hnRNPA2B1 agonists and their combination with chemotherapy agents in tumor prevention and treatment

Abstract

This disclosure relates to heterogeneous nuclear ribonucleic acid A2B1 (hnRNPA2B1) agonist compounds and their use in combination with chemotherapeutic agents for antitumor treatment. Specifically, this disclosure relates to fused ring compounds of formula (I), their cis-trans isomers, their enantiomers, their diastereomers, their racemic mixtures, their solvates, their hydrates, or their pharmaceutically acceptable salts or prodrugs thereof, compositions comprising any of the former, and products thereof, as well as the use of the aforementioned substances as hnRNPA2B1 agonists in the preparation of products for the prevention and / or treatment of tumors, said products may further comprise or be used in combination with chemotherapeutic agents. This disclosure has broad application prospects for the prevention and treatment of tumors.

Description

Technical Field

[0001] This disclosure relates to the fields of medicine and biotechnology. Specifically, this disclosure relates to a class of compounds that are agonists of the nucleoprotein hnRNPA2B1 (i.e., heterogeneous nuclear ribonucleoprotein A2B1), products containing said compounds, and their effects, mechanisms of action, methods of administration, and uses in combination with chemotherapeutic agents in the prevention or treatment of tumor-related diseases or symptoms, tumor-induced damage. Background Technology

[0002] Type I interferon plays a crucial role in the functional regulation and activation of various immune cells and is a key cytokine in regulating tumor immunity. Interferon can directly inhibit the proliferation of human tumor cells and can synergistically interact with various chemotherapeutic drugs. Type I interferon can significantly enhance anti-tumor immune responses by inducing and activating adaptive and innate immune cells, particularly by enhancing T cell activation through the uptake, processing, presentation, and cross-presentation of antigens to T cells by dendritic cells and macrophages. Simultaneously, interferon can also inhibit tumor invasion by regulating the expression of proteins related to tissue remodeling. Therefore, interferon and compounds that can induce interferon production have significant applications in cancer treatment.

[0003] Defects in the interferon activation pathway within tumor cells often lead to tumor immune escape and failure of immune checkpoint blockade therapy. Many small-molecule drugs targeting tumors exert their effects by inducing an increase in free intrinsic nucleic acids, thereby activating interferon. These include DNA intercalators, various epigenetic drugs, and splicesome modulators. Recent studies have shown that the cytoplasmic cGAS-STING pathway can induce type I interferon, making the development of chemotherapeutic agents an important direction in cancer treatment. However, increasing evidence suggests low expression of the cGAS-STING pathway or blocked signaling pathway activation in tumor cells. Furthermore, there is also evidence that cGAS can promote tumor cell survival. Therefore, there is an urgent need in the field of tumor immunotherapy to develop agonists targeting the innate immune receptors that can induce type I interferon.

[0004] Heterogeneous nuclear ribonucleoprotein A2B1 (hnRNP-A2B1) is a newly identified DNA recognition receptor in the cell nucleus, belonging to the hnRNP family. On one hand, hnRNP-A2B1 can sense and recognize the nucleic acid components of DNA viruses (such as HSV-1), and self-activate to form a homodimer. Demethylation is then mediated by the demethylase JMJD6, causing the receptor to translocate from the nucleus to the cytoplasm, thereby activating the TBK1-IRF3 signaling pathway and initiating type I interferon expression to exert its antiviral effect. On the other hand, as an RNA-binding protein, hnRNPA2B1 can also promote m6A modification, nucleoplasmic translocation, and translation on cGAS, IFI16, and STING mRNA, thus ensuring the full induction of type I interferon expression.

[0005] hnRNPA2B1 is overexpressed in various tumors, including lung cancer, liver cancer, breast cancer, pancreatic cancer, and glioblastoma, and is widely considered an important factor promoting tumorigenesis and development. In recent decades, numerous studies have shown that hnRNPA2B1 participates in regulating a variety of fundamental biological functions, such as cell metabolism, migration and invasion, proliferation, and responses to mitochondrial stress. For example, hnRNPA2-mediated activation of the invasive phenotype involves different mechanisms, including alternative splicing of TP53INP2, activation of the CXCL12 / CXCR4 axis, and activation of invasive behavior after mitochondrial DNA depletion. hnRNPA2, as a novel transcriptional coactivator, mediates this process through interactions with NF-κB, NFAT, CREB, and C / EBPδ. Overall, hnRNPA2B1 is broadly involved in the regulation of cancer cell phenotypes such as metabolism, proliferation, apoptosis, migration, and invasion through various molecular mechanisms. Furthermore, hnRNPA2B1 has been reported to induce epithelial-mesenchymal transition (EMT) in various cancer cell lines. Furthermore, hnRNPA2B1 is also involved in regulating fundamental cancer processes such as aerobic glycolysis. It has been reported that hnRNPA2 can regulate the selective splicing of pyruvate kinase isoenzyme M2 (PKM2), activating the metabolic shift of cancer cells towards aerobic glycolysis. hnRNPA2B1 also plays an important role in the regulation of hypoxia. In summary, hnRNPA2B1 is widely involved in the occurrence and development of various tumors. Developing small molecules targeting hnRNPA2B1 holds promise for blocking its pro-tumor effects, which could provide effective support for cancer treatment.

[0006] Clinically, cancer treatment methods are becoming increasingly diverse, including surgery, radiotherapy, chemotherapy, endocrine therapy, and targeted therapy. However, due to tumor heterogeneity and patient tolerance to different therapies, relying on a single treatment method is often insufficient to achieve good results, and drug resistance can develop during treatment. Therefore, combined cancer therapy using two or more methods has become an important strategy. The immune system can recognize and kill tumor cells, but tumor cells can employ various strategies to suppress the body's immune system, achieving immune evasion. Immunotherapy has become a major hope for curing cancer, and combined therapy, including immunotherapy, has become a significant direction in this field. The development of effective drugs to activate the immune system, combined with other therapies, is crucial for more effective tumor killing and elimination.

[0007] In summary, the development of specific agonists targeting hnRNPA2B1 and their combination with other antitumor therapies or drugs holds great potential in cancer treatment. There is an urgent need in this field to develop immunologically active substances and their combination products and methods that can initiate or promote interferon production, enhance antitumor effects, and effectively prevent tumor metastasis. Summary of the Invention

[0008] This disclosure provides fused cyclic compounds having the structure shown in formula (I), related derivatives thereof (e.g., cis-trans isomers, enantiomers, diastereomers, racemic mixtures, solvates, hydrates, or pharmaceutically acceptable salts thereof, or prodrugs thereof), and products comprising said compounds or related derivatives thereof. This disclosure also provides the use of said compounds, derivatives, and products in the prevention and treatment of tumors, and further provides their use in the treatment or prevention of tumors and related diseases or symptoms. The compounds, pharmaceuticals, pharmaceutical compositions, or kits of this disclosure can be used for effective antitumor activity, control of tumor occurrence and / or development.

[0009] In some aspects of this disclosure, fused ring compounds of formula (I), their cis-trans isomers, their enantiomers, their diastereomers, their racemic mixtures, their solvates, their hydrates, or pharmaceutically acceptable salts thereof, or their prodrugs, are provided.

[0010]

[0011] Where L is -(CH2)n-, and n is an integer from 0 to 6;

[0012] X is a halogen;

[0013] R 1 For -NR 4 R 5 , where R 4 R 5Each independently is H or C 1-6 alkyl;

[0014] R 2 and R 3 Each independently is H or C 1-6 alkyl;

[0015] Or R 2 and R 3 Together with the N atoms to which they are attached, they form 5-8 membered heterocyclic alkyl groups containing 1-3 heteroatoms selected from nitrogen, oxygen, or sulfur;

[0016] The C 1-6 The alkyl group and the 5-8 membered heterocyclic alkyl group are either unsubstituted or substituted by one or more substituents selected from the group consisting of: hydroxyl, amino, halogen, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Alkoxy, hydroxy C 1-6 Alkyl, C 1-6 Alkylamino, C 3-6 cycloalkyl, C 3-6 cycloalkyl C 1-6 Alkyl, 3-6 membered heterocyclic alkyl, 3-6 membered heterocyclic alkyl C 1-6 Alkyl, 6-10 aryl, 6-10 aryl C 1-6 Alkyl or 5-10 heteroaryl and 5-10 heteroaryl C 1-6 alkyl.

[0017] Among some aspects of this disclosure, the use of the compounds of the present invention in the preparation of products for the prevention and / or treatment of tumors is provided, the products of which may further comprise chemotherapeutic agents used in combination with the compounds.

[0018] In some aspects of this disclosure, a method for preventing and / or treating tumors is also provided, the method comprising administering to a subject in need a preventive and / or therapeutically effective amount of a compound or product of this disclosure, including further comprising or administering a chemotherapeutic agent in combination with said compound.

[0019] In some aspects of this disclosure, compounds, compositions or products of this disclosure are also provided for the prevention and / or treatment of tumors.

[0020] Among some aspects of this disclosure, the use of the compounds or products of this disclosure in increasing the levels of interferons (such as type I interferons, for example IFN-α and / or IFN-β).

[0021] Those skilled in the art can combine the foregoing technical solutions and features in any way without departing from the inventive concept and protection scope of this disclosure. Other aspects of this disclosure will be apparent to those skilled in the art due to the content of this disclosure. Attached Figure Description

[0022] The present disclosure will be further described below with reference to the accompanying drawings, which are shown only for illustrating the embodiments of the present disclosure and are not intended to limit the scope of the present disclosure.

[0023] Figure 1 The experiment tested the activation effect of the compound on nucleus exit of A549 lung cancer cells hnRNPA2B1.

[0024] Figure 2 The experiment tested the activation effects of the compound on downstream TBK1 and IRF3 in A549 lung cancer cells hnRNPA2B1 after nucleation.

[0025] Figure 3 The ELISA method was used to test the activation effect of the compound on type I interferon (IFN-β) in tumor cells.

[0026] Figure 4 The test compound, in combination with the chemotherapeutic agent oxaliplatin, induced apoptosis in tumor cells.

[0027] Figure 5 The test compound, in combination with the chemotherapeutic agent oxaliplatin, induced DNA breakage and apoptosis in tumor cells. Detailed Implementation

[0028] Through extensive research, development, and experimentation, this application has discovered a novel class of fused-ring compounds that can bind specifically to the hnRNPA2B1 protein with high affinity and can further induce higher levels of IFN-β, and is therefore defined as an hnRNPA2B1 agonist.

[0029] In antitumor tests on different types of tumor cells, the hnRNPA2B1 agonist compounds disclosed herein significantly activated type I interferon in tumor cells, inhibited LLC tumor proliferation, and promoted survival in tumor-bearing mice. Furthermore, since the hnRNPA2B1 agonist compounds disclosed herein can broadly induce higher levels of IFN-β, it is expected that these compounds will exert antitumor effects through type I interferon action. Moreover, when the compounds of this application are used in combination with chemotherapeutic agents, they can induce the production of more type I interferon and significantly kill tumor cells, inhibiting their growth.

[0030] Therefore, this disclosure provides methods and strategies for combining novel antitumor hnRNPA2B1 agonist compounds with chemotherapeutic agents in the prevention and treatment of tumor suppression.

[0031] All numerical ranges provided herein are intended to clearly include all values ​​falling between the endpoints of the range and the range of values ​​between them. Features mentioned in this disclosure or in the embodiments may be combined. All features disclosed in this specification may be used in any compositional form, and each feature disclosed in the specification may be replaced by any alternative feature that provides the same, equivalent, or similar purpose. Therefore, unless otherwise specified, the disclosed features are merely general examples of equivalent or similar features.

[0032] As used in this article, “containing,” “having,” or “including” includes “containing,” “mainly composed of,” “substantially composed of,” and “composed of”; “mainly composed of,” “substantially composed of,” and “composed of” are subordinate concepts of “containing,” “having,” or “including.”

[0033] hnRNPA2B1 agonist compound

[0034] As used herein, the terms "hnRNPA2B1 protein (peptide)" and "hnRNPA2B1" are used interchangeably and refer to the heterogeneous nuclear ribonucleoprotein A2B1. The hnRNPA2B1 protein involved in this disclosure may be a protein encoded by the hnRNPA2B1 gene, its cDNA, or CDS in an animal (e.g., human or mouse), or a homologous sequence of such protein that promotes interferon expression (e.g., homologous sequences of hnRNPA2B1 can be obtained from databases or alignment software known in the art), variant, or modified form.

[0035] As used herein, the terms “hnRNPA2B1 gene,” “hnRNPA2B1 encoding gene,” “hnRNPA2B1 protein encoding gene,” or “nucleic acid molecule encoding hnRNPA2B1” are used interchangeably and refer to a nucleotide sequence encoding the hnRNPA2B1 protein or polypeptide described herein, which may be, for example, the human hnRNPA2B1 gene with Gene ID: 3181, or the mouse hnRNPA2B1 gene with Gene ID: 53379. The term also includes molecules that hybridize with a labeled nucleic acid molecule under stringent conditions, or family gene molecules highly homologous to the aforementioned molecules, whose expression is believed to promote interferon production and its effects.

[0036] This disclosure provides a novel class of compounds that act as hnRNPA2B1 "agonists" (or "promoters"). The terms "agonist" or "hnRNPA2B1 agonist compound" are used interchangeably to refer to a class of novel fused-ring compounds that can increase the level or activity of hnRNPA2B1, specifically bind to the hnRNPA2B1 protein with high affinity, and further broadly induce higher levels of IFN-β, thereby exerting an agonistic effect on hnRNPA2B1.

[0037] The hnRNPA2B1 agonist compound disclosed herein can inhibit tumors, and thus can be further used for the prevention or treatment of tumor-related diseases and / or tumor-related symptoms.

[0038] This application provides a fused ring compound of formula (I), its cis-trans isomer, its enantiomer, its diastereomer, its racemic mixture, its solvate, its hydrate, or a pharmaceutically acceptable salt thereof, or a prodrug thereof.

[0039]

[0040] Where L is -(CH2)n-, and n is an integer from 0 to 6;

[0041] X is a halogen;

[0042] R 1 For -NR 4 R 5 , where R 4 R 5 Each independently is H or C 1-6 alkyl;

[0043] R 2 and R 3 Each independently is H or C 1-6 alkyl;

[0044] Or R 2 and R 3 Together with the N atoms to which they are attached, they form 5-8 membered heterocyclic alkyl groups containing 1-3 heteroatoms selected from nitrogen, oxygen, or sulfur;

[0045] The C 1-6 The alkyl group and the 5-8 membered heterocyclic alkyl group are either unsubstituted or substituted by one or more substituents selected from the group consisting of: hydroxyl, amino, halogen, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Alkoxy, hydroxy C 1-6 Alkyl, C 1-6 Alkylamino, C 3-6 cycloalkyl, C3-6 cycloalkyl C 1-6 Alkyl, 3-6 membered heterocyclic alkyl, 3-6 membered heterocyclic alkyl C 1-6 Alkyl, 6-10 aryl, 6-10 aryl C 1-6 Alkyl or 5-10 heteroaryl and 5-10 heteroaryl C 1-6 alkyl.

[0046] Preferably, X is F or Cl, more preferably, X is F.

[0047] Preferably, L is -(CH2)n-, where n is 1, 2 or 3, more preferably, n is 2.

[0048] Preferably, R 4 R 5 Each can be H, methyl, or ethyl.

[0049] Preferably, R 2 and R 3 Each independently is C 1-3 Alkyl, preferably methyl, ethyl or isopropyl, wherein the C 1-3 The alkyl group is unsubstituted or substituted with a 3-8 membered heterocyclic alkyl group containing 1-2 heteroatoms selected from nitrogen, oxygen, or sulfur; more preferably, R 2 and R 3 Each independently is C 1-3 Alkyl, preferably methyl, ethyl or isopropyl, wherein the C 1-3 The alkyl group is either unsubstituted or substituted with a 4-6 membered heterocyclic alkyl group containing one nitrogen atom; most preferably, R 2 and R 3 Each independently is C 1-3 The alkyl group, preferably methyl, ethyl or isopropyl, wherein the C1-3 alkyl group is unsubstituted or substituted with pyrrolidinyl group.

[0050] Preferably, R 2 and R 3 Together with the N atoms they are attached to, they form a 5-8 membered heterocyclic alkyl group containing two nitrogen atoms, wherein the 5-8 membered heterocyclic alkyl group is bonded to a 3-8 membered heterocyclic alkyl group C. 1-6 Alkyl or hydroxyl C 1-6 Alkyl substitution; more preferably, R 2 and R 3 Together with the N atoms they are attached to, they form a 6-membered heterocyclic alkyl group containing 2 nitrogen atoms, wherein the 6-membered heterocyclic alkyl group is bonded to a 4-6-membered heterocyclic alkyl group C. 1-6 Alkyl or hydroxyl C 1-6 Alkyl substitution; more preferably, R 2 and R 3Together with the N atoms to which they are attached, they form a piperazine group, which is bonded by a 4-6 membered heterocyclic alkyl group. 1-6 Alkyl or hydroxyl C 1-6 Alkyl substitution; most preferably, R 2 and R 3 Together with the N atom to which they are attached, they form a piperazine group, which is bonded to a 5-6 membered heterocyclic alkyl group containing one N atom. 1-6 Alkyl substitution or hydroxyl C 1-6 Alkyl substitution, preferably, the piperazine group is replaced by a pyrrolidinyl C 1-6 Alkyl substitution or hydroxyl C 1-6 Alkyl substitution, preferably, the piperazine group is substituted with pyrrolyl ethyl or hydroxyethyl.

[0051] Preferably, the compound has the following formula (II):

[0052]

[0053] Among them, R a R b Each independently is H or C 1-6 Alkyl, preferably, R a R b Each can be independently H, methyl, or ethyl;

[0054] M is -(CH2)n-, where n is 1, 2 or 3, more preferably n is 2;

[0055] X is F or Cl, more preferably, X is F;

[0056] R c It is a hydroxyl, amino, or 3-8 membered heterocyclic alkyl group containing 1-2 heteroatoms selected from nitrogen, oxygen, or sulfur, more preferably, R c It is a hydroxyl group or a 4-6 membered heterocyclic alkyl group containing one nitrogen atom, more preferably, R c It is hydroxyl or pyrrolidinyl, most preferably, R c It is hydroxyl or 1-pyrrolidinyl.

[0057] Preferably, the compound is selected from the group consisting of compounds or salts thereof (e.g., hydrochlorides):

[0058]

[0059] Unless otherwise stated, the following terms and phrases as used herein are intended to have the following definitions. A particular term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense. When a trade name appears in this document, it is intended to refer to the corresponding product or active ingredient.

[0060] The term “pharmaceutically acceptable” as used herein refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0061] The term "pharmaceutically acceptable salt" refers to a salt of the compounds of this invention, prepared by reacting a compound having specific substituents discovered in this invention with a relatively non-toxic acid or base. When the compounds of this invention contain relatively acidic functional groups, a base addition salt can be obtained by contacting the neutral form of such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts or similar salts. When the compounds of this invention contain relatively basic functional groups, a base addition salt can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Pharmaceutically acceptable examples of acid addition salts include inorganic acid salts, such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid, etc., and organic acid salts, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, octanoic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; salts of amino acids (such as arginine); and salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain both basic and acidic functional groups, and thus can be converted into either a base or an acid addition salt.

[0062] The pharmaceutically acceptable salts of the present invention can be synthesized from parent compounds containing acid radicals or bases by conventional chemical methods. Generally, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture thereof.

[0063] The compounds of this invention can exist in specific geometric or stereoisomeric forms. This invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)- enantiomers, (R)- and (S)- enantiomers, diastereomers, (D)- isomers, (L)- isomers, and racemic mixtures thereof, as well as other mixtures, such as mixtures enriched with enantiomers or diastereomers, all of which are within the scope of this invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of this invention.

[0064] Unless otherwise stated, the terms "enantiomer" or "optical isomer" refer to stereoisomers that are mirror images of each other.

[0065] Unless otherwise stated, the terms "cis-trans isomers" or "geometric isomers" arise because the single bonds of double bonds or cyclic carbon atoms cannot rotate freely.

[0066] Unless otherwise stated, the term "diastereomer" refers to a stereoisomer of a molecule having two or more chiral centers and being a non-mirror image of each other.

[0067] The compounds of this invention can exist in specific forms. Unless otherwise stated, the terms "tautomer" or "tautomer form" refer to isomers of different functional groups in dynamic equilibrium at room temperature, capable of rapidly interconverting into each other. If tautomerization is possible (e.g., in solution), chemical equilibrium of the tautomer may be achieved. For example, proton tautomers (also called prototropic tautomers) include interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions via the recombination of some bonding moieties. A specific example of keto-enol tautomerization is the interconversion between the two tautomers pentane-2,4-dione and 4-hydroxy-3-en-2-one.

[0068] The term "prodrug" generally refers to a compound of general formula (I) that has undergone functional group derivatization, and whose derivative can be readily converted into a compound of general formula (I) in vivo. Suitable prodrug selection and preparation methods are typically described in, for example, Design of Prodrugs, ed. H. Bundgaard, Elsevier, 1985.

[0069] The compounds of the present invention may contain, on one or more atoms constituting the compound, atomic isotopes in non-natural proportions, said isotopes having the same number of atoms but different atomic masses or mass numbers from those that are predominantly found in nature. For example, compounds may be labeled with radioactive isotopes, such as deuterium (…). 2 H), tritium ( 3 H), Iodine-125 ( 125 I) or C-14 14C). All isotopic variations of the compounds of this invention, regardless of radioactivity, are included within the scope of this invention. Isotopic variants may enhance certain therapeutic advantages, such as the use of deuterium to replace hydrogen to form deuterated drugs. The bond between deuterium and carbon is stronger than that between ordinary hydrogen and carbon. Compared to undeuterated drugs, deuterated drugs have advantages such as reduced toxicity, increased drug stability, enhanced efficacy, and prolonged drug biological half-life. Alternatively, they may provide standard compounds that can be used for characterization of biological samples. Isotope-enriched compounds of general formula (I) can be prepared without extensive experimentation using conventional techniques well known to those skilled in the art, or by methods similar to those described in the routes and embodiments of this invention, using appropriate isotope-enriched reagents and / or intermediates.

[0070] "Optional" or "optionally" means that the event or condition described below may occur but is not required to occur, and the description includes both the scenario in which said event or condition occurs and the scenario in which said event or condition does not occur.

[0071] The naming conventions used in this invention are based on the IUPAC system naming generated by ChemDraw software. Any open valence bonds appearing on carbon, oxygen, sulfur, or nitrogen atoms in the structures given in this invention indicate the presence of hydrogen atoms.

[0072] The term "substituted" means that any one or more hydrogen atoms on a specific atom are replaced by a substituent, which can include deuterium and hydrogen variants, provided that the valence state of the specific atom is normal and the substituted compound is stable. When the substituent is oxo (i.e., =O), it means that two hydrogen atoms are replaced; oxygen substitution does not occur on the aromatic group. The term "optionally substituted" means that it may or may not be substituted, unless otherwise specified; the type and number of substituents can be arbitrary on a chemically feasible basis.

[0073] When any variable (e.g., R) appears more than once in the composition or structure of a compound, its definition is independent in each case. Thus, for example, if a group is substituted by 0-2 Rs, the group can optionally be substituted by at most two Rs, and the Rs in each case have independent options. Furthermore, combinations of substituents and / or their variants are only permitted if such combinations produce a stable compound.

[0074] When the number of a linking group is 0, such as -(CRR)0-, it indicates that the linking group is a single bond.

[0075] When one of the variables is selected as a single bond, it means that the two groups it connects to are directly connected. For example, when L in ALZ represents a single bond, it means that its structure is actually AZ.

[0076] When the listed linking groups do not specify their linking direction, the linking direction is arbitrary, for example, The linking group L is at this time The benzene ring and cyclopentyl group can be connected in the same direction as the reading order from left to right to form the structure. Alternatively, the phenyl and cyclopentyl groups can be connected in the reverse order of reading from left to right to form the phenyl group. The combination of linking groups, substituents, and / or their variants is permitted only if such a combination produces a stable compound.

[0077] Unless otherwise specified, the number of atoms in a ring is usually defined as the elemental number of the ring. For example, a “3-7 elemental ring” refers to a “ring” with 3 to 7 atoms arranged around it.

[0078] Unless otherwise specified, the term "halogen" refers to fluorine, chlorine, bromine, and iodine.

[0079] Unless otherwise specified, the term "C" 1-6 "alkyl" is used to denote a straight-chain or branched saturated hydrocarbon group consisting of 1 to 6 carbon atoms. The C 1-6 Alkyl groups include C 1-5 C 1-4 C 1-3 C 1-2 C 2-6 C 2-4 C6 and C5 alkyl groups, etc.; they can be monovalent (e.g., methyl), divalent (e.g., methylene), or polyvalent (e.g., methine). 1-6 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl, etc.

[0080] Unless otherwise specified, the term "C" 1-3 "alkyl" is used to denote a straight-chain or branched saturated hydrocarbon group consisting of 1 to 3 carbon atoms. The C 1-3 Alkyl groups include C 1-2 and C 2-3 Alkyl groups, etc.; they can be monovalent (e.g., methyl), divalent (e.g., methylene), or polyvalent (e.g., methine). C 1-3 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), etc.

[0081] Unless otherwise specified, the term "C" 2-6"Alkenyl" is used to denote a hydrocarbon group consisting of 2 to 6 carbon atoms, either straight-chain or branched, containing at least one carbon-carbon double bond. The carbon-carbon double bond can be located at any position within the group. 2-6 Alkenes include C 2-4 C 2-3 C4, C3, C2 alkenyl groups, etc.; they can be monovalent, divalent, or polyvalent. 2-6 Examples of alkenyl groups include, but are not limited to, vinyl, propenyl, butenyl, pentenyl, hexenyl, butadienyl, and pentadienyl.

[0082] Unless otherwise specified, the term "C" 1-6 "Alkoxy" refers to alkyl groups containing 1 to 6 carbon atoms that are attached to other parts of a molecule by an oxygen atom. The C 1-6 Alkyl groups include C 1-4 C 1-3 C 1-2 C 2-6 C 2-4 C6, C5, C4, and C3 alkoxy groups, etc.; C 1-6 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), butoxy (including n-butoxy, isobutoxy, s-butoxy and t-butoxy), pentoxy (including n-pentoxy, isopentoxy and neopentoxy), hexoxy, etc.

[0083] Unless otherwise specified, the term "C" 1-6 "Alkylamino" refers to alkyl groups containing 1 to 6 carbon atoms that are attached to other parts of a molecule via an amino group. The C 1-6 Alkyl groups include C 1-4 C 1-3 C 1-2 C 2-6 C 2-4 C6, C5, C4, C3, and C2 alkylamino groups, etc.; C 1-6 Examples of alkylamino groups include, but are not limited to, -NHCH3, -N(CH3)2, -NHCH2CH3, -N(CH3)CH2CH3, -N(CH2CH3)2, -NHCH2CH2CH3, -NHCH(CH3)2, -NHCH2CH2CH2CH3, etc.

[0084] Unless otherwise specified, the term "C" 3-6 "Cycloalkyl" refers to a saturated cyclic hydrocarbon group consisting of 3 to 6 carbon atoms, which can be monocyclic or bicyclic. 3-6 Cycloalkyl groups include C 3-5 C 4-5 and C 5-6 Cycloalkyl groups, etc.; they can be monovalent, divalent, or polyvalent. C3-6 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.

[0085] Unless otherwise specified, the term "3-8 membered heterocyclic alkyl" on its own or in combination with other terms refers to a saturated monocyclic group consisting of 3 to 8 ring atoms, wherein 1, 2, 3, or 4 of the ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms may optionally be oxidized (i.e., NO and S(O)). z (z is 1 or 2). Furthermore, regarding the "3-8 membered heterocyclic alkyl", the heteroatom can occupy the connection position between the heterocyclic alkyl group and the rest of the molecule. The 3-8 membered heterocyclic alkyl group includes 4-6, 5-6, 7-8, 4, 5, and 6 membered heterocyclic alkyl groups, etc. Examples of 3-6 membered heterocyclic alkyl groups include, but are not limited to, nitrogen-containing heterocyclic butyl, oxo-heterocyclic butyl, thiocyclic butyl, pyrrolidinyl, pyrazolyl, imidazoalkyl, tetrahydrothiopheneyl, tetrahydrofuranyl (including tetrahydrofuran-2-yl), piperidinyl, tetrahydropyranyl, tetrahydrothiaranyl, piperazinyl, morpholinyl, thiomorpholinyl, etc.

[0086] Unless otherwise specified, the terms "6-10-membered aromatic ring" and "6-10-membered aryl" are used interchangeably, and the term "6-10-aryl" refers to a monovalent aromatic carbon ring system containing 6-10 carbon atoms and having at least one aromatic ring or at least one of the rings being aromatic rings. Examples of aryl groups are, but are not limited to, phenyl, naphthyl, biphenyl, or indenyl.

[0087] Unless otherwise specified, the terms "5-10-membered heteroaryl ring" and "5-10-membered heteroaryl" are used interchangeably in this invention. The term "5-10-membered heteroaryl" refers to a cyclic group consisting of 5 to 10 ring atoms with a conjugated π-electron system, wherein 1, 2, 3, or 4 of the ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms. It can be a monocyclic or fused bicyclic system, wherein at least one ring in the system is aromatic. The nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms may optionally be oxidized (i.e., NO and S(O)). z(z is 1 or 2). 5-10-membered heteroaryl groups can be attached to the rest of the molecule via heteroatoms or carbon atoms, including 5-8-membered, 5-7-membered, 5-6-membered, 5-membered, and 6-membered heteroaryl groups, etc. Examples of the 5-10 membered heteroaryl groups include, but are not limited to, pyrrole (including N-pyrrole, 2-pyrrole, and 3-pyrrole), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl), imidazole (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl, and 5-imidazolyl), oxazolyl (including 2-oxazolyl, 4-oxazolyl, and 5-oxazolyl), triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl, and 4H-1,2,4-triazolyl), tetrazolyl, isoxazolyl (including 3-isooxazolyl, 4-isooxazolyl, and 5-isooxazolyl), and thiazolyl (including 2-thiazolyl, 4-isooxazolyl, 4-isooxazolyl, and 5-isooxazolyl). α-thiazolyl and 5-thiazolyl, etc., furanyl (including 2-furanyl and 3-furanyl), thienyl (including 2-thienyl and 3-thienyl), pyridyl (including 2-pyridyl, 3-pyridyl and 4-pyridyl), pyrazinyl, pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl), benzothiazolyl (including 2-benzothiazolyl), purine, benzimidazolyl (including 2-benzimidazolyl), benzoxazolyl, indazole (including 5-indazole), isoquinolinyl (including 1-isoquinolinyl and 5-isoquinolinyl), quinoxalinyl (including 2-quinoxalinyl and 5-quinoxalinyl), or quinolinyl (including 3-quinolinyl and 6-quinolinyl). etc.

[0088] Unless otherwise specified, the terms "5-6 membered heteroaryl" and "5-6 membered heteroaryl" are used interchangeably in this invention. The term "5-6 membered heteroaryl" refers to a cyclic group consisting of 5 to 6 ring atoms with a conjugated π-electron system, wherein 1, 2, 3, or 4 of the ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms. The nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms may optionally be oxidized (i.e., NO and S(O)). z(z is 1 or 2). The 5-6 membered heteroaryl group can be attached to the rest of the molecule via a heteroatom or carbon atom. The 5-6 membered heteroaryl group includes 5-membered and 6-membered heteroaryl groups, etc. Examples of the 5-6 membered heteroaryl group include, but are not limited to, pyrrole (including N-pyrrole, 2-pyrrole, and 3-pyrrole), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl), imidazole (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl, and 5-imidazolyl), oxazolyl (including 2-oxazolyl, 4-oxazolyl, and 5-oxazolyl), and triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl). And 4H-1,2,4-triazolyl), tetrazolyl, isoxazolyl (including 3-isooxazolyl, 4-isooxazolyl and 5-isooxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4-thiazolyl and 5-thiazolyl, etc.), furanyl (including 2-furanyl and 3-furanyl, etc.), thiophene (including 2-thiophene and 3-thiophene, etc.), pyridyl (including 2-pyridyl, 3-pyridyl and 4-pyridyl, etc.), pyrazinyl, pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl, etc.), etc.

[0089] Unless otherwise specified, C n-n+m Or C n -C n+m This includes any specific case with n to n+m carbons, such as C 1-7 This includes C1, C2, C3, C4, C5, C6, and C7, as well as any range from n to n+m, such as C 1-7 Including C 1-3 C 1-6 C 3-6 C 4-7 and C 5-7 Similarly, n-membered to n+m-membered rings represent the number of atoms in the ring from n to n+m. For example, 3-7-membered rings include 3-membered, 4-membered, 5-membered, 6-membered, and 7-membered rings, and also include any range from n to n+m. For example, 3-7-membered rings include 3-6-membered rings, 4-7-membered rings, 5-7-membered rings, and 6-7-membered rings, etc.

[0090] Preparation method

[0091] This invention also relates to a method for producing compounds of general formula (I) as defined above. In some embodiments, the method for synthesizing the compounds of this application is shown in the figures:

[0092]

[0093] When X is selected from halogens other than Br, such as F, Cl, I, etc., the method includes the following steps:

[0094] a) Using a brominating agent to bromate a compound of formula Ia to generate a compound of formula Ib, preferably, the brominating agent includes, but is not limited to, N-bromosuccinimide (NBS) or 1,3-dibromo-5,5-dimethylhydantoin, preferably, the reaction temperature is 0-30°C.

[0095] b) In the compound of formula Ib, one Br atom is replaced by another halogen atom to obtain a compound of formula Ic. Preferably, step b) uses... The reaction is carried out at low temperature with a strong base of organometallic salt, wherein X is a halogen other than Br, such as F, Cl, I, more preferably, wherein the strong base of organometallic salt is n-butyllithium, and preferably, the reaction temperature is -78°C.

[0096] c) Using an oxidizing agent to oxidize the compound of formula Ic to the compound of formula Id, preferably, the oxidizing agent includes, but is not limited to, K2Cr2O7, and preferably, the reaction temperature is 60-150°C;

[0097] d) Compounds of formula Id with H2N-LR 1 The reaction yields a compound of formula Ie, preferably at a temperature of 50-150°C;

[0098] e) Combine the compound of formula Ie with R 2 R 3 The reaction is carried out by NH to give a compound of formula I. Preferably, the reaction is carried out in the presence of the catalyst bis(triphenylphosphine)palladium(II) chloride. Preferably, the reaction temperature is 20-150°C.

[0099] When X is selected from Br, the method includes:

[0100] a) Using a brominating agent to bromate a compound of formula Ia to generate a compound of formula Ib, preferably, the brominating agent includes, but is not limited to, N-bromosuccinimide (NBS), preferably, the reaction temperature is 0-30°C;

[0101] c) Using an oxidizing agent to oxidize the compound of formula Ib to the compound of formula Id, preferably, the oxidizing agent includes, but is not limited to, K2Cr2O7, and preferably, the reaction temperature is 60-150°C;

[0102] d) The compound of formula Id reacts with H2N-L-R1 to give the compound of formula Ie. Preferably, the reaction temperature is 50-150℃.

[0103] e) Combine the compound of formula Ie with R 2 R 3The reaction is carried out by NH to give a compound of formula I. Preferably, the reaction is carried out in the presence of the catalyst bis(triphenylphosphine)palladium(II) chloride. Preferably, the reaction temperature is 20-150°C.

[0104]

[0105] Alternatively, when X is selected from halogens other than Br, such as F, Cl, I, etc., the method includes the following steps:

[0106] f) Using a brominating agent to bromate the compound of formula Ia to generate the compound of formula Ib, preferably, the brominating agent includes, but is not limited to, N-bromosuccinimide (NBS) or 1,3-dibromo-5,5-dimethylhydantoin, preferably, the reaction temperature is 0-30°C.

[0107] g) In the compound of formula Ib, both Br atoms are replaced by other halogen atoms to obtain the compound of formula Ic. Preferably, step b) uses... The reaction is carried out at low temperature with a strong base of organometallic salt, wherein X is a halogen other than Br, such as F, Cl or I, more preferably, wherein the strong base of organometallic salt is n-butyllithium, and preferably, the reaction temperature is -78°C.

[0108] h) Using an oxidizing agent to oxidize the compounds of formulas I-c' to the compounds of formulas I-d', preferably, the oxidizing agent includes, but is not limited to, K2Cr2O7, and preferably, the reaction temperature is 60-150°C;

[0109] i) Compounds of formula I-d' and H2N-LR 1 The reaction yields compounds of formula I-e', preferably at a reaction temperature of 50-150°C;

[0110] j) The compound of formula I-e' is combined with R 2 R 3 The reaction is carried out by NH to give a compound of formula I. Preferably, the reaction is carried out in the presence of the catalyst bis(triphenylphosphine)palladium(II) chloride. Preferably, the reaction temperature is 20-150°C.

[0111] As is known to those skilled in the art, certain active groups (e.g., -NH2, -OH, etc.) require routine protection and deprotection when necessary, which is familiar to those skilled in the art.

[0112] Chemotherapy drugs

[0113] Based on the disclosure in this application and experimental verification, the compounds in this application can be used in combination with chemotherapeutic agents to enhance their antitumor effects, preferably producing a synergistic effect.

[0114] Chemotherapy agents that can be used in combination with the hnRNPA2B1 agonist of this application include, but are not limited to, one or more selected from the following group: platinum-based chemotherapeutic agents, such as oxaliplatin, cisplatin, and carboplatin; alkylating agents: cyclophosphamide, ifosfamide, nitrosoureas, mitomycin, etc.; antimetabolites: methotrexate, 5-FU, cytarabine, gemcitabine, capecitabine, etc.; antitumor antibiotics: anthracyclines, bleomycin, etc.; antitumor herbal drugs: vinca alkaloids, taxanes, etc.; hormones: adrenocortical hormones, estrogens, androgens and their antagonists, etc.

[0115] In some embodiments, chemotherapeutic agents that can be used in combination with the hnRNPA2B1 agonist of this application include, but are not limited to, one or more selected from the group consisting of: chemotherapeutic agents that interfere with DNA replication, such as platinum-based chemotherapeutic agents (e.g., oxaliplatin, cisplatin, carboplatin), alkylating agents (e.g., cyclophosphamide), and topoisomerase I inhibitors (e.g., irinotecan); chemotherapeutic agents that interfere with RNA transcription, such as anthracyclines (e.g., doxorubicin, epirubicin); chemotherapeutic agents that interfere with microtubules, such as taxanes; and chemotherapeutic agents that inhibit nucleotide synthesis and metabolism, such as antimetabolites (e.g., gemcitabine, methotrexate, 5-FU, capecitabine).

[0116] It should be understood that those skilled in the art, based on the disclosure in this application, can select chemotherapeutic active substances as needed and combine them with the hnRNPA2B1 agonist compound of this application.

[0117] Products and their applications

[0118] This disclosure also provides a product, which may be, for example, a pharmaceutical product, a pharmaceutical composition, or a kit / pharmaceutical kit, containing an effective amount of the hnRNPA2B1 agonist compound of this disclosure, and a pharmaceutically or immunologically acceptable carrier. As used herein, the terms "active substance" and "active substance of this disclosure" are used interchangeably and refer to a hnRNPA2B1 agonist compound having the structural formula (I) or a derivative thereof or product thereof. The product of this disclosure may further comprise a chemotherapeutic agent in combination with the hnRNPA2B1 agonist compound.

[0119] As used herein, the term "pharmaceuticalally acceptable" refers to a substance that is suitable for use in humans and / or animals without excessive adverse effects (such as toxicity, irritation, and allergic reactions), i.e., a reasonable benefit / risk ratio. As used herein, the term "effective amount" refers to an amount that is functional or active in humans and / or animals and is acceptable to humans and / or animals.

[0120] As used herein, the term "pharmaceuticalally acceptable carrier" refers to a carrier used for the administration of therapeutic agents, including various excipients and diluents. This term refers to pharmaceutical carriers that are not essential active ingredients themselves and do not cause excessive toxicity upon administration. Suitable carriers are well known to those skilled in the art, and a thorough discussion of pharmaceutically acceptable excipients can be found in Remington's Pharmaceutical Sciences (Mack Pub. Co., NJ 1991).

[0121] Pharmaceutically acceptable carriers in the composition may contain liquids such as water, saline, glycerol, and ethanol. Additionally, these carriers may contain auxiliary substances such as fillers, disintegrants, lubricants, glidants, effervescent agents, wetting agents or emulsifiers, flavoring agents, pH buffers, etc. Typically, these substances are formulated in a non-toxic, inert, and pharmaceutically acceptable aqueous carrier medium, with a pH usually around 5-8, preferably around 6-8.

[0122] The active substance in the product disclosed herein accounts for 0.001 to 99.9 wt% of the total weight of the composition; preferably 1 to 95 wt% of the total weight of the composition, more preferably 5 to 90 wt%, and more preferably 10 to 80 wt%. The balance consists of pharmaceutically acceptable carriers and other additives.

[0123] hnRNPA2B1 agonist compounds can be combined with chemotherapeutic agents at a ratio of 1:100 to 100:1.

[0124] As used herein, the term "unit dosage form" refers to a dosage form in which the products of this disclosure are prepared for a single dose for ease of administration, including but not limited to various solid dosage forms (such as tablets), liquid dosage forms, capsules, and sustained-release formulations.

[0125] In another preferred embodiment of this disclosure, the product is a single dosage form or multiple dosage forms, and the content of the active substance is 0.01–2000 mg / dose, preferably 0.1–1500 mg / dose, more preferably 1–1000 mg / dose. In another preferred example of this disclosure, 1–6 doses of the composition of this disclosure are administered daily, preferably 1–3 doses; most preferably, the daily dose is 1 dose. Furthermore, when used in combination, the composition may further contain a therapeutically effective amount of a chemotherapeutic agent, or a product containing the compound of this application may be used in combination with a product containing a chemotherapeutic agent.

[0126] It should be understood that the effective dosage of active substances such as hnRNPA2B1 agonist compounds and chemotherapeutic agents can vary depending on the severity of the condition of the patient being treated. The specific dosage is determined based on the individual patient's circumstances (e.g., weight, age, physical condition, and desired outcome), within the judgment of a skilled physician.

[0127] The products disclosed herein may be in solid form (e.g., granules, tablets, lyophilized powder, suppositories, capsules, sublingual tablets) or liquid form (e.g., oral liquid) or other suitable forms. Routes of administration may include, but are not limited to: (1) conventional administration methods, such as gastrointestinal administration (e.g., oral administration), non-gastrointestinal administration (e.g., solution injection, intravenous infusion), such as mucosal administration, transdermal administration, respiratory nebulization, nasal drops, sprays, oral administration, intramuscular injection and / or intravenous administration, etc.; (2) linking hnRNPA2B1 agonists and / or chemotherapeutic agents with transferrin / poly-L-lysine complexes to enhance their biological effects; (3) encapsulating drugs in liposomes to mediate their entry into cells, which facilitates the smooth entry of compound molecules while protecting them from hydrolysis by various extracellular enzymes; (4) using liposomes to transport active substances to their specific target tissues and cells.

[0128] Furthermore, the product disclosed herein may also contain other active substances for improving and treating tumors or may be combined with other tumor prevention and treatment methods. In some embodiments, other active substances that regulate antitumor activity are administered before, simultaneously with, or after the product of this disclosure.

[0129] In some embodiments, the immune system is further modulated to prevent and treat tumors by activating hnRNPA2B1 and chemotherapeutic agents through the compounds or products of this application.

[0130] The term "object" can refer to an animal, including but not limited to primates (e.g., humans), monkeys, cattle, pigs, sheep, goats, horses, dogs, cats, rabbits, rats, or mice. The terms "object" and "patient" are used interchangeably herein, for example, to refer to a mammalian object, such as a human object.

[0131] In the context of treating a disease, condition, or symptom, the terms “treatment,” “management,” and “therapy” are intended to include the reduction or elimination of a disease, condition, or symptom, or one or more symptoms associated with a disease, condition, or symptom; or the slowing of the progression, spread, or worsening of a disease, condition, or symptom, or one or more symptoms thereof. “Cancer treatment” means one or more of the following actions: (1) inhibiting tumor growth to a certain extent, including (i) slowing and (ii) completely stopping growth; (2) reducing the number of tumor cells; (3) maintaining tumor size; (4) reducing tumor size; (5) inhibiting, including (i) reducing, (ii) slowing, or (iii) completely preventing tumor cell infiltration into surrounding organs; (6) inhibiting, including (i) reducing, (ii) slowing, or (iii) completely preventing cancer metastasis; (7) enhancing the antitumor immune response, which may (i) maintain tumor size, (ii) reduce tumor size, (iii) slow tumor growth, (iv) reduce, slow, or prevent invasion, and / or (8) to a certain extent alleviate the severity or number of one or more symptoms associated with the disorder.

[0132] In specific implementations, the disease or condition to be treated is a proliferative disease. In some implementations, the proliferative disease is cancer. In specific implementations, the cancer is selected from: brain and spinal cancer, head and neck cancer, leukemia and blood cancers, skin cancer, reproductive system cancers, gastrointestinal system cancers, liver and bile duct cancers, pancreatic cancer, kidney cancer, prostate and bladder cancer, bone cancer, lung cancer, malignant mesothelioma, sarcoma, lymphoma, adenocarcinoma, thyroid cancer, cardiac tumors, germ cell tumors, malignant neuroendocrine (carcinoid) tumors, midline cancer, and unknown primary cancer (i.e., cancer with metastatic cancer but unknown primary site). In specific implementations, the cancer is present in adult patients. In other implementations, the cancer is present in pediatric patients. In specific implementations, the cancer is associated with AIDS.

[0133] In specific implementation schemes, the cancers are selected from brain and spinal cancers. In specific implementation schemes, the cancers are selected from: anaplastic astrocytoma, glioblastoma, astrocytoma, and sensory neuroblastoma (also known as olfactory blastoma). In specific implementation schemes, the brain cancers are selected from: astrocytomas (e.g., cellular astrocytoma, subependymal giant cell astrocytoma, diffuse astrocytoma, corpus luteum astrocytoma, anaplastic astrocytoma, astrocytoma, giant cell glioblastoma, glioblastoma, secondary glioblastoma, primary adult glioblastoma, and primary pediatric glioblastoma), and maxillary nerve tumors (e.g., oligodendroglioma and anaplastic oligodendroglioma). Oligopica astrocytic tumors (e.g., oliguria astrocytoma and anaplastic follicular astrocytoma), ependymomas (e.g., mucosal papillary ependymomas and anaplastic ependymomas); medulloblastomas, primitive neuroectodermal tumors, schwannomas, meningiomas, atypical meningiomas, anaplastic meningiomas, pituitary adenomas, brainstem gliomas, cerebellar astrocytomas, cerebral aneurysms / malignant gliomas, visual pathway and hypothalamic gliomas, and primary central nervous system lymphomas. In specific examples of these implementation schemes, brain cancers include: gliomas, glioblastomas multiforme, paragangliomas, and epithelial primitive neuroectodermal tumors of the upper urinary tract (sPNET).

[0134] In a specific implementation plan, the cancer is selected from: head and neck cancer, including nasopharyngeal carcinoma, nasal cavity and paranasal sinus cancer, pharyngeal cancer, oral cancer (e.g., squamous cell carcinoma, lymphoma, and sarcoma), lip cancer, oropharyngeal cancer, salivary gland tumors, laryngeal cancer (e.g., laryngeal squamous cell carcinoma, rhabdomyosarcoma), and eye cancer or ocular cancer. In a specific implementation plan, ocular cancer is selected from intraocular melanoma and retinoblastoma.

[0135] In a specific implementation plan, the cancer is selected from leukemia and blood cancers. Specifically, the cancer is selected from: myelodysplastic neoplasms, myelodysplastic syndromes, myelodysplastic / myelodysplastic neoplasms, acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), chronic myeloid leukemia (CML), myelodysplastic neoplasms (MPN), post-MPN AML, post-MDS AML, del(5q)-associated high-risk MDS or AML, blastic chronic myeloid leukemia, angioimmunoblastic lymphoma, acute lymphoblastic leukemia, Langerhans cell histiocytosis, hairy cell leukemia, and plasmacytoma (including plasmacytoma and multiple myeloma). The leukemia mentioned herein can be acute or chronic.

[0136] In a specific implementation, the cancer is selected from skin cancer. Specifically, the skin cancer is selected from melanoma, squamous cell carcinoma, and basal cell carcinoma.

[0137] In specific implementations, the cancer is selected from cancers of the reproductive system. In specific implementations, the cancer is selected from: breast cancer, cervical cancer, vaginal cancer, ovarian cancer, prostate cancer, penile cancer, and testicular cancer. In specific examples of these implementations, the cancer is breast cancer, selected from ductal carcinoma and phyllodes tumor. In specific examples of these implementations, the breast cancer can be male breast cancer or female breast cancer. In specific examples of these implementations, the cancer is cervical cancer, selected from squamous cell carcinoma and adenocarcinoma. In specific examples of these implementations, the cancer is ovarian cancer, selected from epithelial carcinoma.

[0138] In specific implementations, the cancer is selected from cancers of the gastrointestinal system. Specifically, the cancer is selected from: esophageal cancer, gastric cancer (also known as stomach cancer), gastrointestinal carcinoid tumors, pancreatic cancer, gallbladder cancer, colorectal cancer, and anal cancer. In examples of these implementations, the cancer is selected from: esophageal squamous cell carcinoma, esophageal adenocarcinoma, gastric adenocarcinoma, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors, gastric lymphoma, gastrointestinal lymphoma, solid papillary pancreatic tumors, pancreatoblastoma, islet cell tumors, pancreatic cancer (including acinar cell carcinoma and ductal adenocarcinoma), gallbladder adenocarcinoma, colorectal adenocarcinoma, and anal squamous cell carcinoma.

[0139] In specific implementations, the cancer is selected from liver cancer and bile duct cancer. In specific implementations, the cancer is liver cancer (also known as hepatocellular carcinoma). In specific implementations, the cancer is bile duct cancer (also known as cholangiocarcinoma). In examples of these implementations, the bile duct cancer is selected from intrahepatic bile duct cancer and extrahepatic bile duct cancer.

[0140] In a specific implementation, the cancer is selected from kidney cancer and bladder cancer. In a specific implementation, the cancer is kidney cancer, selected from renal cell carcinoma, Wilms' tumor, and transitional cell carcinoma. In a specific implementation, the cancer is bladder cancer, selected from ureteral cancer (transitional cell carcinoma), squamous cell carcinoma, and adenocarcinoma.

[0141] In a specific implementation plan, the cancer is selected from bone cancer. Specifically, the bone cancer is selected from: osteosarcoma, malignant fibrous histiocytoma of bone, Ewing sarcoma, and chordoma (bone cancer along the spine).

[0142] In a specific implementation plan, the cancer is selected from lung cancer. Specifically, the lung cancer is selected from: non-small cell lung cancer, small cell lung cancer, bronchial tumors, and pleural pulmonary blastoma.

[0143] In a specific implementation, the cancer is selected from malignant mesothelioma. In a specific implementation, the cancer is selected from epithelial mesothelioma and sarcomatoid tumor.

[0144] In a specific implementation, the cancer is selected from sarcomas. In a specific implementation, the sarcoma is selected from: central chondrosarcoma, central and periosteal chondroma, fibrosarcoma, clear cell sarcoma of the tendon sheath, and Kaposi's sarcoma.

[0145] In a specific implementation, the cancer is selected from lymphomas. Specifically, the cancer is selected from: Hodgkin lymphoma (e.g., Reed-Stemberg cell), non-Hodgkin lymphoma (e.g., diffuse large B-cell lymphoma, follicular lymphoma, mycosis fungoides, Sezary syndrome, primary central nervous system lymphoma), cutaneous T-cell lymphoma, and primary central nervous system lymphoma.

[0146] In a specific implementation, the cancer is selected from adenocarcinoma. Specifically, the cancer is selected from: adrenocortical carcinoma (also known as adrenocortical carcinoma or adrenocortical cancer), pheochromocytoma, paraganglioma, pituitary adenoma, thymoma, and thymic carcinoma.

[0147] In a specific implementation scheme, the cancer is selected from thyroid cancer. Specifically, the thyroid cancer is selected from medullary thyroid carcinoma, papillary thyroid carcinoma, and follicular thyroid carcinoma.

[0148] In a specific implementation, the cancer is selected from germ cell tumors. In a specific implementation, the cancer is selected from malignant extracranial germ cell tumors and malignant extragonadal germ cell tumors. In specific examples of these implementations, the malignant extragonadal germ cell tumor is selected from non-seminomatous tumors and seminomas.

[0149] In the specific implementation plan, the cancer is selected from cardiac tumors. Specifically, the cardiac tumors are selected from: malignant teratoma, lymphoma, rhabdomyosarcoma, angiosarcoma, chondrosarcoma, infantile fibrosarcoma, and synovial sarcoma.

[0150] In specific implementation schemes, proliferative disorders are selected from: benign papillomas, benign neoplastic diseases, and gestational trophoblastic diseases. In specific implementation schemes, benign neoplastic diseases are selected from: cutaneous papillomas (warts) and genital papillomas. In specific implementation schemes, gestational trophoblastic diseases are selected from: hydatid cystic nevi and gestational trophoblastic growths (e.g., invasive nevi, choriocarcinoma, placental trophoblastic tumors, and epithelioid trophoblastic tumors).

[0151] In some embodiments, the compounds or products of this application are administered prophylactically as a preventative medicine before tumor development to prevent tumor development or reduce the severity of subsequent tumors. In some embodiments, the compounds or products of this application are administered as a therapeutic medicine after tumor development to reduce the severity of the tumor disease, including its metastasis. In some embodiments, the compounds or products of this application are administered both as a preventative and therapeutic medicine, either continuously or intermittently before and after tumor development.

[0152] Those skilled in the art can combine the foregoing technical solutions and features in any way without departing from the inventive concept and protection scope of this invention. Other aspects of this invention will be apparent to those skilled in the art from the disclosure herein.

[0153] Example

[0154] The present disclosure is further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the disclosure. Those skilled in the art can make appropriate modifications and variations to the present disclosure, and such modifications and variations are all within the scope of the present disclosure.

[0155] Experimental methods not specifically described in the following examples can be performed using conventional methods in the art, such as those described in *Molecular Cloning: A Laboratory Manual* (3rd edition, New York: Cold Spring Harbor Laboratory Press, 1989) or according to the conditions recommended by the supplier. DNA sequencing methods are conventional in the art and can also be provided by commercial companies.

[0156] Unless otherwise stated, percentages and parts are by weight. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to one skilled in the art. Furthermore, any methods and materials similar or equivalent to those described herein may be used in the methods of this disclosure. The preferred embodiments and materials described herein are for illustrative purposes only.

[0157] I. Compound Synthesis and Characterization Examples

[0158] Example I.1: Synthesis of compound A (FPC-1):

[0159]

[0160] *eq equivalent

[0161] 1-1. Preparation of Compound 2

[0162]

[0163] NBS (57.5 g, 320 mmol) was added to DMF (120 mL) and cooled to 0 °C. Compound 1 (20 g, 130 mmol) was added, and the resulting suspension was stirred and allowed to warm naturally to room temperature overnight. The reaction solution was filtered, and the filter cake was washed three times with ethanol. The dried filter cake was collected and recrystallized with EA to give the pure product (10.1 g, off-white crystals).

[0164] 1 ¹H NMR (300MHz, chloroform-d) δ 7.79 (d, J = 7.4Hz, 2H), 7.09 (d, J = 8.8Hz, 2H), 3.30 (s, 4H).

[0165] 1-2. Preparation of Compound 3

[0166]

[0167] Compound 2 (6 g, 19.2 mol) was dissolved in 300 mL of anhydrous THF and cooled to -80 °C. Then, n-butyllithium (9.2 mL, 2.5 M) was added dropwise, and the reaction mixture was stirred at the same temperature for at least one hour. After the slow addition of N-fluorobis(benzenesulfonamide) (9 g, 28.8 mmol) in 100 mL of anhydrous THF, the mixture was stirred at -80 °C for another 60 minutes. The reaction mixture was then heated to room temperature overnight and poured into an NH4Cl solution. The aqueous phase was then extracted with DCM (3 × 50 mL), the organic phases were combined, and dried over MgSO4. The solvent was removed under low pressure. The product was purified by column chromatography using n-hexane as the eluent to give compound 3 (4.1 g, white solid).

[0168] 1 ¹H NMR (300MHz, chloroform-d) δ 7.66 (d, J = 7.4Hz, 1H), 7.21–7.09 (m, 3H), 3.35 (s, 4H).

[0169] 1-3. Preparation of Compound 4

[0170]

[0171] Compound 3 (4.5 g, 17.9 mmol) was added to a solution of potassium dichromate (24 g, 82.4 mmol) in glacial acetic acid (50 mL). The solution was heated under reflux for 16 hours and then poured into ice water. The resulting precipitate was collected by filtration and washed with water. The filter cake was air-dried, added to DCM and refluxed for 30 minutes, filtered, washed with DCM, and the filter cake was added to DCM and refluxed for 15 minutes, filtered, and washed with DCM. All filtrates were combined and concentrated under vacuum to give compound 4 (4.1 g, crude product, yellow solid).

[0172] 1-4. Preparation of INT-1

[0173]

[0174] N-Boc-ethylenediamine (3.3 g, 20.8 mmol) was added to a 100 mL solution of compound 4 (4.1 g, 13.9 mmol) in EtOH, and the mixture was heated under reflux for 1 hour. The reaction mixture was concentrated, and the residue was purified by column chromatography (PE / EA = 5 / 1) to give a white solid (2.8 g).

[0175] 1 H NMR (300MHz, chloroform-d) δ8.65(dd,J=8.2,4.6Hz,1H),8.43(d,J=8.0Hz,1H),8.08(d,J=8.0Hz,1H), 7.50(dd,J=12.1,8.1Hz,1H),4.89(s,1H),4.39-4.28(m,2H),3.58-3.44(m,2H),1.26(s,9H).

[0176] 1-5. Preparation of FPC-1-1

[0177]

[0178] Cesium carbonate (5.55 g, 17 mmol) and 1-(2-pyridone ethyl)piperazine (1.05 g, 5.7 mmol) were added to a toluene solution (175 mL) of INT-1 (2.5 g, 5.7 mol). Bis(triphenylphosphine)palladium(II) chloride (400 mg, 0.57 mmol) was added to the mixture under argon protection. The reaction was carried out at 80 °C for 16 hours. The reaction solution was cooled to room temperature, water was added, and the mixture was stirred. The organic phase was separated, and the aqueous phase was extracted once with DCM. The combined organic phases were dried over anhydrous magnesium sulfate. The crude product was purified by Prep-TLC (DCM / MeOH = 10 / 1 + NH3·H2O) by vacuum concentration and column chromatography to give 240 mg of a pure yellow solid.

[0179] 1 ¹H NMR (400MHz, chloroform-d) δ 8.56 (dd, J = 7.7, 4.3Hz, 1H), 8.51 (d, J = 8.3Hz, 1H), 7.34–7.27 (m, 1H), 7.17 (d, J = 8.2Hz, 1H), 4.99 (s, 1H), 4.31 (s, 2H), 3.79–2.45 (m, 18H), 2.18 (s, 4H), 1.28 (s, 9H).

[0180] 1-6. Preparation of FPC-1

[0181]

[0182] Compound FPC-1-1 (240 mg, 0.44 mmol) was dissolved in 11 mL of DCM / MeOH (10 / 1). 5 M HCl / 1,4-dioxane (5 mL) was added to the mixture and the mixture was stirred at room temperature for 2 hours. The mixture was then concentrated under vacuum. The residue was dissolved in a small amount of methanol, precipitated with diethyl ether, centrifuged, washed with diethyl ether, centrifuged again, and dried under vacuum to give 240 mg of the pure product (yellow solid).

[0183] 1 ¹H NMR (300MHz, methanol-d⁴) δ 8.67–8.58 (m, 2H), 7.56 (dd, J = 13.2, 8.3 Hz, 1H), 7.47 (d, J = 8.3 Hz, 1H), 4.49–4.42 (m, 2H), 4.02–3.69 (m, 10H), 3.69–3.38 (m, 8H), 2.18 (s, 4H).

[0184] MS(ESI)m / z[M+H] + =440.3.

[0185] Example I.2: Synthesis of Compound B (FPC-2)

[0186]

[0187] 2-1. Preparation of FPC-2-1

[0188]

[0189] 10 mL of ethylene glycol monomethyl ether was added to a single-necked flask, followed by INT-1 (110 mg, 0.25 mmol) and 3-(piperazin-1-yl)prop-1-ol (44 mg, 0.3 mmol). The reaction mixture was stirred at 100 °C. After 2 hours, the solvent was removed under vacuum, and the residue was purified by TLC to give 20 mg of a yellow solid.

[0190] 1¹H NMR (400MHz, chloroform-d) δ 8.63–8.49 (m, 2H), 7.31 (dd, J = 12.8, 8.3Hz, 1H), 7.18 (d, J = 8.3Hz, 1H), 4.97 (s, 1H), 4.42–4.23 (m, 2H), 3.87 (t, J = 5.0Hz, 2H), 3.60–3.28 (m, 8H), 2.92 (s, 4H), 1.89 (s, 2H), 1.29 (s, 9H).

[0191] 2-2. Preparation of FPC-2

[0192]

[0193] Compound FPC-2-1 (20 mg, mmol) was dissolved in 2 mL of DCM / MeOH (1 / 1). 5 M HCl / 1,4-dioxane (2 mL) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was then concentrated under vacuum, and the residue was dissolved in a small amount of methanol, precipitated with diethyl ether, centrifuged, washed with diethyl ether, centrifuged again, and dried under vacuum to give 5 mg of the pure product (yellow solid).

[0194] 1 ¹H NMR (400MHz, methanol-d⁴) δ 8.72–8.46 (m, 2H), 7.50 (ddd, J = 26.1, 12.4, 8.3 Hz, 2H), 4.45 (t, J = 5.6 Hz, 2H), 3.93–3.69 (m, 6H), 3.45 (dt, J = 15.0, 9.8 Hz, 5H), 3.34 (s, 3H), 2.07 (dt, J = 12.2, 6.1 Hz, 2H).

[0195] MS(ESI)m / z[M+H] + =401.47.

[0196] Example I.3: Synthesis of Compound C (FPC-3)

[0197]

[0198] 3-1. Preparation of FPC-3-1

[0199]

[0200] Cesium carbonate (1.78 g, 5.48 mmol) and N-ethyl-2-(pyrrolidone-1-yl)ethyl-1-amine (286 mg, 2.01 mmol) were added to a toluene solution (70 mL) of INT-1 (800 mg, 1.82 mmol). Bis(triphenylphosphine)palladium(II) chloride (128 mg, 0.18 mmol) was added to the mixture under argon protection. The reaction was carried out at 80 °C for 16 hours. The reaction solution was cooled to room temperature, water was added, and the mixture was stirred. The organic phase was separated, and the aqueous phase was extracted once with DCM. The obtained organic phases were combined and dried over anhydrous magnesium sulfate. The crude product was purified by Prep-TLC (DCM / MeOH = 10 / 1 + NH3·H2O) by vacuum concentration and column purification to give 30 mg of a pure yellow solid.

[0201] 1 H NMR (400MHz, chloroform-d) δ8.63-8.52(m,2H),7.37-7.28(m,2H),4.96(s,1H),4.32(t,J=5.5Hz,2H),3.99(s,2H),3.93-3.75(m,2H),3.49(s,2H) ),3.44(d,J=6.8Hz,2H),3.36-3.16(m,2H),2.87-2.62(m,2H),2.29-2.12(m,2H),2.12-1.91(m,2H),1.28(s,9H),1.10(t,J=7.1Hz,3H).

[0202] 3-2. Preparation of FPC-3

[0203]

[0204] Compound FPC-3-1 (30 mg, 0.06 mmol) was dissolved in 4.4 mL LCM / MeOH (10 / 1). 5 M HCl / 1,4-dioxane (2 mL) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was then concentrated under vacuum, and the residue was dissolved in a small amount of methanol, precipitated with diethyl ether, centrifuged, washed with diethyl ether, centrifuged again, and dried under vacuum to give 25 mg of the pure product (yellow solid).

[0205] 1H NMR (300MHz, methanol-d4) δ8.67-8.54(m,2H),7.58-7.46(m,2H),4.51-4.41(m,2H),3.87-3.77(m,2H),3.77-3.65(m ,2H),3.51(q,J=6.6Hz,4H),3.34(d,J=1.8Hz,1H),3.20-3.05(m,2H),2.20-1.92(m,5H),1.10(t,J=7.0Hz,3H).

[0206] MS(ESI)m / z[M+H] + =399.47.

[0207] Example I.4: Synthesis of Compound D (FPC-5)

[0208]

[0209] 4-1. Preparation of FPC-5-1

[0210]

[0211] Cesium carbonate (460 mg, 1.41 mmol) and N,2-dimethylpropyl-1-amine (49 mg, 0.56 mmol) were added to a toluene solution (20 mL) of INT-1 (205 mg, 0.47 mmol). Bis(triphenylphosphine)palladium(II) chloride (32 mg, 0.045 mmol) was added to the mixture under argon protection. The reaction was carried out at 80 °C for 16 hours. The reaction solution was cooled to room temperature, water was added and stirred, and the organic phase was separated. The aqueous phase was extracted once with DCM. The combined organic phases were dried over anhydrous magnesium sulfate. The crude product was purified by vacuum concentration and column chromatography (DCM / MeOH = 10 / 1 + NH3·H2O) to give 10 mg of a pure yellow solid.

[0212] 1 H NMR (300MHz, chloroform-d) δ8.56(dd,J=8.2,4.9Hz,1H),8.45(d,J=8.5Hz,1H),7.24-7.18(m,1H),7.07(d,J=8.5Hz,1H),5.05(s,1H),4.39-4.2 6(m,2H),3.51(dd,J=11.9,5.2Hz,2H),3.20-3.13(m,1H),3.05(d,J=4.0Hz,3H),2.17-2.05(m,2H),1.31(s,9H),0.87(d,J=6.6Hz,6H).

[0213] 4-2. Preparation of FPC-5

[0214]

[0215] Compound FPC-5-1 (10 mg, 0.022 mmol) was dissolved in 1.1 mL of DCM / MeOH (10 / 1). 5 M HCl / 1,4-dioxane (0.5 mL) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was then concentrated under vacuum, and the residue was dissolved in a small amount of methanol, precipitated with diethyl ether, centrifuged, washed with diethyl ether, centrifuged again, and dried under vacuum to give 9 mg of the pure product (yellow solid).

[0216] 1 H NMR (400MHz, methanol-d4) δ8.61-8.52(m,1H),8.44(d,J=8.3Hz,1H),7.38(dd,J=12.5,8.5Hz,1H),7.25(d,J=8.4Hz,1H),4.44(t,J=5.1Hz ,2H),3.22(dd,J=19.7,7.1Hz,4H),3.10(d,J=3.1Hz,3H),2.16(dp,J=13.3,7.2Hz,1H),1.31(d,J=7.7Hz,1H),0.87(d,J=6.4Hz,6H).

[0217] MS(ESI)m / z[M+H] + =344.42.

[0218] Example I.5: Synthesis of compound E (AIR-2)

[0219]

[0220] 5-1. Preparation of Compound 5

[0221]

[0222] Compound 3 (31.90 g, 127.04 mmol) was dissolved in anhydrous THF (800 mL). The reaction mixture was purged three times with argon and then cooled to -78 °C. Then, n-BuLi (60.98 mL, 2.5 M / L, 152.45 mmol) was added dropwise, and the reaction mixture was stirred at the same temperature for at least 10 minutes. N-fluorobis(benzenesulfonamide) (NFSI) (60.09 g, 190.57 mmol) in 200 mL of anhydrous THF was added dropwise, and the mixture was stirred at -78 °C for 1 hour. The reaction mixture was then allowed to warm to room temperature naturally and stirred overnight. The reaction mixture was quenched with NH4Cl solution and then extracted twice with EA. The combined organic layers were washed with water and brine, dried over Na2SO4, and filtered. The filtrate was concentrated to give the crude product, which was added to DCM and stirred. The precipitate was filtered, the filter cake was washed twice with DCM, all filtrates were combined and concentrated under vacuum to obtain a crude product, which was then purified by column chromatography (PE) to give compound 5 as a white solid (15.42 g, 63.82%).

[0223] 1 H NMR (300MHz, Chloroform-d) δ7.16 (d, J = 7.6Hz, 2H), 7.12–7.04 (m, 2H), 3.37 (s, 4H).

[0224] 5-2. Preparation of INT-2

[0225]

[0226] Compound 5 (15.42 g, 81.08 mmol) was added to a solution of potassium dichromate (109.72 g, 372.96 mmol) in glacial acetic acid (220 mL). The solution was stirred at 80 °C for 12 hours. The reaction mixture was quenched with EA and water, and extracted twice with EA. The combined organic layers were washed with water and brine, dried over Na₂SO₄, and filtered. The filtrate was concentrated to give the crude product, which was then added to PE and an ultrasonic dispersion. The resulting precipitate was filtered and dried to obtain crude INT-2 (10.55 g) as a brownish-yellow solid, which was ready for use without further purification.

[0227] 5-3. Preparation of AIR-2-1

[0228]

[0229] INT-2 (5 g, 21.35 mmol) was dissolved in EtOH (120 mL). The reaction system was purged three times with argon gas, and then heated to 50 °C. N2 was added. 1 N 1-Dimethylethane-1,2-diamine (2.79 mL, 25.62 mmol) was heated to 80 °C (reflux) for 4 hours. TLC showed the reaction was complete. The reaction mixture was concentrated, and the residue obtained was purified by column chromatography (DCM:MeOH = 30:1 to 10:1) to give a brownish-yellow solid AIR-2-1 (1.81 g, 27.86%). MS (ESI), m / z: 305.4 [M+H] + .

[0230] 1 H NMR (300MHz, Chloroform-d) δ8.65(t,J=2.5Hz,1H),8.62(t,J=2.5Hz,1H),7.47–7.39(m,2H),4.38(t,J=6.7Hz,2H),2.89–2.81(m,2H),2.49(s,6H).

[0231] 5-4. Preparation of AIR-2

[0232]

[0233] AIR-2-1 (1.81 g, 5.95 mmol) was dissolved in 1,4-dioxane (150 mL), and the reaction system was purged three times with argon. 1-(2-pyridone-ethyl)piperazine (compound A) (1.20 g, 6.54 mmol) was added, and the mixture was stirred overnight at room temperature. The reaction mixture was concentrated, and the residue was purified by column chromatography (DCM:MeOH = 15:1 to 5:1) to give the pure product dissolved in DCM. Then, 5 M HCl / 1,4-dioxane (1.5 mL) was added and stirring continued, followed by precipitation with diethyl ether. The precipitate was filtered, washed with diethyl ether, and dried to give a yellow solid AIR-2 (1.21 g). MS (ESI), m / z: 468.47 [M+H] + .

[0234] 1 H NMR(300MHz,D2O)δ8.36(dd,J=8.3,4.7Hz,1H),8.29(d,J=8.3Hz,1H),7.40(dd,J=13.1,8.3Hz,1H),7.27(d,J=8.4Hz,1 H),4.42(t,J=6.0Hz,2H),3.90–3.54(m,12H),3.47(t,J=6.0Hz,2H),3.38-3.12(m,4H),2.99(s,6H),2.21–2.02(m,4H).

[0235] Example I.6: Synthesis of compound F (AIR-3)

[0236]

[0237] 6-1. Preparation of Compound 2

[0238]

[0239] NBS (499.72 g, 2.81 mol) was added to DMF (1000 mL) and stirred until most of the solid dissolved. The reaction system was purged twice with argon and then cooled to 9 °C. Compound 1 (176 g, 1.14 mol) was added, and the mixture was stirred at 9 °C for 2 hours. The reaction mixture was then allowed to warm to room temperature naturally and stirred over the weekend. The reaction solution was filtered, and the filter cake was washed twice with PE / EA = 1 / 1. The dried filter cake was collected and recrystallized from EA to give pure compound 2 (102.57 g, 28.80%) as off-white crystals.

[0240] 1 H NMR (300 MHz, Chloroform-d) δ 7.79 (d, J = 7.4 Hz, 2H), 7.09 (d, J = 8.8 Hz, 2H), 3.30 (s, 4H).

[0241] 6-2. Preparation of Compound 3

[0242]

[0243] Compound 2 (50 g, 160.26 mmol) was dissolved in anhydrous THF (950 mL). The reaction mixture was purged three times with argon and then cooled to -78 °C. Then, n-BuLi (76.92 mL, 2.5 M / L, 192.31 mmol) was added dropwise, and the reaction mixture was stirred at the same temperature for at least 20 minutes. N-fluorobis(benzenesulfonamide) (75.8 g, 240.38 mmol) was added dropwise to 250 mL of anhydrous THF, and the mixture was stirred at -78 °C for 1.5 hours. The reaction mixture was then allowed to warm to room temperature naturally and stirred overnight. The reaction mixture was quenched with NH4Cl solution and then extracted twice with EA. The combined organic layers were washed with water and brine, dried over Na2SO4, and filtered. The filtrate was concentrated to give the crude product, which was added to DCM and stirred. The precipitate obtained by filtration was washed twice with DCM, all filtrates were combined and concentrated under vacuum to obtain crude product, which was then purified by column chromatography (PE:EA = 50:1) to obtain compound 3 (31.90 g, 79.27%) as a white solid.

[0244] 1H NMR (300MHz, Chloroform-d) δ7.66 (d, J = 7.4Hz, 1H), 7.21–7.09 (m, 3H), 3.35 (s, 4H).

[0245] 6-3. Preparation of Compound 4

[0246]

[0247] Compound 3 (9.29 g, 37.00 mmol) was added to a solution of potassium dichromate (50.07 g, 170.19 mmol) in glacial acetic acid (100 mL). The solution was heated under reflux for 16 hours and then poured into ice water. The precipitate was collected by filtration and washed with water. The filter cake was air-dried, added to DCM and refluxed for 30 minutes, filtered, washed with DCM, and the filter cake was added to DCM and refluxed for 15 minutes, filtered, and washed with DCM. All filtrates were concentrated to give crude compound 3 (6.72 g), which could be used without further purification.

[0248] 6-4. Preparation of AIR-3-1

[0249]

[0250] To a 20 mL solution of compound 4 (400 mg, 1.36 mmol) in EtOH, DIPEA (358.48 μL, 2.17 mmol) and tert-butyl (2-aminoethyl)(methyl)carbamate hydrochloride (428.46 mg, 2.03 mmol) were added. The reaction mixture was purged with argon five times and then stirred (refluxed) at 80 °C for 2 hours. TLC showed that the reaction was complete. The reaction mixture was concentrated, and the residue was purified by column chromatography (PE:EA = 10:1 to 4:1) to give a yellow solid AIR-3-1 (244 mg, 39.88%).

[0251] 6-5. Preparation of AIR-3-2

[0252]

[0253] To a 15 mL solution (244 mg, 540.68 mmol) of AIR-3-1 in toluene, NaOt-Bu (103.92 mg, 1.08 mol), 1-(2-pyridone-ethyl)piperazine (118.92 mg, 648.81 mmol), and BINAP (101.00 mg, 162.20 mmol) were added. The reaction mixture was purged with argon five times, followed by the addition of Pd(OAc)₂ (24.28 mg, 108.14 mmol). The reaction mixture was purged with argon three times, and the reaction mixture was stirred at 90 °C for 12 hours under argon protection. The reaction mixture was quenched with water and extracted twice with EA. The combined organic layers were washed with water and brine, dried over Na₂SO₄, and filtered. The filtrate was concentrated to obtain the crude product, which was purified by column chromatography (DCM:MeOH = 100:1 to 10:1, with the addition of NH3·H2O) and Prep-TLC (DCM:MeOH = 10:1, with the addition of NH3·H2O) to obtain AIR-3-2 (40 mg, 13.36%).

[0254] 1 H NMR(300MHz,Chloroform-d)δ8.56(dd,J=8.2,4.7Hz,1H),8.51(d,J=8.3Hz,1H),7.29(dd,J=7.7,5.1Hz,1H),7.15(d,J=8.3Hz, 1H),4.36–4.29(m,2H),3.62-3.55(m,2H),3.45–3.11(m,6H),2.95(s,3H),2.82–2.62(m,10H),1.82-1.89(m,4H),1.14(s,9H).

[0255] 6-6. Preparation of AIR-3

[0256]

[0257] AIR-3-2 (40 mg, 72.24 mmol) in a DCM / MeOH = 10 / 1 solution (3 mL) was purged three times with argon, followed by the addition of 5 M HCl / 1,4-dioxane (1.5 mL). The resulting mixture was stirred at room temperature for 1.5 hours. TLC showed the reaction was complete. The reaction mixture was precipitated with diethyl ether, centrifuged, and the precipitate was washed with diethyl ether, centrifuged, and dried to give 33 mg of crude product. The crude product was further purified by Prep-TLC (DCM:MeOH = 20:1, with the addition of NH3·H2O) to give AIR-3 (10 mg) as a yellow solid. MS (ESI), m / z: 454.58 [M+H] +.HPLC 92.439% (220nm), HPLC 96.712% (254nm).

[0258] 1 H NMR(300MHz,D2O)δ8.35(dd,J=8.4,4.7Hz,1H),8.28(d,J=8.3Hz,1H),7.39(dd,J=13.2,8.5Hz,1H),7.26(d,J=8.3Hz,1H),4. 39–4.31(m,2H),3.86-3.62(m,14H),3.37–3.31(m,2H),3.25–3.15(m,2H),2.70(s,3H),2.24–2.13(m,2H),2.08-1.98(m,2H).

[0259] Example II. Activity Test

[0260] Example II.1: Testing the activation of the hnRNPA2B1 and TBK1-IRF3 pathways in tumor cells by the compound.

[0261] A549 lung cancer cells (cell density of 1 x 10^5 cells per well) were stimulated with 20 μM test compound E (prepared as in Example I.5, dissolved in physiological saline) for 30 minutes. Subcellular localization of hnRNPA2B1 was detected by immunofluorescence, and activation of TBK1 and IRF3 was detected by SDS-PAGE electrophoresis and immunoblotting.

[0262] The activation status of hnRNPA2B1, TBK1, and IRF3 is as follows: Figure 1 , 2 As shown.

[0263] The results showed that compound E significantly activated the nuclear export of tumor cells hnRNPA2B1 and the activation of downstream TBK1 and IRF3. Furthermore, similar results were obtained when other compounds prepared in Example I were used in the same experiment.

[0264] The above results demonstrate that the tested compound activates the hnRNPA2B1 pathway and is an hnRNPA2B1 agonist.

[0265] Example II.2: Testing the broad induction of type I interferon production in various tumor types by the compound

[0266] A549 (non-small cell lung cancer), HepG2 (liver cancer), LLC (Lewis lung cancer), and B16F10 (melanoma) tumor cells were stimulated with a 20 μM concentration of test compound E (cell density 1 x 10^4 cells per well); after 18 hours of stimulation, the cell culture supernatant was collected, and the IFN-β protein level was detected by ELISA.

[0267] Test the activation of type I interferon by the compound, such as Figure 3 As shown.

[0268] The results showed that compound E significantly activated the expression of type I interferon in various cells. Furthermore, similar results were obtained when other compounds prepared in Example I were used in the same experiment.

[0269] The above results demonstrate that the tested compound can induce the production of type I interferon by activating the hnRNPA2B1 pathway, and is therefore a biologically active hnRNPA2B1 agonist. Thus, this disclosure defines such novel compounds as hnRNPA2B1 agonists.

[0270] Example II.3: Synergistic killing of tumor cells by hnRNPA2B1 agonist combined with chemotherapy agents

[0271] A549 tumor cells were stimulated with a combination of hnRNPA2B1 agonist compound E (5 μM) and the chemotherapeutic agent oxaliplatin (62.5 μM, purchased from Selleck, catalog number S1224). Apoptosis of the tumor cells was detected by flow cytometry after 36 hours.

[0272] The results are as follows Figure 4 As shown.

[0273] The results showed that the hnRNPA2B1 agonist compound E could synergistically exert a stronger tumor-killing effect with chemotherapeutic drugs. Furthermore, similar results were obtained when the same experiments were performed using other compounds prepared in Example I.

[0274] Example II.4: Synergistic killing of tumor cells by hnRNPA2B1 agonist combined with chemotherapy agents

[0275] A549 tumor cells were stimulated with a combination of hnRNPA2B1 agonist compound E (5 μM) and the chemotherapeutic agent oxaliplatin (62.5 μM, purchased from Selleck, catalog number S1224). After 16 hours, total cellular protein was collected, and Western blotting was used to detect caspase 3 cleavage and DNA fragmentation (γH2AX) in the tumor cells.

[0276] The results are as follows Figure 5 As shown.

[0277] The results showed that the hnRNPA2B1 agonist compound E synergistically promoted DNA breakage and apoptosis with chemotherapeutic drugs, thus exerting a stronger tumor-killing effect. Furthermore, similar results were obtained when other compounds prepared in Example I were used in the same experiments.

[0278] Bioevaluation

[0279] The compounds disclosed herein are defined as hnRNPA2B1 agonists by: (i) binding to the hnRNPA2B1 protein, as demonstrated by Biacore assays of the high affinity of the compounds to the hnRNPA2B1 protein at concentrations as low as 20 μM; and (ii) demonstrated by cellular analysis that the compounds can broadly induce higher levels of IFN-β at concentrations as low as 20 μM.

[0280] (i) Biacore T200 detection of the binding of the compound to hnRNPA2B1

[0281] The S-series CM5 chip was used; amino coupling reagents (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide EDC and N-hydroxysuccinimide NHS);

[0282] Coupling buffer: 10mM sodium acetate, pH 4.0, pH 4.5;

[0283] Run buffer: 10×PBS-P+;

[0284] Analytical grade DMSO, deionized water (0.22μm membrane filtration);

[0285] hnRNPA2B1 recombinant protein (concentration must be greater than 0.5 mg / mL, total protein amount must be at least 20 μg);

[0286] Compound: Mother liquor concentration greater than 20 mM, volume greater than 30 μL, purity greater than 90%, dissolved in 100% DMSO;

[0287] Other consumables: 96-well plates and sealing film, 1.5mL centrifuge tubes without caps, and type 2 rubber bottle caps.

[0288] Experimental steps:

[0289] 1. Place the running buffer 1×PBS and waste bottle in the designated locations, and then place the chip.

[0290] 2. Protein coupling. Select the immobilization program in the Wizard template, check flow cell 2 or 4, select amine as the method, select specify contact time and flow rate to achieve high coupling, and enter contact time 900s and flow rate 10μL / min.

[0291] 3. Prepare the required protein solution according to the instructions. Dilute the protein with sodium acetate at pH 4.0 to the required volume, with a final concentration of 20 μg / mL. Place 100 μL EDC, 100 μL NHS, and 140 μL ethanolamine into the corresponding positions.

[0292] 4. The system will automatically coat the chip surface with the target amount of hnRNPA2B1 protein and automatically generate a coupling report.

[0293] 5. Prepare the running buffer for interaction. For small molecule samples, use 1×PBS-P+ containing 5% DMSO: Dilute 105 mL of 10×PBS-P+ with deionized water to 1 L to prepare 1.05×PBS-P+.

[0294] Solvent-corrected stock solution containing 4.5% DMSO: 1.05×PBS-P + 9.5 mL + 0.45 mL DMSO

[0295] Solvent correction stock solution containing 5.8% DMSO: 1.05×PBS-P + 9.5mL + 0.58mL DMSO

[0296] Run buffer containing 5% DMSO: 1.05×PBS-P + 950mL + 50mL DMSO

[0297] Dilute the 10mM small molecule stock solution with DMSO-free 1.05×PBS-P+ buffer: 0.7μL small molecule stock solution + 6.3μL DMSO + 133μL 1.05×PBS-P+ buffer, for a total volume of 140μL and a final concentration of 50μM. Add to 96-well plates and seal with film.

[0298] After the test is completed, binding affinity is determined using Kinetics or Affinity mode.

[0299] (ii) Total RNA from cells can be extracted using TRIzol or a rapid RNA extraction kit.

[0300] 24-well plate cells (approximately 2-3 × 10⁻⁶) 5Rapid RNA extraction (cells / well): Discard the culture medium, add 500 μL of RA2 lysis buffer to each well, lyse by pipetting several times, transfer to the inner sleeve of a dedicated adsorption tube, and centrifuge at 12000 rpm for 1 min at room temperature. Discard the filtrate, add 500 μL of wash buffer, and centrifuge at 12000 rpm for 1 min at room temperature. Repeat the washing process once. Discard the filtrate, replace the inner sleeve, and centrifuge at 12000 rpm for 2 min at room temperature without adding wash buffer. Transfer the inner sleeve to a new 1.5 mL centrifuge tube, add 25 μL of Elution Buffer to the center of the membrane, incubate at room temperature for 5 min, and centrifuge at 12000 rpm for 1 min at room temperature. The resulting filtrate is the RNA; determine the RNA concentration using Nanodrop One.

[0301] qRT-PCR: After determining the concentration of each sample, take 1 μg of total RNA and use ReverTra... The qRT-PCR RTMaster Mix reverse transcribes RNA into cDNA. The reverse transcription system and reaction conditions are as follows:

[0302] Reverse transcription reaction system (20 μL system)

[0303]

[0304] Reverse transcription reaction conditions (20 μL system)

[0305]

[0306] Add 60 μL of deionized water to the cDNA sample obtained by reverse transcription, mix well, and then centrifuge briefly. Primer sequence information used for qRT-PCR is shown in the table. The relative quantification of gene mRNA levels was performed using SYBR Green Realtime PCR Master Mix.

[0307] qRT-PCR reaction system (20 μL)

[0308]

[0309] qRT-PCR reaction conditions

[0310]

[0311] qRT-PCR primers

[0312]

[0313]

[0314] All documents mentioned in this disclosure are incorporated herein by reference as if each document were individually incorporated herein by reference. Furthermore, it should be understood that after reading the foregoing teachings of this disclosure, those skilled in the art can make various alterations or modifications to this disclosure, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A product selected from: pharmaceutical compositions, pharmaceutical products, kits, or cassettes, comprising a fused cyclic compound of formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically or immunologically acceptable carrier or excipient. in, L is -(CH2)n-, and n is 1, 2, or 3; X is a halogen; R 1 For -NR 4 R 5 , where R 4 R 5 Each independently is H or C 1-6 alkyl; R 2 and R 3 Each independently is C 1-3 Alkyl, the C 1-3 The alkyl group is either unsubstituted or substituted with a 4-6 membered heterocyclic alkyl group containing one nitrogen atom; Or R 2 and R 3 Together with the N atoms to which they are attached, they form a piperazine group, which is bonded by a 4-6 membered heterocyclic alkyl group. 1-6 Alkyl or hydroxy C 1-6 Alkyl substitution, The product may also contain or be used in combination with chemotherapy agents; The product is used to induce type I interferon production in the subject and / or to prevent and / or treat tumors in the subject.

2. The product as described in claim 1, wherein, The chemotherapeutic agents are selected from: chemotherapeutic agents that interfere with DNA replication; chemotherapeutic agents that interfere with RNA transcription; chemotherapeutic agents that interfere with microtubules; and chemotherapeutic agents that inhibit nucleotide synthesis and metabolism.

3. The product as described in claim 1, wherein, The chemotherapeutic agents are selected from platinum-based chemotherapeutic agents, alkylating agents, topoisomerase I inhibitors, anthracyclines, taxanes, and antimetabolites.

4. The product as described in claim 3, wherein, The platinum-based chemotherapy agents are selected from oxaliplatin, cisplatin, and carboplatin; the alkylating agent is selected from cyclophosphamide; the topoisomerase I inhibitor is selected from irinotecan; the anthracyclines are selected from doxorubicin and epirubicin; and the antimetabolites are selected from gemcitabine, methotrexate, 5-FU, and capecitabine.

5. The product as described in claim 1, in, X is either F or Cl; L is -(CH2)n-, where n is 1 or 2; R 4 R 5 Each can be H, methyl, or ethyl.

6. The product as described in claim 1, in, X is F; L is -(CH2)n-, where n is 2; R 4 R 5 Each can be H, methyl, or ethyl.

7. The product as described in claim 1, in, X is F; L is -(CH2)n-, where n is 2; R 4 R 5 Each can be independently H, methyl, or ethyl; R 2 and R 3 Each is independently methyl, ethyl, or isopropyl, wherein the methyl, ethyl, or isopropyl groups are either unsubstituted or substituted with pyrrolidinyl groups; Or R 2 and R 3 Together with the N atoms to which they are attached, they form a piperazine group, which is substituted with a pyrrolidinyl ethyl or a hydroxyethyl.

8. The product of claim 1, wherein the compound has the following formula (II): in, R a R b Each independently is H or C 1-6 alkyl; M is -(CH2)n-, where n is 1, 2 or 3; X is either F or Cl; R c It is a hydroxyl group containing 1-2 heteroatoms selected from nitrogen, oxygen or sulfur, and is a 4-6 membered heterocyclic alkyl group.

9. The product of claim 8, wherein R c It is a hydroxyl group or a 4-6 membered heterocyclic alkyl group containing one nitrogen atom.

10. The product of claim 8, wherein R c It is hydroxyl or pyrroleyl.

11. The product of claim 8, wherein R c It is hydroxyl or 1-pyrrolidinyl.

12. The product of claim 1, wherein the compound is selected from the group consisting of: Or its hydrochloride salt.

13. The use of the fused cyclic compound of formula (I) or a pharmaceutically acceptable salt thereof in the preparation of products for use in combination with chemotherapeutic agents to induce type I interferon production in a subject, or for use in combination with chemotherapeutic agents to prevent and / or treat tumors in a subject. in, L is -(CH2)n-, and n is 1, 2, or 3; X is a halogen; R 1 For -NR 4 R 5 , where R 4 R 5 Each independently is H or C 1-6 alkyl; R 2 and R 3 Each independently is C 1-3 Alkyl, the C 1-3 The alkyl group is either unsubstituted or substituted with a 4-6 membered heterocyclic alkyl group containing one nitrogen atom; Or R 2 and R 3 Together with the N atoms to which they are attached, they form a piperazine group, which is bonded by a 4-6 membered heterocyclic alkyl group. 1-6 Alkyl or hydroxy C 1-6 Alkyl substitution.

14. The application as described in claim 13, wherein, The object is a person; and / or Wherein, the production of type I interferon is mediated by hnRNPA2B1; and / or Wherein, the tumor prevention and / or treatment of the subject can benefit from the production or increased levels of type I interferon in the body; and / or The tumor is selected from one or more tumors in the group consisting of: solid tumors; non-solid tumors; and / or The product is selected from: pharmaceutical compositions, pharmaceutical products, reagent kits or medicine boxes; and / or The chemotherapeutic agents are selected from: chemotherapeutic agents that interfere with DNA replication; chemotherapeutic agents that interfere with RNA transcription; chemotherapeutic agents that interfere with microtubules; chemotherapeutic agents that inhibit nucleotide synthesis metabolism; and / or The product is used to induce type I interferon production in subjects and / or to prevent and / or treat tumors in subjects.

15. The application as described in claim 14, wherein, The tumor is one or more tumors selected from the group consisting of: liver cancer, lung cancer, melanoma, breast cancer, ovarian cancer, prostate cancer, kidney cancer, colorectal cancer, head and neck cancer, skin cancer, bladder cancer, pancreatic cancer, hematologic malignancies, and nervous system tumors; and / or The chemotherapeutic agents are selected from platinum-based chemotherapeutic agents, alkylating agents, topoisomerase I inhibitors, anthracyclines, taxanes, and antimetabolites.

16. The application as described in claim 14, wherein, The tumor is one or more tumors selected from the group consisting of: leukemia, glioma; and / or The platinum-based chemotherapy agents are selected from oxaliplatin, cisplatin, and carboplatin; the alkylating agent is selected from cyclophosphamide; the topoisomerase I inhibitor is selected from irinotecan; the anthracyclines are selected from doxorubicin and epirubicin; and the antimetabolites are selected from gemcitabine, methotrexate, 5-FU, and capecitabine.

17. The application as described in claim 14, wherein, The tumor is one or more tumors selected from the group consisting of: non-small cell lung cancer, liver cancer, Lewis lung cancer, melanoma, breast cancer, and colon cancer.

18. The application as described in claim 13, in, X is either F or Cl; L is -(CH2)n-, where n is 1 or 2; R 4 R 5 Each can be H, methyl, or ethyl.

19. The application as described in claim 13, in, X is F; L is -(CH2)n-, where n is 2; R 4 R 5 Each can be H, methyl, or ethyl.

20. The application as described in claim 13, in, X is F; L is -(CH2)n-, where n is 2; R 4 R 5 Each can be independently H, methyl, or ethyl; R 2 and R 3 Each is independently methyl, ethyl, or isopropyl, wherein the methyl, ethyl, or isopropyl groups are either unsubstituted or substituted with pyrrolidinyl groups; Or R 2 and R 3 Together with the N atoms to which they are attached, they form a piperazine group, which is substituted with a pyrrolidinyl ethyl or a hydroxyethyl.

21. The application as described in claim 13, wherein the compound has the following formula (II): in, R a R b Each independently is H or C 1-6 alkyl; M is -(CH2)n-, where n is 1, 2 or 3; X is either F or Cl; R c It is a hydroxyl group containing 1-2 heteroatoms selected from nitrogen, oxygen or sulfur, and is a 4-6 membered heterocyclic alkyl group.

22. The application as described in claim 21, wherein R c It is a hydroxyl group or a 4-6 membered heterocyclic alkyl group containing one nitrogen atom.

23. The application as described in claim 21, wherein R c It is hydroxyl or pyrroleyl.

24. The application as described in claim 21, wherein R c It is hydroxyl or 1-pyrrolidinyl.

25. The application as described in claim 13, wherein the compound is selected from the group consisting of: Or its hydrochloride salt.

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