1.5 h, followed by TLC analysis (PE:EtOAc = 1:1, R f(起始原料) =0.68, product R f(产物)=0.50) indicates the reaction is complete. The reaction mixture was diluted with water (30 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with water (30 mL x 3) and brine (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (SiO2, PE:EtOAc = 4:1 to 2:1) to give N-[4-methylsulfonyl-2-(trifluoromethyl)phenyl]-N-prop-2-ynyl-carbamate tert-butyl ester (1.7 g, 4.50 mmol, 76.43% yield) as a colorless gel.

[0637] Synthesis of 4-(methanesulfonyl)-N-(prop-2-yn-1-yl)-2-(trifluoromethyl)aniline: A solution of N-[4-methanesulfonyl-2-(trifluoromethyl)phenyl]-N-prop-2-yn-yl-carbamate tert-butyl ester (1.7 g, 4.50 mmol, 1 equivalent) in HCl / EtOAc (4 M, 34 mL, 30.19 equivalents) was stirred at 25 °C for 1 h, followed by TLC analysis (PE:EtOAc = 1:1, R f(起始原料) =0.49,R f(产物) =0.27) indicates that the reaction is complete. The reaction was directly concentrated to give 4-methylsulfonyl-N-prop-2-ynyl-2-(trifluoromethyl)aniline (1.1 g, 3.97 mmol, 88.07% yield), a white solid.

[0638] Example A9: Preparation of 2-chloro-4-(methanesulfonyl)-N-(prop-2-yn-1-yl)aniline.

[0639]

[0640] Synthesis of (3-chloro-4-nitrophenyl)(methyl)thion: Under N2 at 0 °C, NaSMe (39.93 g, 113.93 mmol, 36.30 mL, 20% purity, 2 equivalents) was added in one step to a mixture of 2-chloro-4-fluoro-1-nitro-benzene (10 g, 56.97 mmol, 1 equivalent) in DMF (120 mL). The mixture was stirred at 25 °C for 60 min, followed by TLC analysis (PE:EtOAc = 10:1, Rm). f1 =0.66,R f2=0.55) indicates the reaction is complete. The reaction mixture was diluted with water (100 mL) and extracted with EtOAc (100 mL x 3). The combined organic layers were washed with semi-saturated brine (100 mL x 5), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (SiO2, PE:EtOAc = 5:1) to give 2-chloro-4-methylthioalkyl-1-nitrobenzene (4.3 g, 21.12 mmol, 37.07% yield) as a yellow solid.

[0641] Synthesis of 2-chloro-4-(methylsulfonyl)-1-nitrobenzene: Under N2 at 0 °C, potassium persulfate (25.96 g, 42.23 mmol, 2 equivalents) was added in one step to a mixture of 2-chloro-4-methylthioalkyl-1-nitrobenzene (4.3 g, 21.12 mmol, 1 equivalent) in toluene (25 mL), MeOH (2.5 mL), and water (25 mL). The mixture was stirred at 25 °C for 60 min, followed by TLC (PE:EtOAc = 1:1, R f(sm) =0.63,R f(pdt) =0.51) indicates that the reaction is complete. The reaction was quenched with saturated Na2S2O3 (200 mL) and extracted with EtOAc (100 mL x 3). The combined organic layers were washed with brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated to give crude 2-chloro-4-methylsulfonyl-1-nitrobenzene (5.0 g, crude product) as a yellow solid.

[0642] Synthesis of 2-chloro-4-(methanesulfonyl)aniline: A mixture of 2-chloro-4-methanesulfonyl-1-nitrobenzene (4.5 g, 19.10 mmol, 1 equivalent) in EtOH (40 mL) and saturated NH4Cl solution (10 mL) was heated to 90 °C, and then Fe (3.20 g, 57.29 mmol, 3 equivalents) was added in one step. The reaction mixture was stirred at 90 °C for 1 hour, and subsequent TLC analysis (PE:EtOAc = 1:1, R f(起始原料) =0.7,R f(产物) =0.31) indicates the reaction is complete. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (100 mL x 3). The combined organic layers were washed with water (50 mL x 3) and brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, PE:EtOAc = 4:1 to 2:1) to give 2-chloro-4-methylsulfonyl-aniline (3.7 g, 17.99 mmol, 94.21% yield) as a pale white solid.

[0643] Synthesis of tert-butyl (2-chloro-4-(methylsulfonyl)phenyl)carbamate: DMAP (2.20 g, 17.99 mmol, 1 equivalent) was added in a single batch to a mixture of 2-chloro-4-methylsulfonyl-aniline (3.7 g, 17.99 mmol, 1 equivalent) and (Boc)₂O (4.71 g, 21.59 mmol, 4.96 mL, 1.2 equivalent) in THF (50 mL). The mixture was stirred at 70 °C for 12 hours, followed by TLC (PE:EtOAc = 1:1, R f(sm) =0.45,R f(pdt) =0.66) indicates that some initial primary amine remains in the mixture. The reaction mixture was diluted with water (30 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with water (30 mL x 3) and brine (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (SiO2, PE:EtOAc = 4:1 to 2:1) to give N-(2-chloro-4-methylsulfonyl-phenyl)carbamate tert-butyl ester (2.7 g, 8.83 mmol, 49.08% yield) as a white solid.

[0644] Synthesis of tert-butyl (2-chloro-4-(methylsulfonyl)phenyl)(prop-2-yn-1-yl)carbamate: Cs₂CO₃ (7.67 g, 23.55 mmol, 3 equivalents) and propargyl bromide (2.80 g, 23.55 mmol, 2.03 mL, 3 equivalents) were added to a mixture of N-(2-chloro-4-methylsulfonyl-phenyl)carbamate (2.4 g, 7.85 mmol, 1 equivalent) in DMF (24 mL). The reaction mixture was stirred at 25 °C for 1.5 h, followed by TLC analysis (PE:EtOAc = 1:1, R0). f(起始原料) =0.68, R f(产物) =0.60) indicates the reaction is complete. The reaction mixture was diluted with water (30 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with water (30 mL x 3) and brine (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (SiO2, PE:EtOAc = 4:1 to 3:1) to give N-(2-chloro-4-methylsulfonyl-phenyl)-N-prop-2-ynyl-carbamate tert-butyl ester (1.7 g, 4.94 mmol, 62.99% yield) as a colorless gel.

[0645] Synthesis of 2-chloro-4-(methanesulfonyl)-N-(prop-2-yn-1-yl)aniline: A solution of N-(2-chloro-4-methanesulfonyl-phenyl)-N-prop-2-yn-yl-carbamate tert-butyl ester (300 mg, 872.54 mmol, 1 equivalent) in HCl / EtOAc (4 M, 6.59 L, 30.19 equivalents) was stirred at 25 °C for 1 h. Subsequent LC-MS analysis indicated that the reaction was complete. The reaction mixture was directly concentrated to give crude 2-chloro-4-methanesulfonyl-N-prop-2-yn-yl-aniline (180 mg, crude), as a brown solid.

[0646] Example A10: Preparation of 3-methoxy-N-methyl-4-(prop-2-yn-1-ylamino)benzamide.

[0647]

[0648] A mixture of 3-methoxy-4-(prop-2-yn-1-ylamino)benzoic acid (50 mg, 207.11 μmol, 1 equivalent), HATU (94.50 mg, 248.53 μmol, 1.2 equivalent), and DIPEA (53.53 mg, 414.21 μmol, 72.15 μL, 2 equivalent) in DMF (3 mL) was stirred at 25 °C for 15 min, and NH2Me (20.97 mg, 310.66 μmol, 1.5 equivalent) was added. The mixture was stirred for 3.75 h, and subsequent LC-MS analysis indicated that the reaction was complete. The reaction mixture was quenched by adding water (40 mL), and the resulting mixture was extracted with EtOAc (10 mL x 4). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was analyzed by preparative TLC (SiO2, EtOAc:PE = 2:1, R f Purification with 0.25 g yielded 3-methoxy-N-methyl-4-(prop-2-yn-1-ylamino)benzamide (45 mg, 185.57 μmol, 89.60% yield), which was a pale yellow oil.

[0649] Example A11: Preparation of N,N-bis(2-hydroxyethyl)-3-methoxy-4-(prop-2-yn-1-ylamino)benzenesulfonamide.

[0650]

[0651] Synthesis of N,N-bis(2-hydroxyethyl)-3-methoxy-4-nitrobenzenesulfonamide: A solution of 3-methoxy-4-nitrobenzenesulfonyl chloride (1 g, 3.97 mmol, 1 equivalent) in DCM (5 mL) was added to a mixture of diethanolamine (835.59 mg, 7.95 mmol, 766.59 μL, 2 equivalents) and TEA (804.23 mg, 7.95 mmol, 1.11 mL, 2 equivalents) in DCM (10 mL) at 0 °C. The reaction was heated to 25 °C with stirring for 1 hour, and subsequent TLC analysis (PE:EtOAc = 1:2, R f =0.3) indicates the reaction is complete. The reaction mixture was diluted with water (20 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was subjected to column chromatography (SiO2, PE:EtOAc = 1:2, R... f Purification was performed at 0.3 g to give N,N-bis(2-hydroxyethyl)-3-methoxy-4-nitrobenzenesulfonamide (1.2 g, 3.0 mmol, 75.42% yield) as a yellow solid.

[0652] Synthesis of 4-amino-N,N-bis(2-hydroxyethyl)-3-methoxybenzenesulfonamide: Fe (836.92 mg, 15 mmol, 5 equivalents) was added to a mixture of N,N-bis(2-hydroxyethyl)-3-methoxy-4-nitrobenzenesulfonamide (1.2 g, 3 mmol, 1 equivalent) and NH4Cl (801.55 mg, 15 mmol, 523.89 μL, 5 equivalents) in EtOH (20 mL) and water (4 mL) at 70 °C. The mixture was stirred at 70 °C for 1 h, and subsequent LC-MS analysis indicated the completion of the reaction. The reaction mixture was diluted with water (20 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was subjected to column chromatography (SiO2, EtOAc, R...) f Purification with 0.28 g yielded 4-amino-N,N-bis(2-hydroxyethyl)-3-methoxybenzenesulfonamide (0.8 g, 2.20 mmol, 73.55% yield) as a yellow oil.

[0653] Synthesis of N,N-bis(2-hydroxyethyl)-3-methoxy-4-(prop-2-yn-1-ylamino)benzenesulfonamide: A mixture of 4-amino-N,N-bis(2-hydroxyethyl)-3-methoxybenzenesulfonamide (0.1 g, 275.54 μmol, 1 equivalent), propyne bromide (49.17 mg, 413.32 μmol, 35.63 μL, 1.5 equivalent) in DMF (2 mL) was stirred at 50 °C for 12 h. Subsequent LC-MS analysis showed that the desired product was present in the mixture. The reaction mixture was diluted with water (20 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was analyzed by preparative TLC (SiO2, EtOAc:PE = 1:1, R f =0.31) Purification yielded N,N-bis(2-hydroxyethyl)-3-methoxy-4-(prop-2-yn-1-ylamino)benzenesulfonamide (0.05 g, 121.81 μmol, 44.21% yield), as a yellow oil.

[0654] Example A12: Preparation of 2-methoxy-4-((4-methylpiperazin-1-yl)sulfonyl)-N-(prop-2-yn-1-yl)aniline.

[0655]

[0656] Synthesis of 1-((4-fluoro-3-methoxyphenyl)sulfonyl)-4-methylpiperazine: TEA (315.32 mg, 3.12 mmol, 345.65 μL, 2 equivalents) was added to a solution of N-methylpiperazine (312.12 mg, 3.12 mmol, 345.65 μL, 2 equivalents) in DCM (2 mL). The resulting solution was then added dropwise to a solution of 4-fluoro-3-methoxybenzenesulfonyl chloride (350 mg, 1.56 mmol, 1 equivalent) in DCM (4 mL). The reaction mixture was heated to 25 °C with stirring for 2 hours, followed by TLC analysis (PE:EtOAc = 1:1, R f =0.40) indicates the reaction is complete. The reaction mixture was quenched at 25°C by adding water (60 mL) and extracted with EtOAc (20 mL x 4). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by column chromatography (SiO2, PE:EtOAc = 1:0 to 1:1) to give 1-((4-fluoro-3-methoxyphenyl)sulfonyl)-4-methylpiperazine (410 mg, 1.35 mmol, 86.70% yield) as a pale yellow solid. MS (ES) +,m / z):288.9.

[0657] Synthesis of 2-methoxy-4-((4-methylpiperazin-1-yl)sulfonyl)-N-(prop-2-yn-1-yl)aniline: In a sealed tube, a mixture of 1-((4-fluoro-3-methoxyphenyl)sulfonyl)-4-methylpiperazine (100 mg, 329.47 μmol, 1 equivalent), propyneamine (1.72 g, 31.23 mmol, 2 mL, 94.78 equivalent), K2CO3 (91.07 mg, 658.95 μmol, 2 equivalent) and KF (38.28 mg, 658.95 μmol, 15.44 μL, 2 equivalent) was stirred at 100 °C for 12 h. Subsequent TLC analysis (PE:EtOAc = 1:1, R f A new compound was detected (p = 0.23). The reaction mixture was quenched by adding water (40 mL) and extracted with EtOAc (10 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO2, PE:EtOAc = 1:1) in two runs to give 2-methoxy-4-((4-methylpiperazin-1-yl)sulfonyl)-N-(prop-2-yn-1-yl)aniline (25 mg, 69.57 μmol, 21.12% yield) as a pale yellow solid.

[0658] Example A13: Preparation of 2-(5-(methylsulfonyl)-2-(prop-2-yn-1-ylamino)phenoxy)acetamide.

[0659]

[0660] Synthesis of 5-(methylthio)-2-nitrophenol: NaSMe (66.93 g, 190.98 mmol, 60.85 mL, 6 equivalents) was added to a solution of 5-fluoro-2-nitrophenol (5 g, 31.83 mmol, 1 equivalent) in DMF (50 mL) at 0 °C. The mixture was heated to 50 °C for 5 hours, and subsequent HPLC and LC-MS analyses indicated the reaction was complete. The residue was decanted into a saturated aqueous solution of NH4Cl (300 mL), and the aqueous phase was extracted with EtOAc (200 mL x 3). The combined organic layers were washed with brine (200 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give 5-(methylthio)-2-nitrophenol (5.20 g, crude product) as a yellow solid.

[0661] Synthesis of 5-(methylsulfonyl)-2-nitrophenol: Potassium persulfate (8.30 g, 13.50 mmol, 2.50 equivalents) was added to a solution of 5-(methylthio)-2-nitrophenol (1 g, 5.40 mmol, 1 equivalent) in acetone (10 mL), water (10 mL), and MeOH (1 mL) at 0 °C. The mixture was heated to 20 °C and stirred for 5 hours. Subsequent LC-MS analysis indicated that the reaction was complete. The residue was decanted into a saturated aqueous solution of Na₂SO₃ (50 mL), and 12N HCl (20 mL) was added to adjust the pH of the solution to less than 7. The aqueous phase was extracted with EtOAc (50 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give 5-(methylsulfonyl)-2-nitrophenol (1.10 g, crude product) as a yellow solid.

[0662] Synthesis of 2-(5-(methanesulfonyl)-2-nitrophenoxy)acetamide: K₂CO₃ (953.65 mg, 6.90 mmol, 3 equivalents), 2-chloroacetamide (537.68 mg, 5.75 mmol, 2.50 equivalents), and KI (382.14 mg, 2.30 mmol, 1 equivalent) were added to a mixture of 5-(methanesulfonyl)-2-nitrophenol (500 mg, 2.30 mmol, 2.50 equivalents) and DMF (10 mL). The mixture was stirred at 50 °C for 2 hours. Subsequent HPLC analysis showed a reactant-to-product ratio of 1:1. The second portion of 2-chloroacetamide (215.07 mg, 2.30 mmol, 1 equivalent), K₂CO₃ (476.82 mg, 3.45 mmol, 1.50 equivalent), and KI (190.90 mg, 1.15 mmol, 0.50 equivalent) was added to the reaction mixture, and the resulting mixture was stirred at 50 °C for 2 hours. HPLC analysis showed that the reactant to product ratio was 1:5. The residue was poured into water (30 mL), and the aqueous phase was extracted with EtOAc (30 mL x 3). The combined organic layers were washed with brine (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give 2-(5-(methanesulfonyl)-2-nitrophenoxy)acetamide (380 mg, crude product) as a yellow solid.

[0663] Synthesis of 2-(2-amino-5-(methanesulfonyl)phenoxy)acetamide: NH4Cl (74.12 mg, 1.39 mmol, 48.44 μL, 1 equivalent) was added to a solution of 2-(5-(methanesulfonyl)-2-nitrophenoxy)acetamide (380 mg, 1.39 mmol, 1 equivalent) in EtOH (3 mL). The mixture was heated to 70 °C, and Fe (773.86 mg, 13.86 mmol, 10 equivalent) was added. The reaction mixture was stirred at 70 °C for 1 hour, and subsequent HPLC analysis indicated that the reaction was complete. The mixture was poured into water (50 mL), filtered through diatomaceous earth, and extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (50 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give 2-(2-amino-5-(methylsulfonyl)phenoxy)acetamide (270 mg, crude product), a dark brown solid.

[0664] Synthesis of 2-(5-(methanesulfonyl)-2-(prop-2-yn-1-ylamino)phenoxy)acetamide: K₂CO₃ (407.38 mg, 2.95 mmol, 3 equivalents) and 3-bromoprop-1-yne (584.40 mg, 4.91 mmol, 423.48 μL, 5 equivalents) were added to a solution of 2-(2-amino-5-(methanesulfonyl)phenoxy)acetamide (240 mg, 982.52 μmol, 1 equivalent) in DMF (8 mL). The mixture was stirred at 70 °C for 3 h. Subsequent HPLC analysis showed 29.5% of the starting primary amine remaining and 25.5% of the desired compound was detected. The percentage values ​​refer to peak area. The reaction mixture was decanted into water (20 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residues were analyzed by preparative TLC (DCM:MeOH = 10:1, R...). f Purification with 0.40 g yielded 2-(5-(methylsulfonyl)-2-(prop-2-yn-1-ylamino)phenoxy)acetamide (80 mg, 276.68 μmol, 28.16% yield), a pale red solid. MS (ES) + ,m / z):283.0.

[0665] Example A14: Preparation of N-(isoxazo-3-yl)-3-methoxy-4-(prop-2-yn-1-ylamino)benzenesulfonamide.

[0666]

[0667] Synthesis of N-(isoxazo-3-yl)-3-methoxy-4-nitrobenzenesulfonamide: Isoxazol-3-amine (801.86 mg, 9.54 mmol, 703.39 μL, 1.2 equivalents) was added to a solution of 3-methoxy-4-nitrobenzenesulfonyl chloride (2 g, 7.95 mmol, 1 equivalent) in pyridine (10 mL). The mixture was stirred at 20 °C for 2 h, followed by TLC analysis (EtOAc:DCM:PE:TEA = 1:1:3:0.5, R0). f(起始原料) =0.40,R f(产物) =0.04) indicates that the starting material was consumed. The reaction mixture was decanted into water (20 mL) and extracted with EtOAc (20 mL x 2, 10 mL x 1). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to N-(isoxazo-3-yl)-3-methoxy-4-nitrobenzenesulfonamide (6.5 g, 21.72 mmol, 91.10% yield) as a dark brown oil. MS (ES) + ,m / z):300.0.

[0668] Synthesis of N-(isoxazo-3-yl)-3-methoxy-4-nitro-N-((2-(trimethylsilyl)ethoxy)methyl)benzenesulfonamide: Under N2 and at 0 °C, NaH (534.60 mg, 13.37 mmol, 60% in mineral oil, 2 equivalents) was added to a solution of N-(isoxazo-3-yl)-3-methoxy-4-nitrobenzenesulfonamide (2 g, 6.68 mmol, 1 equivalent) in THF (20 mL). The mixture was stirred at 0 °C for 30 min, and (2-(chloromethoxy)ethyl)trimethylsilane (SEMCl) (1.67 g, 10.02 mmol, 1.77 mL, 1.5 equivalents) was added. The resulting mixture was stirred at 0 °C for 1 h, followed by TLC analysis (PE:EtOAc = 3:1, R f(起始原料) =0.60,R f(产物) =0.35) indicates the reaction is complete. The residue was poured into water (100 mL), and the aqueous phase was extracted with EtOAc (40 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (PE:EtOAc = 1:0 to 10:1) to give N-(isoxazo-3-yl)-3-methoxy-4-nitro-N-((2-(trimethylsilyl)ethoxy)methyl)benzenesulfonamide (1.8 g, 3.98 mmol, 59.57% yield) as a dark brown oil.

[0669] Synthesis of 4-amino-N-(isoxazo-3-yl)-3-methoxy-N-((2-(trimethylsilyl)ethoxy)methyl)benzenesulfonamide: A saturated NH4Cl solution (0.5 mL) was added to a solution of N-(isoxazo-3-yl)-3-methoxy-4-nitro-N-((2-(trimethylsilyl)ethoxy)methyl)benzenesulfonamide (1.8 g, 3.98 mmol, 1 equivalent) in EtOH (10 mL). The mixture was heated to 70 °C, and Fe (2.22 g, 39.81 mmol, 10 equivalent) was added. The mixture was further stirred at 70 °C for 2 hours. TLC analysis (PE:EtOAc = 3:1, R f =0.35) indicates the reaction is complete. The mixture was poured into a saturated NaHCO3 aqueous solution (100 mL), filtered with diatomaceous earth, and extracted with EtOAc (60 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (PE:EtOAc = 1:0 to 3:1) to give 4-amino-N-(isoxazo-3-yl)-3-methoxy-N-((2-(trimethylsilyl)ethoxy)methyl)benzenesulfonamide (1 g, 2.25 mmol, 56.58% yield) as a dark brown oil.

[0670] Synthesis of N-(isoxazo-3-yl)-3-methoxy-4-(prop-2-yn-1-ylamino)-N-((2-(trimethylsilyl)ethoxy)methyl)benzenesulfonamide: DIEA (655.04 mg, 5.07 mmol, 882.80 μL, 5 equivalents) was added to a mixture of 4-amino-N-(isoxazo-3-yl)-3-methoxy-N-((2-(trimethylsilyl)ethoxy)methyl)benzenesulfonamide (450 mg, 1.01 mmol, 1 equivalent) in CHCl3 (10 mL). The mixture was heated to 70 °C, and propargyl bromide (241.17 mg, 2.03 mmol, 174.76 μL, 2 equivalents) was added. The mixture was further stirred for 12 hours. LC-MS and HPLC analysis showed approximately 58% of the initial primary amine remaining, and 11% of the product was detected. Percentages refer to peak area. Another part of propargyl bromide (602.93 mg, 5.07 mmol, 436.91 μL, 5 equivalents) was added, and the mixture was further stirred at 70 °C for 6 hours. LC-MS and HPLC analysis showed approximately 15% of the initial primary amine residue and 45% of the desired product was detected, percentages referring to peak area. The mixture was decanted into water (30 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (PE:EtOAc = 1:0 to 10:1) to give N-(isoxazo-3-yl)-3-methoxy-4-(prop-2-yn-1-ylamino)-N-((2-(trimethylsilyl)ethoxy)methyl)benzenesulfonamide (340 mg, 568.29 μmol, 56% yield) as a yellow oil.

[0671] Synthesis of N-(isoxazo-3-yl)-3-methoxy-4-(prop-2-yn-1-ylamino)benzenesulfonamide: TFA (1 mL) was added to a solution of N-(isoxazo-3-yl)-3-methoxy-4-(prop-2-yn-1-ylamino)-N-((2-(trimethylsilyl)ethoxy)methyl)benzenesulfonamide (280 mg, 468.01 μmol, 1 equivalent) in DCM (4 mL). The mixture was stirred at 20 °C for 12 h, and subsequent LC-MS and HPLC analyses indicated the reaction was complete. The mixture was poured into a saturated aqueous solution of NaHCO3 (40 mL) and extracted with EtOAc (25 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (PE:EtOAc = 1:0 to 3:1) to give N-(isoxazo-3-yl)-3-methoxy-4-(prop-2-yn-1-ylamino)benzenesulfonamide (150 mg, 379.50 μmol, 81% yield) as a yellow solid.

[0672] Example A15: Preparation of N-(2-hydroxyethyl)-3-methoxy-N-methyl-4-(prop-2-yn-1-ylamino)benzenesulfonamide.

[0673]

[0674] Synthesis of N-(2-hydroxyethyl)-3-methoxy-N-methyl-4-nitrobenzenesulfonamide. TEA (5.03 g, 49.65 mmol, 6.91 mL, 5 equivalents) was added to a mixture of 3-methoxy-4-nitrobenzenesulfonyl chloride (2.5 g, 9.93 mmol, 1 equivalent) and 2-(methylamino)ethanol-1-ol (969.59 mg, 12.91 mmol, 1.04 mL, 1.3 equivalents) in DCM (25 mL) at 25 °C. The mixture was stirred at 25 °C for 12 hours, and subsequent LC-MS analysis indicated that the reaction was complete. The mixture was poured into water (100 mL) and extracted with DCM (80 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain N-(2-hydroxyethyl)-3-methoxy-N-methyl-4-nitrobenzenesulfonamide (2.1 g, crude product), a dark brown oil. MS(ES) + ,m / z):291.1.

[0675] Synthesis of 4-amino-N-(2-hydroxyethyl)-3-methoxy-N-methylbenzenesulfonamide: N-(2-hydroxyethyl)-3-methoxy-N-methyl-4-nitrobenzenesulfonamide (2.1 g, 7.23 mmol, 1 equivalent) was dissolved in AcOH (20 mL), and the mixture was heated to 70 °C. Fe (4.04 g, 72.34 mmol, 10 equivalent) was then added, and the mixture was further stirred at 70 °C for 2 hours. Subsequent LC-MS analysis indicated that the reaction was complete. The residue was decanted into a saturated NaHCO3 aqueous solution (500 mL), filtered with diatomaceous earth, and extracted with EtOAc (300 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography to give 4-amino-N-(2-hydroxyethyl)-3-methoxy-N-methylbenzenesulfonamide (1.8 g, 6.22 mmol, 86% yield) as a dark brown solid. MS (ES) + ,m / z):261.2.

[0676] Synthesis of N-(2-hydroxyethyl)-3-methoxy-N-methyl-4-(prop-2-yn-1-ylamino)benzenesulfonamide. DIPEA (1.12 g, 8.64 mmol, 1.51 mL, 5 equivalents) was added to a mixture of 4-amino-N-(2-hydroxyethyl)-3-methoxy-N-methylbenzenesulfonamide (500 mg, 1.73 mmol, 1 equivalent) in CHCl3 (5 mL). The mixture was heated to 70 °C, and propargyl bromide (1.03 g, 8.64 mmol, 745.10 μL, 5 equivalents) was added. The mixture was stirred at 70 °C for 12 h. Subsequent HPLC and LC-MS analysis showed 12.6% of the starting material remaining, 68.1% of the product detected, and 7.7% of byproducts detected (percentages refer to peak area). The mixture was decanted into water (60 mL) and extracted with EtOAc (60 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel), concentrated, dissolved in PE:EtOAc = 1:1 (20 mL), and heated to 70 °C. Additional EtOAc (5 mL) was added to dissolve any remaining solids, and the mixture was stirred further for 1 hour. The mixture was cooled to 25 °C, and the resulting solid precipitate was filtered. The mother liquor underwent two rounds of preparative HPLC, and the two fractions were then combined to give N-(2-hydroxyethyl)-3-methoxy-N-methyl-4-(prop-2-yn-1-ylamino)benzenesulfonamide (0.28 g, 50.0% yield) as a yellow solid. MS (ES) + ,m / z):299.1.

[0677] =Example A16: Preparation of 3-methoxy-N-(5-methylisoxazol-3-yl)-4-(prop-2-yn-1-ylamino)benzenesulfonamide.

[0678]

[0679] 3-Methoxy-N-(5-methylisoxazol-3-yl)-4-(prop-2-yn-1-ylamino)benzenesulfonamide was prepared by a procedure similar to that used in the synthesis of N-(isoxazol-3-yl)-3-methoxy-4-(prop-2-yn-1-ylamino)benzenesulfonamide according to Example A14, but using 5-methylisoxazol-3-amine instead of isoxazol-3-amine.

[0680] Example A17: Preparation of 2-(2-(dimethylamino)ethoxy)-4-(methanesulfonyl)-N-(prop-2-yn-1-yl)aniline hydrochloride.

[0681]

[0682] Synthesis of N,N-dimethyl-2-(5-(methanesulfonyl)-2-nitrophenoxy)ethyl-1-amine: KI (29.96 mg, 180.48 μmol, 0.196 equivalents), 2-chloro-N,N-dimethyl-ethylamine (213.54 mg, 1.48 mmol, 1.61 equivalents, HCl) and Cs₂CO₃ (738.05 mg, 2.27 mmol, 2.46 equivalents) were added to a mixture of 5-(methanesulfonyl)-2-nitrophenol (prepared according to the first two steps of Example A13) in THF (10 mL). The mixture was stirred at 70 °C for 16 hours, and subsequent HPLC analysis showed a reactant:product ratio of 4:1. The mixture was poured into water (40 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give N,N-dimethyl-2-(5-(methanesulfonyl)-2-nitrophenoxy)ethyl-1-amine (300 mg, crude) as a yellow oil. MS (ES) + ,m / z):288.9.

[0683] Synthesis of 2-(2-(dimethylamino)ethoxy)-4-(methanesulfonyl)aniline: Pd / C (50 mg, 227.18 mmol, 15% purity, 131 equivalents) was added to a solution of N,N-dimethyl-2-(5-(methanesulfonyl)-2-nitrophenoxy)ethyl-1-amine (0.5 g, 1.73 mmol, 1 equivalent) in MeOH (50 mL). The mixture was degassed and purged with H2 (349.59 μg, 173.42 μmol, 2.33 e⁻² μL), and stirred at 20 °C under H2 (50 psi) for 12 h. Subsequent LC-MS analysis indicated that the reaction was complete. The mixture was poured into MeOH (100 mL), filtered through diatomaceous earth, and concentrated to give 2-(2-(dimethylamino)ethoxy)-4-(methylsulfonyl)aniline (450 mg, crude product), as a dark brown oil. MS (ES) + ,m / z):259.1.

[0684] Synthesis of tert-butyl (tert-butyl)carbamate (2-(2-(dimethylamino)ethoxy)-4-(methanesulfonyl)phenyl)carbamate: Boc₂O (1.01 g, 4.65 mmol, 1.07 mL, 6 equivalents) and DMAP (94.58 mg, 774.18 μmol, 1 equivalent) were added to a solution of 2-(2-(dimethylamino)ethoxy)-4-(methanesulfonyl)aniline (200 mg, 774.18 μmol, 1 equivalent) in dioxane (7 mL). The reaction was then stirred at 110 °C for 16 h, and subsequent LC-MS analysis indicated that the reaction was complete. The mixture was poured into water (50 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with brine (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give tert-butyl (tert-butyloxycarbonyl)(2-(2-(dimethylamino)ethoxy)-4-(methanesulfonyl)phenyl)carbamate (550 mg, crude), as a dark brown oil. MS (ES) + ,m / z):459.1.

[0685] Synthesis of tert-butyl (2-(2-(dimethylamino)ethoxy)-4-(methanesulfonyl)phenyl)carbamate: K₂CO₃ (497.29 mg, 3.60 mmol, 3 equivalents) was added to a solution of (tert-butoxycarbonyl)(2-(2-(dimethylamino)ethoxy)-4-(methanesulfonyl)phenyl)carbamate (550 mg, 1.20 mmol, 1 equivalent) in MeOH (10 mL). The resulting mixture was stirred at 40 °C for 4 hours, and subsequent LC-MS analysis indicated that the reaction was complete. The mixture was concentrated under vacuum, diluted with EtOAc (30 mL) and water (30 mL), and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with brine (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by column chromatography (SiO2, DCM to DCM:MeOH = 10:1) to give tert-butyl (2-(2-(dimethylamino)ethoxy)-4-(methylsulfonyl)phenyl)carbamate (210 mg, 535.18 μmol, 44.6% yield) as a yellow oil. MS (ES) + ,m / z):359.1.

[0686] Synthesis of tert-butyl (2-(2-(dimethylamino)ethoxy)-4-(methanesulfonyl)phenyl)(prop-2-yn-1-yl)carbamate: At 0 °C, NaH (10.19 mg, 254.85 μmol, 60% in mineral oil, 2 equivalents) was added to a mixture of tert-butyl (2-(2-(dimethylamino)ethoxy)-4-(methanesulfonyl)phenyl)carbamate (50 mg, 127.42 μmol, 1 equivalent) in 1.5 mL of DMF. The mixture was stirred at 0 °C for 0.5 h, and then a solution of propargyl bromide (22.74 mg, 191.13 μmol, 16.48 μL, 1.5 equivalents) in 0.5 mL of DMF was added dropwise. The mixture was further stirred at 0 °C for 1 h, and then analyzed by TLC (DCM:MeOH = 20:1, R f(起始原料) =0.23,R f(产物) =0.17) New spots were observed. The mixture was poured into a saturated NH4Cl aqueous solution (10 mL), EtOAc (10 mL) was added, and the resulting mixture was extracted with EtOAc (10 mL x 3). The combined organic layers were washed with brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by preparative TLC to give tert-butyl (2-(2-(dimethylamino)ethoxy)-4-(methanesulfonyl)phenyl)(prop-2-yn-1-yl)carbamate (35 mg, 80.26 μmol, 31.5% yield) as a yellow oil. MS (ES) + ,m / z):397.4.

[0687] Synthesis of 2-(2-(dimethylamino)ethoxy)-4-(methanesulfonyl)-N-(prop-2-yn-1-yl)aniline hydrochloride: A solution of tert-butyl (2-(2-(dimethylamino)ethoxy)-4-(methanesulfonyl)phenyl)(prop-2-yn-1-yl)carbamate (35 mg, 80.26 μmol, 1 equivalent) in HCl / EtOAc (4 M, 20.06 μL) was stirred at 15 °C for 1 hour. Subsequent HPLC and LC-MS analysis indicated that the reaction was complete. The mixture was concentrated under vacuum to give 2-(2-(dimethylamino)ethoxy)-4-(methanesulfonyl)-N-(prop-2-yn-1-yl)aniline hydrochloride (30 mg, crude product) as a yellow oil. MS (ES) + ,m / z):296.9.

[0688] Example A18: Preparation of 2-methoxy-4-(morpholinosulfonyl)-N-(prop-2-yn-1-yl)aniline.

[0689]

[0690] 2-Methoxy-4-(morpholinosulfonyl)-N-(prop-2-yn-1-yl)aniline was prepared by a procedure similar to that used in the synthesis of N-(2-hydroxyethyl)-3-methoxy-N-methyl-4-(prop-2-yn-1-ylamino)benzenesulfonamide according to Example A15, using morpholine instead of 2-(methylamino)ethanol. MS(ES) + ,m / z):311.1.

[0691] Example A19: Preparation of 1-(4-((3-methoxy-4-(prop-2-yn-1-ylamino)phenyl)sulfonyl)piperazin-1-yl)ethyl-1-one.

[0692]

[0693] 1-(4-((3-methoxy-4-(prop-2-yn-1-ylamino)phenyl)sulfonyl)piperazin-1-yl)ethyl-1-one was prepared by a procedure similar to that used in the synthesis of N-(2-hydroxyethyl)-3-methoxy-N-methyl-4-(prop-2-yn-1-ylamino)benzenesulfonamide according to Example A15, using N-acetylpiperazine instead of 2-(methylamino)ethyl-1-ol.

[0694] Example A20: Preparation of N-(2,3-dihydroxypropyl)-3-methoxy-4-(prop-2-yn-1-ylamino)benzenesulfonamide.

[0695]

[0696] N-(2,3-dihydroxypropyl)-3-methoxy-4-(prop-2-yn-1-ylamino)benzenesulfonamide was prepared by a procedure similar to that used in the synthesis of N-(2-hydroxyethyl)-3-methoxy-N-methyl-4-(prop-2-yn-1-ylamino)benzenesulfonamide according to Example A15, using (rac)-3-aminopropane-1,2-diol instead of 2-(methylamino)ethanol.

[0697] Example A21: Preparation of 2-(fluoromethoxy)-4-(methanesulfonyl)-N-(prop-2-yn-1-yl)aniline.

[0698]

[0699] Under N2, K2CO3 (662.61 mg, 4.79 mmol, 3 equivalents) and bromo(fluoro)methane (360.95 mg, 3.20 mmol, 2 equivalents) were added in one step to a solution of 5-(methanesulfonyl)-2-(prop-2-yn-1-ylamino)phenol (400 mg, 1.60 mmol, 1 equivalent) in DMF (8 mL). The mixture was stirred at 40 °C for 60 min, and TLC and LC-MS analysis indicated that the reaction was complete. The reaction mixture was diluted with water (30 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with water (30 mL x 3) and brine (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The crude residue was purified by column chromatography (SiO2, PE:EtOAc = 1.5:1 to 1:1) to give the desired product (320 mg, 1.24 mmol, 77.83% yield) as a pink solid. MS (ES) + ,m / z):258.0.

[0700] Example A22: Preparation of methyl 2-(5-(methylsulfonyl)-2-(prop-2-yn-1-ylamino)phenoxy)acetate

[0701]

[0702] A solution of 5-(methylsulfonyl)-2-(prop-2-yn-1-ylamino)phenol (0.3 g, 1.3 mmol, 1 equivalent) in acetonitrile (5 mL) was added with K₂CO₃ (552.18 mg, 4 mmol, 3 equivalents) and methyl 2-bromoacetate (1.5 equivalents). The mixture was then stirred at 40 °C under N₂ for 0.5 h. TLC analysis indicated that the reaction was complete. The reaction was quenched with water (20 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by preparative TLC (SiO₂, DCM:MeOH = 1:0) to give the desired product as a yellow solid. 75.8% yield, MS (ES) + ,m / z):298.1.

[0703] Example A23: Preparation of 6-(methanesulfonyl)-3-(prop-2-yn-1-yl)benzo[d]oxazol-2(3H)-one.

[0704]

[0705] CDI (777.40 mg, 4.79 mmol, 1.2 equivalents) was added to a solution of 5-(methanesulfonyl)-2-(prop-2-yn-1-ylamino)phenol (0.9 g, 4 mmol, 1 equivalent) in DMF (9 mL) at 25 °C. The mixture was stirred at 25 °C for 1 hour, and then at 80 °C for 1 hour. LC-MS analysis showed complete consumption of the starting material and a main peak with the desired product mass was detected. The mixture was decanted into water (50 mL) and extracted with DCM (20 mL x 3). The combined organic layers were washed with water (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the residue. The crude residue was lyophilized to give the desired product (1 g, crude) as a yellow solid. MS (ES) + ,m / z):252.0.

[0706] Example A24: Synthesis of (3R,4R)-3-methoxy-N-(prop-2-yn-1-yl)tetrahydro-2H-pyran-4-amine.

[0707]

[0708] K₂CO₃ (0.5 g, 3.62 mmol, 6 equivalents) and 3-bromoprop-1-yne (56.77 mg, 477.23 μmol, 41.14 μL, 0.8 equivalents) were added to a solution of (3R)-3-methoxytetrahydropyran-4-amine (0.1 g, 596.54 μmol, 1 equivalent, HCl) in CH₃CN (2 mL). The mixture was stirred at 25 °C for 1 h. LC-MS analysis indicated that the reaction was complete. The reaction was concentrated under reduced pressure and purified by preparative TLC (SiO₂, DCM:MeOH = 20:1) to give the desired product (0.029 g, 145.67 μmol, 24.4% yield) as a yellow oil. MS (ES) + ,m / z):170.2.

[0709] Example A25: Synthesis of 2-fluoro-5-methoxy-4-(prop-2-yn-1-ylamino)benzoic acid.

[0710]

[0711] Preparation of 2-fluoro-5-methoxy-4-nitrobenzoic acid: A solution of KOH (4.14 g, 73.86 mmol, 3 equivalents) in MeOH (20 mL) was added dropwise to a solution of 2,5-difluoro-4-nitrobenzoic acid (5 g, 24.62 mmol, 1 equivalent) in MeOH (60 mL). The mixture was heated under reflux for 2 hours (oil bath temperature: 80 °C). The resulting mixture was stirred at 80 °C for 2 hours. LC-MS analysis indicated that the reaction was complete. 2N HCl was added to the reaction mixture at 20 °C to adjust the pH of the mixture to 2. The mixture was then concentrated to remove MeOH. The residue was extracted with water (100 mL) and EtOAc (100 mL x 3). The organic layer was washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product (5.2 g, crude product) as a yellow solid.

[0712] Preparation of methyl 2-fluoro-5-methoxy-4-nitrobenzoate: A solution of 2-fluoro-5-methoxy-4-nitrobenzoic acid (0.3 g, 1.39 mmol, 1 equivalent) in HCl / MeOH (10 mL) was stirred at 25 °C for 3 hours until a yellow solid was formed. LC-MS analysis indicated that the reaction was complete. The reaction was concentrated under reduced pressure to give the desired product (0.3 g, 1.24 mmol, 89.2% yield) as a yellow solid.

[0713] Preparation of methyl 4-amino-2-fluoro-5-methoxybenzoate: Fe (347.26 mg, 6.22 mmol, 5 equivalents) was added to a mixture of methyl 2-fluoro-5-methoxy-4-nitrobenzoate (0.3 g, 1.24 mmol, 1 equivalent) in EtOH (3 mL) and saturated NH4Cl aqueous solution (1 mL) at 90 °C. The mixture was stirred at 90 °C for 1 hour. TLC analysis showed that the reaction was complete. The mixture was extracted with EtOAc (20 mL x 3). The organic layer was washed with brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO2, PE:EtOAc = 3:1) to give the desired product (0.22 g, 994.08 μmol, 79.93% yield) as an orange solid.

[0714] Preparation of methyl 4-((tert-Butoxycarbonyl)amino)-2-fluoro-5-methoxybenzoate: A solution of methyl 4-amino-2-fluoro-5-methoxybenzoate (200 mg, 903.71 μmol, 1 equivalent) in di-tert-butyl dicarbonate (4.75 g, 21.76 mmol, 5 mL, 24.08 equivalent) was stirred for 6 hours at 110 °C. LC-MS analysis showed some residual starting material. The reaction mixture was concentrated under reduced pressure and purified by preparative TLC (SiO2, PE:EtOAc = 4:1) to give the desired product (0.23 g, 691.63 μmol, 76.53% yield) as a white solid. MS (ES) + ,m / z):300.2.

[0715] Preparation of methyl 4-((tert-butoxycarbonyl)(prop-2-yn-1-yl)amino)-2-fluoro-5-methoxybenzoate: Cs₂CO₃ (587.86 mg, 1.80 mmol, 3 equivalents) and 3-bromoprop-1-yne (143.09 mg, 1.20 mmol, 103.69 μL, 2 equivalents) were added to a solution of methyl 4-((tert-butoxycarbonyl)amino)-2-fluoro-5-methoxybenzoate (0.2 g, 601.42 μmol, 1 equivalent) in DMF (4 mL). The reaction mixture was stirred at 40 °C for 1 hour. LC-MS analysis indicated that the reaction was complete. The reaction mixture was poured into EtOAc (15 mL) and extracted with EtOAc (15 mL x 3). The combined organic layers were washed with brine (15 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO2, PE:EtOAc = 5:1) to obtain the desired product (0.18 g, 480.22 μmol, 79.85% yield), which was a white oil.

[0716] Preparation of 4-((tert-Butoxycarbonyl)(prop-2-yn-1-yl)amino)-2-fluoro-5-methoxybenzoic acid: Lithium hydroxide hydrate (53.74 mg, 1.28 mmol, 3 equivalents) was added to a solution of methyl 4-((tert-Butoxycarbonyl)(prop-2-yn-1-yl)amino)-2-fluoro-5-methoxybenzoate (0.16 g, 426.87 μmol, 1 equivalent) in THF (1 mL), MeOH (1 mL), and water (1 mL). The mixture was stirred at 25 °C for 1 h. TLC analysis indicated that the reaction was complete. 1 M HCl was added to adjust the pH of the reaction mixture to 2. The mixture was extracted with EtOAc (20 mL x 3). The organic layer was washed with brine (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give crude product (0.14 g, crude product) as a white solid. The crude product was used without purification.

[0717] Preparation of 2-fluoro-5-methoxy-4-(prop-2-yn-1-ylamino)benzoic acid: A solution of 4-((tert-butoxycarbonyl)(prop-2-yn-1-yl)amino)-2-fluoro-5-methoxybenzoic acid (0.15 g, 463.94 μmol, 1 equivalent) in 4N HCl / EtOAc (6 mL, 51.73 equivalents) was stirred at 25 °C for 1 hour. LC-MS analysis indicated that the reaction was complete. The reaction was concentrated under reduced pressure to give a crude product (0.1 g, crude product, HCl) as a yellow solid. The crude product was used without purification. MS (ES) + ,m / z):222.0.

[0718] Example A26: Preparation of (3S)-3-methoxy-N-(prop-2-yn-1-yl)tetrahydro-2H-pyran-4-amine.

[0719]

[0720] K₂CO₃ (1.58 g, 11.44 mmol, 3 equivalents) was added to a solution of (3S)-3-methoxytetrahydro-2H-pyran-4-amine (0.5 g, 3.81 mmol, 1 equivalent) in CH₃CN (8 mL). The mixture was stirred at 25 °C, and 3-bromoprop-1-yne (362.76 mg, 3.05 mmol, 262.87 μL, 0.8 equivalents) was added to the solution. The resulting reaction mixture was stirred at 25 °C for 3 hours. TLC analysis indicated that the reaction was complete, with some starting material remaining. The reaction mixture was filtered and concentrated under reduced pressure. The residue was purified by column chromatography (SiO₂, PE:EtOAc = 10:1 to 5:1) to give the desired product (0.33 g, 1.76 mmol, 46.04% yield) as a yellow oil. MS (ES) + ,m / z):170.1.

[0721] Example A27: Synthesis of 2-(COOR)-4-(methylsulfonyl)-N-(prop-2-yn-1-yl)aniline.

[0722]

[0723] A solution of 5-(methanesulfonyl)-2-(prop-2-yn-1-ylamino)phenol (1 equivalent) in DCM (5 mL) was added with pyridine (1 equivalent) and R-anhydride (1 equivalent). The mixture was stirred at 25 °C for 2 hours. LC-MS analysis showed the desired product. The reaction mixture was quenched by adding water (100 mL) at 0 °C and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was analyzed by preparative TLC (SiO2, PE:EtOAc = PE:EtOAc = 1:1, R-anhydride). f =0.5) Purification yields the desired product.

[0724] Example A28: Synthesis of 4-(methylsulfonyl)-N-(prop-2-yn-1-yl)-2-(trifluoromethoxy)aniline.

[0725]

[0726] Preparation of 4-(methylsulfonyl)-2-(trifluoromethoxy)aniline: L-proline (1.12 g, 9.76 mmol, 0.5 equivalence) and CuI (1.49 g, 7.81 mmol, 0.4 equivalence) were added to a mixture of 4-bromo-2-(trifluoromethoxy)aniline (5 g, 19.53 mmol, 2.96 mL, 1 equivalent) and sodium methyl sulfate (5.98 g, 58.59 mmol, 3 equivalent) in DMSO (50 mL). The reaction mixture was stirred at 100 °C for 16 h under N2. TLC analysis showed some starting material residue. The mixture was stirred at 20 °C for 1 h and extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (50 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, PE:EtOAc = 10:1 to 1:1) to give the desired product (2.6 g, 10.19 mmol, 52.16% yield) as a white solid.

[0727] Preparation of tert-butyl (4-(methanesulfonyl)-2-(trifluoromethoxy)phenyl)carbamate: Boc₂O (461.78 mg, 2.12 mmol, 486.09 μL, 1.2 equivalents) and DMAP (258.49 mg, 2.12 mmol, 1.2 equivalents) were added to a solution of tert-butyl (4-(methanesulfonyl)-2-(trifluoromethoxy)phenyl)carbamate (500 mg, 1.76 mmol, 1 equivalent) in THF (10 mL). The reaction mixture was stirred at 70 °C for 1 hour. TLC and LC-MS analysis indicated that the reaction was complete. The reaction mixture was diluted with water (20 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was purified by column chromatography (SiO2, PE:EtOAc = 10:1 to 1:1) to give tert-butyl 4-(methanesulfonyl)-2-(trifluoromethoxy)phenyl)carbamate (900 mg, 1.98 mmol, 112.07% yield) and N,N-di(tert-butoxycarbonyl)-4-(methanesulfonyl)-2-(trifluoromethoxy)aniline (900 mg, 2.53 mmol, 143.65% yield) as white solids.

[0728] A mixture of tert-butyl (4-(methylsulfonyl)-2-(trifluoromethoxy)phenyl)carbamate (700 mg, 1.54 mmol, 1 equivalent), N,N-di(tert-butoxycarbonyl)-4-(methylsulfonyl)-2-(trifluoromethoxy)aniline (700 mg, 1.97 mmol, 1.28 equivalent) and K₂CO₃ (637.25 mg, 4.61 mmol, 3 equivalent) in MeOH (18 mL) was stirred at 40 °C for 2 hours. LC-MS analysis indicated that the reaction was complete. The reaction mixture was filtered and concentrated under reduced pressure to give a crude product (1 g) as a pale yellow solid.

[0729] Preparation of tert-butyl (4-(methylsulfonyl)-2-(trifluoromethoxy)phenyl)(prop-2-yn-1-yl)carbamate: Cs₂CO₃ (2.20 g, 6.75 mmol, 3 equivalents) and 3-bromoprop-1-yne (803.49 mg, 6.75 mmol, 582.24 μL, 3 equivalents) were added to a solution of tert-butyl (4-(methylsulfonyl)-2-(trifluoromethoxy)phenyl)carbamate (0.8 g, 2.25 mmol, 1 equivalent) in DMF (20 mL) at 25 °C. The mixture was stirred for 1 hour. TLC and LC-MS indicated the reaction was complete. The reaction mixture was diluted with water (20 mL) and extracted with EtOAc (10 mL x 3). The combined organic layers were washed with brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The crude residue was purified by column chromatography (SiO2, DCM:MeOH = 10:1) to obtain the desired product (0.65 g, 1.49 mmol, 66.05% yield) as a yellow solid.

[0730] Preparation of 4-(methanesulfonyl)-N-(prop-2-yn-1-yl)-2-(trifluoromethoxy)aniline: A solution of tert-butyl (4-(methanesulfonyl)-2-(trifluoromethoxy)phenyl)(prop-2-yn-1-yl)carbamate (650 mg, 1.49 mmol, 1 equivalent) in HCl / EtOAc (4 M, 13.50 mL, 36.31 equivalents) was stirred at 25 °C for 0.5 h. LC-MS analysis indicated that the reaction was complete. The reaction mixture was diluted with EtOAc (10 mL) and concentrated under vacuum. The desired product (340 mg, crude product, HCl) was obtained as a yellow solid. MS (ES) + ,m / z):291.9.

[0731] Example A29: Synthesis of 2-methyl-4-(methanesulfonyl)-N-(prop-2-yn-1-yl)aniline

[0732]

[0733] K₂CO₃ (2.24 g, 16.19 mmol, 3 equivalents) and 3-bromoprop-1-yne (642.18 mg, 5.40 mmol, 465.35 μL, 1 equivalent) were added to a solution of 2-methyl-4-(methanesulfonyl)aniline (1 g, 5.40 mmol, 1 equivalent) in DMF (10 mL) at 70 °C. The mixture was stirred at 70 °C for 12 h. TLC analysis showed some starting material residue. The reaction mixture was decanted into water (20 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by column chromatography (SiO₂, PE:EtOAc = 10:1 to 1:1) to give the desired product (0.5 g, 2.02 mmol, 37.33% yield) as a yellow solid. MS (ES) + ,m / z):224.1.

[0734] Example A30: Synthesis of 2-fluoro-5-methoxy-N-methyl-4-(prop-2-yn-1-ylamino)benzamide

[0735]

[0736] Preparation of 2-fluoro-5-methoxy-4-nitrobenzoic acid: A solution of 2,5-difluoro-4-nitrobenzoic acid (4 g, 19.69 mmol, 1 equivalent) in MeOH (64 mL) was added dropwise to a solution of KOH (3.31 g, 59.08 mmol, 3 equivalents). The resulting mixture was stirred at 80 °C for 2 hours. HPLC analysis indicated that the reaction was complete. 2NHCl was added to the solution at 20 °C to adjust the pH of the mixture to 2. The mixture was concentrated to remove MeOH, and the residue was extracted with water (30 mL) and EtOAc (40 mL x 3). The organic layer was washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product (4 g, 18.59 mmol, 94.41% yield) was given as a yellow solid, which was used without purification.

[0737] Preparation of 4-amino-2-fluoro-5-methoxybenzoic acid: A mixture of 2-fluoro-5-methoxy-4-nitrobenzoic acid (4 g, 18.59 mmol, 1 equivalent) and Pd / C (2 g, 1.88 mmol, 10% purity, 1.01e-1 equivalent) in MeOH (50 mL) was degassed, purged three times with N2, and stirred at 20 °C for 5 h under H2 (15 Psi). LC-MS and HPLC analysis indicated that the reaction was complete. The mixture was filtered through silica gel, and the filtrate was concentrated. A crude residue (3.5 g, 17.01 mmol, 91.50% yield) was given as a yellow solid, which was used without purification. MS (ES) + ,m / z):184.2.

[0738] Preparation of 4-amino-2-fluoro-5-methoxy-N-methylbenzamide: A mixture of 4-amino-2-fluoro-5-methoxybenzoic acid (2 g, 10.80 mmol, 1 equivalent), methylamine hydrochloride (1.46 g, 21.60 mmol, 2 equivalents), HOBt (2.19 g, 16.20 mmol, 1.5 equivalents), EDCI (3.11 g, 16.20 mmol, 1.5 equivalents), and TEA (4.37 g, 43.21 mmol, 6 mL, 4 equivalents) in DCM (30 mL) was degassed and purged three times with N2. The mixture was stirred at 20 °C under N2 for 2 hours. LC-MS analysis indicated that the reaction was complete. The mixture was extracted with water (30 mL) and DCM (50 mL x 5). The organic layer was washed with brine (40 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (SiO2, PE:EtOAc = 3:1) to give the desired product (1.1 g, 5 mmol, 46.24% yield) as a white solid. MS (ES) + ,m / z):199.1.

[0739] Preparation of 2-fluoro-5-methoxy-N-methyl-4-(prop-2-yn-1-ylamino)benzamide: A mixture of 4-amino-2-fluoro-5-methoxy-N-methylbenzamide (0.7 g, 3.18 mmol, 1 equivalent), 3-bromoprop-1-yne (2.27 g, 19.07 mmol, 1.64 mL, 6 equivalents), and K₂CO₃ (1.32 g, 9.54 mmol, 3 equivalents) in DMF (10 mL) was degassed and purged three times with N₂. The mixture was stirred at 105 °C for 12 h under N₂. TLC analysis showed that the starting material was consumed. The reaction mixture was extracted with water (60 mL) and EtOAc (40 mL x 3). The organic layer was washed with brine (15 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by column chromatography (SiO2, PE:EtOAc = 5:1 to 3:1) to obtain the desired product (0.6 g, 1.78 mmol, 55.93% yield) as a yellow solid.

[0740] Example A31: Synthesis of 2-amino-N-((3-methoxy-4-(prop-2-yn-1-ylamino)phenyl)sulfonyl)acetamide.

[0741]

[0742] Preparation of tert-butyl (2-((3-methoxy-4-(prop-2-yn-1-ylamino)phenyl)sulfinylamino)-2-oxoethyl)carbamate: HATU (949.47 mg, 2.50 mmol, 2 equivalents) and TEA (252.68 mg, 2.50 mmol, 347.57 μL, 2 equivalents) were added to a solution of (tert-butoxycarbonyl)glycine (437.45 mg, 2.50 mmol, 70.16 μL, 2 equivalents) in DCM (6 mL). The mixture was stirred at 25 °C for 0.5 h. Then, 3-methoxy-4-(prop-2-yn-1-ylamino)benzenesulfonamide (300 mg, 1.25 mmol, 1 equivalent) was added to the reaction mixture, and the mixture was stirred at 25 °C for 2 h. TLC analysis showed 40% of the starting material remaining. The second part of 3-methoxy-4-(prop-2-yn-1-ylamino)benzenesulfonamide (437.45 mg, 2.50 mmol, 2 equivalents), HATU (949.47 mg, 2.50 mmol, 2 equivalents), and TEA (252.68 mg, 2.50 mmol, 347.57 μL, 2 equivalents) were added to the reaction mixture, and the mixture was further stirred at 25 °C for 10 hours. TLC analysis showed that the starting material was consumed. The mixture was poured into water (10 mL), and the aqueous phase was extracted with DCM (10 mL x 3). The combined organic layers were washed with brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (PE:EtOAc = 1:1 to 1:2) to give the desired product (560 mg, 845.40 μmol, 67.71% yield) as a colorless oil.

[0743] Preparation of 2-amino-N-((3-methoxy-4-(prop-2-yn-1-ylamino)phenyl)sulfonyl)acetamide: tert-butyl (2-((3-methoxy-4-(prop-2-yn-1-ylamino)phenyl)sulfonylamino)-2-oxoethyl)carbamate (490 mg, 739.72 μmol, 1 equivalent) was dissolved in 4N HCl in EtOAc (5 mL), and the solution was stirred at 25 °C for 1 hour. LC-MS analysis indicated that the reaction was complete. The residue was concentrated under vacuum to give the crude product (350 mg, crude product), a white solid. MS (ES) + ,m / z):298.1.

[0744] Example A32: Synthesis of 4-methoxy-N-prop-2-ynyl-pyridine-3-amine.

[0745]

[0746] Preparation of N-tert-butoxycarbonyl-N-(4-methoxy-3-pyridyl)carbamate tert-butyl ester: LiHMDS (1M, 399.55μL, 2.48 equivalents) was added to a solution of 4-methoxypyridin-3-amine (810 mg, 6.52 mmol, 1 equivalent) in THF (25 mL). The solution was purged three times with N2, and the mixture was stirred at 0 °C for 30 min under N2. Then, Boc2O (2.85 g, 13.04 mmol, 3 mL, 2 equivalents) was added to the reaction mixture, and the mixture was stirred at 0 °C for 2 h under N2. TLC analysis showed that the starting material was partially consumed and a spot of the desired product was detected. The reaction mixture was poured into a saturated NH4Cl solution (100 mL) and extracted with EtOAc (50 mL x 1, then 25 mL x 2). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain N-tert-butoxycarbonyl-N-(4-methoxy-3-pyridyl)carbamate tert-butyl ester (1.5 g, crude product), which was a yellow oil.

[0747] Preparation of N-(4-methoxy-3-pyridyl)carbamate tert-butyl ester: A mixture of N-tert-butoxycarbonyl-N-(4-methoxy-3-pyridyl)carbamate tert-butyl ester (1.50 g, 4.62 mmol, 1 equivalent) and K₂CO₃ (639.13 mg, 4.62 mmol, 1 equivalent) in MeOH (2 mL) was degassed and purged three times with N₂. The mixture was stirred at 50 °C under N₂ for 16 h. TLC analysis showed that the starting material was consumed and a spot of the desired product was observed. The reaction mixture was poured into water (100 mL) and extracted with EtOAc (50 mL x 1, then 25 mL x 2). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by column chromatography to obtain N-(4-methoxy-3-pyridyl)carbamate tert-butyl ester (1 g, 4.01 mmol, 86.87% yield), which was a yellow oil.

[0748] Preparation of N-(4-methoxy-3-pyridyl)-N-prop-2-ynyl-tert-butyl carbamate: A mixture of N-(4-methoxy-3-pyridyl)-tert-butyl carbamate (500 mg, 2.23 mmol, 1 equivalent) and NaH (160.56 mg, 6.69 mmol, 3 equivalent, 60% in mineral oil) in THF (25 mL) was stirred at 0 °C for 1 h under N2. Then, 3-bromoprop-1-yne (530.46 mg, 4.46 mmol, 384.39 μL, 2 equivalent) was added, and the resulting mixture was stirred at 0 °C for 1 h under N2. TLC analysis showed that the starting material was consumed and a new spot of the desired product was observed. The reaction mixture was poured into water (100 mL) and extracted with EtOAc (50 mL x 1, then 25 mL x 2). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by column chromatography (PE:EtOAc = 2:1) to give N-(4-methoxy-3-pyridyl)-N-prop-2-ynyl-carbamate tert-butyl ester (200 mg, 762.49 μmol, 34.19% yield) as a yellow oil.

[0749] Preparation of 4-methoxy-N-prop-2-ynyl-pyridine-3-amine: HCl / EtOAc (4M, 190.62 μL, 1 equivalent) was added to a solution of N-(4-methoxy-3-pyridyl)-N-prop-2-ynyl-carbamate tert-butyl ester (200 mg, 762.49 μmol, 1 equivalent) in EtOH (10 mL). The solution was purged three times with N2 and stirred at 25 °C for 2 h under N2. TLC analysis showed that the starting material was consumed and a spot of the desired product was observed. The mixture was concentrated under reduced pressure to give 4-methoxy-N-prop-2-ynyl-pyridine-3-amine (150 mg, 755.10 μmol, 99.03% yield, HCl) as a yellow solid. The desired product was used without further purification.

[0750] Example A33: Synthesis of 2-methoxy-N-prop-2-ynyl-4-(trifluoromethyl)aniline.

[0751]

[0752] Synthesis of 2-methoxy-1-nitro-4-(trifluoromethyl)benzene: At 80 °C, a solution of KOH (18.51 g, 329.99 mmol, 3 equivalents) in MeOH (100 mL) was added to a solution of 2-fluoro-1-nitro-4-(trifluoromethyl)benzene (23 g, 110 mmol, 1 equivalent) in MeOH (350 mL). The resulting mixture was stirred at 80 °C for 2 hours. TLC analysis (R) was performed. f(产物)=0.6, PE:EtOAc = 5:1) indicates that the starting material was consumed and a new spot was formed. 2N HCl was added to the reaction mixture to adjust the pH of the mixture to 2. The solution was then concentrated. The crude residue was washed with water (150 mL) and extracted with EtOAc (300 mL x 2). The organic layer was washed with brine (15 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give 2-methoxy-1-nitro-4-(trifluoromethyl)benzene (22.5 g, 91.57 mmol, 83.25% yield) as a yellow solid. The crude residue was used directly without any purification.

[0753] Synthesis of 2-methoxy-4-(trifluoromethyl)aniline: Fe (27.05 g, 484.32 mmol, 5 equivalents) was added in portions over 10 minutes to a solution of 2-methoxy-1-nitro-4-(trifluoromethyl)benzene (23.8 g, 96.86 mmol, 1 equivalent) in EtOH (300 mL) and saturated NH4Cl (100 mL) at 70 °C. The resulting mixture was stirred at 70 °C for 0.5 hours. TLC analysis (R) was performed. f(产物) =0.50, PE:EtOAc = 5:1) indicates the reaction is complete. The reaction mixture was poured into EtOAc (1500 mL), and the resulting mixture was washed with water (500 mL) and extracted with EtOAc (300 mL x 2). The organic layer was washed with brine (200 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give 2-methoxy-4-(trifluoromethyl)aniline (19 g, 89.46 mmol, 92.35% yield) as a yellow oil. The residue was used directly without any purification.

[0754] Synthesis of 2-methoxy-N-prop-2-ynyl-4-(trifluoromethyl)aniline: A mixture of 2-methoxy-4-(trifluoromethyl)aniline (1 g, 5.23 mmol, 1 equivalent), 3-bromoprop-1-yne (3.11 g, 26.16 mmol, 2.25 mL, 5 equivalents), and K₂CO₃ (2.17 g, 15.69 mmol, 3 equivalents) was prepared in DMF (10 mL). The mixture was degassed and purged three times with N₂, and stirred at 105 °C for 8 h under N₂ atmosphere. TLC analysis showed that the starting material was consumed (PE:EtOAc = 5:1). The mixture was washed with water (60 mL) and extracted with EtOAc (40 mL x 3). The combined organic layers were washed with brine (15 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by column chromatography (SiO2, PE:EtOAc = 25:1 to 15:1) to give 2-methoxy-N-prop-2-ynyl-4-(trifluoromethyl)aniline (0.8 g, 2.44 mmol, 46.70% yield) as a yellow oil.

[0755] Example A34: Synthesis of ((4-((tert-butoxycarbonyl)(prop-2-yn-1-yl)amino)-3-methoxyphenyl)sulfonyl)methyl acetate.

[0756]

[0757] Preparation of (3-methoxy-4-nitrophenyl)(methyl)thion: Sodium methanethiol (5.32 g, 15.19 mmol, 4.84 mL, 20% purity, 1.3 equivalents) was added to a solution of 4-fluoro-2-methoxy-1-nitrobenzene (2 g, 11.69 mmol, 1 equivalent) in DMF (20 mL). The mixture was stirred at 20 °C for 2 hours. TLC analysis indicated that the reaction was complete. The reaction mixture was diluted by adding saturated NH4Cl solution (100 mL). The mixture was filtered and concentrated under reduced pressure to give (3-methoxy-4-nitrophenyl)(methyl)thion (2.4 g, crude product) as a yellow solid.

[0758] Preparation of ((3-methoxy-4-nitrophenyl)thio)methyl acetate: Add phenyl-λ to a solution of (3-methoxy-4-nitrophenyl)(methyl)thione (1.4 g, 7.03 mmol, 1 equivalent) in DCE (15 mL). 3- Iodoalkyl diacetate (3.40 g, 10.54 mmol, 1.5 equivalents) and Pd(OAc)₂ (473.30 mg, 2.11 mmol, 0.3 equivalents). The mixture was stirred at 100 °C for 6 hours under N₂. TLC analysis indicated that the reaction was complete. The reaction mixture was quenched with water (300 mL) and extracted with EtOAc (100 mL x 3). The combined organic layers were washed with brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO₂, PE:EtOAc = 8:1 to 3:1) to give ((3-methoxy-4-nitrophenyl)thio)methyl acetate (1 g, 3.89 mmol, 55.31% yield) as a yellow solid.

[0759] Preparation of ((3-methoxy-4-nitrophenyl)sulfonyl)methyl acetate: Potassium persulfate (oxone) (6.45 g, 10.50 mmol, 3 equivalents) was added to a solution of ((3-methoxy-4-nitrophenyl)thio)methyl acetate (0.9 g, 3.50 mmol, 1 equivalent) in a mixture of acetone (4 mL), water (0.4 mL), and MeOH (4 mL). The mixture was stirred at 25 °C for 5 hours. LC-MS analysis indicated that the reaction was complete. The reaction mixture was diluted with saturated Na₂S₂O₃ solution (200 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give ((3-methoxy-4-nitrophenyl)sulfonyl)methyl acetate (1 g, crude) as a yellow solid. The crude product was used for the next step without purification. MS (ES) + ,m / z):311.9.

[0760] Preparation of ((4-amino-3-methoxyphenyl)sulfonyl)methyl acetate: A saturated NH4Cl solution (166.43 mg, 3.11 mmol, 1 equivalent) and Fe (521.26 mg, 9.33 mmol, 3 equivalent) were added to a solution of ((3-methoxy-4-nitrophenyl)sulfonyl)methyl acetate (0.9 g, 3.11 mmol, 1 equivalent) in EtOH (8 mL). The mixture was stirred at 60 °C for 2 hours. TLC analysis indicated that the reaction was complete. The reaction mixture was filtered, diluted with water (100 mL), and extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was purified by column chromatography (SiO2, PE:EtOAc = 10:1 to 4:1) to give ((4-amino-3-methoxyphenyl)sulfonyl)methyl acetate (580 mg, 2.24 mmol, 71.90% yield) as a yellow solid.

[0761] Preparation of ((4-((tert-Butoxycarbonyl)amino)-3-methoxyphenyl)sulfonyl)methyl acetate: A solution of ((4-amino-3-methoxyphenyl)sulfonyl)methyl acetate (0.49 g, 1.89 mmol, 1 equivalent) in tert-butoxycarbonyl tert-butyl carbonate (20.62 g, 94.49 mmol, 21.71 mL, 50 equivalent) was stirred at 130 °C for 4 hours. LC-MS analysis indicated that the reaction was complete. The reaction mixture was filtered, diluted with water (100 mL), and extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was purified by column chromatography (SiO2, PE:EtOAc = 10:1 to 4:1) to give ((4-((tert-butoxycarbonyl)amino)-3-methoxyphenyl)sulfonyl)methyl acetate (0.45 g, 1.25 mmol, 66.25% yield) as a white oil.

[0762] Preparation of ((4-((tert-butoxycarbonyl)(prop-2-yn-1-yl)amino)-3-methoxyphenyl)sulfonyl)methyl acetate: 3-bromoprop-1-yne (217.22 mg, 1.46 mmol, 157.41 μL, 1.5 equivalents) and Cs₂CO₃ (634.61 mg, 1.95 mmol, 2 equivalents) were added to a solution of ((4-((tert-butoxycarbonyl)amino)-3-methoxyphenyl)sulfonyl)methyl acetate (0.35 g, 973.86 μmol, 1 equivalent) in DMF (4 mL). The mixture was stirred at 25 °C for 1 h under N₂. LC-MS analysis indicated that the reaction was complete. The reaction mixture was diluted with water (100 mL) and extracted with EtOAc (50 mL x 2). The combined organic layers were washed with water (50 mL x 2) and brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was purified by preparative TLC (SiO2, PE:EtOAc = 1:1) to give ((4-((tert-butoxycarbonyl)(prop-2-yn-1-yl)amino)-3-methoxyphenyl)sulfonyl)methyl acetate (0.3 g, 754.83 μmol, 77.51% yield), as a yellow oil. MS (ES) + ,m / z):342.0.

[0763] Example A35: Synthesis of tert-butyl (5-fluoro-2-methoxy-4-(methylcarbamoyl)phenyl)(prop-2-yn-1-yl)carbamate.

[0764]

[0765] Preparation of 2-fluoro-5-methoxy-4-nitrobenzoic acid: A mixture of 2,5-difluoro-4-nitrobenzoic acid (20 g, 98.47 mmol, 1 equivalent) in MeOH (200 mL) was added dropwise to KOH (16.57 g, 295.42 mmol, 3 equivalents) in MeOH (50 mL) at 80 °C. The mixture was stirred at 80 °C for 1 hour. TLC analysis (SiO2, DCM:MeOH:AcOH = 400:20:1, R f =0.6) indicates that the starting material was completely consumed. 6M HCl was added dropwise to the mixture to adjust the pH of the solution to <2. The mixture was then concentrated under reduced pressure to remove MeOH. The mixture was diluted with water (200 mL) and EtOAc (200 mL) and extracted with EtOAc (200 mL x 3). The combined organic layers were washed with brine (200 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude product. The crude product was used directly, unpurified, for the next step. 1H NMR (400MHz, DMSO-d6) δ 8.02-7.99 (d, J = 9.6 Hz, 1H), 7.67-7.66 (d, J = 5.6 Hz, 1H), 3.956 (s, 3H).

[0766] Preparation of methyl 2-fluoro-5-methoxy-4-nitrobenzoate: A mixture of 2-fluoro-5-methoxy-4-nitrobenzoic acid (19.5 g, 90.64 mmol, 1 equivalent) in HCl / MeOH (4 M, 195 mL, 8.61 equivalents) was stirred at 25 °C for 8 hours. TLC (SiO2, PE:EtOAc = 2:1, R f =0.5) indicates that the starting material was completely consumed. The reaction mixture was concentrated under reduced pressure to remove the solvent, yielding a residue. A crude product (19 g) was obtained as a yellow solid and was used in the next step without purification. 1 H NMR (400MHz, DMSO-d6) δ8.21-8.20 (d, J = 4.0 Hz, 1H), 7.50-7.48 (d, J = 8.0 Hz, 1H), 4.01 (s, 3H), 3.71 (s, 3H).

[0767] Preparation of methyl 4-amino-2-fluoro-5-methoxy-benzoate: Fe (13.89 g, 248.73 mmol, 3 equivalents) was added to a solution of methyl 2-fluoro-5-methoxy-4-nitro-benzoate (19 g, 82.91 mmol, 1 equivalent) and NH4Cl (26.61 g, 497.46 mmol, 6 equivalents) in EtOH (200 mL) and water (40 mL) at 90 °C, and the resulting mixture was stirred for 1 h. LC-MS analysis showed 23% nitro starting material residue, observed several new peaks, and 22% of the desired compound was detected. Fe (9.26 g, 165.82 mmol, 2 equivalents) was added to the mixture, and the mixture was further stirred at 90 °C for 2 h. TLC analysis showed complete consumption of the starting material. The mixture was diluted with EtOH (200 mL) and filtered through a diatomaceous earth mat. The filtrate was concentrated under reduced pressure to obtain the crude product, which was then subjected to column chromatography (SiO2, PE:EtOAc = 30:1 to PE:EtOAc:DCM = 30:2:3, R). f =0.5) Purification. Methyl 4-amino-2-fluoro-5-methoxy-benzoate (17 g, 80.23 mmol, 53.27% yield) was obtained as a pale yellow solid. 1H NMR (400MHz, DMSO-d6) δ7.16-7.15 (d, J = 2.0 Hz, 1H), 6.02-6.01 (d, J = 6.4 Hz, 1H), 3.74 (s, 3H), 3.41 (s, 3H). MS(ES+,m / z):199.1.

[0768] Preparation of methyl 4-(tert-Butoxycarbonylamino)-2-fluoro-5-methoxybenzoate: A mixture of methyl 4-amino-2-fluoro-5-methoxybenzoate (16 g, 80.33 mmol, 1 equivalent) and Boc₂O (152 g, 696.46 mmol, 160 mL, 8.67 equivalent) was stirred at 110 °C for 6 hours. TLC analysis (SiO₂, PE:EtOAc = 4:1, R f =0.6) indicates 10% of the starting material remains, and a major new spot with a lower polarity than the starting material was detected. The reaction mixture was concentrated under reduced pressure to remove the solvent, yielding the residue. The residue was subjected to column chromatography (SiO2, PE:EtOAc = 60:1 to 50:1, R... f =0.6) Purification. Methyl 4-(tert-Butoxycarbonylamino)-2-fluoro-5-methoxybenzoate (17 g, 51.12 mmol, 63.64% yield) was obtained as a pale yellow solid. 1 H NMR (400MHz, DMSO-d6) δ8.45(s,1H),7.62(d,J=6.0Hz,1H),7.45(d,J=5.6Hz,1H),3.90(s,3H),3.80(s,3H),1.48(s,6H).

[0769] Preparation of methyl 4-((tert-butoxycarbonyl)(prop-2-yn-1-yl)amino)-2-fluoro-5-methoxybenzoate: Propylene bromide (10.73 g, 90.21 mmol, 7.78 mL, 2 equivalents) was added to a mixture of methyl 4-(tert-butoxycarbonylamino)-2-fluoro-5-methoxybenzoate (15 g, 45.11 mmol, 1 equivalent) and Cs₂CO₃ (29.39 g, 90.21 mmol, 2 equivalents) in DMF (110 mL). The mixture was stirred at 25 °C for 1 hour. TLC (SiO₂, PE:EtOAc = 8:1, Rf = 0.5) indicated complete consumption of the starting material. The mixture was diluted with water (500 mL). The mixture was extracted with EtOAc (200 mL x 3). The combined organic layers were washed with saturated brine (200 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The residue was purified by column chromatography (SiO2, PE:EtOAc = 0:1) to give methyl 4-((tert-butoxycarbonyl)(prop-2-yn-1-yl)amino)-2-fluoro-5-methoxybenzoate. ¹H NMR (400 MHz, DMSO-d6) δ 7.45 (d, J = 2.4 Hz, 1H), 7.30 (d, J = 5.6 Hz, 1H), 7.35 (s, 2H), 3.90 (s, 3H), 3.80 (s, 3H), 3.20 (s, 1H), 1.35 (s, 6H).

[0770] Preparation of 4-((tert-Butoxycarbonyl)(prop-2-yn-1-yl)amino)-2-fluoro-5-methoxybenzoic acid: NaOH (474.26 mg, 11.86 mmol, 2 equivalents) was added to a solution of methyl 4-((tert-Butoxycarbonyl)(prop-2-yn-1-yl)amino)-2-fluoro-5-methoxybenzoate (2 g, 5.93 mmol, 1 equivalent) in THF (5 mL), MeOH (5 mL), and water (5 mL). The mixture was stirred at 40 °C for 0.5 h. TLC analysis indicated that the reaction was complete. The reaction was quenched with water (50 mL), and the pH of the mixture was adjusted to 3 using 1 N HCl. The resulting mixture was filtered and concentrated to give 4-((tert-Butoxycarbonyl)(prop-2-yn-1-yl)amino)-2-fluoro-5-methoxybenzoic acid as a pale yellow solid.

[0771] Preparation of tert-butyl (5-fluoro-2-methoxy-4-(methylcarbamoyl)phenyl)(prop-2-yn-1-yl)carbamate: HOBt (1.42 g, 10.52 mmol, 2 equivalents), EDIC (2.02 g, 10.52 mmol, 2 equivalents), DIPEA (2.04 g, 15.77 mmol, 2.75 mL, 3 equivalents), and methylamine (1.07 g, 15.77 mmol, 3 equivalents, HCl salt) were added to a solution of 4-((tert-butoxycarbonyl)(prop-2-yn-1-yl)amino)-2-fluoro-5-methoxybenzoic acid (1.7 g, 5.26 mmol, 1 equivalent) in DMF (15 mL). The mixture was stirred at 25 °C for 1 hour under N2. TLC analysis indicated that the reaction was complete. The reaction was quenched with water (50 mL) and extracted with EtOAc (30 mL). The combined organic layers were washed with brine (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The crude residue was purified by column chromatography (SiO2, PE:EtOAc = 4:1 to 2:1) to give tert-butyl (5-fluoro-2-methoxy-4-(methylcarbamoyl)phenyl)(prop-2-yn-1-yl)carbamate (1.6 g, 4.76 mmol, 90.47% yield) as a yellow oil.

[0772] Example A36: Synthesis of 3-methoxy-N,N-dimethyl-4-(prop-2-yn-1-ylamino)benzenesulfonamide.

[0773]

[0774] Preparation of 3-methoxy-N,N-dimethyl-4-nitrobenzenesulfonamide: TEA (241.27 mg, 2.38 mmol, 331.87 μL, 2 equivalents) was added to a solution of dimethylamine (145.82 mg, 1.79 mmol, 1.5 equivalents) in DCM (1 mL). The resulting mixture was then added dropwise to a solution of 3-methoxy-4-nitrobenzenesulfonyl chloride (300 mg, 1.19 mmol, 1 equivalent) in DCM and stirred at 25 °C for 2 hours. TLC analysis (PE:EtOAc = 3:1, R0) was performed. f =0.40) indicates the reaction is complete. The mixture was quenched with water (40 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was purified by preparative TLC (SiO2, PE:EtOAc = 3:1) to give 3-methoxy-N,N-dimethyl-4-nitrobenzenesulfonamide (270 mg, 933.66 μmol, 78.32% yield) as a pale yellow solid.

[0775] Preparation of 4-amino-3-methoxy-N,N-dimethylbenzenesulfonamide: Fe (482.78 mg, 8.64 mmol, 10 equivalents) was added to a solution of 3-methoxy-N,N-dimethyl-4-nitrobenzenesulfonamide (250 mg, 864.50 μmol, 1 equivalent) and solid NH4Cl (231.22 mg, 4.32 mmol, 5 equivalents) in EtOH (5 mL) and water (1 mL) at 70 °C. The mixture was stirred for 2 hours. TLC analysis (PE:EtOAc = 1:1, R f =0.24) indicates that the reaction is complete. The mixture was quenched with water (60 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give 4-amino-3-methoxy-N,N-dimethylbenzenesulfonamide (210 mg, crude product) as a pale yellow solid.

[0776] Preparation of 3-methoxy-N,N-dimethyl-4-(prop-2-yn-1-ylamino)benzenesulfonamide: A solution of 4-amino-3-methoxy-N,N-dimethylbenzenesulfonamide (330 mg, 1.43 μmol, 1 equivalent) in CHCl3 (10 mL) was added to a mixture of 3-bromoprop-1-yne (340.94 mg, 2.87 μmol, 247.06 μL, 2 equivalents) and DIPEA (926.02 mg, 7.17 mmol, 1.25 mL, 5 equivalents) in CHCl3 (3 mL). The mixture was stirred at 70 °C for 16 hours. TLC analysis (PE:EtOAc = 1:1, R f =0.43) indicates the reaction is complete. The mixture was quenched with water (40 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO2, PE:EtOAc = 1:1) to give 3-methoxy-N,N-dimethyl-4-(prop-2-yn-1-ylamino)benzenesulfonamide (210 mg, 47.5% yield) as a pale yellow solid. MS (ES) + ,m / z):269.2.

[0777] Example A37: Synthesis of tert-butyl (2-fluoro-6-methoxy-4-(methylsulfonyl)phenyl)(prop-2-yn-1-yl)carbamate.

[0778]

[0779] Preparation of 2-fluoro-6-methoxy-4-(methanesulfonyl)aniline: DL-proline (117.73 mg, 1.02 mmol, 0.5 equivalence), CuI (389.49 mg, 2.05 mmol, 1 equivalence), and NaOH (81.80 mg, 2.05 mmol, 1 equivalence) were added to a solution of 4-bromo-2-fluoro-6-methoxyaniline (626.34 mg, 6.14 mmol, 3 equivalents) in DMSO (15 mL). The reaction mixture was stirred at 90 °C for 16 hours under N2. TLC analysis (PE:EtOAc = 2:1, R f =0.5) indicates that the starting material was completely consumed, and a major new spot with a lower polarity than the starting material was detected. The mixture was diluted with saturated EDTA solution (100 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with brine (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was purified by column chromatography (SiO2, PE:EtOAc = 10:1 to 2:1) to give the desired product (1.2 g, 5.47 mmol, 89.22% yield) as a white solid.

[0780] Preparation of 2-fluoro-6-methoxy-N,N-di(tert-butyloxycarbonyl)-4-(methylsulfonyl)aniline: DMAP (60.18 mg, 492.63 μmol, 0.1 equivalent) and TEA (1.99 g, 19.71 mmol, 4.53 mL, 4 equivalent) were added to a mixture of 2-fluoro-6-methoxy-4-(methylsulfonyl)aniline (1.2 g, 4.93 mmol, 1 equivalent) and Boc₂O (4.30 g, 19.71 mmol, 4.53 mL, 4 equivalent) in 1,4-dioxane (12 mL). The reaction mixture was stirred at 110 °C for 6 hours. TLC analysis (PE:EtOAc = 2:1, R f =0.5) indicates that the starting material was completely consumed, and a major new spot with a lower polarity than the starting material was detected. The mixture was diluted with water (100 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with brine (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, PE:EtOAc = 20:1 to 2:1) to give the desired product (1.9 g, 4.08 mmol, 82.75% yield) as a yellow oil.

[0781] Preparation of tert-butyl (2-fluoro-6-methoxy-4-(methylsulfonyl)phenyl)carbamate: A mixture of 2-fluoro-6-methoxy-N,N-di(tert-butyloxycarbonyl)-4-(methylsulfonyl)aniline (900 mg, 1.93 mmol, 1 equivalent) and K₂CO₃ (1.33 g, 9.66 mmol, 5 equivalent) in MeOH (10 mL) was stirred at 25 °C for 2 hours. The mixture was then heated to 40 °C and stirred for another 2 hours. TLC analysis (PE:EtOAc = 2:1, R f =0.4) indicates that the starting material was completely consumed, and a major new spot with a polarity greater than that of the starting material was detected. The reaction mixture was concentrated under reduced pressure. The crude residue was diluted with water (200 mL) and extracted with EtOAc (70 mL x 3). The combined organic layers were washed with brine (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue (1.6 g, crude product) was obtained as a pale yellow solid and used directly in the next step.

[0782] Preparation of tert-butyl (2-fluoro-6-methoxy-4-(methylsulfonyl)phenyl)(prop-2-yn-1-yl)carbamate: A mixture of tert-butyl (2-fluoro-6-methoxy-4-(methylsulfonyl)phenyl)carbamate (1.5 g, 4.23 mmol, 1 equivalent) and Cs₂CO₃ (2.75 g, 8.45 mmol, 2 equivalents) in DMF (16 mL) was stirred at 25 °C for 1 hour. TLC analysis (PE:EtOAc = 2:1, R f =0.4) indicates that the starting material was completely consumed, and a major new spot with a lower polarity than the starting material was detected. The mixture was diluted with water (100 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with brine (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was purified by column chromatography (SiO2, PE:EtOAc = 20:1 to 4:1) to give tert-butyl (2-fluoro-6-methoxy-4-(methanesulfonyl)phenyl)(prop-2-yn-1-yl)carbamate (1.9 g, 3.99 mmol, 94.32% yield) as a pale yellow oil.

[0783] Example A38: Synthesis of 3-methoxy-N,N-dimethyl-4-(prop-2-yn-1-ylamino)benzenesulfonamide.

[0784]

[0785] Preparation of 3-methoxy-N,N-dimethyl-4-nitrobenzenesulfonamide: A solution of N-methylmethylamine (194.43 mg, 2.38 mmol, 218.46 μL, 1.2 equivalents, HCl) in DCM (15 mL) and Et3N (1.01 g, 9.93 mmol, 1.38 mL, 5 equivalents) was prepared at 0 °C under N2. A solution of 3-methoxy-4-nitrobenzenesulfonyl chloride (500 mg, 1.99 mmol, 1 equivalent) in DCM (5 mL) was added dropwise to the mixture, and the mixture was stirred at 20 °C for 2 hours. TLC analysis (PE:EtOAc = 1:1, R0) was performed. f =0.42) indicates that the reaction is complete. The mixture was concentrated under vacuum and subjected to column chromatography (SiO2, PE:EtOAc = 20:1 to 0:1, R f =0.42) purification yielded the desired product (500 mg, 1.86 mmol, 93.69% yield) as a yellow solid. MS (ES) + ,m / z):261.1.

[0786] Preparation of 4-amino-3-methoxy-N,N-dimethylbenzenesulfonamide: Under N2 conditions, NH4Cl (448.09 mg, 8.38 mmol, 292.87 μL, 5 equivalents) was added to a solution of 3-methoxy-N,N-dimethyl-4-nitrobenzenesulfonamide (450 mg, 1.68 mmol, 1 equivalent) in EtOH (15 mL) and water (5 mL). Fe (467.85 mg, 8.38 mmol, 5 equivalents) was added to the mixture at 90 °C, and the resulting mixture was stirred at 90 °C for 0.5 h. TLC analysis (PE:EtOAc = 1:1, R f =0.39) indicates the reaction is complete. The reaction mixture is subjected to hot filtration, followed by filtration and vacuum concentration. The crude residue is then subjected to column chromatography (SiO2, PE:EtOAc = 10:1 to 0:1, R... f =0.39) purification yielded the desired product (380 mg, 1.54 mmol, 91.99% yield) as a pale yellow solid. MS (ES) + ,m / z):231.0.

[0787] Preparation of 3-methoxy-N,N-dimethyl-4-(prop-2-yn-1-ylamino)benzenesulfonamide: DIPEA (865.91 mg, 6.70 mmol, 1.17 mL, 5 equivalents) and 3-bromoprop-1-yne (797.03 mg, 6.70 mmol, 577.56 μL, 5 equivalents) were added to a mixture of 4-amino-3-methoxy-N,N-dimethylbenzenesulfonamide in CHCl3 (10 mL). The mixture was degassed and purged three times with N2 at 20 °C, and then stirred at 70 °C for 10 h. Then, DIPEA (346.37 mg, 2.68 mmol, 466.81 μL, 2 equivalents) and 3-bromoprop-1-yne (318.81 mg, 2.68 mmol, 231.02 μL, 2 equivalents) were added to the mixture, and the resulting mixture was further stirred at 70 °C for 10 h. LC-MS and TLC analysis (PE:EtOAc = 1:1, R f =0.50) indicates 20% of the starting material remains, and a major new spot was found. The mixture was concentrated under vacuum and subjected to column chromatography (SiO2, PE:EtOAc = 10:1 to 0:1, R f =0.5) purification yielded the desired product (120 mg, 290.69 μmol, 21.69% yield), a pale yellow solid. MS (ES) + ,m / z):268.9.

[0788] Example A41: Synthesis of 4-methoxy-6-(methanesulfonyl)-N-(prop-2-yn-1-yl)pyridine-3-amine.

[0789]

[0790] Preparation of 4-methoxy-2-(methylthio)-5-nitropyridine: NaSMe (3.34 g, 47.70 mmol, 3.04 mL, 6 equivalents) was added to a solution of 2-chloro-4-methoxy-5-nitropyridine (1.50 g, 7.95 mmol, 1 equivalent) in DMF (20 mL). The mixture was stirred at 15 °C for 2 h. LC-MS analysis showed complete consumption of the starting material and a main peak with the desired product mass was detected. The reaction mixture was separated by adding water (50 mL) and EtOAc (50 mL). The organic phase was separated, washed with brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the desired product (1.30 g, 6.49 mmol, 81.68% yield) as a brown solid. MS (ES) + ,m / z):200.8.

[0791] Preparation of 4-methoxy-2-(methylsulfonyl)-5-nitropyridine: Potassium persulfate (oxone) (11.98 g, 19.48 mmol, 3 equivalents) was added to a solution of 4-methoxy-2-(methylthio)-5-nitropyridine (1.30 g, 6.49 mmol, 1 equivalent) in acetone (20 mL), MeOH (2 mL), and water (20 mL). The mixture was stirred at 0–15 °C for 2 hours. LC-MS analysis showed complete consumption of the starting material and a main peak with the desired mass was detected. The reaction mixture was separated into layers using saturated Na₂S₂O₄ solution (100 mL) and EtOAc (100 mL). The organic phase was separated, washed with brine (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the desired product (1.45 g, 5.68 mmol, 87.56% yield) as a yellow solid. MS (ES) + ,m / z):233.1.

[0792] Preparation of 4-methoxy-6-(methanesulfonyl)pyridine-3-amine: Fe (2.41 g, 43.10 mmol, 10 equivalents) was added to a solution of 4-methoxy-2-methanesulfonyl-5-nitro-pyridine (1 g, 4.31 mmol, 1 equivalent) in AcOH (20 mL). The mixture was stirred at 50 °C for 2 h. LC-MS analysis showed complete consumption of the starting material and a main peak with the desired mass was detected. The reaction mixture was separated by adding water (100 mL) and EtOAc (100 mL). The organic phase was separated, washed with brine (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, PE:EtOAc = 10:1 to 1:1) to give the desired product (510 mg, 2.52 mmol, 58.51% yield) as a brown solid. MS (ES) + ,m / z):202.8.

[0793] Preparation of tert-butyl (4-methoxy-6-(methanesulfonyl)pyridin-3-yl)carbamate: Boc₂O (4.20 g, 19.26 mmol, 4.42 mL, 6 equivalents) was added to a solution of 4-methoxy-6-(methanesulfonyl)pyridin-3-amine (650 mg, 3.21 mmol, 1 equivalent) in dioxane (10 mL). The mixture was stirred at 110 °C for 14 h. LC-MS analysis showed complete consumption of the starting material and a main peak with the desired quality was detected. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO₂, PE:EtOAc = 10:1 to 1:1) to give the desired product (0.65 g, 2.15 mmol, 66.97% yield) as a yellow oil. MS (ES) + ,m / z):302.9.

[0794] Preparation of tert-butyl (4-methoxy-6-(methanesulfonyl)pyridin-3-yl)(prop-2-yn-1-yl)carbamate: Tert-butyl (400 mg, 1.32 mmol, 1 equivalent) of (4-methoxy-6-(methanesulfonyl)pyridin-3-yl)carbamate was added to a mixture of NaH (529.20 mg, 13.23 mmol, 60% in mineral oil, 10 equivalents) in DMF (4 mL). The mixture was stirred at 0 °C for 30 min, and 3-bromoprop-1-yne (236.07 mg, 1.98 mmol, 171.07 μL, 1.50 equivalents) was added to the mixture. LC-MS analysis showed complete consumption of the starting material and a main peak with the desired mass was detected. The reaction mixture was separated into layers by adding water (40 mL) and EtOAc (40 mL). The organic phase was separated, washed with brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, PE:EtOAc = 10:1 to 1:1) to give the desired product (250 mg, 734.44 μmol, 55.51% yield) as a yellow oil. MS (ES) + ,m / z):341.2.

[0795] Preparation of 4-methoxy-6-(methanesulfonyl)-N-(prop-2-yn-1-yl)pyridin-3-amine: HCl / EtOAc (4M, 2.02 mL, 16.17 equivalents) was added to a solution of (4-methoxy-6-(methanesulfonyl)pyridin-3-yl)(prop-2-yn-1-yl)carbamate tert-butyl ester (170 mg, 499.42 μmol, 1 equivalent). The mixture was stirred at 15 °C for 1 hour. LC-MS analysis showed complete consumption of the starting material and a main peak with the desired mass was detected. The reaction mixture was concentrated under reduced pressure to give the desired product (100 mg, 361.35 μmol, 72.35% yield, HCl) as a brown solid. MS (ES) + ,m / z):241.1.

[0796] Example A42: Synthesis of 2-(4-methoxy-5-(prop-2-yn-1-ylamino)pyridin-2-yl)-2-methylpropionitrile.

[0797] Route 1:

[0798]

[0799] Preparation of 2-(4-methoxy-5-nitropyridin-2-yl)-2-methylpropionitrile: Under N2 at 0 °C, KHMDS (1M, 53.03 mL, 5 equivalents) was added dropwise to a solution of 2-chloro-4-methoxy-5-nitropyridinium (2 g, 10.61 mmol, 1 equivalent) in THF (5 mL). Then, isobutyronitrile (2.20 g, 31.82 mmol, 3 equivalents) was added, and the resulting mixture was stirred at 0 °C for 2 h. TLC analysis (PE:EtOAc = 1:1) indicated complete consumption of the starting material. The reaction was quenched by slow addition of ice, and the mixture was extracted with EtOAc (10 mL x 2). The combined organic layers were washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by column chromatography (SiO2, PE:EtOAc = 10:1 to 1:1) to give 2-(4-methoxy-5-nitropyridin-2-yl)-2-methylpropionitrile (0.66 g, 2.98 mmol, 28.13% yield) as a yellow solid.

[0800] Preparation of 2-(5-amino-4-methoxypyridin-2-yl)-2-methylpropionitrile: Under N2 and at 90 °C, NH4Cl (423.16 mg, 7.91 mmol, 276.57 μL, 5 equivalents) and Fe (441.83 mg, 7.91 mmol, 5 equivalents) were added sequentially to a solution of 2-(4-methoxy-5-nitropyridin-2-yl)-2-methylpropionitrile (0.35 g, 1.58 mmol, 1 equivalent) in EtOH (5 mL) and water (1 mL). The mixture was heated to 90 °C and stirred for 1 hour. TLC analysis indicated that the reaction was complete. The mixture was filtered and concentrated under reduced pressure. The residue was poured into a mixture of DCM and water (w / w = 1:1) (20 mL) and stirred for 30 minutes. The aqueous phase was extracted with DCM (5 mL x 2). The combined organic layers were washed with brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by preparative TLC (SiO2, PE:EtOAc = 1:1) to give the desired product (0.22 g, 1.15 mmol, 72.71% yield) as a yellow solid.

[0801] Preparation of 2-(4-methoxy-5-(prop-2-yn-1-ylamino)pyridin-2-yl)-2-methylpropionitrile: Under N2 at 25 °C, K2CO3 (520.36 mg, 3.77 mmol, 3 equivalents) was added in one step to a mixture of 2-(5-amino-4-methoxypyridin-2-yl)-2-methylpropionitrile (0.24 g, 1.26 mmol, 1 equivalent) and 3-bromoprop-1-yne (746.50 mg, 6.28 mmol, 540.94 μL, 5 equivalents) in DMF (5 mL). The mixture was stirred at 70 °C for 12 hours. LC-MS and TLC analysis (PE:EtOAc = 1:1, R f =0.45) indicates the reaction is complete. Pour the mixture into water (50 mL) and stir for 2 minutes. Extract the aqueous phase with EtOAc (20 mL x 3). Wash the combined organic layers with brine (20 mL), dry over anhydrous sodium sulfate, filter, and concentrate under vacuum. Purify the crude residue by silica gel chromatography (PE:EtOAc = 10:1 to 0:1) to give the desired product (0.255 g, 889.75 μmol, 70.89% yield) as a yellow solid. MS (ES) + ,m / z):230.0.

[0802] Route 2:

[0803]

[0804] Preparation of tert-butyl (6-(2-cyanoprop-2-yl)-4-methoxypyridin-3-yl)carbamate: Under N2 and at 25 °C, (Boc)2O (855.96 mg, 3.92 mmol, 901.01 μL, 5 equivalents) and DMAP (191.66 mg, 1.57 mmol, 2 equivalents) were added in one step to a solution of 2-(5-amino-4-methoxypyridin-2-yl)-2-methylpropionitrile (0.15 g, 784.40 μmol, 1 equivalent) in dioxane (5 mL). The mixture was stirred at 110 °C for 12 h. LC-MS analysis indicated that the reaction was complete and the di-Boc byproduct was detected. The mixture was cooled to 25 °C and concentrated under reduced pressure at 50 °C. The residue was added to a solution of 500 mg solid Na₂CO₃ in MeOH (10 mL) to convert the di-Boc byproduct into the desired mono-Boc-protected product. The mixture was stirred at 40 °C for 2 hours. The mixture was cooled to 25 °C and concentrated under reduced pressure at 40 °C. The crude residue was purified by silica gel chromatography (PE:EtOAc = 30:1 to 3:1) to give the desired product (0.18 g, 586.93 μmol, 74.83% yield) as a colorless oil. MS (ES) + ,m / z):291.9. =

[0805] Preparation of tert-butyl (6-(2-cyanopropyl-2-yl)-4-methoxypyridin-3-yl)(prop-2-yn-1-yl)carbamate: NaH (37.07 mg, 926.74 μmol, 60% in mineral oil, 1.5 equivalents) was added in a single addition to a mixture of tert-butyl (6-(2-cyanopropyl-2-yl)-4-methoxypyridin-3-yl)carbamate (0.18 g, 617.82 μmol, 1 equivalent) in 2 mL of DMF under N2 at 0 °C. The mixture was stirred at 0 °C for 30 min, and then 3-bromoprop-1-yne (88.20 mg, 741.39 μmol, 63.91 μL, 1.2 equivalents) was added in a single addition under N2 at 0 °C. The mixture was stirred at 0 °C for 1.5 h. LC-MS analysis indicated that the reaction was complete. Pour the mixture into water (20 mL) and stir for 2 minutes. Extract the aqueous phase with EtOAc (10 mL x 2). Wash the combined organic layers with brine (10 mL), dry over anhydrous sodium sulfate, filter, and concentrate under vacuum. Purify the crude residue by silica gel chromatography (PE:EtOAc = 30:1 to 3:1) to give the desired product (0.15 g, 432.61 μmol, 70.02% yield) as a colorless oil. MS (ES) + ,m / z):329.9.

[0806] Preparation of 2-(4-methoxy-5-(prop-2-yn-1-ylamino)pyridin-2-yl)-2-methylpropionitrile: A solution of (6-(2-cyanoprop-2-yl)-4-methoxypyridin-3-yl)(prop-2-yn-1-yl)carbamate tert-butyl ester (120 mg, 364.31 μmol, 1 equivalent) in HCl / EtOAc (5 mL) was prepared at 0 °C under N2 conditions and stirred at 0 °C for 2 hours. TLC analysis (PE:EtOAc = 3:1, R f =0) indicates the reaction is complete. Pour the mixture into a saturated Na₂CO₃ solution (50 mL) and stir for 2 minutes. Extract the aqueous phase with EtOAc (10 mL x 3). Wash the combined organic layers with brine (10 mL), dry over anhydrous sodium sulfate, filter, and concentrate under vacuum. Analyze the crude residue by preparative TLC (PE:EtOAc = 3:1, R... f =0.4) purification yielded the desired product (100 mg, 417.88 μmol), a white solid. MS (ES) + ,m / z):230.3.

[0807] Example A43: Synthesis of 3-methoxy-4-(prop-2-yn-1-ylamino)benzoic acid.

[0808]

[0809] A solution of 4-[tert-butoxycarbonyl(prop-2-ynyl)amino]-3-methoxybenzoic acid (1.1 g, 3.60 mmol, 1 equivalent) in 4N HCl / EtOAc (50 mL) was stirred at 20 °C for 2 hours. TLC analysis (PE:EtOAc = 1:1, Rm) was performed. f =0.5) indicates that the starting material was consumed. The mixture was concentrated to give a crude product (0.8 g, 3.51 mmol, 97.39% yield) as a yellow solid. The crude product was used without purification.

[0810] Example A44: Synthesis of methyl 3-methoxy-4-(prop-2-yn-1-ylamino)benzoate.

[0811]

[0812] Preparation of methyl 3-methoxy-4-(prop-2-yn-1-ylamino)benzoate: The solution of methyl 4-((tert-butoxycarbonyl)(prop-2-yn-1-yl)amino)-3-methoxybenzoate in EtOAc was degassed in 4N HCl (20 mL) and purged three times with N2. The mixture was then stirred at 20 °C for 1 hour under N2. TLC analysis (PE:EtOAc = 3:1, Rm) f=0.55) indicates that the starting material was consumed and a new spot was formed. The reaction mixture was quenched by adding saturated NaHCO3 solution (30 mL) and extracted with EtOAc (40 mL x 3). The combined organic layers were washed with brine (25 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product (0.58 g, 2.12 mmol, 67.59% yield) was given as a yellow solid, which was used without purification.

[0813] Preparation of 3-methoxy-4-(prop-2-yn-1-ylamino)benzoic acid: A solution of methyl 3-methoxy-4-(prop-2-yn-1-ylamino)benzoate in MeOH and water (10 mL, MeOH:water = 1:3) was degassed and purged three times with N2. The solution was stirred at 20 °C for 1 hour under N2. TLC analysis (PE:EtOAc = 3:1, R f =0) indicates residual starting material, and a major new spot was detected. The reaction mixture was extracted with EtOAc (50 mL x 2), and the pH of the mixture was adjusted to 3–4 by adding 2 M HCl. The organic layer was washed with brine (50 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the desired product (0.6 g, 2.34 mmol, 42.73% yield) as a yellow solid.

[0814] Example A45: Synthesis of 3-methoxy-4-(prop-2-yn-1-ylamino)benzamide.

[0815]

[0816] Preparation of 4-amino-3-methoxybenzoic acid: Under N2 at 25°C, a solution of methyl 4-amino-3-methoxybenzoate (4.5 g, 23.59 mmol, 1 equivalent) in MeOH (45 mL), water (15 mL), and THF (15 mL) was added in one step to LiOH (4.95 g, 117.97 mmol, 5 equivalents). The mixture was stirred at 25°C for 12 hours. TLC analysis (PE:EtOAc = 3:1, R f =0) indicates the reaction is complete. The mixture was concentrated under reduced pressure at 40°C. The residue was poured into water (50 mL) and stirred for 1 minute. The aqueous phase was extracted with EtOAc (30 mL x 3). 2N HCl was added to the aqueous phase to adjust the pH of the solution to 2. The aqueous phase was filtered and concentrated under vacuum to give the desired product (4 g, 22.73 mmol, 96.35% yield) as a pale yellow solid.

[0817] Preparation of 4-amino-3-methoxybenzoamide: Under N2 and at 25 °C, NH4OAc (8.76 g, 113.66 mmol, 5 equivalents), DIPEA (29.38 g, 227.32 mmol, 39.60 mL, 10 equivalents), and HATU (17.29 g, 45.46 mmol, 2 equivalents) were added in one step to a solution of 4-amino-3-methoxybenzoic acid (4 g, 22.73 mmol, 1 equivalent, 95% purity) in DMF (50 mL). The mixture was stirred at 25 °C for 2 hours. TLC analysis (PE:EtOAc = 0:1, R f =0.30) indicates the reaction is complete. Pour the mixture into water (800 mL) and stir for 2 minutes. Extract the aqueous phase with EtOAc (300 mL x 3). Wash the combined organic layers with brine (300 mL x 1), dry over anhydrous sodium sulfate, filter, and concentrate under vacuum. Purify the residue by silica gel chromatography (SiO2, PE:EtOAc = 100:1 to 0:1) to give the desired product (5 g, 18.05 mmol, 79.42% yield) as a yellow oil.

[0818] Preparation of 3-methoxy-4-(prop-2-yn-1-ylamino)benzamide: Under N2 at 25 °C, K2CO3 (7.49 g, 54.16 mmol, 3 equivalents) was added in one step to a mixture of 4-amino-3-methoxybenzamide (5 g, 18.05 mmol, 1 equivalent, 60% purity) and 3-bromoprop-1-yne (4.52 g, 36.11 mmol, 3.28 mL, 2 equivalents, 95% purity) in DMF (50 mL). The mixture was stirred at 70 °C for 4 hours. TLC analysis (PE:EtOAc = 0:1, R f =0.40) indicates the reaction is complete. The mixture was cooled to 25°C, and the residue was poured into water (500 mL) and stirred for 2 minutes. The aqueous phase was extracted with EtOAc (200 mL x 3). The combined organic layers were washed with brine (300 mL x 1), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (SiO2, PE:EtOAc = 100:1 to 0:1) to give the desired product (3.32 g, 12.19 mmol, 67.54% yield) as a yellow solid.

[0819] Example A46: Synthesis of 2-((3-methoxy-4-(prop-2-yn-1-ylamino)phenyl)sulfonyl)ethane-1-ol.

[0820]

[0821] Preparation of 2-((3-methoxy-4-nitrophenyl)thio)ethanol-1-ol: A solution of 4-fluoro-2-methoxy-1-nitrobenzene (1.24 g, 11.69 mmol, 1 equivalent) and 2-mercaptoethanol-1-ol (1.83 g, 23.37 mmol, 1.63 mL, 2 equivalents) was degassed and purged three times with N2. The mixture was stirred at 25 °C for 19 hours under N2. TLC analysis (PE:EtOAc = 5:1, R f =0.05; DCM:MeOH=10:1,R f =0.45) indicates the reaction is complete. Pour the residue into water (200 mL) and stir for 30 minutes. Filter the mixture and concentrate under vacuum. Pour the residue into water (200 mL) and extract the mixture with EtOAc (100 mL x 3). Wash the combined organic layers with brine (80 mL x 3), filter, and concentrate under vacuum. A crude product (2.8 g, crude) is given as a yellow solid, which is used without purification.

[0822] Preparation of 2-((3-methoxy-4-nitrophenyl)sulfonyl)ethane-1-ol: Potassium persulfate (oxone) (15.02 g, 24.43 mmol, 2 equivalents) was added to a solution of 2-((3-methoxy-4-nitrophenyl)thio)ethane-1-ol (2.8 g, 12.21 mmol, 1 equivalent) in acetone (40 mL), water (40 mL), and MeOH (4 mL). The mixture was stirred at 25 °C for 2 hours. TLC analysis (DCM:MeOH = 10:1, R f =0.4) indicates that the starting material was completely consumed and a new spot was detected. The residue was poured into a saturated Na₂SO₃ (300 mL) solution and stirred for 30 minutes. The mixture was filtered and concentrated under vacuum. The residue was poured into a saturated Na₂SO₃ solution (300 mL) and stirred for 30 minutes, then extracted with EtOAc (100 mL x 3). The combined organic layers were washed with brine (80 mL x 3), filtered, and concentrated under vacuum. A crude product (3 g, crude) was obtained as a white solid, which was used without purification.

[0823] Preparation of 2-((4-amino-3-methoxyphenyl)sulfonyl)ethane-1-ol: NH4Cl (3.69 g, 68.90 mmol, 6 equivalents) and Fe (1.92 g, 34.45 mmol, 3 equivalents) were added to a solution of 2-((3-methoxy-4-nitrophenyl)sulfonyl)ethane-1-ol (3 g, 11.48 mmol, 1 equivalent) in EtOH (20 mL) and water (4 mL) at 90 °C. The mixture was stirred at 90 °C for 0.5 h. TLC analysis (DCM:MeOH = 10:1, R f=0.5) indicates 50% of the starting material remains, and two major new spots with greater polarity than the starting material were detected. An additional portion of Fe (1.28 g, 22.97 mmol, 2 equivalents) was added to the mixture, and the mixture was stirred at 90 °C for 1 hour. TLC analysis (DCM:MeOH = 10:1, R0.5) was performed. f =0.5) indicates that the starting material was completely consumed, and a major new spot with a polarity greater than that of the starting material was detected. The residue was diluted with EtOAc (400 mL). The mixture was diluted with water (400 mL) and extracted with EtOAc (200 mL x 3). The combined organic layers were washed with brine (200 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. A crude product (2.8 g, crude product) was given as a white oil. MS (ES) + ,m / z):232.0.1H NMR(400MHz,DMSO-d6)δppm 3.27-3.35(m,2H)3.60-3.68(m,2H)3.80-3.88(m,3H)4.78-4.87(m,1H) 5.67-5.81(m,2H)6.69-6.77(m,1H)7.13-7.18(m,1H)7.18-7.24(m,1H).

[0824] Preparation of 2-((3-methoxy-4-(prop-2-yn-1-ylamino)phenyl)sulfonyl)ethane-1-ol: K₂CO₃ (179.3 mg, 1.30 mmol, 2 equivalents) and 3-bromoprop-1-yne (67.51 mg, 454.02 μmol, 48.92 μL, 0.7 equivalents) were added to a solution of 2-((4-amino-3-methoxyphenyl)sulfonyl)ethane-1-ol (0.15 g, 648.6 μmol, 1 equivalent) in DMF (1 mL). The mixture was degassed and purged three times with N₂, and then stirred at 50 °C for 19 hours under N₂. TLC analysis (PE:EtOAc = 1:2, R f=0.5) indicated 10% of the starting material remained, and a major new spot with a lower polarity than the starting material was detected. The residue was diluted with water (50 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with brine (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO2, PE:EtOAc = 0:1) to give the desired product (0.053 g, 177.12 μmol, 27.31% yield) as a yellow oil. 1HNMR (400MHz, DMSO-d6) δppm 3.09 (s, 1H) 3.33-3.37 (m, 2H) 3.63 (q, J = 6.44Hz, 2H) 3.87 (s, 3H) 4.00 (br d,J=4.16Hz,2H)4.84(t,J=5.56Hz,1H)6.26-6.38(m,1H)6.69-6.80(m,1H)7.11-7.24(m,1H)7.35(br d,J=8.19Hz,1H).

[0825] Example A47: Synthesis of 3-methoxy-N-methyl-4-(prop-2-yn-1-ylamino)benzenesulfonamide and 3-methoxy-N,N-dimethyl-4-(prop-2-yn-1-ylamino)benzenesulfonamide.

[0826]

[0827] A solution of 3-methoxy-4-(prop-2-yn-1-ylamino)benzenesulfonamide (1 g, 4.16 mmol, 1 equivalent) in acetone (10 mL) was treated with K₂CO₃ (1.15 g, 8.32 mmol, 2 equivalents) and CH₃I (708.87 mg, 4.99 mmol, 310.91 μL, 1.2 equivalents). The mixture was stirred at 50 °C for 6 hours. LC-MS analysis showed 27% of the starting material remaining and the desired compound was detected. The reaction mixture was diluted with water (300 mL) and extracted with EtOAc (100 mL x 2). The combined organic layers were washed with water (200 mL x 2) and brine (200 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO₂, PE:EtOAc = 1:1) to give the desired product as a yellow oil. 3-Methoxy-N-methyl-4-(prop-2-yn-1-ylamino)benzenesulfonamide (0.7 g, 2.75 mmol), 66.08% yield, MS (ES) +m / z): 255.1; 3-methoxy-N,N-dimethyl-4-(prop-2-yn-1-ylamino)benzenesulfonamide (0.2 g, 745.35 μmol), 17.91% yield, MS (ES) + ,m / z):269.1.

[0828] Example A48: Synthesis of 5-((tert-butoxycarbonyl)(prop-2-yn-1-yl)amino)thiophene-2-carboxylic acid.

[0829]

[0830] Preparation of methyl 5-((tert-Butoxycarbonyl)amino)thiophene-2-carboxylate: A mixture of methyl 5-aminothiophene-2-carboxylate (1 g, 6.36 mmol, 1 equivalent) and Boc₂O (4.17 g, 19.09 mmol, 4.38 mL, 3 equivalent) was degassed and purged three times with N₂, and then stirred at 110 °C for 1.5 h under N₂. TLC analysis (PE:EtOAc = 3:1, R f =0.50) indicated the presence of residual starting material, and a major new spot was detected. The reaction mixture was quenched by adding PE (200 mL) and the mixture was stirred for 1 hour. The mixture was filtered and concentrated under reduced pressure to give the residue. A crude product (1.6 g, 5.60 mmol, 87.97% yield) was given as a yellow solid, which was used without further purification. MS (ES) + ,m / z):258.1.

[0831] Preparation of methyl 5-((tert-butoxycarbonyl)(prop-2-yn-1-yl)amino)thiophene-2-carboxylate: A mixture of methyl 5-(tert-butoxycarbonylamino)thiophene-2-carboxylate (1.5 g, 5.25 mmol, 1 equivalent), 3-bromoprop-1-yne (686.56 mg, 5.77 mmol, 497.51 μL, 1.1 equivalent), and Cs₂CO₃ (5.13 g, 15.74 mmol, 3 equivalent) in DMF (20 mL) was degassed and purged three times with N₂. The mixture was then stirred at 25 °C for 4 hours under N₂. TLC analysis (PE:EtOAc = 3:1, R f =0.60) indicated the presence of residual starting material, and a major new spot was detected. The reaction mixture was quenched by adding water (200 mL) and extracted with EtOAc (100 mL x 3). The combined organic layers were washed with brine (30 mL x 4), filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, PE:EtOAc = 1:0 to 40:1) to give the desired product (1.1 g, 3.35 mmol, 63.89% yield) as a yellow solid.

[0832] Preparation of 5-((tert-Butoxycarbonyl)(prop-2-yn-1-yl)amino)thiophen-2-carboxylic acid: A mixture of methyl 5-((tert-Butoxycarbonyl)(prop-2-yn-1-yl)amino)thiophen-2-carboxylic acid (0.5 g, 1.69 mmol, 1 equivalent) and Cs₂CO₃ (5.52 g, 16.93 mmol, 10 equivalent) in MeOH (5 mL), water (5 mL), and THF (5 mL) was degassed and purged three times with N₂. The mixture was stirred at 25 °C for 4 hours under N₂ atmosphere. TLC analysis (EtOAc = 1, R f =0.3) indicates residual starting material, and a major new spot was detected. The reaction mixture was concentrated under reduced pressure to remove THF and MeOH. The residue was diluted with water (10 mL) and 2N HCl was added to adjust the pH of the mixture to 5. The mixture was then extracted with EtOAc (30 mL x 3). The combined organic layers were washed with brine (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product (0.43 g, 1.36 mmol, 80.36% yield) was given as a yellow solid, which was used without purification. MS (ES) + ,m / z):282.0.

[0833] Example A49: Synthesis of ethyl 3-methoxy-4-(prop-2-yn-1-ylamino)benzoate.

[0834]

[0835] Preparation of ethyl 3-methoxy-4-(prop-2-yn-1-ylamino)benzoate: Iodoethane (570.02 mg, 3.65 mmol, 292.32 μL, 1.5 equivalent) and K₂CO₃ (1.01 g, 7.31 mmol, 3 equivalent) were added to a solution of 3-methoxy-4-(prop-2-yn-1-ylamino)benzoic acid (500 mg, 2.44 mmol, 1 equivalent) in DMF (6 mL). The mixture was stirred at 50 °C for 1 hour. TLC analysis indicated that the reaction was complete. The reaction mixture was quenched by adding water (80 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with brine (50 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The desired product (540 mg, crude) was given as a yellow solid, which was used without purification.

[0836] Example A50: Synthesis of 3-hydroxy-4-(prop-2-yn-1-ylamino)benzenesulfonamide.

[0837]

[0838] Under N2, at 0 °C, boron tribromide (886.24 mg, 3.54 mmol, 340.86 μL, 5 equivalents) was added to a solution of 3-methoxy-4-(prop-2-yn-1-ylamino)benzenesulfonamide (0.2 g, 707.51 μmol, 1 equivalent) in DCM (2 mL). The mixture was stirred at 0–25 °C for 1 h. TLC analysis indicated that the reaction was complete. The reaction mixture was quenched at 0 °C by adding water (100 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was purified by preparative TLC (SiO2, PE:EtOAc = 1:3) to give the desired product (0.12 g, 424.31 μmol, 59.97% yield) as a yellow oil.

[0839] B. Compounds having a 2-ethynyl-N-(alkyl)-1H-indole-4-amine core.

[0840] Example B1: Synthesis of compounds 6A, 7A, 8A and 9A.

[0841]

[0842] Preparation of 1-(2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)-3-(1-methylpiperidin-4-yl)urea and N-(2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)-4-methylpiperazin-1-carboxamide: Triphosgene (441.07 μmol, 1 equivalent) was added to a solution of 2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-amine (1 equivalent) and DIPEA (12 equivalents) in DCM (6 mL). The mixture was stirred at 0 °C for 0.5 h. Then, 1-methylpiperazine or 1-methylpiperidin-4-amine (1.2 equivalents) was added to the mixture, and the resulting mixture was stirred at 0 °C for another 0.5 h. The reaction mixture was poured into a saturated aqueous solution of Na₂CO₃ (20 mL) and extracted with DCM (20 mL x 3). The combined organic layers were concentrated under reduced pressure to obtain a residue. The crude residue was dissolved in toluene and concentrated (10 mL x 2) to give the desired product.

[0843] Preparation of N-[2-iodo-1-(2,2,2-trifluoroethyl)indol-4-yl]-4-methylpiperazine-1-carboxamide: Triphosgene (131 mg, 441.07 μmol, 1 equivalent) was added to a solution of 2-iodo-1-(2,2,2-trifluoroethyl)indol-4-amine (150 mg, 441.07 μmol, 1 equivalent) and DIPEA (684 mg, 5.29 mmol, 921.92 μL, 12 equivalent) in DCM (2 mL). The mixture was stirred at 0 °C for 0.5 h. 1-Methylpiperazine (53 mg, 529.28 μmol, 59 μL, 1.2 equivalent) was added to the mixture and the mixture was stirred at 0 °C for 0.5 h. TLC analysis (DCM:MeOH = 20:1, R f =0.2) indicates that the starting material was completely consumed and a new spot was detected. The reaction mixture was poured into a saturated Na2CO3 aqueous solution (20 mL) and extracted with DCM (20 mL x 3). The combined organic layers were concentrated under reduced pressure to give the residue. The residue was diluted with toluene (10 mL) and the mixture was concentrated under reduced pressure to give the residue. After repeating the toluene dilution step twice, the residue was diluted with toluene (10 mL) and filtered to give the desired product. N-[2-iodo-1-(2,2,2-trifluoroethyl)indol-4-yl]-4-methyl-piperazin-1-carboxamide (150 mg, 294.06 μmol, 66.67% yield). LC-MS (ES+, m / z): 467.0 [(M+H)] + ].

[0844] Preparation of the final product: Add i-Pr₂NH (10-30 equivalents), CuI (1-2 equivalents), 1-(2-iodo-1-(2-trifluoroethyl)-1H-indol-4-yl)-3-(1-methylpiperidin-4-yl)urea or N-(2-iodo-1-(2-trifluoroethyl)-1H-indol-4-yl)-4-methylpiperazin-1-carboxamide (1 equivalent) and Pd(PPh₃)₄ (0.20-0.50 equivalents) to a mixture of 3-methoxy-4-(prop-2-yn-1-ylamino)benzamide or 2-methoxy-4-(methanesulfonyl)-N-(prop-2-yn-1-yl)aniline (1-2 equivalents) in DMSO (2 mL). Stir the mixture under N₂ at 20-40 °C for 1-3 hours. LC-MS or TLC analysis confirmed the completion of the reaction. The mixture was poured into a saturated EDTA solution (20 mL), stirred for 1 hour, and the aqueous phase was extracted with EtOAc (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by TLC, preparative-TLC, or preparative-HPLC to obtain the desired product.

[0845] 3-Methoxy-4-{[3-(4-{[(1-methylpiperidin-4-yl)carbamoyl]amino}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}benzamide, MS(ES) + MS(ES) : 557.1; 3-(2-{3-[(4-methanesulfonyl-2-methoxyphenyl)amino]prop-1-yn-1-yl}-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)-1-(1-methylpiperidin-4-yl)urea, MS(ES) + ,m / z):592.1;N-(2-{3-[(4-carbamoyl-2-methoxyphenyl)amino]prop-1-yn-1-yl}-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)-4-methylpiperazin-1-carboxamide, MS(ES + ,m / z):543.1; and N-(2-{3-[(4-methanesulfonyl-2-methoxyphenyl)amino]prop-1-yn-1-yl}-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)-4-methylpiperazin-1-carboxamide, MS(ES + ,m / z):578.3.

[0846] Example B2: Synthesis of compounds 13A, 15A, 16A and 17A.

[0847]

[0848] Preparation of tert-butyl piperidine-1-carboxylate 1-(2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)ureoyl)piperidin-1-carboxylate: Triphosgene (698.07 mg, 2.35 mmol, 1 equivalent) was added to a solution of 2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-amine (800 mg, 2.35 mmol) and DIPEA (3.65 g, 28.23 mmol, 4.92 mL, 12 equivalents) in DCM (10 mL). The mixture was then stirred at 0 °C for 0.5 h. 4-Aminopiperidine-1-carboxylic acid tert-butyl ester (565.35 mg, 2.82 mmol, 1.2 equivalents) was added to the mixture, and the resulting mixture was stirred at 0 °C for 0.5 hours. TLC analysis (PE:EtOAc = 1:1, Rm) was performed. f=0.16) indicates that the starting material was completely consumed. A saturated NaHCO3 solution (30 mL) was added to the reaction mixture, and the aqueous phase was extracted with EtOAc (20 mL x 3). The combined organic layers were concentrated under vacuum to obtain the crude product. MS (ES) + ,m / z):566.8.

[0849] Preparation of 1-(2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)-3-(piperidin-4-yl)urea: TFA (4.59 g, 40.26 mmol, 2.98 mL, 30 equivalents) was added to a solution of 4-(3-(2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)urea (0.8 g, 1.34 mmol, 1 equivalent) in DCM (3 mL). The reaction mixture was stirred at 25 °C for 0.5 h. LC-MS analysis showed complete consumption of the starting material. The reaction mixture was washed with saturated NaHCO3 solution (30 mL) and extracted with EtOAc (5 mL x 3). The combined organic layers were washed with brine (80 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude product as a brown solid (350 mg, 55.9% yield). MS (ES) + ,m / z):467.0.

[0850] Preparation of 2-iodo-NC(O)R-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine: K₂CO₃ (59.29 mg, 428.96 μmol, 2 equivalents) was added to a mixture of 1-(2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-4-yl)-3-(piperidin-4-yl)urea (100 mg, 214.48 μmol, 1 equivalent) and R-Br (428.96 μmol, 2 equivalents) in DMF (3 mL). The mixture was stirred at 25 °C for 4 hours. LC-MS or TLC analysis indicated that the reaction was complete. The reaction mixture was quenched by adding water (40 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC to give the desired product as a brown oil.

[0851] 2-(4-(3-(2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)ureido)piperidin-1-yl)acetamide, 36% yield, MS (ES) +m / z): 524.0; 1-(1-(2-hydroxyethyl)piperidin-4-yl)-3-(2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-4-yl)urea, 34% yield, MS (ES) + m / z): 511.0; 1-(2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)-3-(1-(2-methoxyethyl)piperidin-4-yl)urea, MS(ES + m / z): 525.0; 1-(1-(2,3-dihydroxypropyl)piperidin-4-yl)-3-(2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-4-yl)urea, 37% yield, MS (ES) + ,m / z):541.0.

[0852] Preparation of 2-(3-((2-methoxy-4-(methanesulfonyl)phenyl)amino)prop-1-yn-1-yl)-NC(O)R-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine: 2-iodo-NC(O)R-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine (45 mg, 74.96 μmol, 1 equivalent), CuI (1 equivalent), N-isopropylprop-2-amine (1 equivalent), and Pd(PPh3)4 (0.02 equivalent) were added to a solution of 2-methoxy-4-(methanesulfonyl)-N-(prop-2-yn-1-yl)aniline (26.90 mg, 112.44 μmol, 1.5 equivalent) in DMSO (3 mL). The mixture was stirred at 45 °C for 1 hour. LC-MS or TLC analysis indicated that the reaction was complete. The reaction mixture was quenched at 25°C by adding saturated EDTA solution (40 mL), and the mixture was extracted with EtOAc (20 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the residue. The residue was analyzed by preparative TLC (SiO2, DCM:MeOH = 10:1, R... f =0.24) and preparation-HPLC purification yielded the desired product, a pale yellow solid.

[0853] 2-(4-{[(2-{3-[(4-methanesulfonyl-2-methoxyphenyl)amino]prop-1-yn-1-yl}-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)carbamoyl]amino}piperidin-1-yl)acetamide, MS(ES) + ,m / z):635.5; 1-[1-(2-hydroxyethyl)piperidin-4-yl]-3-(2-{3-[(4-methanesulfonyl-2-methoxyphenyl)amino]prop-1-yn-1-yl}-1-(2,2,2-trifluoroethyl)-1H-indole-4-yl)urea, MS(ES +,m / z):622.3; 3-(2-{3-[(4-methanesulfonyl-2-methoxyphenyl)amino]prop-1-yn-1-yl}-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)-1-[1-(2-methoxyethyl)piperidin-4-yl]urea, MS(ES + ,m / z):636.1; and 1-[1-(2,3-dihydroxypropyl)piperidin-4-yl]-3-(2-{3-[(4-methanesulfonyl-2-methoxyphenyl)amino]prop-1-yn-1-yl}-1-(2,2,2-trifluoroethyl)-1H-indole-4-yl)urea, MS(ES + ,m / z):652.2.

[0854] Example B3: Synthesis of 3-(2-{3-[(4-methanesulfonyl-2-methoxyphenyl)amino]prop-1-yn-1-yl}-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)-1-[1-(oxan-4-yl)piperidin-4-yl]urea (compound 14A).

[0855]

[0856] Preparation of 1-(2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)-3-(1-(tetrahydro-2H-pyran-4-yl)piperidin-4-yl)urea: 1-(2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)-3-(piperidin-4-yl)urea (107.37 mg, 1.07 mmol, 98.50 μL, 5 oz) was prepared. A mixture of tetrahydro-4H-pyran-4-one (100 mg, 214.48 μmol, 1 equivalent), AcOH (12.88 μg, 2.14e-1 μmol, 1.23e-2 μL, 0.001 equivalent) and NaBH3CN (26.96 mg, 428.96 μmol, 2 equivalent) in MeOH (2 mL) was stirred at 25 °C for 2 hours. The reaction was confirmed by LC-MS analysis. The reaction mixture was quenched by adding saturated NH4HCO3 solution (40 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the residue. The crude residue was subjected to preparative TLC (SiO2, DCM:MeOH = 10:1, R f =0.24) Purification yielded the desired product as a brown solid. 29% yield, MS (ES) + ,m / z):551.0.

[0857] Preparation of 3-(2-{3-[(4-methanesulfonyl-2-methoxyphenyl)amino]prop-1-yn-1-yl}-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)-1-[1-(oxan-4-yl)piperidin-4-yl]urea: 1-(2-iod-1-yl)-[2-iod-1-yl]urea was added to a solution of 2-methoxy-4-(methanesulfonyl)-N-(prop-2-yn-1-yl)aniline (22.17 mg, 92.67 μmol, 1.5 equivalents) in DMSO (3 mL). (2,2,2-trifluoroethyl)-1H-indol-4-yl)-3-(1-(tetrahydro-2H-pyran-4-yl)piperidin-4-yl)urea (40 mg, 61.78 μmol, 1 equivalent), CuI (11.77 mg, 61.78 μmol, 1 equivalent), N-isopropylpropyl-2-amine (6.25 mg, 61.78 μmol, 8.73 μL, 1 equivalent), and Pd(PPh3)4 (1.43 mg, 1.24 μmol, 0.02 equivalent). The mixture was stirred at 45 °C for 1 hour. LC-MS analysis indicated that the reaction was complete. The reaction mixture was quenched at 25 °C by adding saturated EDTA solution (40 mL), and the mixture was extracted with EtOAc (20 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by preparative HPLC to give the desired product (17.2 mg, 25.89 μmol, 41.91% yield) as a pale yellow solid. 3-(2-{3-[(4-methanesulfonyl-2-methoxyphenyl)amino]prop-1-yn-1-yl}-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)-1-[1-(oxan-4-yl)piperidin-4-yl]urea, MS (ES) + ,m / z):662.3.

[0858] Example B4: Synthesis of compounds 10A, 11A and 12A.

[0859]

[0860] The general procedure for the preparation of 1-(4-(dimethylamino)cyclohexyl)-3-(2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-4-yl)urea and 1-(2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-4-yl)-3-(pyridin-4-yl)urea is as follows: Triphosgene (1 equivalent) is added to a solution of 2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine (1 equivalent) and DIPEA (12 equivalents) in DCM. The mixture is stirred at 0°C for 0.5 hours, and N... 1 N 1Dimethylcyclohexane-1,4-diamine or pyridine-4-amine (1.2 equivalents) was added to the reaction mixture. The resulting reaction mixture was further stirred at 0°C for 0.5 hours. TLC analysis showed that the starting material was completely consumed. The reaction mixture was quenched by adding saturated Na₂CO₃ solution. The reaction mixture was separated into layers by adding EtOAc, and the aqueous phase was extracted with EtOAc (x³). The organic phase was washed with brine (x³), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product. The residue was purified by preparative TLC to give the desired product, a brown solid.

[0861] Preparation of 1-(4-(dimethylamino)cyclohexyl)-3-(2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)urea and 1-(2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)-3-(pyridin-4-yl)urea: 2-methoxy-4-(methanesulfonyl)-N-(prop-2-yn-1-yl)aniline (1.2 equivalents) was reacted with DMS at 25 °C. The solution of O was supplemented with i-Pr₂NH (10 equivalents), CuI (1 equivalent), 1-(4-(dimethylamino)cyclohexyl)-3-(2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)urea or 1-(2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)-3-(pyridin-4-yl)urea (1 equivalent), and Pd(PPh₃)₄ (0.2 equivalents). The mixture was stirred at 25°C for 1–2 hours under N₂. LC-MS and TLC analyses indicated that the reaction was complete. The reaction mixture was quenched by adding saturated EDTA solution and stirred at 25°C for 2 hours. The reaction mixture was separated into layers by adding EtOAc, and the aqueous phase was extracted with EtOAc. The organic phase was washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by preparative TLC and preparative HPLC to obtain a solution of the desired product. The solution was freeze-dried to obtain the desired product, which was a yellow solid.

[0862] 3-(2-{3-[(4-methanesulfonyl-2-methoxyphenyl)amino]prop-1-yn-1-yl}-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)-1-[(1R,4R)-4-(dimethylamino)cyclohexyl]urea, MS(ES) + ,m / z):620.3; 3-(2-{3-[(4-methanesulfonyl-2-methoxyphenyl)amino]prop-1-yn-1-yl}-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)-1-[(1S,4S)-4-(dimethylamino)cyclohexyl]urea, MS(ES +,m / z):620.3; and 1-(2-(3-((2-methoxy-4-(methylsulfonyl)phenyl)amino)prop-1-yn-1-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)-3-(pyridin-4-yl)urea, MS(ES + ,m / z):572.3.

[0863] Example B5: Preparation of compound 2-{3-[(4-methanesulfonyl-2-methoxyphenyl)amino]prop-1-yn-1-yl}-N,N-dimethyl-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine (compound 1A).

[0864]

[0865] Preparation of tert-butyl (3-(4-(dimethylamino)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl)(2-methoxy-4-(methylsulfonyl)phenyl)carbamate: Brettphos was added to a mixture of tert-butyl (3-(4-bromo-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl)(2-methoxy-4-(methylsulfonyl)phenyl)carbamate (150 mg, 238.8 μmol, 1 equivalent) and dimethylamine hydrochloride (31 mg, 262.73 μmol, 1.1 equivalent) in DMF (2 mL). Pd(G4) (15 mg, 16.72 μmol, 0.07 equivalents), Cs2CO3 (233 mg, 716.54 μmol, 3 equivalents), and RuPhos (15 mg, 33.44 μmol, 0.14 equivalents). The reaction mixture was degassed and purged with N2. The mixture was heated to 90 °C and stirred for 2 hours, followed by TLC (PE:EtOAc = 1:1, R f =0.45) and LC-MS analysis indicated that the reaction was complete. The mixture was poured into a saturated EDTA solution (10 mL) and stirred for 1 hour. The mixture was extracted with EtOAc (20 mL x 3), and the combined organic layers were washed with brine (10 mL), dried over anhydrous sodium sulfate, and concentrated. The crude residue was purified by column chromatography (SiO2, PE:EtOAc = 8:1 to 3:1) to give tert-butyl (3-(4-(dimethylamino)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl)(2-methoxy-4-(methanesulfonyl)phenyl)carbamate (100 mg, 141.47 μmol, 72% yield).

[0866] Preparation of the final product: HCl / EtOAc (4M, 10mL, 231.85 equivalents) was added to a solution of (3-(4-(dimethylamino)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl)(2-methoxy-4-(methanesulfonyl)phenyl)carbamate (0.1 g, 172.52 μmol, 1 equivalent) in EtOAc (2 mL). The mixture was stirred at 25 °C for 1 hour, followed by TLC analysis (PE:EtOAc = 1:1, R0). f =0.3) indicates that the reaction is complete. The reaction mixture was filtered and concentrated under vacuum. The residue was purified by preparative HPLC to give 2-{3-[(4-methanesulfonyl-2-methoxyphenyl)amino]prop-1-yn-1-yl}-N,N-dimethyl-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine (27.6 mg, 57.56 μmol, 33.36% yield) as a pale yellow solid.

[0867] Example B6: Preparation of compound 4-[(3-{4-[(1,5-dihydroxypent-3-yl)amino]-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]-3-methoxybenzene-1-sulfonamide (compound 4A).

[0868]

[0869] Synthesis of 3-((2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-4-yl)amino)pentan-1,5-diol: 1,5-dihydroxypentan-3-one (69.47 mg, 588.09 μmol, 40.50 μL, 2 equivalents) and SnCl₂-2H₂O (13.27 mg, 58.81 μmol, 4.90 μL, 0.20 equivalents) were added to a mixture of 2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine (0.1 g, 294.05 μmol, 1 equivalent) in MeOH (5 mL), followed by the addition of PMHS (70.57 mg, 1.18 mmol, 4 equivalents). The resulting mixture was stirred at 70 °C for 3 h, and subsequent LC-MS analysis showed that a substance of the desired quality had been formed. The mixture was concentrated under reduced pressure to obtain a crude residue, which was then subjected to preparative TLC (SiO2, EtOAc:PE = 2:1, R...). f =0.16) Purification. 3-((2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino)pentane-1,5-diol (0.05 g, 96.11 μmol, 32.68% yield) was obtained as a yellow oil. MS (ES) + ,m / z):443.1.

[0870] Synthesis of the final product: CuI (18.30 mg, 96.11 μmol, 1 equivalent) was added to a mixture of 3-methoxy-4-(prop-2-yn-1-ylamino)benzenesulfonamide (40.75 mg, 144.16 μmol, 1.5 equivalent) and 3-((2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino)pent-1,5-diol (0.05 g, 96.11 μmol, 1 equivalent) in DMSO (2 mL), followed by Pd(PPh3)4 (11.11 mg, 9.61 μmol, 0.10 equivalent) and N-isopropylpropyl-2-amine (9.73 mg, 96.11 μmol, 13.58 μL, 1 equivalent). The reaction mixture was stirred at 30°C for 1 hour under N2, followed by TLC analysis (PE:EtOAc = 1:2, Rm). f =0.30) indicates the reaction is complete. The reaction mixture was quenched by adding saturated EDTA solution (30 mL). EtOAc (10 mL) was added, and the resulting mixture was stirred at 25 °C for 1 hour, and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was subjected to preparative TLC (SiO2, PE:EtOAc = 1:2, R f =0.30) Purified and further purified by preparative HPLC, yielding 4-[(3-{4-[(1,5-dihydroxypentan-3-yl)amino]-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]-3-methoxybenzene-1-sulfonamide (0.007 g, 12.47 μmol, 12.98% yield), as a white solid. MS (ES) + ,m / z):555.2.

[0871] Example B7: Synthesis of 2-[(2-{3-[(2-methoxy-4-aminosulfonylphenyl)amino]prop-1-yn-1-yl}-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino]acetamide (compound 3A).

[0872]

[0873] Synthesis of ethyl (2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-4-yl)glycine: Under N2 at 25 °C, DIPEA (1.28 mL, 4.41 mmol, 3 equivalents) was added in a single batch to a mixture of 2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine (0.5 g, 1.47 mmol, 1 equivalent) and ethyl 2-bromoacetate (2.46 g, 14.70 mmol, 1.63 mL, 10 equivalents) in 5 mL of THF. The mixture was stirred at 25 °C for 96 hours, and subsequent TLC analysis (PE:EtOAc = 5:1, R f =0.4) indicates that the reaction is complete. The mixture was concentrated under vacuum at 45 °C, and the residue was purified by silica gel chromatography (SiO2, PE:EtOAc = 100:1 to 5:1) to give (2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)glycine ethyl ester (400 mg, 844.73 μmol, 57.46% yield) as a yellow solid.

[0874] Synthesis of (2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)glycine: Under N2 at 25 °C, ethyl ester of (2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)glycine (400 mg, 938.59 μmol, 1 equivalent) in a mixture of MeOH (3 mL), THF (9 mL), and water (3 mL) was added in one step to LiOH·H2O (196.93 mg, 4.69 mmol, 5 equivalents). The mixture was stirred at 25 °C for 12 h, and subsequent LC-MS analysis indicated that the reaction was complete. The mixture was concentrated under reduced pressure at 40 °C, and the residue was decanted into water (10 mL) and stirred for 1 min. The aqueous phase was extracted with EtOAc (10 mL x 2), the pH was adjusted to 2 by adding 2N HCl, and the mixture was extracted again with EtOAc (10 mL x 2). The combined organic layers were washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain (2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-4-yl)glycine (280 mg, 668.14 μmol, 71.19% yield), as a yellow solid.

[0875] Synthesis of 2-((2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino)acetamide: Under N2, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI) (115.56 mg, 602.84 μmol, 2 equivalents), HOBt (81.46 mg, 602.84 μmol, 2 equivalents), DIPEA (155.82 mg, 1.21 mmol, 210.01 μL, 4 equivalents) and NH4Cl (32.25 mg, 602.84 μmol, 21.08 μL, 2 equivalents) were added to a mixture of (2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)glycine (120 mg, 301.42 μmol, 1 equivalent) in DMF (5 mL). The mixture was stirred at 25 °C for 12 hours, and subsequent LC-MS analysis indicated that the reaction was complete. The mixture was poured into water (30 mL) and stirred for 2 minutes. The aqueous phase was extracted with EtOAc (20 mL x 5). The combined organic layers were washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was washed with DCM (10 mL x 3), filtered, and concentrated under vacuum to give 2-((2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino)acetamide (70 mg, 96.66 μmol, 32.07% yield) as a pale yellow solid.

[0876] Preparation of the final product: Under N2, i-Pr2NH (127.40 mg, 1.26 mmol, 177.93 μL, 10 equivalents), CuI (23.98 mg, 125.90 μmol, 1 equivalent), 2-((2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino)acetamide (50 mg, 125.90 μmol, 1 equivalent), and Pd(PPh3)4 (29.10 mg, 25.18 μmol, 0.2 equivalents) were added to a mixture of 3-methoxy-4-(prop-2-yn-1-ylamino)benzenesulfonamide (36.30 mg, 151.08 μmol, 1.2 equivalents) in DMSO (2 mL) were added. The mixture was stirred at 45 °C for 1 h, followed by LC-MS and TLC analysis (PE:EtOAc = 0:1, R f=0.32) indicates the reaction is complete. Add EtOAc (10 mL) and pour the mixture into a saturated EDTA solution (40 mL), stirring for 15 minutes. Extract the aqueous phase with EtOAc (40 mL x 2), and pour the organic layer back into a saturated EDTA solution (40 mL), stirring for 1 hour. Extract the aqueous phase again with EtOAc (40 mL x 3). Wash the combined organic layers with brine (40 mL x 3), dry with anhydrous sodium sulfate, treat with activated carbon, filter, and concentrate under vacuum. Analyze the residue by preparative TLC (PE:EtOAc = 0:1, R f =0.32) was purified, and further purified by preparative HPLC to give 2-[(2-{3-[(2-methoxy-4-aminosulfonylphenyl)amino]prop-1-yn-1-yl}-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino]acetamide (5.1 mg, 9.04 μmol, 7.18% yield), as a pale yellow solid. MS (ES) + ,m / z):510.1.

[0877] Table 2 shows compounds having a 2-ethynyl-N-(alkyl)-1H-indole-4-amine core.

[0878] Table 2

[0879]

[0880]

[0881]

[0882]

[0883]

[0884] C. Compounds having a 2-ethynyl-N-(cycloalkyl)-1H-indole-4-amine core.

[0885] Example C1: Synthesis of compounds 31A, 32A, 33A, 34A and 35A.

[0886]

[0887] The general procedure for the preparation of 4-((2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino)-1-methylcyclohexane-1-ol and 4-((2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino)cyclohexane-1-carboxylonitrile is as follows: 4-hydroxy-4-methylcyclohexane-1-one or 4-oxocyclohexane-1-carboxylonitrile (5 equivalents) and Ti(OEt)4 (5 equivalents) are added to a solution of 2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-amine (1 equivalent) in EtOH. The reaction mixture is stirred at 50 °C for 3–5 hours. Then, NaBH3CN (5 equivalents) is added to the reaction mixture at 0 °C under N2, and the mixture is stirred for 5 minutes. The reaction mixture is then heated to 50 °C and stirred further for 1 hour. TLC analysis showed that the starting material was completely consumed. The solution was dried under vacuum to obtain a crude residue, which was purified by column chromatography or preparative TLC to give 4-((2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino)-1-methylcyclohexane-1-ol and 4-((2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino)cyclohexane-1-carboxynitrile, as a yellow or brown oil.

[0888] Preparation of the final product: At 25 °C, i-Pr₂NH (10 equivalents), CuI (1 equivalent), 4-((2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino)-1-methylcyclohexane-1-ol or 4-((2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino)cyclohexane-1-nitrile (1 equivalent) and Pd(PPh₃)₄ (0.2 equivalents) were added to a mixture of 3-methoxy-N-methyl-4-(prop-2-yn-1-yl)amino)cyclohexane-1-nitrile (1 equivalent) and Pd(PPh₃)₄ (0.2 equivalents). The mixture was stirred under N₂ for 1–2 hours. LC-MS and TLC analyses indicated that the reaction was complete. The reaction mixture was quenched at 25 °C by adding a saturated EDTA solution and stirred for 2 hours. The reaction mixture was separated into layers by adding EtOAc and the aqueous phase was extracted with EtOAc. The organic phase was washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The crude residue was purified by column chromatography or preparative TLC, followed by further purification by preparative HPLC to obtain a solution of the desired product. The solution was lyophilized to obtain the desired product as a yellow solid.

[0889] 3-Methoxy-N-methyl-4-{[3-(4-{[(1R,4R)-4-hydroxy-4-methylcyclohexyl]amino}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}benzamide, MS(ES) +,m / z):543.2; 3-methoxy-N-methyl-4-{[3-(4-{[(1S,4S)-4-hydroxy-4-methylcyclohexyl]amino}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}benzamide, MS(ES + ,m / z):543.2; 4-[(3-{4-[(4-cyanocyclohexyl)amino]-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]-3-methoxy-N-methylbenzamide, MS(ES + ,m / z):538.2; 3-Methoxy-N-methyl-4-{[3-(4-{[(1R,4R)-4-cyanocyclohexyl]amino}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}benzamide, MS(ES + ,m / z):538.2; and 3-methoxy-N-methyl-4-{[3-(4-{[(1S,4S)-4-cyanocyclohexyl]amino}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}benzamide, MS(ES + ,m / z):538.2.

[0890] Example C2: Synthesis of compounds 279A and 280A.

[0891]

[0892] To a solution of 3-methoxy-4-(prop-2-yn-1-ylamino)benzoic acid (52.68 mg, 231.06 μmol, 1.5 equivalents) in DMSO (3 mL), N-(4-(2-oxa-6-azaspiro[3.3]hept-6-yl)cyclohexyl)-2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine (100 mg, 154.04 μmol, 1 equivalent), CuI (29.34 mg, 154.04 μmol, 1 equivalent), N-isopropylpropyl-2-amine (15.59 mg, 154.04 μmol, 21.77 μL, 1 equivalent) and Pd(PPh3)4 (3.56 mg, 3.08 μmol, 0.02 equivalents) were added. The resulting mixture was stirred at 45 °C for 1 hour. TLC analysis (DCM:MeOH = 10:1, R f=0.24) indicates the reaction is complete. The reaction mixture was quenched at 25°C with 40 mL of saturated EDTA solution and extracted with EtOAc (20 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was purified by preparative TLC (SiO2, DCM:MeOH = 10:1) and preparative HPLC to give the desired product as a yellow solid.

[0893] 4-((3-(4-(((1R,4R)-4-(2-oxa-6-azaspiro[3.3]hept-6-yl)cyclohexyl)amino)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl)amino)-3-methoxybenzoic acid (26.6 mg, 44.27 μmol, 28.74% yield), MS (ES) + ,m / z):597.2; and 4-((3-(4-(((1S,4S)-4-(2-oxa-6-azaspiro[3.3]hept-6-yl)cyclohexyl)amino)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl)amino)-3-methoxybenzoic acid (26.3 mg, 40.51 μmol, 26.30% yield), MS (ES + ,m / z):597.2.

[0894] Example C3: Synthesis of 3-methoxy-N-methyl-4-{[3-(4-{[(1S,4S)-4-aminocyclohexyl]amino}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}benzamide (compound 25A).

[0895]

[0896] Preparation of tert-butyl 4-((2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-4-yl)amino)cyclohexyl)carbamate: Tert-butyl 4-(4-oxocyclohexyl)carbamate (940.68 mg, 4.41 mmol, 940.68 μL, 5 equivalents) and Ti(OEt)4 (1.01 g, 4.41 mmol, 914.66 μL, 5 equivalents) were added to a solution of 2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine (300 mg, 882.14 μmol, 1 equivalent) in EtOH (3 mL). The reaction mixture was stirred at 50 °C for 3 hours. Then, NaBH3CN (184.78 mg, 2.94 mmol, 5 equivalents) was added to the reaction mixture at 0 °C under N2, and the reaction mixture was stirred further for 5 minutes. The reaction mixture was heated to 50°C and stirred for another 1 hour. TLC analysis showed that the starting material was completely consumed. The solution was dried under vacuum, and the crude residue was purified by column chromatography or preparative TLC to give the desired product (300 mg, crude) as a yellow oil or solid. MS (ES) + ,m / z):552.1.

[0897] Preparation of tert-butyl (4-((2-(3-((2-methoxy-4-(methylcarbamoyl)phenyl)amino)prop-1-yn-1-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino)cyclohexyl)carbamate: Under N2, 3-methoxy-N-methyl-4-(prop-2-yn-1-ylamino)benzamide (73.11 mg, 334.97 μmol, 2 equivalents) and (4-((2-iodo-1-(2,2,2-trifluoro))carbamate were reacted with tert-butyl (2-((2-iodo-1-(2,2,2-trifluoro)) tert-butyl carbamate (100 mg, 167.49 μmol, 1 equivalent) was added to a mixture of ethyl(1H-indol-4-yl)amino(cyclohexyl)carbamate (tert-butyl) ester (100 mg, 167.49 μmol, 1 equivalent) in DMSO (3 mL), along with i-Pr₂NH (16.95 mg, 167.49 μmol, 23.67 μL, 1 equivalent), Pd(PPh₃)₄ (3.87 mg, 3.35 μmol, 0.02 equivalent) and CuI (31.90 mg, 167.49 μmol, 1 equivalent). The reaction mixture was stirred at 45 °C for 1 hour. TLC analysis indicated that the reaction was complete. The reaction mixture was quenched at 25 °C by adding saturated EDTA solution (40 mL), and the mixture was extracted with EtOAc (20 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the residue. The crude residue was purified by preparative TLC to obtain (4-((2-(3-((2-methoxy-4-(methylcarbamoyl)phenyl)amino)prop-1-yn-1-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino)cyclohexyl)tert-butyl carbamate (70% yield), as a pale yellow solid.

[0898] Preparation of 3-methoxy-N-methyl-4-{[3-(4-{[(1S,4S)-4-aminocyclohexyl]amino}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}benzamide: A solution of tert-butyl carbamate (80 mg, 114.71 μmol, 1 equivalent) in a 1:1 mixture of DCM (0.5 mL) and TFA (0.5 mL) was stirred at 25 °C for 1 hour. TLC analysis indicated that the reaction was complete. The reaction mixture was concentrated under reduced pressure and purified by preparative TLC and preparative HPLC to give the desired product (13.4 mg, 24.51 μmol, 21.37% yield) as a pale yellow solid. MS (ES) + ,m / z):528.2.

[0899] Example C4: Synthesis of compounds 269A, 270A, 396A and 397A.

[0900]

[0901] General procedure: At 25°C, i-Pr₂NH (10 equivalents), CuI (0.5 equivalents), N-(4-(2-oxa-6-azaspiro[3.3]hept-6-yl)cyclohexyl)-2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine or N-(4-(2-oxa-7-azaspiro[3.5]non-7-yl)cyclohexyl)-2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine (1 equivalent) and Pd(PPh₃)₄ (0.20 equivalents) were added to a mixture of 4-(ethylsulfonyl)-2-methoxy-N-(prop-2-yn-1-yl)aniline (1.3 equivalents) in DMSO. The mixture was stirred for 1 hour. LC-MS analysis indicated that the reaction was complete. EtOAc was poured into the reaction vessel, and the resulting mixture was then poured into a saturated EDTA solution and stirred for 1 hour. The aqueous phase was extracted with EtOAc (3x). The combined organic layers were washed with brine (3x), dried over anhydrous sodium sulfate, mixed with activated carbon to remove color, and concentrated under vacuum. The crude residue was purified by preparative TLC and preparative HPLC to obtain the desired product as a pale yellow solid.

[0902] N-((1R,4R)-4-(2-oxa-6-azaspiro[3.3]hept-6-yl)cyclohexyl)-2-(3-((4-(ethylsulfonyl)-2-methoxyphenyl)amino)prop-1-yn-1-yl)-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine, MS(ES) + ,m / z):645.2;N-((1S,4S)-4-(2-oxa-6-azaspiro[3.3]hept-6-yl)cyclohexyl)-2-(3-((4-(ethylsulfonyl)-2-methoxyphenyl)amino)prop-1-yn-1-yl)-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine,MS(ES + ,m / z):645.2;N-((1R,4R)-4-(2-oxa-7-azaspiro[3.5]non-7-yl)cyclohexyl)-2-(3-((4-(ethylsulfonyl)-2-methoxyphenyl)amino)prop-1-yn-1-yl)-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine,MS(ES +,m / z):673.2; and N-((1S,4S)-4-(2-oxa-7-azaspiro[3.5]non-7-yl)cyclohexyl)-2-(3-((4-(ethylsulfonyl)-2-methoxyphenyl)amino)prop-1-yn-1-yl)-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine, MS(ES + ,m / z):673.2.

[0903] Example C5: Synthesis of 3-methoxy-4-{[3-(4-{[(1R,4R)-4-(pyrrolidin-1-yl)cyclohexyl]amino}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}benzene-1-sulfonamide (compound 422A).

[0904]

[0905] The general procedure for the preparation of 2-iodo-N-(4-(pyrrolidin-1-yl)cyclohexyl)-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine is as follows: Pyrrolidine (5 equivalents) and Ti(OEt)4 (5 equivalents) are added to a solution of 4-((2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-4-yl)amino)cyclohexane-1-one (1 equivalent) in EtOH. The reaction mixture is stirred at 50 °C for 3 hours. Then, NaBH3CN (5 equivalents) is added to the reaction mixture at 0 °C under N2, and the resulting mixture is stirred for 5 minutes. The reaction mixture is heated to 50 °C and stirred for 1 hour. TLC and LC-MS analysis indicate that the starting material is completely consumed. The solution was dried under vacuum, and the crude residue was purified by column chromatography (SiO2) to obtain 2-iodo-N-(4-(pyrrolidine-1-yl)cyclohexyl)-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine.

[0906] The general procedure for the preparation of 3-methoxy-4-((3-(4-(((1R,4R)-4-(pyrrolidine-1-yl)cyclohexyl)amino)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl)amino)benzenesulfonamide is as follows: At 25–45 °C, i-Pr₂NH (10 equivalents), CuI (1 equivalent), 2-iodo-N-(4-(pyrrolidine-1-yl)cyclohexyl)-1-(2,2,2-trifluoroethyl)-1H-indol-4-amine (1 equivalent) and Pd(PPh₃)₄ (0.2 equivalents) are added to a mixture of 3-methoxy-4-(prop-2-yn-1-yl)amino)benzenesulfonamide (1.2 equivalents) in DMSO. The mixture is stirred under N₂ for 1 hour. LC-MS and TLC analyses indicate that the reaction is complete. The reaction mixture was quenched by adding a saturated EDTA solution and stirred at 25°C for 2 hours. EtOAc was added to the reaction mixture, and the aqueous phase was extracted with EtOAc. The organic phase was washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product. The crude residue was purified by preparative TLC and preparative HPLC to give a solution of the desired product. The solution was lyophilized to give 3-methoxy-4-{[3-(4-{[(1R,4R)-4-(pyrrolidin-1-yl)cyclohexyl]amino}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}benzene-1-sulfonamide. MS(ES) + ,m / z):604.2.

[0907] Example C6: Synthesis of compounds 331A, 332A, 333A and 334A.

[0908]

[0909] At 25 °C, i-Pr2NH (9.98 mg, 98.65 μmol, 13.94 μL, 1 equivalent), CuI (626.26 mg, 3.29 mmol, 2 equivalent), and N-(4-(7-oxa-2-azaspiro[3]) were added to a solution of 3-methoxy-4-(prop-2-yn-1-ylamino)benzamide (35.88 mg, 147.97 μmol, 1.5 equivalent) in DMSO (3 mL). [3.5]Non-2-yl)cyclohexyl)-2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine or N-(4-(6-oxa-2-azaspiro[3.5]non-2-yl)cyclohexyl)-2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine (60 mg, 98.65 μmol, 1 equivalent) and Pd(PPh3)4 (2.28 mg, 1.97 μmol, 0.02 equivalent). The mixture was stirred at 25 °C for 1 hour under N2. The mixture was poured into a saturated EDTA solution (20 mL), stirred at 25 °C for 1 hour, and extracted with EtOAc (20 mL x 3). The combined organic layers were concentrated under reduced pressure to obtain the residue, which was purified by preparative TLC and preparative HPLC to obtain the desired product.

[0910] 3-Methoxy-4-{[3-(4-{[(1R,4R)-4-{7-oxa-2-azaspiro[3.5]non-2-yl}cyclohexyl]amino}-1-(2,2,2-trifluoroethyl)-1H-indol...

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1. A compound, wherein the compound is: Or, or a pharmaceutically acceptable salt thereof.

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