Hnrnpa2b1 agonist compound and use thereof in tumor prevention and treatment

Abstract

The present disclosure relates to a heterogeneous nuclear ribosomal protein A2B1(hnRNPA2B1)hnRNPA2B1 agonist compound and an anti-tumor use thereof. Specifically, the present disclosure relates to a condensed cyclic compound as represented by formula (I), a cis-trans isomer thereof, an enantiomer thereof, a diastereoisomer thereof, a racemate thereof, a solvate thereof, a hydrate or a pharmaceutically acceptable salt thereof or a prodrug thereof, a composition and product comprising any one of the foregoing, and a use of the described substances as an hnRNPA2B1 agonist in the preparation of a product for tumor prevention and / or treatment. The present disclosure can be used for broad-spectrum tumor prevention and treatment, and has a wide application prospect.

Description

hnRNPA2B1 agonist compounds and their applications in tumor prevention and treatment Technical Field

[0001] The present disclosure relates to the fields of medicine and biotechnology. Specifically, the present disclosure relates to a class of compounds that act as agonists of the nuclear protein hnRNPA2B1 (i.e., heterogeneous nuclear ribonucleoprotein A2B1), products containing the compounds, and their effects, mechanisms of action, methods of implementation, and uses in preventing or treating tumor-related diseases or symptoms, and tumor-induced damage. Background Art

[0002] Type I interferon is crucial for 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 synergize with various chemotherapy drugs. Type I interferon can significantly enhance anti-tumor immune responses by inducing the activation of adaptive and innate immune cells, especially by enhancing the uptake, processing, presentation, and cross-presentation of antigens by dendritic cells and macrophages to T cells, thereby enhancing T cell activation. At the same time, 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 important uses in tumor treatment.

[0003] Defects in the interferon activation pathway within tumor cells often lead to tumor immune escape and failure of immune checkpoint blockade therapy. A variety of small molecule drugs targeting tumors work by inducing an increase in free autologous nucleic acids, thereby activating interferon production. These drugs include DNA intercalators, various epigenetic drugs, and spliceosome regulators. Recent studies have shown that the cGAS-STING pathway in the cytoplasm can induce type I interferon, making the development of STING agonists an important direction in tumor treatment. However, increasing evidence indicates that the cGAS-STING pathway is underexpressed in tumor cells or that signaling pathway activation is blocked. Furthermore, there is also evidence that cGAS can promote tumor cell survival. Therefore, in the field of tumor immunotherapy, there is an urgent need to develop agonists for 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 nucleus, belonging to the hnRNP family. hnRNP-A2B1 can sense and recognize the nucleic acid components of DNA viruses (such as HSV-1) and self-activate to form homodimers. Demethylation is mediated by the demethylase JMJD6, leading to translocation from the nucleus to the cytoplasm, where it activates the TBK1-IRF3 signaling pathway and initiates the expression of type I interferons, exerting an antiviral effect. Furthermore, as an RNA-binding protein, hnRNP-A2B1 promotes m6A modification, nucleocytoplasmic translocation, and translation of cGAS, IFI16, and STING mRNAs, thereby ensuring sufficient induction of type I interferon expression.

[0005] hnRNPA2B1 is overexpressed in a variety of tumors, including lung cancer, liver cancer, breast cancer, pancreatic cancer, and glioblastoma, and is widely considered to be a key factor in promoting tumorigenesis and progression. In recent decades, numerous studies have demonstrated that hnRNPA2B1 is involved in regulating a variety of fundamental biological functions, such as cellular metabolism, migration and invasion, proliferation, and responses to mitochondrial stress. For example, hnRNPA2-mediated activation of the invasive phenotype involves diverse mechanisms, including alternative splicing of TP53INP2, activation of the CXCL12 / CXCR4 axis, and activation of invasive behavior following 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 widely involved in regulating cancer cell phenotypes, including metabolism, proliferation, apoptosis, migration, and invasion, through diverse molecular mechanisms. Furthermore, hnRNPA2B1 has been reported to induce epithelial-mesenchymal transition (EMT) in multiple cancer cell lines. Furthermore, hnRNPA2B1 is involved in regulating fundamental cancer processes such as aerobic glycolysis. It has been reported that hnRNPA2 regulates the alternative splicing of the pyruvate kinase isoenzyme M2 (PKM2), activating the metabolic switch of cancer cells to aerobic glycolysis. hnRNPA2B1 also plays a key role in regulating hypoxia. Overall, hnRNPA2B1 is widely involved in the occurrence and progression of various tumors. The development of small molecules targeting hnRNPA2B1 has the potential to block the tumor-promoting effects of hnRNPAB1 and provide effective support for cancer treatment.

[0006] In summary, the development of specific agonists targeting hnRNPA2B1 has great potential in the treatment of tumors. There is an urgent need in this field to develop immunologically active substances that can activate or promote interferon production, enhance anti-tumor effects, and effectively prevent tumor metastasis.

[0007] Summary of the Invention

[0008] The present disclosure provides fused ring compounds having a structure shown in formula (I), their related derivatives (e.g., their cis-trans isomers, their enantiomers, their diastereomers, their racemates, their solvates, their hydrates, or their pharmaceutically acceptable salts or prodrugs thereof), and products comprising the compounds or their related derivatives. The present disclosure also provides uses of the compounds, derivatives, and products in tumor prevention and treatment, and further provides uses of the compounds, derivatives, and products in treating or preventing tumors and related diseases or symptoms. The compounds, drugs, pharmaceutical compositions, or kits disclosed herein can be used to effectively fight tumors and control the occurrence and / or metastasis of tumors.

[0009] In some aspects of the present disclosure, provided are fused ring compounds represented by formula (I), their cis-trans isomers, their enantiomers, their diastereomers, their racemates, their solvates, their hydrates, or their pharmaceutically acceptable salts or their prodrugs.

[0010] Wherein, L is -(CH2)n-, and n is an integer from 0 to 6;

[0011] X is a halogen;

[0012] R 1 -NR 4 R 5 , where R 4 、R 5 Each independently is H or C 1-6 alkyl;

[0013] R 2 and R 3 Each independently is H or C 1-6 alkyl;

[0014] or R 2 and R 3 Together with the nitrogen atom to which they are attached, they form a 5-8 membered heterocycloalkyl group containing 1-3 heteroatoms selected from nitrogen, oxygen or sulfur;

[0015] The C 1-6 The alkyl group and the 5-8 membered heterocycloalkyl group are unsubstituted or substituted by one or more substituents selected from the group consisting of hydroxy, amino, halogen, C 1-6 Alkyl, C 2-6 Alkenyl, C2-6 Alkynyl, 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 heterocycloalkyl, 3-6 membered heterocycloalkyl C 1-6 Alkyl, 6-10 membered aryl, 6-10 membered aryl C 1-6 Alkyl or 5-10 membered heteroaryl and 5-10 membered heteroaryl C 1-6 alkyl.

[0016] In some aspects of the present disclosure, provided is the use of the compound of the present invention in preparing a product for preventing and / or treating tumors.

[0017] In some aspects of the present disclosure, a method for preventing and / or treating tumors is also provided, comprising administering a preventively and / or therapeutically effective amount of a compound or product of the present disclosure to a subject in need thereof.

[0018] In some aspects of the present disclosure, the compounds or products of the present disclosure are also provided for use in preventing and / or treating tumors.

[0019] In some aspects of the present disclosure, there is provided use of a compound or product of the present disclosure for increasing the level of an interferon (eg, type I interferon, such as IFN-α and / or IFN-β).

[0020] Those skilled in the art may arbitrarily combine the aforementioned technical solutions and technical features without departing from the inventive concept and protection scope of the present disclosure. Other aspects of the present disclosure will be apparent to those skilled in the art from the disclosure herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] The present disclosure will be further described below in conjunction with the accompanying drawings, wherein these drawings are only for illustrating the embodiments of the present disclosure and are not intended to limit the scope of the present disclosure.

[0022] Figure 1: Activation effect of test compounds on hnRNPA2B1 nuclear export in A549 lung cancer cells.

[0023] Figure 2: Effects of test compounds on the activation of downstream TBK1 and IRF3 after hnRNPA2B1 nuclear export in A549 lung cancer cells.

[0024] Figure 3: Effects of test compounds on activation of type I interferon in tumor cells.

[0025] Figure 4: Inhibitory effects of test compounds on tumor cells.

[0026] Figure 5: Killing effect of test compound E on tumor cells.

[0027] Figure 6: Test compounds inhibit tumor growth.

[0028] Figure 7: Test compounds prolong the survival of tumor-bearing mice.

[0029] Figures 8 and 9: Test compounds inhibit experimental lung metastasis of B16F10 tumors. DETAILED DESCRIPTION

[0030] Through extensive research, development, and testing, this application discovered a new class of fused ring compounds that can specifically bind to the hnRNPA2B1 protein with high affinity and can further widely induce the production of higher levels of IFN-β, and are thus defined as hnRNPA2B1 agonists.

[0031] In anti-tumor tests on various tumor cell types, the hnRNPA2B1 agonist compounds disclosed herein significantly activated type I interferon in tumor cells, inhibited LLC tumor proliferation, and promoted the survival of 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 may exert anti-tumor effects through the action of type I interferon.

[0032] Thus, the present disclosure provides methods and strategies for applying novel anti-tumor hnRNPA2B1 agonist compounds in the prevention and treatment of tumors.

[0033] All numerical ranges provided herein are intended to expressly include all values ​​falling between the endpoints of the ranges and the ranges therebetween. Features disclosed herein or in the embodiments may be combined. All features disclosed herein may be used in any combination, and each feature disclosed herein may be replaced by any alternative feature that serves the same, equivalent, or similar purpose. Therefore, unless otherwise specified, the features disclosed herein are merely general examples of equivalent or similar features.

[0034] As used herein, “containing,” “having,” or “including” encompasses “comprising,” “mainly consisting of,” “substantially consisting of,” and “consisting of”; “mainly consisting of,” “substantially consisting of,” and “consisting of” are subordinate concepts of “containing,” “having,” or “including.”

[0035] hnRNPA2B1 agonist compounds

[0036] As used herein, the terms "hnRNPA2B1 protein (polypeptide)" and "hnRNPA2B1" are used interchangeably to refer to the heterogeneous nuclear riboprotein A2B1. The hnRNPA2B1 protein referred to in the present disclosure can be a protein encoded by the hnRNPA2B1 gene, cDNA, or CDS thereof, in an animal (e.g., human or mouse), or a homologous sequence of such protein that promotes interferon expression (e.g., a homologous sequence of hnRNPA2B1 can be obtained through databases or alignment software known in the art), a variant, or a modified form thereof.

[0037] As used herein, the terms "hnRNPA2B1 gene," "hnRNPA2B1-encoding gene," "hnRNPA2B1 protein-encoding gene," or "hnRNPA2B1-encoding nucleic acid molecule" are used interchangeably to refer to a nucleotide sequence encoding the hnRNPA2B1 protein or polypeptide described herein, such as the human hnRNPA2B1 gene (Gene ID: 3181) or the mouse hnRNPA2B1 gene (Gene ID: 53379). The term also encompasses molecules that hybridize under stringent conditions to a signed nucleic acid molecule or molecules of family genes that are highly homologous to such molecules. Expression of such genes is believed to promote the production and effects of interferon.

[0038] This disclosure provides a novel class of compounds that act as hnRNPA2B1 "agonists" (or "promoters"). The terms "agonist" and "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. These compounds specifically bind to the hnRNPA2B1 protein with high affinity and can further induce the production of higher levels of IFN-β, thereby exerting an hnRNPA2B1 agonist effect.

[0039] The hnRNPA2B1 agonist compounds disclosed herein can inhibit tumors and thus can be further used to prevent or treat tumor-related diseases and / or tumor-induced related symptoms.

[0040] The present application provides a fused ring compound represented by formula (I), its cis-trans isomers, its enantiomers, its diastereomers, its racemates, its solvates, its hydrates, or its pharmaceutically acceptable salts or prodrugs thereof.

[0041] Wherein, L is -(CH2)n-, and n is an integer from 0 to 6;

[0042] X is a halogen;

[0043] R 1 -NR 4 R5 , where R 4 、R 5 Each independently is H or C 1-6 alkyl;

[0044] R 2 and R 3 Each independently is H or C 1-6 alkyl;

[0045] or R 2 and R 3 Together with the nitrogen atom to which they are attached, they form a 5-8 membered heterocycloalkyl group containing 1-3 heteroatoms selected from nitrogen, oxygen or sulfur;

[0046] The C 1-6 The alkyl group and the 5-8 membered heterocycloalkyl group are unsubstituted or substituted by one or more substituents selected from the group consisting of hydroxy, amino, halogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, 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 heterocycloalkyl, 3-6 membered heterocycloalkyl C 1-6 Alkyl, 6-10 membered aryl, 6-10 membered aryl C 1-6 Alkyl or 5-10 membered heteroaryl and 5-10 membered heteroaryl C 1-6 alkyl.

[0047] Preferably, X is F or Cl, more preferably, X is F.

[0048] Preferably, L is -(CH2)n-, wherein n is 1, 2 or 3, more preferably, n is 2.

[0049] Preferably, R 4 、R 5 Each is independently H, methyl or ethyl.

[0050] Preferably, R 2 and R 3 Each independently is C 1-3 Alkyl, preferably methyl, ethyl or isopropyl, said C 1-3 The alkyl group is unsubstituted or substituted with a 3-8 membered heterocycloalkyl 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, said C1-3 The alkyl group is unsubstituted or substituted with a 4-6 membered heterocycloalkyl group containing one nitrogen heteroatom. Most preferably, R 2 and R 3 Each independently is C 1-3 The alkyl group is preferably methyl, ethyl or isopropyl, and the C1-3 alkyl group is unsubstituted or substituted by pyrrolidinyl.

[0051] Preferably, R 2 and R 3 Together with the nitrogen atom to which they are attached, they form a 5-8 membered heterocycloalkyl group containing 2 nitrogen atoms, wherein the 5-8 membered heterocycloalkyl group is surrounded by a 3-8 membered heterocycloalkyl group C 1-6 Alkyl or hydroxy C 1-6 Alkyl substituted; more preferably, R 2 and R 3 Together with the nitrogen atom to which they are attached, they form a 6-membered heterocycloalkyl group containing 2 nitrogen atoms, wherein the 6-membered heterocycloalkyl group is surrounded by a 4-6-membered heterocycloalkyl group C 1-6 Alkyl or hydroxy C 1-6 Alkyl substituted; further preferably, R 2 and R 3 Together with the nitrogen atom to which they are attached, they form a piperazinyl group which is substituted by a 4-6 membered heterocycloalkyl C 1-6 Alkyl or hydroxy C 1-6 Alkyl substituted; most preferably, R 2 and R 3 Together with the nitrogen atom to which they are attached, they form a piperazinyl group, which is substituted by a 5-6 membered heterocycloalkyl C containing one nitrogen atom. 1-6 Alkyl substituted or hydroxyl C 1-6 Alkyl is substituted, preferably, the piperazinyl is substituted with pyrrolidinyl C 1-6 Alkyl substituted or hydroxyl C 1-6 Alkyl substituted, more preferably, the piperazinyl is substituted by pyrrolidinylethyl or hydroxyethyl.

[0052] Preferably, the compound has the following formula (II):

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

[0054] M is -(CH2)n-, wherein 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 is hydroxy, amino, or a 3-8 membered heterocycloalkyl group containing 1-2 heteroatoms selected from nitrogen, oxygen, or sulfur. More preferably, R c is hydroxy or a 4-6 membered heterocycloalkyl group containing one nitrogen heteroatom, more preferably, R c is hydroxy or pyrrolidinyl, most preferably, R c is hydroxy or 1-pyrrolidinyl.

[0057] Preferably, the compound is selected from the following group of compounds or their salts (eg, hydrochloride):

[0058] Unless otherwise indicated, the following terms and phrases used herein are intended to have the following definitions. The absence of a specific definition for a particular term or phrase should not be construed as indefinite or unclear, but rather as having its ordinary meaning. When a trade name appears in this document, it is intended to refer to the corresponding commercial product or active ingredient.

[0059] The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0060] The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention, prepared by reacting the compounds of the present invention with relatively nontoxic acids or bases, having specific substituents. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of base in neat solution or in 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 the present invention contain relatively basic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent. Examples of pharmaceutically acceptable 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, bisulfate, hydroiodic acid, phosphorous acid, and the like, and organic acid salts such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid, as well as salts of amino acids (such as arginine), and organic acids such as glucuronic acid. Certain specific compounds of the present invention contain basic and acidic functional groups and can be converted into either base or acid addition salts.

[0061] Pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. 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 of the two.

[0062] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are encompassed within the scope of the present invention.

[0063] Unless otherwise indicated, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of one another.

[0064] Unless otherwise indicated, the term "cis-trans isomers" or "geometric isomers" arises from the inability to rotate freely about double bonds or single bonds forming ring carbon atoms.

[0065] Unless otherwise indicated, the term "diastereomer" refers to stereoisomers that have two or more chiral centers and that are not mirror images of each other.

[0066] The compounds of the present invention may exist in specific forms. Unless otherwise indicated, the term "tautomer" or "tautomeric form" refers to the fact that at room temperature, different functional group isomers are in dynamic equilibrium and can quickly convert into each other. If tautomerism is possible (such as in solution), it is possible to reach a chemical equilibrium of tautomers. For example, proton tautomers (also known as prototropic tautomers) include interconversions carried out by proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions carried out by the reorganization of some bonding monomers. A specific example of keto-enol tautomerization is the interconversion between two tautomers of pentane-2,4-dione and 4-hydroxy-3-ene-2-one.

[0067] The term "prodrug" generally refers to a compound of formula (I) that has been derivatized with a functional group, and the derivative can be readily converted into the compound of formula (I) in vivo. Suitable prodrug selection and preparation can generally be found in, for example, Design of Prodrugs, ed. H. Bundgaard, Elsevier, 1985.

[0068] The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms comprising the compound, said isotopes having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number predominantly found in nature. For example, the compounds may be labeled with radioactive isotopes, such as deuterium ( 2 H), tritium ( 3 H), iodine-125 ( 125 I) or C-14( 14 C). All isotopic variations of the compounds of the present invention, whether radioactive or not, are encompassed by the present invention. Isotopic variations may provide certain therapeutic advantages. For example, deuterated drugs can be formed by replacing hydrogen with heavy hydrogen. The bond formed by deuterium to carbon is stronger than the bond formed by ordinary hydrogen to carbon. Compared to non-deuterated drugs, deuterated drugs have advantages such as reduced toxic side effects, increased drug stability, enhanced therapeutic efficacy, and prolonged biological half-life. Alternatively, they may provide standard compounds useful for characterizing biological samples. Isotopically enriched compounds within Formula (I) can be prepared without undue experimentation by conventional techniques well known to those skilled in the art, or by methods similar to those described in the schemes and examples of the present invention, using appropriate isotopically enriched reagents and / or intermediates.

[0069] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0070] The nomenclature used in the present invention is based on the IUPAC system generated by ChemDraw software. Any open valence bonds appearing on carbon, oxygen, sulfur or nitrogen atoms in the structures given in the present invention indicate the presence of hydrogen atoms.

[0071] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include deuterium and hydrogen variants, as long as the valence state of the particular 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, and oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may be substituted or unsubstituted, and unless otherwise specified, the type and number of substituents can be any on the basis of chemical achievable.

[0072] When any variable (e.g., R) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 0-2 Rs, the group may be optionally substituted with up to two Rs, with each occurrence of R being an independent choice. Furthermore, combinations of substituents and / or their variants are permissible only if such combinations result in stable compounds.

[0073] When the number of a linking group is 0, such as -(CRR)0-, it means that the linking group is a single bond.

[0074] When one of the variables is selected from a single bond, it means that the two groups it connects are directly connected. For example, when L in ALZ represents a single bond, it means that the structure is actually AZ.

[0075] When the linking group is listed without specifying its linking direction, its linking direction is arbitrary, for example, The connecting group L is at this time The benzene ring and cyclopentyl group can be connected in the same direction as reading from left to right to form It is also possible to connect phenyl and cyclopentyl groups in the opposite direction of reading from left to right to form Combinations of linkers, substituents, and / or variations thereof are permissible only if such combinations result in stable compounds.

[0076] Unless otherwise specified, the number of atoms in a ring is generally defined as the number of members of the ring, for example, a "3-7 membered ring" refers to a "ring" having 3-7 atoms arranged around it.

[0077] Unless otherwise specified, the term "halogen" refers to fluorine, chlorine, bromine and iodine.

[0078] Unless otherwise specified, the term “C 1-6 "Alkyl" is used to represent a straight or branched chain saturated hydrocarbon group consisting of 1 to 6 carbon atoms. 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, etc.; which can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1-6Examples 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, and the like.

[0079] Unless otherwise specified, the term “C 1-3 "Alkyl" is used to represent a straight or branched chain saturated hydrocarbon group consisting of 1 to 3 carbon atoms. 1-3 Alkyl groups include C 1-2 and C 2-3 Alkyl, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1-3 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like.

[0080] Unless otherwise specified, the term “C 2-6 "Alkenyl" is used to indicate a linear or branched hydrocarbon group consisting of 2 to 6 carbon atoms containing at least one carbon-carbon double bond. The carbon-carbon double bond may be located at any position of the group. 2-6 Alkenyl groups include C 2-4 、C 2-3 , C4, C3, C2 alkenyl, etc.; which may be monovalent, divalent or polyvalent. 2-6 Examples of alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, pentenyl, hexenyl, butadienyl, piperyl, and the like.

[0081] Unless otherwise specified, the term “C 1-6 "Alkoxy" refers to an alkyl group containing 1 to 6 carbon atoms which is attached to the rest of the molecule via an oxygen atom. 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, 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), hexyloxy, and the like.

[0082] Unless otherwise specified, the term “C 1-6 "Alkylamino" refers to those alkyl groups containing 1 to 6 carbon atoms which are linked to the rest of the molecule through an amino group. 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, 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, and the like.

[0083] Unless otherwise specified, the term “C 3-6 "Cycloalkyl" means a saturated cyclic hydrocarbon group consisting of 3 to 6 carbon atoms, which is a monocyclic or bicyclic system. 3-6 Cycloalkyl groups include C 3-5 、C 4-5 and C 5-6 Cycloalkyl, etc.; it may be monovalent, divalent or polyvalent. 3-6 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

[0084] Unless otherwise specified, the term "3-8 membered heterocycloalkyl" by itself or in combination with other terms means a saturated monocyclic radical consisting of 3 to 8 ring atoms, 1, 2, 3 or 4 of which are heteroatoms independently selected from O, S and N, and the remainder being carbon atoms, wherein the nitrogen atom is optionally quaternized and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O)). z , z is 1 or 2). In addition, with respect to such "3-8 membered heterocycloalkyl", a heteroatom may occupy the position at which the heterocycloalkyl is attached to the rest of the molecule. Such 3-8 membered heterocycloalkyls include 4-6 membered, 5-6 membered, 7-8 membered, 4 membered, 5 membered and 6 membered heterocycloalkyls, and examples of 3-6 membered heterocycloalkyls include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl, tetrahydrofuranyl (including tetrahydrofuran-2-yl), piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperazinyl, morpholinyl, thiomorpholinyl, and the like.

[0085] Unless otherwise specified, the terms "6-10 membered aromatic ring" and "6-10 membered aryl" are used interchangeably. The term "6-10 aryl" refers to a monovalent aromatic carbocyclic ring system containing 6-10 carbon atoms and having at least one aromatic ring or multiple fused rings in which at least one ring is aromatic. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, biphenyl, and indanyl.

[0086] Unless otherwise specified, the terms "5-10 membered heteroaromatic ring" and "5-10 membered heteroaryl" are used interchangeably. 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 ring atoms are heteroatoms independently selected from O, S and N, and the rest 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 are optionally oxidized (i.e., NO and S(O) z , z is 1 or 2). The 5-10 membered heteroaryl group can be attached to the rest of the molecule through a heteroatom or a carbon atom, and the 5-10 membered heteroaryl group includes 5-8 membered, 5-7 membered, 5-6 membered, 5 membered and 6 membered heteroaryl groups, etc. Examples of the 5-10 membered heteroaryl group include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl), imidazolyl (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-isoxazolyl, 4-isoxazolyl and 5-isoxazolyl), thiazolyl (including 2-thiazolyl, 4 1-thiazolyl and 5-thiazolyl, etc.), furyl (including 2-furyl and 3-furyl, etc.), thienyl (including 2-thienyl and 3-thienyl, etc.), pyridyl (including 2-pyridyl, 3-pyridyl and 4-pyridyl, etc.), pyrazinyl, pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl, etc.), benzothiazolyl (including 2-benzothiazolyl, etc.), purinyl, benzimidazolyl (including 2-benzimidazolyl, etc.), benzoxazolyl, indazolyl (including 5-indazolyl, etc.), isoquinolyl (including 1-isoquinolyl and 5-isoquinolyl, etc.), quinoxalinyl (including 2-quinoxalinyl and 5-quinoxalinyl, etc.) or quinolyl (including 3-quinolyl and 6-quinolyl, etc.), etc.

[0087] Unless otherwise specified, the terms "5-6 membered heteroaromatic ring" and "5-6 membered heteroaryl" are used interchangeably in the present 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 ring atoms are heteroatoms independently selected from O, S and N, and the rest are carbon atoms. The nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O) z, z is 1 or 2). 5-6 yuan of heteroaryl can be connected to the rest of the molecule through a heteroatom or a carbon atom, and the 5-6 yuan of heteroaryl includes 5 yuan and 6 yuan of heteroaryl etc. The example of the 5-6 yuan of heteroaryl includes but is not limited to pyrrolyl (including N-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl etc.), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl and 5-imidazolyl etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl and 5-oxazolyl etc.), 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-isoxazolyl, 4-isoxazolyl and 5-isoxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4-thiazolyl and 5-thiazolyl, etc.), furyl (including 2-furyl and 3-furyl, etc.), thienyl (including 2-thienyl and 3-thienyl, etc.), pyridyl (including 2-pyridyl, 3-pyridyl and 4-pyridyl, etc.), pyrazinyl, pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl, etc.), etc.

[0088] Unless otherwise specified, C n-n+m or C n -C n+m Any specific case including n to n+m carbons, such as C 1-7 Including C1, C2, C3, C4, C5, C6 and C7, and also including 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 means that the number of atoms in the ring is n to n+m, for example, a 3-7-membered ring includes a 3-membered, 4-membered, 5-membered, 6-membered and 7-membered ring, and also includes any range from n to n+m, for example, a 3-7-membered ring includes a 3-6-membered ring, a 4-7-membered ring, a 5-7-membered ring and a 6-7-membered ring, etc.

[0089] Preparation method

[0090] The present invention also relates to a method for producing the compound of formula (I) defined above. In some embodiments, the synthesis method of the compound of the present application is shown in the figure:

[0091] When X is selected from halogens other than Br, such as F, Cl, I, etc., the method comprises the following steps:

[0092] a) brominating the compound of formula Ia using a brominating agent to produce 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.;

[0093] b) one Br atom in the compound of formula Ib is replaced by another halogen atom to obtain a compound of formula Ic. Preferably, step b) uses and a metal organic salt strong base at low temperature, wherein X is a halogen other than Br, such as F, Cl, I, more preferably, wherein the metal organic salt strong base is n-butyl lithium, preferably, the reaction temperature is -78 ° C;

[0094] c) oxidizing the compound of formula Ic to a compound of formula Id using an oxidizing agent, preferably, the oxidizing agent includes but is not limited to K2Cr2O7, preferably, the reaction temperature is 60-150°C;

[0095] d) Compound of formula Id and H2N-LR 1 Reaction to obtain a compound of formula Ie, preferably, the reaction temperature is 50-150° C.;

[0096] e) reacting a compound of formula Ie with R 2 R 3 NH reaction to obtain a compound of formula I, preferably, the reaction is carried out in the presence of a catalyst bis(triphenylphosphine)palladium(II) chloride, preferably, the reaction temperature is 20-150°C.

[0097] When X is selected from Br, the method comprises:

[0098] a) brominating the compound of formula Ia using a brominating agent 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.;

[0099] c) oxidizing the compound of formula Ib to a compound of formula Id using an oxidizing agent, preferably, the oxidizing agent includes but is not limited to K2Cr2O7, preferably, the reaction temperature is 60-150°C;

[0100] d) reacting the compound of formula Id with H2N-L-R1 to obtain a compound of formula Ie, preferably at a reaction temperature of 50-150°C;

[0101] e) reacting a compound of formula Ie with R 2 R 3NH reaction to obtain a compound of formula I, preferably, the reaction is carried out in the presence of a catalyst bis(triphenylphosphine)palladium(II) chloride, preferably, the reaction temperature is 20-150°C.

[0102] Alternatively, when X is selected from halogens other than Br, such as F, Cl, I, etc., the method comprises the following steps:

[0103] f) brominating the compound of formula Ia using a brominating agent to produce 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.;

[0104] g) All two Br atoms in the compound of formula Ib are replaced by other halogen atoms to obtain a compound of formula Ic. Preferably, step b) uses and a metal organic salt strong base at low temperature, wherein X is a halogen other than Br, such as F, Cl or I, more preferably, wherein the metal organic salt strong base is n-butyl lithium, preferably, the reaction temperature is -78 ° C;

[0105] h) oxidizing the compound of formula I-c' to a compound of formula I-d' using an oxidizing agent, preferably, the oxidizing agent includes but is not limited to K2Cr2O7, preferably, the reaction temperature is 60-150°C;

[0106] i) Compound of formula I-d' and H2N-LR 1 Reaction to obtain a compound of formula I-e', preferably, the reaction temperature is 50-150 ° C;

[0107] j) reacting a compound of formula I-e' with R 2 R 3 NH reaction to obtain a compound of formula I, preferably, the reaction is carried out in the presence of a catalyst bis(triphenylphosphine)palladium(II) chloride, preferably, the reaction temperature is 20-150°C.

[0108] As known to those skilled in the art, certain reactive groups (eg, -NH2, -OH, etc.) require conventional protection and deprotection when necessary, which is familiar to those skilled in the art.

[0109] Products and Applications

[0110] The present disclosure also provides a product, which can be, for example, a medicament, a pharmaceutical composition, or a kit, comprising an effective amount of a hnRNPA2B1 agonist compound of the present disclosure and a pharmaceutically or immunologically acceptable carrier. As used herein, the terms "active substance" or "active substance of the present disclosure" are used interchangeably to refer to a hnRNPA2B1 agonist compound having the structural formula (I) or a derivative or product thereof.

[0111] As used herein, the term "pharmaceutically acceptable" refers to a substance that is suitable for use in humans and / or animals without excessive adverse reactions (such as toxicity, irritation, and allergic reactions), i.e., a substance with a reasonable benefit / risk ratio. As used herein, the term "effective amount" refers to an amount that can produce a function or activity in humans and / or animals and is acceptable to humans and / or animals.

[0112] As used herein, the term "pharmaceutically acceptable carrier" refers to a carrier for administering a therapeutic agent, including various excipients and diluents. The term refers to pharmaceutical carriers that are not essential active ingredients themselves and are not unduly toxic after administration. Suitable carriers are well known to those of ordinary skill in the art, and a comprehensive discussion of pharmaceutically acceptable excipients can be found in Remington's Pharmaceutical Sciences (Mack Pub. Co., NJ 1991).

[0113] The pharmaceutically acceptable carrier in the composition may contain a liquid such as water, saline, glycerol, and ethanol. In addition, these carriers may also contain auxiliary substances such as fillers, disintegrants, lubricants, glidants, effervescent agents, wetting agents or emulsifiers, flavoring agents, pH buffering substances, etc. Generally, these substances can be formulated in a non-toxic, inert, and pharmaceutically acceptable aqueous carrier medium, wherein the pH is generally about 5-8, preferably, about 6-8.

[0114] The active substance in the product of the present disclosure accounts for 0.001-99.9 wt% of the total weight of the composition, preferably 1-95 wt%, more preferably 5-90 wt%, and even more preferably 10-80 wt%, with the remainder being pharmaceutically acceptable carriers and other additives.

[0115] As used herein, the term "unit dosage form" refers to a dosage form for single administration of the product of the present disclosure for the convenience of administration, including but not limited to various solid dosage forms (such as tablets), liquid dosage forms, capsules, and sustained-release dosage forms.

[0116] In another preferred embodiment of the present disclosure, the product is in unit dosage form or multi-dose form, and the active substance content is 0.01 to 2000 mg / dose, preferably 0.1 to 1500 mg / dose, and more preferably 1 to 1000 mg / dose. In another preferred embodiment of the present disclosure, 1 to 6 doses of the composition of the present disclosure are administered daily, preferably 1 to 3 doses; most preferably, the daily dose is 1 dose.

[0117] It should be understood that the effective dosage of an active substance, such as a hnRNPA2B1 agonist compound, may vary depending on the severity of the condition being administered or treated. The specific dosage is determined based on the individual circumstances of the subject (e.g., weight, age, physical condition, and desired effect), which is within the judgment of a skilled physician.

[0118] The products disclosed herein may be in solid form (e.g., granules, tablets, lyophilized powders, suppositories, capsules, sublingual tablets) or liquid form (e.g., oral solution) or other suitable forms. The routes of administration may include, but are not limited to: (1) traditional administration methods, such as gastrointestinal administration (e.g., oral administration), parenteral administration (e.g., solution injection, intravenous infusion), such as mucosal administration, transdermal administration, respiratory tract atomization inhalation, nasal drops, spray, oral administration, intramuscular injection, and / or intravenous administration; (2) linking the hnRNPA2B1 agonist to a transferrin / poly-L-lysine complex to enhance its biological effect; (3) encapsulating the drug with liposomes to mediate its entry into cells, which is beneficial for the smooth entry of the compound molecules and protects them from the hydrolysis of various extracellular enzymes; and (4) transporting active substances with liposomes to enable their specific transport to target tissues and target cells.

[0119] In addition, the products of the present disclosure 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 anti-tumor effects are administered before, simultaneously with, or after administration of the products of the present disclosure.

[0120] In some embodiments, the compounds or products of the present application activate hnRNPA2B1 to further regulate the immune system to prevent and treat tumors.

[0121] The term "subject" can refer to an animal, including but not limited to a primate (e.g., human), monkey, cow, pig, sheep, goat, horse, dog, cat, rabbit, rat, or mouse. The terms "subject" and "patient" are used interchangeably herein, e.g., to refer to a mammalian subject, e.g., a human subject.

[0122] In the context of treating a disease, disorder or condition, the terms "treat," "treat," and "therapy" are intended to include alleviating or eliminating the disease, disorder or condition or one or more symptoms associated with the disease, disorder or condition; or slowing the progression, spread, or worsening of the disease, disorder or condition or one or more symptoms thereof. "Cancer treatment" refers to one or more of the following effects: (1) inhibiting, to some extent, tumor growth, including (i) slowing and (ii) complete arrest of 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 an anti-tumor immune response, which can (i) maintain tumor size, (ii) reduce tumor size, (iii) slow tumor growth, (iv) reduce, slow, or prevent invasion, and / or (8) reducing, to some extent, the severity or number of one or more symptoms associated with the disorder.

[0123] The compounds disclosed herein can be hnRNPA2B1 agonists. These compounds can potentially be used to treat diseases or conditions, including but not limited to cell proliferative disorders. Cell proliferative disorders include but are not limited to cancer, benign papillomatosis, gestational trophoblastic disease, and benign neoplastic diseases, such as skin papillomas (warts) and genital papillomas.

[0124] In a specific embodiment, the disease or condition to be treated is a cell proliferative disorder. In certain embodiments, the cell proliferative disorder is cancer. In a specific embodiment, the cancer is selected from: brain and spinal cancer, head and neck cancer, leukemia and blood cancer, skin cancer, reproductive system cancer, gastrointestinal system cancer, liver and bile duct cancer, pancreatic cancer, kidney cancer, prostate cancer and bladder cancer, bone cancer, lung cancer, malignant mesothelioma, sarcoma, lymphoma, adenocarcinoma, thyroid cancer, heart tumor, germ cell tumor, malignant neuroendocrine (carcinoid) tumor, midline tract cancer and unknown primary cancer (i.e., cancer with metastatic cancer but unknown original cancer site). In a specific embodiment, cancer is present in adult patients. In another embodiment, cancer is present in pediatric patients. In a specific embodiment, cancer is relevant to AIDS.

[0125] In a specific embodiment, the cancer is selected from the group consisting of brain and spinal cancer. In a specific embodiment, the cancer is selected from the group consisting of: anaplastic astrocytoma, glioblastoma, astrocytoma, and sensory neuroblastoma (also known as olfactory blastoma). In a specific embodiment, the brain cancer is selected from the group consisting of: astrocytoma (e.g., cellular astrocytoma, subependymal giant cell astrocytoma, diffuse astrocytoma, multiform luteal astrocytoma, anaplastic astrocytoma, astrocytoma, giant cell glioblastoma, glioblastoma, secondary glioblastoma, primary adult glioblastoma, and primary pediatric glioblastoma), maxillary neuroma (e.g., oligodendroglioma and anaplastic oligodendroglioma), oligotrophic astrocytic tumor (e.g., oligotrophic astrocytoma), and esthesia neuroblastoma (e.g., olfactory astrocytoma). In some embodiments, the present invention relates to a brain cancer comprising: a glioma, a glioblastoma, a paraganglioma, a glioma of the upper tract, a glioma of the lower limb ...

[0126] In specific embodiments, the cancer is selected from the group consisting of head and neck cancer, including nasopharyngeal cancer, nasal cavity and paranasal sinus cancer, hypopharyngeal 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 eye cancer. In specific embodiments, the eye cancer is selected from the group consisting of intraocular melanoma and retinoblastoma.

[0127] In a specific embodiment, the cancer is selected from leukemia and hematological cancer. In a specific embodiment, the cancer is selected from: myeloproliferative neoplasms, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasms, acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), chronic myeloid leukemia (CML), myeloproliferative neoplasms (MPN), AML after MPN, AML after MDS, high-risk MDS or AML associated with del (5q), blast stage chronic myeloid leukemia, angioimmunoblastic lymphoma, acute lymphocytic leukemia, Langerhans cell histiocytosis, hairy cell leukemia and plasmacytoma (including plasmacytoma and multiple myeloma). The leukemia mentioned herein can be acute or chronic.

[0128] In a specific embodiment, the cancer is selected from skin cancer. In a specific embodiment, the skin cancer is selected from melanoma, squamous cell carcinoma, and basal cell carcinoma.

[0129] In specific embodiments, the cancer is selected from cancers of the reproductive system. In specific embodiments, 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 embodiments, the cancer is breast cancer selected from ductal carcinoma and phyllodes tumor. In specific examples of these embodiments, the breast cancer can be male breast cancer or female breast cancer. In specific examples of these embodiments, the cancer is cervical cancer selected from squamous cell carcinoma and adenocarcinoma. In specific examples of these embodiments, the cancer is ovarian cancer selected from epithelial carcinoma.

[0130] In specific embodiments, the cancer is selected from cancers of the gastrointestinal system. In specific embodiments, the cancer is selected from the group consisting of 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 embodiments, the cancer is selected from the group consisting of 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.

[0131] In a specific embodiment, the cancer is selected from liver cancer and bile duct cancer. In a specific embodiment, the cancer is liver cancer (also known as hepatocellular carcinoma). In a specific embodiment, the cancer is bile duct cancer (also known as cholangiocarcinoma). In examples of these embodiments, bile duct cancer is selected from intrahepatic bile duct cancer and extrahepatic bile duct cancer.

[0132] In a specific embodiment, the cancer is selected from kidney cancer and bladder cancer. In a specific embodiment, the cancer is kidney cancer selected from renal cell carcinoma, Wilms tumor, and transitional cell carcinoma. In a specific embodiment, the cancer is bladder cancer selected from ureteral cancer (transitional cell carcinoma), squamous cell carcinoma, and adenocarcinoma.

[0133] In a specific embodiment, the cancer is selected from bone cancer. In a specific embodiment, the bone cancer is selected from the group consisting of osteosarcoma, malignant fibrous histiocytoma of bone, Ewing's sarcoma, chordoma (bone cancer along the spine).

[0134] In a specific embodiment, the cancer is selected from lung cancer. In a specific embodiment, the lung cancer is selected from non-small cell lung cancer, small cell lung cancer, bronchogenic carcinoma, and pleuropulmonary blastoma.

[0135] In a specific embodiment, the cancer is selected from malignant mesothelioma.In a specific embodiment, the cancer is selected from epithelial mesothelioma and sarcomatoid tumor.

[0136] In a specific embodiment, the cancer is selected from a sarcoma. In a specific embodiment, the sarcoma is selected from the group consisting of: central chondrosarcoma, central and periosteal chondromas, fibrosarcoma, clear cell sarcoma of the tendon sheath, and Kaposi's sarcoma.

[0137] In a specific embodiment, the cancer is selected from lymphoma. In a specific embodiment, 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, Sézary syndrome, primary central nervous system lymphoma), cutaneous T-cell lymphoma, primary central nervous system lymphoma.

[0138] In a specific embodiment, the cancer is selected from adenocarcinoma. In a specific embodiment, the cancer is selected from adrenocortical carcinoma (also known as adrenocortical carcinoma or adrenocortical carcinoma), pheochromocytoma, paraganglioma, pituitary tumor, thymoma and thymic carcinoma.

[0139] In a specific embodiment, the cancer is selected from thyroid cancer.In a specific embodiment, the thyroid cancer is selected from medullary thyroid cancer, papillary thyroid cancer, and follicular thyroid cancer.

[0140] In specific embodiments, the cancer is selected from a germ cell tumor. In specific embodiments, the cancer is selected from a malignant extracranial germ cell tumor and a malignant extragonadal germ cell tumor. In specific examples of these embodiments, the malignant extragonadal germ cell tumor is selected from a non-seminoma and a seminoma.

[0141] In a specific embodiment, the cancer is selected from cardiac tumors. In a specific embodiment, the cardiac tumor is selected from the group consisting of malignant teratoma, lymphoma, rhabdomyosarcoma, angiosarcoma, chondrosarcoma, infantile fibrosarcoma, and synovial sarcoma.

[0142] In a specific embodiment, the cell proliferative disorder is selected from the group consisting of benign papillomatosis, benign neoplastic disease, and gestational trophoblastic disease. In a specific embodiment, the benign neoplastic disease is selected from the group consisting of cutaneous papillomas (warts) and genital papillomas. In a specific embodiment, the gestational trophoblastic disease is selected from the group consisting of hydatid cystic nevi and gestational trophoblastic neoplasms (e.g., invasive nevi, choriocarcinoma, placental site trophoblastic tumor, and epithelioid trophoblastic tumor).

[0143] In some embodiments, the compounds or products of the present application are administered as preventive drugs before a tumor develops to prevent the onset of the tumor or reduce the severity of subsequent tumors. In some embodiments, the compounds or products of the present application are administered as therapeutic drugs after a tumor develops to reduce the severity of the tumor disease, including its metastasis. In some embodiments, the compounds or products of the present application are administered as both preventive drugs and therapeutic drugs, either continuously or intermittently before and after the onset of a tumor.

[0144] Those skilled in the art may arbitrarily combine the above technical solutions and technical features without departing from the inventive concept and protection scope of the present invention. Other aspects of the present invention will be obvious to those skilled in the art due to the disclosure herein.

[0145] Example

[0146] The present disclosure will be further described below in conjunction with specific examples. It should be understood that these examples are intended only to illustrate the present disclosure and are not intended to limit the scope of the present disclosure. Those skilled in the art may make appropriate modifications and variations to the present disclosure, and these modifications and variations are within the scope of the present disclosure.

[0147] For experimental procedures in the following examples where specific conditions are not specified, conventional methods in the art may be employed, for example, as described in Molecular Cloning: A Laboratory Manual (3rd ed., Cold Spring Harbor Laboratory Press, New York, 1989) or according to the conditions recommended by the supplier. DNA sequencing methods are conventional in the art and are also available from commercial companies.

[0148] Unless otherwise indicated, percentages and parts are by weight. Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be used in the disclosed methods. The preferred embodiments and materials described herein are for illustrative purposes only.

[0149] I. Synthesis and Characterization Examples of Compounds

[0150] Example I.1: Synthesis of Compound A (FPC-1):

[0151] *eq equivalent

[0152] 1-1. Preparation of Compound 2

[0153] 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, allowed to naturally warm to room temperature, and stirred overnight. The reaction solution was filtered, and the filter cake was washed three times with ethanol. The dried filter cake was collected and recrystallized from EA to obtain the pure product (10.1 g, off-white crystals).

[0154] 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).

[0155] 1-2. Preparation of Compound 3

[0156] Compound 2 (6 g, 19.2 mol) was dissolved in 300 mL of anhydrous THF and cooled to -80 ° C. Then n-butyl lithium (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 slowly adding N-fluorobisbenzenesulfonamide (NFSI) (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 then poured into 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 eluent. Compound 3 (4.1 g, white solid) was obtained.

[0157] 1 H NMR (300 MHz, chloroform-d) δ 7.66 (d, J=7.4 Hz, 1H), 7.21-7.09 (m, 3H), 3.35 (s, 4H).

[0158] 1-3. Preparation of Compound 4

[0159] Compound 3 (4.5g, 17.9mmol) is joined in the glacial acetic acid solution (50mL) of potassium dichromate (24g, 82.4mmol).Solution is heated to reflux 16 hours, then poured into frozen water.Gained precipitate is collected by filtration and washed with water.Filter cake is air-dried, joined among the DCM and refluxed 30 minutes, filtered, washed with DCM, filter cake is joined among the DCM and refluxed 15 minutes, filtered and washed with DCM.Merge all filtrates and vacuum concentration, obtain compound 4 (4.1g, crude product, yellow solid).

[0160] 1-4. Preparation of INT-1

[0161] N-Boc-ethylenediamine (3.3 g, 20.8 mmol) was added to a solution of compound 4 (4.1 g, 13.9 mmol) in EtOH (100 mL), and the mixture was heated at reflux for 1 hour. The reaction mixture was concentrated, and the resulting residue was purified by column chromatography (PE / EA = 5 / 1). An off-white solid (2.8 g) was obtained.

[0162] 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).

[0163] 1-5. Preparation of FPC-1-1

[0164] Cesium carbonate (5.55 g, 17 mmol) and 1-(2-pyridoneethyl)piperazine (1.05 g, 5.7 mmol) were added to a toluene solution (175 mL) of INT-1 (2.5 g, 5.7 mol), and bis(triphenylphosphine)palladium chloride (II) (400 mg, 0.57 mmol) was added to the mixture under argon protection. The mixture was reacted at 80° C. for 16 hours, the reaction solution was cooled to room temperature, water was added to the reaction solution and stirred, the organic phase was separated, the aqueous phase was extracted once with DCM, the organic phases were combined, and dried over anhydrous magnesium sulfate. The crude product was purified by Prep-TLC (DCM / MeOH=10 / 1+NH 3 ·H 2 O) to give 240 mg of a pure yellow solid.

[0165] 1 H NMR (400 MHz, CHLOROFORM-d) δ 8.56 (dd, J = 7.7, 4.3 Hz, 1H), 8.51 (d, J = 8.3 Hz, 1H), 7.34-7.27 (m, 1H), 7.17 (d, J = 8.2 Hz, 1H), 4.99 (s, 1H), 4.31 (s, 2H), 3.79-2.45 (m, 18H), 2.18 (s, 4H), 1.28 (s, 9H).

[0166] 1-6. Preparation of FPC-1

[0167] 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 stirred at room temperature for 2 hours. The mixture was concentrated under vacuum, and the residue was dissolved in a small amount of methanol, precipitated with ether, and centrifuged. The precipitate was washed with ether, centrifuged, and dried under vacuum to give 240 mg of the pure product (yellow solid).

[0168] 1 H NMR (300 MHz, 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).

[0169] MS (ESI) m / z [M+H] + =440.3.

[0170] Example I.2: Synthesis of Compound B (FPC-2)

[0171] Preparation of FPC-2-1

[0172] In a single-necked flask, 10 mL of ethylene glycol monomethyl ether was added, followed by INT-1 (110 mg, 0.25 mmol) and 3-(piperazin-1-yl)propan-1-ol (44 mg, 0.3 mmol), and the reaction mixture was stirred at 100° C. After 2 hours, the solvent was removed in vacuo, and the residue was purified by TLC to give 20 mg of a yellow solid.

[0173] 1 H NMR (400 MHz, CHLOROFORM-d) δ 8.63-8.49 (m, 2H), 7.31 (dd, J = 12.8, 8.3 Hz, 1H), 7.18 (d, J = 8.3 Hz, 1H), 4.97 (s, 1H), 4.42-4.23 (m, 2H), 3.87 (t, J = 5.0 Hz, 2H), 3.60-3.28 (m, 8H), 2.92 (s, 4H), 1.89 (s, 2H), 1.29 (s, 9H).

[0174] 2-2. Preparation of FPC-2

[0175] 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 concentrated in vacuo and the residue was dissolved in a small amount of methanol, precipitated with ether, centrifuged, and the precipitate was washed with ether, centrifuged, and dried in vacuo to give 5 mg of the pure product (yellow solid).

[0176] 1 H NMR (400 MHz, 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).

[0177] MS (ESI) m / z [M+H] + =401.47.

[0178] Example I.3: Synthesis of Compound C (FPC-3)

[0179] Preparation of FPC-3-1

[0180] Cesium carbonate (1.78 g, 5.48 mmol) and N-ethyl-2-(pyrrolidin-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 chloride (II) (128 mg, 0.18 mmol) was added to the mixture under argon protection. The mixture was reacted at 80° C. for 16 hours. The reaction solution was cooled to room temperature, water was added to the reaction solution and stirred, the organic phase was separated, 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+NH 3 · H 2 O) to give 30 mg of a pure yellow solid.

[0181] 1H NMR (400MHz, chloroform-d) δ 8.63-8.52 (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).

[0182] 3-2. Preparation of FPC-3

[0183] Compound FPC-3-1 (30 mg, 0.06 mmol) was dissolved in 4.4 mL of DCM / 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 concentrated in vacuo, and the residue was dissolved in a small amount of methanol, precipitated with ether, and centrifuged. The precipitate was washed with ether, centrifuged, and dried in vacuo to obtain 25 mg of the pure product (yellow solid).

[0184] 1 H 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).

[0185] MS (ESI) m / z [M+H] + =399.47.

[0186] Example I.4: Synthesis of Compound D (FPC-5)

[0187] 4-1. Preparation of FPC-5-1

[0188] Cesium carbonate (460 mg, 1.41 mmol) and N, 2-dimethylpropane-1-amine (49 mg, 0.56 mmol) were added to a toluene solution (20 mL) of INT-1 (205 mg, 0.47 mmol). Under argon protection, bis(triphenylphosphine)palladium chloride (II) (32 mg, 0.0.045 mmol) was added to the mixture. The reaction was carried out at 80° C. for 16 hours. The reaction solution was cooled to room temperature, water was added to the reaction solution and stirred, the organic phase was separated, the aqueous phase was extracted once with DCM, the organic phases were combined, and dried over anhydrous magnesium sulfate. The crude product was purified by vacuum concentration and column purification through TLC (DCM / MeOH=10 / 1+NH 3 ·H 2 O) to give 10 mg of a pure yellow solid.

[0189] 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).

[0190] 4-2. Preparation of FPC-5

[0191] 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 concentrated in vacuo, and the residue was dissolved in a small amount of methanol, precipitated with ether, and centrifuged. The precipitate was washed with ether, centrifuged, and dried in vacuo to give 9 mg of the pure product (yellow solid).

[0192] 1H NMR (400MHz, methanol-d4) δ 8.61-8.52 (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).

[0193] MS (ESI) m / z [M+H] + =344.42.

[0194] Example I.5: Synthesis of Compound E (AIR-2)

[0195] 5-1. Preparation of Compound 5

[0196] Compound 3 (31.90 g, 127.04 mmol) was dissolved in anhydrous THF (800 mL), and the reaction system was replaced with argon three times 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 for at least 10 minutes at the same temperature. N-fluorobisbenzenesulfonamide (NFSI) (60.09 g, 190.57 mmol) in 200 mL of anhydrous THF was added dropwise, and after stirring for 1 hour at -78 ° C, the reaction mixture was naturally warmed to room temperature and stirred overnight. The reaction mixture was quenched with NH4Cl solution and then extracted twice with EA. The combined organic layer was washed with water and brine, dried and filtered with Na2SO4. The filtrate was concentrated to give a crude product, added to DCM and stirred. The resulting precipitate was filtered, the filter cake was washed twice with DCM, and all filtrates were combined and concentrated in vacuo to give the crude product, which was then purified by column chromatography (PE) to afford compound 5 (15.42 g, 63.82%) as a white solid.

[0197] 1 H NMR (300MHz, Chloroform-d) δ 7.16 (d, J=7.6Hz, 2H), 7.12-7.04 (m, 2H), 3.37 (s, 4H).

[0198] 5-2. Preparation of INT-2

[0199] Compound 5 (15.42 g, 81.08 mmol) was added to a glacial acetic acid solution (220 mL) of potassium dichromate (109.72 g, 372.96 mmol). 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 layer was washed with water and brine, dried over Na2SO4 and filtered. The filtrate was concentrated to give a crude product, which was then added with PE and ultrasonic dispersion. The resulting precipitate was filtered and dried to obtain a crude INT-2 (10.55 g) as a brown solid, which was used without further purification.

[0200] 5-3. Preparation of AIR-2-1

[0201] INT-2 (5 g, 21.35 mmol) was dissolved in EtOH (120 mL), the reaction system was replaced with argon three times, and then heated to 50°C. N 1 , N 1 -dimethylethane-1,2-diamine (2.79 mL, 25.62 mmol) was added, and the mixture was heated to 80°C (reflux) for 4 hours. TLC showed that the reaction was complete. The reaction mixture was concentrated, and the obtained residue was purified by column chromatography (DCM: MeOH = 30: 1 to 10: 1) to give AIR-2-1 (1.81 g, 27.86%) as a brown solid. MS (ESI), m / z: 305.4 [M+H] + .

[0202] 1 H NMR (300MHz, Chloroform-d) δ 8.65 (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).

[0203] 5-4. Preparation of AIR-2

[0204] AIR-2-1 (1.81 g, 5.95 mmol) was dissolved in 1,4-dioxane (150 mL), and the reaction system was replaced with argon three times. 1-(2-pyridone-ethyl)piperazine (Compound A) (1.20 g, 6.54 mmol) was added, and the mixture was stirred at room temperature overnight. The reaction mixture was concentrated, and the resulting residue was purified by column chromatography (DCM: MeOH = 15: 1 to 5: 1) to give the pure product dissolved in DCM. 5M HCl / 1,4-dioxane (1.5 mL) was then added and stirring was continued, and the mixture was precipitated with diethyl ether. The resulting precipitate was filtered, washed with diethyl ether, and dried to give AIR-2 (1.21 g) as a yellow solid. MS (ESI), m / z: 468.47 [M+H] + .

[0205] 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).

[0206] Example I.6: Synthesis of Compound F (AIR-3)

[0207] 6-1. Preparation of Compound 2

[0208] 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 with argon twice and then cooled to 9°C. Compound 1 (176 g, 1.14 mol) was added and stirred at 9°C for 2 hours. The reaction mixture was then allowed to warm to room temperature 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 obtain pure compound 2 (102.57 g, 28.80%) as off-white crystals.

[0209] 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).

[0210] 6-2. Preparation of Compound 3

[0211] Compound 2 (50 g, 160.26 mmol) was dissolved in anhydrous THF (950 mL), the reaction system was replaced with argon three times, 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-fluorobisbenzenesulfonamide (NFSI) (75.8 g, 240.38 mmol) in 250 mL of anhydrous THF was added dropwise, and after stirring at -78 ° C for 1.5 hours, the reaction mixture was naturally warmed to room temperature and stirred overnight. The reaction mixture was quenched with NH4Cl solution and then extracted twice with EA. The combined organic layer was washed with water and brine, dried and filtered with Na2SO4. The filtrate was concentrated to give a crude product, added to DCM and stirred. The resulting precipitate was filtered, the filter cake was washed twice with DCM, and all filtrates were combined and concentrated in vacuo to give the crude product, which was then purified by column chromatography (PE:EA=50:1) to give compound 3 (31.90 g, 79.27%) as a white solid.

[0212] 1 H NMR (300MHz, Chloroform-d) δ7.66 (d, J=7.4Hz, 1H), 7.21-7.09 (m, 3H), 3.35 (s, 4H).

[0213] 6-3. Preparation of Compound 4

[0214] Compound 3 (9.29 g, 37.00 mmol) was added to a glacial acetic acid solution (100 ml) of potassium dichromate (50.07 g, 170.19 mmol). The solution was heated at 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 concentrated to give crude compound 3 (6.72 g), which can be used without further purification.

[0215] 6-4. Preparation of AIR-3-1

[0216] To a solution of compound 4 (400 mg, 1.36 mmol) in EtOH (20 mL) was added DIPEA (358.48 uL, 2.17 mmol) and tert-butyl (2-aminoethyl)(methyl)carbamate hydrochloride (428.46 mg, 2.03 mmol). The reaction system was replaced 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 obtained residue was purified by column chromatography (PE: EA = 10: 1 to 4: 1) to give AIR-3-1 (244 mg, 39.88%) as a yellow solid.

[0217] 6-5. Preparation of AIR-3-2

[0218] To a toluene solution (15 mL) of AIR-3-1 (244 mg, 540.68 mmol) was added 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). The reaction system was purged with argon 5 times, and then Pd(OAc) 2 (24.28 mg, 108.14 mmol) was added. The reaction system was purged with argon 3 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 Na 2 SO 4 and filtered. The filtrate was concentrated to give a crude product, which was purified by column chromatography (DCM:MeOH=100:1 to 10:1, with addition of NH3·H2O) and Prep-TLC (DCM:MeOH=10:1, with addition of NH3·H2O) to give AIR-3-2 (40 mg, 13.36%).

[0219] 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).

[0220] 6-6. Preparation of AIR-3

[0221] A solution of AIR-3-2 (40 mg, 72.24 mmol) in DCM / MeOH = 10 / 1 (3 mL) was replaced with argon three times, and then 5M HCl / 1,4-dioxane (1.5 mL) was added. The resulting mixture was stirred at room temperature for 1.5 hours. TLC showed that 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, NH3·H2O was added) 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).

[0222] 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).

[0223] Example II. Activity Test

[0224] Example II.1: Activation of hnRNPA2B1 and TBK1-IRF3 pathways in tumor cells by test compounds

[0225] After stimulating A549 lung cancer cells (cell density of 1×10 5 cells per well) with 20 μM test compound E (prepared as in Example 1.5, dissolved in saline) for 30 minutes, the subcellular localization of hnRNPA2B1 was detected by immunofluorescence, and the activation of TBK1 and IRF3 was detected by SDS-PAGE electrophoresis and immunoblotting.

[0226] The activation of hnRNPA2B1, TBK1, and IRF3 are shown in Figures 1 and 2 .

[0227] The results showed that the test compound E could significantly activate the nuclear export of hnRNPA2B1 in tumor cells and the activation of downstream TBK1 and IRF3. In addition, similar results were obtained by performing the same experiment using other compounds prepared in Example 1.

[0228] The above results demonstrate that the test compound has an activating effect on the hnRNPA2B1 pathway and is a hnRNPA2B1 agonist.

[0229] Example II.2: Broad induction of type I interferon production by test compounds in various tumor types

[0230] A549 (non-small cell lung cancer), HepG2 (liver cancer), LLC (Lewis lung cancer), and B16F10 (melanoma) tumor cells were stimulated with test compound E at concentrations of 20 and 40 μM (cell density of 1x10^4 cells per well); cell culture supernatants were collected 18 and 24 hours after stimulation, and IFN-β protein levels were detected by ELISA.

[0231] The activation of type I interferon by the test compounds is shown in FIG3 .

[0232] The results showed that the test compound E could significantly activate multiple cells to express type I interferon. In addition, similar results were obtained by performing the same experiment using other compounds prepared in Example 1.

[0233] The above results demonstrate that the test compound can induce the production of type I interferon by activating the hnRNPA2B1 pathway and is a biologically active hnRNPA2B1 agonist. Therefore, this disclosure defines such novel compounds as hnRNPA2B1 agonists.

[0234] Example II.3: Inhibitory effects of test compounds on proliferation of various tumor cell types

[0235] A549 (non-small cell lung cancer), HepG2 (liver cancer), LLC (Lewis lung cancer), and B16F10 (melanoma) tumor cells (cell density of 1x 10^4 cells per well) were stimulated with test compound E at a concentration of 20 μM; cells were lysed 24 and 48 hours after stimulation, and the effect of compound E on tumor cell activity was detected by the CCK-8 method.

[0236] The inhibitory effect of test compound E on tumor cells is shown in FIG4 .

[0237] The results showed that the test compound E could significantly inhibit the proliferation of various tumor cells. In addition, similar results were obtained by performing the same experiment using other compounds prepared in Example 1.

[0238] The above results demonstrate that the test compound can significantly inhibit the proliferation of tumor cells by targeting hnRNPA2B1.

[0239] Example II.4: Killing effect of test compounds on proliferation of various types of tumor cells

[0240] LLC (Lewis lung cancer) tumor cells (cell density of 1x10^5 cells per well) were stimulated with test compound E at concentrations of 20 and 40 μM. After 36 hours of stimulation, cell morphology was observed under a microscope, and the killing effect of compound E on tumor cells was detected by flow cytometry after cell digestion.

[0241] The killing effect of test compound E on tumor cells is shown in FIG5 .

[0242] The results showed that the test compound E could significantly induce apoptosis of tumor cells. In addition, the same experiment was carried out using other compounds prepared in Example 1, and similar results were obtained.

[0243] The above results demonstrate that the test compound can effectively kill tumor cells by targeting hnRNPA2B1.

[0244] Example II.5: hnRNPA2B1 agonist inhibits LLC tumor proliferation and promotes survival of tumor-bearing mice

[0245] Mice (6-8 week old male C57BL / 6J mice) were subcutaneously inoculated with 5x10^5 LLC tumor cells. Starting from the 10th day after inoculation, compound E (15 mg / kg mouse body weight) was injected via the tail vein once every four days for a total of four doses. The mice were continuously observed for tumor growth. Tumor size = length x width 2 / 2, and observe the survival time of mice.

[0246] The test results are shown in Figures 6 and 7.

[0247] The results showed that the hnRNPA2B1 agonist compound E could inhibit LLC tumor proliferation and promote the survival of tumor-bearing mice. In addition, similar results were obtained by performing the same experiment using other compounds prepared in Example 1.

[0248] These results suggest that hnRNPA2B1 agonists can effectively exert anti-tumor effects.

[0249] Example II.6: hnRNPA2B1 agonist inhibits B16F10 tumor proliferation and promotes survival of tumor-bearing mice

[0250] Mice (6-8 week old male C57BL / 6J mice) were subcutaneously inoculated with 5x10^5 B16-F10 melanoma cells. Starting on day 10 of inoculation, compound E (15 mg / kg mouse body weight) was administered via tail vein injection every four days for a total of four doses. The mice were continuously observed for tumor growth. Tumor size = length x width 2 / 2.

[0251] The results showed that the hnRNPA2B1 agonist could inhibit the proliferation of B16F10 tumors. In addition, the same experiment was carried out using other compounds prepared in Example 1, and similar results were obtained.

[0252] These results suggest that hnRNPA2B1 agonists can effectively exert anti-tumor effects.

[0253] Example II.7: hnRNPA2B1 agonist inhibits experimental lung metastasis of B16F10 tumors and promotes survival of tumor-bearing mice

[0254] Mice (6-8 week old male C57BL / 6J mice) were injected with 5x10^4 B16-F10 melanoma cells via the tail vein. Starting on day 6 post-injection, Compound E (10 mg / kg mouse body weight) was administered via tail vein injection every three days for a total of three doses. On day 13 post-administration, lung tumors were observed using a small animal in vivo imaging device and quantitatively compared. On day 15, the lungs of the treated mice were isolated and the tumors were observed.

[0255] The test results are shown in Figures 8 and 9.

[0256] The results showed that the hnRNPA2B1 agonist could inhibit the experimental lung metastasis of B16F10 tumor. In addition, the same experiment was carried out using other compounds prepared in Example 1, and similar results were obtained.

[0257] The above results suggest that hnRNPA2B1 agonists can effectively exert anti-tumor metastasis effects.

[0258] Biological evaluation

[0259] The compounds of the present disclosure are defined as hnRNPA2B1 agonists by: (i) binding to the hnRNPA2B1 protein, as demonstrated by high affinity of the compounds to the hnRNPA2B1 protein at concentrations as low as 20 μM as determined by Biacore; and (ii) demonstrating by cell-based assays that the compounds can broadly induce higher levels of IFN-β production at concentrations as low as 20 μM.

[0260] (i) Biacore T200 detection of the binding between the compound and hnRNPAB

[0261] Using S series CM5 chip; amino coupling reagents (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide EDC and N-hydroxysuccinimide NHS);

[0262] Coupling buffer: 10 mM sodium acetate pH 4.0, pH 4.5;

[0263] Running buffer: 10× PBS-P+;

[0264] Analytical grade DMSO, deionized water (0.22 μm membrane filtration);

[0265] hnRNPA2B1 recombinant protein (concentration must be greater than 0.5 mg / mL, total protein volume must be at least 20 μg);

[0266] Compound: Stock solution concentration greater than 20 mM, volume greater than 30 μL, purity greater than 90%, dissolved in 100% DMSO;

[0267] Other consumables: 96-well plates and sealing film, 1.5 mL centrifuge tubes without caps, and type 2 rubber bottle caps.

[0268] Experimental steps:

[0269] 1. Place the running buffer 1× PBS and waste liquid bottle in the designated positions and place the chip.

[0270] 2. Protein coupling. Select the immobilization program in the Wizard template, check flow cell 2 or 4, select amine as the method, and specify contact time and flow rate to achieve high coupling. Enter a contact time of 900 s and a flow rate of 10 μL / min.

[0271] 3. Prepare the required protein solution according to the instructions. Dilute the protein to the required volume with sodium acetate (pH 4.0) to a final concentration of 20 μg / mL. Place 100 μL of EDC, 100 μL of NHS, and 140 μL of ethanolamine in the corresponding positions.

[0272] 4. The system will automatically coat the target conjugated amount of hnRNPA2B1 protein on the chip surface and automatically generate a conjugation report.

[0273] 5. Prepare the running buffer for the interaction. For small molecule samples, use 1× PBS-P+ containing 5% DMSO as the running buffer: dilute 105 mL of 10× PBS-P+ to 1 L with deionized water to make 1.05× PBS-P+.

[0274] Solvent correction stock solution containing 4.5% DMSO: 1.05×PBS-P + 9.5mL + 0.45mL DMSO

[0275] Solvent correction stock solution containing 5.8% DMSO: 1.05×PBS-P + 9.5mL + 0.58mL DMSO

[0276] Running buffer containing 5% DMSO: 1.05×PBS-P + 950 mL + 50 mL DMSO

[0277] Dilute a 10 mM small molecule stock solution in 1.05× PBS-P+ buffer without DMSO: 0.7 μL small molecule stock solution + 6.3 μL DMSO + 133 μL 1.05× PBS-P+ buffer, for a total volume of 140 μL, for a final concentration of 50 μM. Add to a 96-well plate and seal with film.

[0278] After the test, use Kinetics or Affinity mode to measure binding affinity.

[0279] (ii) Total cellular RNA can be extracted using TRIzol or a rapid RNA extraction kit.

[0280] 24-well plate cells (about 2-3×10 5 Rapid RNA extraction (100 cells / well): Discard the culture medium, add 500 μL RA2 lysis buffer to each well, pipette and lyse the cells several times, transfer the cells to the inner tube of the dedicated adsorption tube, and centrifuge at 12,000 rpm for 1 minute at room temperature. Discard the filtrate, add 500 μL wash buffer, and centrifuge at 12,000 rpm for 1 minute at room temperature. Repeat the wash operation once. Discard the filtrate, replace the inner tube, centrifuge at 12,000 rpm for 2 minutes at room temperature without adding wash buffer. Transfer the inner tube to a new 1.5 mL centrifuge tube, add 25 μL Elution Buffer to the center of the membrane, let it stand at room temperature for 5 minutes, and centrifuge at 12,000 rpm for 1 minute at room temperature. The resulting filtrate is RNA, and the RNA concentration is measured using Nanodrop One.

[0281] qRT-PCR: After the concentration of each sample was determined, 1 μg of total RNA was taken and ReverTra qRT-PCR RT Master Mix reverse transcribes RNA into cDNA. The reverse transcription system and reaction conditions are as follows:

[0282] Reverse transcription reaction system (20 μL system)

[0283] Reverse transcription reaction conditions (20 μL system)

[0284] Dilute the reverse-transcribed cDNA sample with 60 μL of deionized water, mix thoroughly, and centrifuge briefly. See the table for primer sequences used for qRT-PCR. Use SYBR Green Realtime PCR Master Mix for relative quantification of gene mRNA levels.

[0285] qRT-PCR reaction system (20 μL system)

[0286] qRT-PCR reaction conditions

[0287] qRT-PCR primers

[0288] All documents mentioned in this disclosure are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of this disclosure, those skilled in the art may make various changes or modifications to this disclosure, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. A fused ring compound represented by formula (I), its cis-trans isomers, its enantiomers, its diastereoisomers, its racemates, its solvates, its hydrates or its pharmaceutically acceptable salts or its prodrugs, Among them, L is -(CH2)n-, where n is an integer from 0 to 6; X is a halogen; R 1 is -NR 4 R 5 , where R 4 and R 5 are each independently H or C 1-6 alkyl; R 2 and R 3 each independently is H or C 1-6 alkyl; or R 2 and R 3 together with the N atom to which they are attached form a 5- to 8-membered heteroalkyl group containing 1 to 3 heteroatoms selected from nitrogen, oxygen or sulfur; The C 1-6 alkyl group and the 5- to 8-membered heterocycloalkyl group are unsubstituted or substituted by one or more substituent groups selected from the group consisting of: hydroxy, amino, halogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, 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- to 6-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl C 1-6 alkyl, 6- to 10-membered aryl, 6- to 10-membered aryl C 1-6 alkyl or 5- to 10-membered heteroaryl and 5- to 10-membered heteroaryl C 1-6 alkyl.

2. The compound according to claim 1, its cis-trans isomers, its enantiomers, its diastereoisomers, its racemates, its solvates, its hydrates or its pharmaceutically acceptable salts or its prodrugs, Among them, X is F or Cl; L is -(CH2)n-, where n is 1, 2 or 3; R 4 、R 5 Each independently is H, methyl or ethyl; R 2 and R 3 each independently is C 1-3 alkyl, and the C 1-3 alkyl is unsubstituted or substituted by a 3- to 8-membered heterocyclic alkyl group containing 1 to 2 heteroatoms selected from nitrogen, oxygen, or sulfur; or R 2 and R 3 together with the N atom to which they are attached form a 5- to 8-membered heterocycloalkyl group containing 2 nitrogen atoms, said 5- to 8-membered heterocycloalkyl group being substituted by a 3- to 8-membered heterocycloalkyl C 1-6 alkyl or hydroxy C 1-6 alkyl substituted.

3. The compound according to claim 1, its cis-trans isomers, its enantiomers, its diastereoisomers, its racemates, its solvates, its hydrates or its pharmaceutically acceptable salts or its prodrugs, Among them, X is F; L is -(CH2)n-, where n is 2; R 4 、R 5 Each independently is H, methyl or ethyl; R 2 and R 3 each independently is C 1-3 alkyl, and the C 1-3 alkyl is unsubstituted or substituted by a 4- to 6-membered heteroalkyl containing 1 nitrogen heteroatom; or R 2 and R 3 together with the N atom to which they are attached form a piperazinyl group, said piperazinyl group being substituted by a 4- to 6-membered heterocycloalkyl C 1-6 alkyl or hydroxy C 1-6 alkyl substituted.

4. The compound according to claim 1, its cis-trans isomers, its enantiomers, its diastereoisomers, its racemates, its solvates, its hydrates or its pharmaceutically acceptable salts or its prodrugs, Among them, X is F; L is -(CH2)n-, where n is 2; R 4 、R 5 are each independently H, methyl or ethyl; R 2 and R 3 each independently is methyl, ethyl or isopropyl, and the C 1-3 alkyl group is unsubstituted or unsubstituted or substituted by pyrrolidinyl; or R 2 and R 3 together with the N atom to which they are attached form a piperazinyl group, which is substituted by pyrrolidinoethyl or by hydroxyethyl.

5. The compound according to claim 1, its cis-trans isomers, its enantiomers, its diastereoisomers, its racemates, its solvates, its hydrates or its pharmaceutically acceptable salts or its prodrugs, said compound having the following formula (II): Among them, 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 F or Cl; R c is a hydroxyl group, an amino group, a 3- to 8-membered heterocycloalkyl group containing 1 to 2 heteroatoms selected from nitrogen, oxygen or sulfur, more preferably R c is a hydroxyl group or a 4- to 6-membered heterocycloalkyl group containing 1 nitrogen heteroatom, more preferably, R c is a hydroxyl group or a pyrrolidinyl group, most preferably, R c is a hydroxyl group or 1-pyrrolidinyl group.

6. The compound according to claim 1, its cis-trans isomers, its enantiomers, its diastereoisomers, its racemates, its solvates, its hydrates or its pharmaceutically acceptable salts or its prodrugs, wherein the compound is selected from the group consisting of: or its hydrochloride.

7. A product comprising the compound according to any one of claims 1 to 6, and a pharmaceutically or immunologically acceptable carrier or excipient.

8. The product according to claim 7, wherein: The product is selected from: pharmaceutical compositions, medicaments, kits or cartridges; and / or wherein the product is for inducing the production of type I interferon in a subject and / or for preventing and / or treating tumors in a subject.

9. Use of the compound according to any one of claims 1 to 6 in the preparation of a product for inducing the production of type I interferon in a subject or a product for preventing and / or treating tumors in a subject.

10. The application according to claim 9, wherein The subject is a human or a non-human mammal, such as a non-human primate (gorilla, chimpanzee), a pet (such as a pet cat, dog, guinea pig, rabbit), a livestock animal (such as a cow, horse, donkey, mule, camel, pig, sheep, chicken, duck, goose, rabbit, deer, mink, otter, musk, etc.); and / or wherein the production of the type I interferon is mediated by hnRNPA2B1; and / or wherein the prevention and / or treatment of tumors in the subject can benefit from the production or increased level of type I interferon in vivo; and / or wherein the tumor is one or more tumors selected from the following group: solid tumors, such as liver cancer, lung cancer, melanoma, breast cancer, ovarian cancer, prostate cancer, kidney cancer, intestinal cancer, head and neck cancer, skin cancer, bladder cancer, pancreatic cancer; non-solid tumors, such as hematological tumors (such as leukemia), nervous system tumors (such as glioma); preferably, non-small cell lung cancer, liver cancer, Lewis lung cancer, melanoma, breast cancer, colon cancer; and / or wherein the product is selected from: pharmaceutical compositions, medicaments, kits or cartridges; and / or wherein the product is for inducing the production of type I interferon in a subject and / or for preventing and / or treating tumors in a subject.

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