HnRNPA2B1 agonist and tumor prevention and treatment application of hnRNPA2B1 agonist combined with STING agonist

Abstract

The invention relates to a heterogeneity cell nuclear glycoprotein A2B1 (hnRNPA2B1) hnRNPA2B1 agonist compound and application of the heterogeneity cell nuclear glycoprotein A2B1 (hnRNPA2B1) hnRNPA2B1 agonist compound to tumor resistance in combination with an STING agonist. Specifically, the invention relates to a fused ring compound as shown in a formula (I), cis-trans isomers, enantiomers, diastereoisomers, racemes, solvates, hydrates or pharmaceutically acceptable salts or prodrugs thereof, and a composition and a product containing any one of the former, the invention also discloses an application of the substance as an hnRNPA2B1 agonist in preparation of a product for preventing and / or treating tumors. The invention can be used for broad-spectrum prevention and treatment of tumors, and has a wide application prospect. # imgabs0 #

Description

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 are agonists of nuclear protein hnRNPA2B1 (i.e., heterogeneous nuclear ribonucleoprotein A2B1), products containing the compounds, and their effects, mechanisms of action, implementation methods, and uses in combination with STING agonists 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 a variety of immune cells and is a key cytokine that regulates tumor immunity. Interferon can directly inhibit the proliferation of human tumor cells and can synergize with a variety of chemotherapeutic drugs. Type I interferon can significantly enhance the anti-tumor immune response 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 to enhance 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 in 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 self-nucleic acids and then activating interferon, including DNA intercalators, various epigenetic therapeutic drugs, splicesome regulators, etc. Studies in recent years have shown that the cGAS-STING pathway in the cytoplasm can induce type I interferon, and the development of STING agonists has become an important direction in the field of tumor treatment. However, there is increasing evidence that the cGAS-STING pathway is underexpressed in tumor cells or that signaling pathway activation is blocked. In addition, there is also evidence that cGAS can promote tumor cell survival. Therefore, in the field of tumor immunotherapy, it is urgent to develop agonists for natural 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 in recent years, belonging to the hnRNP family. On the one hand, hnRNP-A2B1 can sense and recognize the nucleic acid components of DNA viruses (such as HSV-1, etc.), and self-activate to form homodimers, which are demethylated by the demethylase JMJD6 and translocated from the nucleus to the cytoplasm, thereby activating the TBK1-IRF3 signaling pathway and initiating the expression of type I interferon to exert an antiviral effect. On the other hand, as an RNA-binding protein, hnRNPA2B1 can also promote m6A modification, nuclear-cytoplasmic transfer and translation on cGAS, IFI16 and STING mRNA, thereby ensuring the full 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 generally considered to be an important factor promoting tumorigenesis and development. In recent decades, many studies have shown that hnRNPA2B1 is involved in regulating a variety of basic biological functions, such as regulating cell metabolism, migration and invasion, proliferation, and response to mitochondrial stress. For example, hnRNPA2-mediated activation of the invasive phenotype involves different mechanisms, including alternative splicing of TP53INP2, activation of the CXCL12 / CXCR4 axis, and activation of invasive behavior after mitochondrial DNA depletion, among which 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 the regulation of cancer cell phenotypes such as metabolism, proliferation, apoptosis, migration, and invasion through different molecular mechanisms. In addition, hnRNPA2B1 has been reported to induce epithelial-mesenchymal transition (EMT) in a variety of cancer cell lines. In addition, hnRNPA2B1 is also involved in regulating basic cancer processes such as aerobic glycolysis. It is reported that hnRNPA2 can regulate the alternative splicing of pyruvate kinase isozyme M2 (PKM2) and activate the metabolic conversion of cancer cells to aerobic glycolysis. hnRNPA2B1 also plays an important role in the regulation of hypoxia. In general, hnRNPA2B1 is widely involved in the occurrence and development of various tumors. The development of small molecules targeting hnRNPA2B1 will hopefully block the tumor-promoting effects of hnRNPAB1 and provide effective support for tumor treatment.

[0006] In clinical practice, there are more and more methods for treating tumors, such as surgical treatment, radiotherapy, chemotherapy, endocrine therapy, targeted therapy, etc. However, due to the heterogeneity of tumors and the tolerance of patients to different therapies, it is difficult to achieve good results by relying solely on a certain treatment method. At the same time, problems such as drug resistance will arise during the treatment process. Therefore, combined tumor treatment with two or more methods has become an important strategy. The immune system can recognize and kill tumor cells, while tumor cells can adopt different strategies to suppress the human immune system and achieve immune escape. Immunotherapy has become a major hope for curing tumors, and combined treatments including immunotherapy have become a major direction of traditional Chinese medicine in this field. It is necessary to invent effective drugs to activate the immune system and cooperate with other therapies to kill and clear tumors more effectively.

[0007] The cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) signaling pathway is a cytoplasmic DNA sensing pathway that plays a key role in the innate immune response. When tumor-derived DNA is recognized by the DNA receptor cGAS in the cytoplasm, cGAS catalyzes the connection of two nucleotides to form cyclic dinucleotides (CDNs, such as 2',3'-cGAMP), after which CDNs bind to STING as a second messenger and activate it. Signaling downstream of STING leads to the activation of antigen-presenting cells (APCs) and the production of inflammatory cytokines, which in turn promotes the priming and recruitment of T cells. Based on the important role of STING in promoting anti-tumor immunity, a large number of teams have initiated the development of STING agonists. Scientists believe that exogenous STING agonists may have the potential to trigger innate immune activation, thereby leading to adaptive immune responses that can control tumor growth when used as a single agent or in combination with checkpoint blockade therapy.

[0008] In summary, the development of specific agonists targeting hnRNPA2B1 and their combination with other anti-tumor therapies or drugs has great potential in treating tumors. There is an urgent need in this field to develop an immunologically active substance that can activate or promote interferon production, enhance anti-tumor effects, and effectively prevent tumor metastasis, as well as its combined products and methods. Summary of the invention

[0009] The present disclosure provides a fused ring compound having a structure shown in formula (I), its related derivatives (e.g., its cis-trans isomers, its enantiomers, its diastereomers, its racemates, its solvates, its hydrates or its pharmaceutically acceptable salts or its prodrugs), and products containing the compound or its related derivatives. The present disclosure also provides the use of the compound, derivatives and products in tumor prevention and treatment, and further provides their use in treating or preventing tumors and related diseases or symptoms. The compounds, drugs, pharmaceutical compositions or kits disclosed in the present disclosure can be used to effectively fight tumors and control the occurrence and / or metastasis of tumors.

[0010] In some aspects of the present disclosure, provided are fused ring compounds represented by formula (I), cis-trans isomers, enantiomers, diastereomers, racemates, solvates, hydrates, or pharmaceutically acceptable salts or prodrugs thereof.

[0011]

[0012] Wherein, L is -(CH2)n-, and n is an integer of 0-6;

[0013] X is a halogen;

[0014] R 1 For-NR 4 R 5 , where R 4 , R 5 Each independently is H or C 1-6 alkyl;

[0015] R 2 and R 3 Each independently is H or C 1-6 alkyl;

[0016] or R 2 and R 3 Together with the N atom to which they are attached, they form a 5-8 membered heterocycloalkyl group containing 1-3 heteroatoms selected from nitrogen, oxygen or sulfur;

[0017] 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 hydroxyl, amino, halogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, hydroxyl C 1-6 Alkyl, C 1-6 Alkylamino, C 3-6 Cycloalkyl, C 3-6 Cycloalkyl C 1-6Alkyl, 3-6 membered heterocycloalkyl, 3-6 membered heterocycloalkylC 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.

[0018] 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, wherein the product may further comprise a STING agonist used in combination with the compound.

[0019] In some aspects of the present disclosure, a method for preventing and / or treating tumors is also provided, comprising administering to a subject in need thereof an effective amount of a compound or product of the present disclosure for prevention and / or treatment, which further comprises or administers a STING agonist in combination with the compound.

[0020] In some aspects of the present disclosure, the compounds, compositions or products of the present disclosure are also provided, which are used for preventing and / or treating tumors.

[0021] In some aspects of the present disclosure, there is provided use of a compound or product of the present disclosure in increasing the level of an interferon (eg, type I interferon, such as IFN-α and / or IFN-β).

[0022] 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 disclosure. Other aspects of the present disclosure are obvious to those skilled in the art due to the disclosure of this article. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] 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 rather than for limiting the scope of the present disclosure.

[0024] Figure 1 : The test compounds activated the nuclear export of hnRNPA2B1 in A549 lung cancer cells.

[0025] Figure 2 :The test compounds activated the downstream TBK1 and IRF3 after hnRNPA2B1 was exported from the nucleus of A549 lung cancer cells.

[0026] Figure 3 : The ELISA method was used to detect the activation effect of compounds on type I interferon (IFN-β) in tumor cells.

[0027] Figure 4 :Western blotting method was used to detect the upregulation effect of compounds on STING protein levels.

[0028] Figure 5 : Test compounds combined with STING agonists induce stronger type I interferon production.

[0029] Figure 6 : The test compound synergistically inhibits tumor growth in combination with a STING agonist. DETAILED DESCRIPTION

[0030] In the present application, through a large amount of research, development and experiments, a new type of condensed ring compound is discovered, which can specifically bind to the hnRNPA2B1 protein with high affinity and can further widely induce the production of higher levels of IFN-β, and is thus defined as a hnRNPA2B1 agonist.

[0031] In anti-tumor tests of different types of tumor cells, the hnRNPA2B1 agonist compounds disclosed herein can significantly activate type I interferon in tumor cells, inhibit LLC tumor proliferation, and promote the survival of tumor-bearing mice. In addition, since the hnRNPA2B1 agonist compounds disclosed herein can widely induce the production of higher levels of IFN-β, it can be expected that these compounds can play an anti-tumor role through the action of type I interferon. In addition, when the compounds of the present application are used in combination with STING agonists, more type I interferon can be induced and tumor growth can be significantly inhibited.

[0032] Thus, the present disclosure provides methods and strategies for combining novel anti-tumor hnRNPA2B1 agonist compounds with STING agonists for use in tumor prevention and treatment.

[0033] All numerical ranges provided herein are intended to clearly include all values ​​falling between the range endpoints and the numerical ranges therebetween. Features mentioned in the present disclosure or features mentioned in the embodiments may be combined. All features disclosed in this specification may be used in combination with any form of composition, and each feature disclosed in the specification may be replaced by any alternative feature that can provide the same, equal or similar purpose. Therefore, unless otherwise specified, the disclosed features are only general examples of equal or similar features.

[0034] As used herein, “containing,” “having,” or “including” include “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 and refer to heterogeneous nuclear ribonucleoprotein A2B1. The hnRNPA2B1 protein involved in the present disclosure may be a protein encoded by the hnRNPA2B1 gene, its cDNA or CDS in an animal (such as a human or mouse, etc.), or a homologous sequence of these proteins that promotes the expression of interferon (for example, the homologous sequence of hnRNPA2B1 can be obtained through a database or comparison software known in the art), a variant or a modified form.

[0037] As used herein, the terms "hnRNPA2B1 gene", "hnRNPA2B1 encoding gene", "hnRNPA2B1 protein encoding gene" or "nucleic acid molecule encoding hnRNPA2B1" are used interchangeably and refer to a nucleotide sequence encoding the hnRNPA2B1 protein or polypeptide described in the present disclosure, which can be, for example, the human hnRNPA2B1 gene of Gene ID: 3181, the mouse hnRNPA2B1 gene of Gene ID: 53379. The term also includes molecules that hybridize with the signed nucleic acid molecule under stringent conditions, or family gene molecules that are highly homologous to the above molecules, and it is believed that the expression of the gene has a certain promoting effect on the production and effect of interferon.

[0038] The present disclosure provides a class of novel compounds that are hnRNPA2B1 "agonists" (or "promoters"). The terms "agonist" or "hnRNPA2B1 agonist compound" are used interchangeably and refer to a class of novel condensed ring compounds that can increase the level or activity of hnRNPA2B1, which can specifically bind to the hnRNPA2B1 protein with high affinity, and can further widely 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 its prodrugs,

[0041]

[0042] Wherein, L is -(CH2)n-, and n is an integer of 0-6;

[0043] X is a halogen;

[0044] R 1For-NR 4 R 5 , where R 4 , R 5 Each independently is H or C 1-6 alkyl;

[0045] R 2 and R 3 Each independently is H or C 1-6 alkyl;

[0046] or R 2 and R 3 Together with the N atom to which they are attached, they form a 5-8 membered heterocycloalkyl group containing 1-3 heteroatoms selected from nitrogen, oxygen or sulfur;

[0047] 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 hydroxyl, amino, halogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, hydroxyl 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 heterocycloalkylC 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.

[0048] Preferably, X is F or Cl, more preferably, X is F.

[0049] Preferably, L is -(CH2)n-, wherein n is 1, 2 or 3, more preferably, n is 2.

[0050] Preferably, R 4 , R 5 Each is independently H, methyl or ethyl.

[0051] Preferably, R 2 and R 3 Each independently is C 1-3 Alkyl, preferably methyl, ethyl or isopropyl, the 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 1-3Alkyl, preferably methyl, ethyl or isopropyl, the C 1-3 The alkyl group is unsubstituted or substituted with a 4-6 membered heterocycloalkyl group containing 1 nitrogen atom. 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.

[0052] 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 hydroxyl 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 hydroxyl 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 hydroxyl C 1-6 Alkyl substituted; most preferably, R 2 and R 3 Together with the N atom to which they are attached, they form a piperazinyl group, which is substituted by a 5-6 membered heterocycloalkyl C containing one N atom. 1-6 Alkyl substituted or hydroxyl C 1-6 Alkyl is substituted, preferably, the piperazinyl is substituted with a pyrrolidinyl C 1-6 Alkyl substituted or hydroxyl C 1-6 Alkyl substituted, more preferably, the piperazinyl is substituted by pyrrolidinylethyl or hydroxyethyl.

[0053] Preferably, the compound has the following formula (II):

[0054]

[0055] 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;

[0056] M is -(CH2)n-, wherein n is 1, 2 or 3, more preferably, n is 2;

[0057] X is F or Cl, more preferably, X is F;

[0058] R c is hydroxyl, amino, 3-8 membered heterocycloalkyl containing 1-2 heteroatoms selected from nitrogen, oxygen or sulfur, more preferably, R c is hydroxyl or a 4-6 membered heterocycloalkyl group containing 1 nitrogen atom, more preferably, R c is hydroxyl or pyrrolidinyl, most preferably, R c is hydroxy or 1-pyrrolidinyl.

[0059] Preferably, the compound is selected from the following group of compounds or their salts (eg, hydrochloride):

[0060]

[0061] Unless otherwise indicated, the following terms and phrases used herein are intended to have the following definitions. A particular term or phrase should not be considered to be uncertain or unclear in the absence of a special definition, but should be understood according to its ordinary meaning. When a trade name appears in this article, it is intended to refer to its corresponding commercial product or active ingredient.

[0062] The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, 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.

[0063] The term "pharmaceutically acceptable salt" refers to a salt of a compound of the invention, prepared from a compound having a specific substituent discovered by the invention and a relatively nontoxic acid or base. When the compound of the invention contains a relatively acidic functional group, a base addition salt can be obtained by contacting the neutral form of such compound with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium calcium, ammonium, organic amine or magnesium salts or similar salts. When the compound of the invention contains a relatively basic functional group, a base addition salt can be obtained by contacting the neutral form of such compound with a sufficient amount of acid in a pure solution or 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, hydrogen sulfate, hydroiodic acid, phosphorous acid, etc., and organic acid salts, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid and methanesulfonic acid, and salts of amino acids (such as arginine, etc.), and salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain basic and acidic functional groups, and can be converted into any base or acid addition salt.

[0064] Pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. Generally, the preparation method of such salts is: in water or an organic solvent or a mixture of the two, these compounds in free acid or base form are reacted with a stoichiometric amount of an appropriate base or acid to prepare.

[0065] 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. All of these isomers and their mixtures are included within the scope of the present invention.

[0066] Unless otherwise indicated, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of one another.

[0067] Unless otherwise indicated, the term "cis-trans isomers" or "geometric isomers" arises from the inability of a ring to rotate freely about double bonds or single bonds of ring carbon atoms.

[0068] 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.

[0069] The compounds of the present invention may exist in specific forms. Unless otherwise indicated, the term "tautomer" or "tautomeric form" means that at room temperature, different functional group isomers are in dynamic equilibrium and can quickly convert to each other. If tautomers are possible (such as in solution), it is possible to reach a chemical equilibrium of tautomers. For example, proton tautomers (proton tautomers) (also known as prototropic tautomers) include interconversions carried out by proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence isomers (valencetautomers) include interconversions carried out by the reorganization of some bonding monads. Among them, 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.

[0070] The term "prodrug" generally refers to a compound of formula (I) that has been derivatized with a functional group, and its derivative can be easily converted into a compound of formula (I) in vivo. Generally, the selection and preparation of suitable prodrugs can be described in, for example, Design of Prodrugs, ed. H. Bundgaard, Elsevier, 1985.

[0071] The compounds of the invention may contain unnatural proportions of atomic isotopes on one or more of the atoms constituting the compound, said isotopes having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number that predominantly occurs 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( 14C). All isotopic variations of the compounds of the present invention, whether radioactive or not, are included within the scope of the present invention. Isotopic variants may enhance certain therapeutic advantages, such as the substitution of heavy hydrogen for hydrogen to form deuterated drugs, the bond between deuterium and carbon being stronger than the bond between ordinary hydrogen and carbon, and the deuterated drugs having advantages such as reduced toxic side effects, increased drug stability, enhanced efficacy, and prolonged drug biological half-life compared to undeuterated drugs, or providing standard compounds that can be used as characterization of biological samples. Isotopically enriched compounds within the general formula (I) may 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 routes and embodiments of the present invention, using appropriate isotopically enriched reagents and / or intermediates.

[0072] "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.

[0073] The naming convention used in the present invention is based on the IUPAC system nomenclature generated by ChemDraw software.Any open valence bonds occurring on carbon, oxygen, sulfur or nitrogen atoms in the structures given in the present invention indicate the presence of hydrogen atoms.

[0074] The term "substituted" means that any one or more hydrogen atoms on a specific atom are replaced by a substituent, which may include deuterium and hydrogen variants, as long as the valence state of the specific atom is normal and the substituted compound is stable. When the substituent is oxo (i.e., =O), it means that two hydrogen atoms are replaced, and oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may be substituted or not substituted, and unless otherwise specified, the type and number of substituents can be arbitrary on the basis of chemical realization.

[0075] 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, and each occurrence of R is an independent choice. In addition, combinations of substituents and / or variants thereof are permitted only if such combinations result in stable compounds.

[0076] When the number of a linking group is 0, such as -(CRR)0-, it means that the linking group is a single bond.

[0077] 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.

[0078] When the listed linking group does not indicate its linking direction, its linking direction is arbitrary, for example, The connecting group L is at this time You can connect the benzene ring and the cyclopentyl group 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.

[0079] Unless otherwise specified, the number of atoms in a ring is generally defined as the ring member number, for example, "3-7 membered ring" refers to a "ring" having 3-7 atoms arranged around it.

[0080] Unless otherwise specified, the term "halogen" refers to fluorine, chlorine, bromine and iodine.

[0081] Unless otherwise specified, the term “C 1-6 "Alkyl" is used to refer to 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.; it can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1-6 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl, and the like.

[0082] Unless otherwise specified, the term “C 1-3 "Alkyl" is used to refer to 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 may be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1-3 Examples of the alkyl group include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like.

[0083] Unless otherwise specified, the term “C 2-6"Alkenyl" is used to refer to a straight or branched hydrocarbon group consisting of 2 to 6 carbon atoms containing at least one carbon-carbon double bond. The carbon-carbon double bond can be located at any position of the group. 2-6 Alkenyl includes 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.

[0084] Unless otherwise specified, the term “C 1-6 "Alkoxy" refers to those alkyl groups containing 1 to 6 carbon atoms which are attached to the rest of the molecule through 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.

[0085] Unless otherwise specified, the term “C 1-6 "Alkylamino" refers to those alkyl groups containing 1 to 6 carbon atoms which are attached to the rest of the molecule via 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.

[0086] 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 includes C 3-5 , C 4-5 and C 5-6 Cycloalkyl, etc.; it may be monovalent, divalent or polyvalent.3-6 Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

[0087] Unless otherwise specified, the term "3-8 membered heterocycloalkyl" by itself or in combination with other terms refers to 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 rest are 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, for the "3-8 membered heterocycloalkyl", a heteroatom may occupy the position where the heterocycloalkyl is connected to the rest of the molecule. The 3-8 membered heterocycloalkyl includes 4-6 membered, 5-6 membered, 7-8 membered, 4 membered, 5 membered and 6 membered heterocycloalkyls, etc. Examples of 3-6 membered heterocycloalkyls include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothienyl, tetrahydrofuranyl (including tetrahydrofuran-2-yl), piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperazinyl, morpholinyl, thiomorpholinyl, etc.

[0088] Unless otherwise specified, the terms "6-10 membered aromatic ring" and "6-10 membered aryl" can be used interchangeably, and the term "6-10 aryl" means a monovalent aromatic carbocyclic ring system containing 6-10 carbon atoms and having at least one aromatic ring or multiple condensed rings in which at least one ring is an aromatic ring. Examples of aryl are, but not limited to, phenyl, naphthyl, biphenyl or indanyl, etc.

[0089] 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 with a conjugated π electron system consisting of 5 to 10 ring atoms, 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 and 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 may be attached to the rest of the molecule via a heteroatom or a carbon atom, including 5-8 membered, 5-7 membered, 5-6 membered, 5 membered and 6 membered heteroaryl groups, etc. Examples of the 5-10 membered heteroaryl group include, but are 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 1-thiazolyl and 5-thiazolyl, etc.), furanyl (including 2-furanyl and 3-furanyl, 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.

[0090] 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 with a conjugated π electron system consisting of 5 to 6 ring atoms, 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). The 5-6 membered heteroaryl may be attached to the rest of the molecule via a heteroatom or a carbon atom, and the 5-6 membered heteroaryl includes 5-membered and 6-membered heteroaryl, etc. Examples of the 5-6 membered heteroaryl include, but are 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.), furanyl (including 2-furanyl and 3-furanyl, 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.

[0091] 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 Etc.; similarly, n-membered to n+m-membered means that the number of atoms in the ring is n to n+m, for example, 3-7-membered ring includes 3-membered, 4-membered, 5-membered, 6-membered and 7-membered rings, and also includes any range from n to n+m, for example, 3-7-membered ring includes 3-6-membered ring, 4-7-membered ring, 5-7-membered ring and 6-7-membered ring, etc.

[0092] Preparation method

[0093] The present invention also relates to a method for producing the compound of the general formula (I) defined above. In some embodiments, the synthesis method of the compound of the present application is as follows:

[0094]

[0095] When X is selected from halogens other than Br, such as F, Cl, I, etc., the method comprises the following steps:

[0096] 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) or 1,3-dibromo-5,5-dimethylhydantoin, preferably, the reaction temperature is 0-30° C.;

[0097] 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;

[0098] c) oxidizing the compound of formula Ic to the compound of formula Id using an oxidant, preferably, the oxidant includes but is not limited to K2Cr2O7, preferably, the reaction temperature is 60-150°C;

[0099] 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.;

[0100] 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.

[0101] When X is selected from Br, the method comprises:

[0102] 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;

[0103] c) oxidizing the compound of formula Ib to a compound of formula Id using an oxidant, preferably, the oxidant includes but is not limited to K2Cr2O7, preferably, the reaction temperature is 60-150°C;

[0104] 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;

[0105] 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.

[0106]

[0107] Alternatively, when X is selected from halogens other than Br, such as F, Cl, I, etc., the method comprises the following steps:

[0108] f) 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) or 1,3-dibromo-5,5-dimethylhydantoin, preferably, the reaction temperature is 0-30° C.;

[0109] 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;

[0110] h) oxidizing the compound of formula I-c' to a compound of formula I-d' using an oxidant, preferably, the oxidant includes but is not limited to K2Cr2O7, preferably, the reaction temperature is 60-150°C;

[0111] 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;

[0112] 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.

[0113] 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.

[0114] STING agonists

[0115] According to the contents disclosed in the present application and experimental verification, the compounds of the present application can be used in combination with STING agonists to enhance their anti-tumor effects, preferably to produce a synergistic effect.

[0116] Based on the important role of STING in promoting anti-tumor immunity, a large number of teams have initiated the research and development of STING agonists. Scientists believe that exogenous STING agonists may have the potential to trigger innate immune activation, thereby leading to adaptive immune responses that can control tumor growth when used as a single drug or in combination with checkpoint blockade therapy.

[0117] The STING agonists that can be used in combination with the hnRNPA2B1 agonist of the present application include, but are not limited to, one or more selected from the following group: SR-717, MSA2, MK-1454, ADU-S100, BMS-986301, TAK-676, E7766, GSK3745417, SB11285, IMSA101, SR-8314, IACS-8803, AN3005, OX401, TTI-10001, VTX-001, IACS-8779, SOMCL-18-202, and MSA-2.

[0118] As of September 2020, some STING agonists in clinical and preclinical development are summarized in the table below. It should be understood that a person of ordinary skill in the art can select STING agonist active substances as needed based on the content disclosed in this application and use them in combination with the hnRNPA2B1 agonist compound of this application.

[0119]

[0120]

[0121] Products and Applications

[0122] The present disclosure also provides a product, which may be, for example, a drug, a pharmaceutical composition or a kit / medicine box, containing an effective amount of the 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 and refer to a hnRNPA2B1 agonist compound having the structural formula (I) or a derivative or product thereof. The product of the present disclosure may further include a STING agonist used in combination with the hnRNPA2B1 agonist compound.

[0123] As used herein, the term "pharmaceutically acceptable" ingredients are suitable for humans and / or animals without excessive adverse reactions (such as toxicity, irritation and allergic reactions), i.e., substances with a reasonable benefit / risk ratio. As used herein, the term "effective amount" refers to an amount that can produce a function or activity to humans and / or animals and can be accepted by humans and / or animals.

[0124] As used herein, the term "pharmaceutically acceptable carrier" refers to a carrier for the administration of a therapeutic agent, including various excipients and diluents. The term refers to such pharmaceutical carriers: they are not necessary active ingredients themselves and are not excessively toxic after administration. Suitable carriers are well known to those of ordinary skill in the art, and a full discussion of pharmaceutically acceptable excipients can be found in Remington's Pharmaceutical Sciences (Mack Pub. Co., NJ 1991).

[0125] The pharmaceutically acceptable carrier in the composition may contain a liquid such as water, saline, glycerol and ethanol. In addition, auxiliary substances such as fillers, disintegrants, lubricants, glidants, effervescent agents, wetting agents or emulsifiers, flavoring agents, pH buffer substances, etc. may also be present in these carriers. 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, the pH is about 6-8.

[0126] The active substance in the product disclosed herein accounts for 0.001-99.9 wt% of the total weight of the composition, preferably 1-95 wt%, more preferably 5-90 wt%, and more preferably 10-80 wt%, and the remainder is pharmaceutically acceptable carriers and other additives.

[0127] The hnRNPA2B1 agonist compound and the STING agonist can be used in combination at a ratio of 1:100 to 100:1.

[0128] As used herein, the term "unit dosage form" means that the product of the present disclosure is prepared into a dosage form required for a single dose for the convenience of taking, including but not limited to various solid doses (such as tablets), liquid doses, capsules, and sustained-release preparations.

[0129] In another preferred embodiment of the present disclosure, the product is in unit dosage form or multiple dosage form, and the content of the active substance therein 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 example of the present disclosure, 1 to 6 doses of the composition of the present disclosure are administered per day, preferably 1 to 3 doses; most preferably, the daily dose is 1 dose. Moreover, when used in combination, the composition may further comprise a therapeutically effective amount of a STING agonist, or a product comprising the compound of the present application may be used in combination with a product comprising a STING agonist.

[0130] It should be understood that the effective dosage of the hnRNPA2B1 agonist compound and the active substance such as the STING agonist used may vary depending on the severity of the subject to be administered or treated. The specific situation is determined according to the individual situation of the subject (e.g., the subject's weight, age, physical condition, and the desired effect), which is within the scope of judgment of a skilled physician.

[0131] The products disclosed herein may be solid (such as granules, tablets, lyophilized powders, suppositories, capsules, sublingual tablets) or liquid (such as oral liquids) or other suitable shapes. The routes of administration may include, but are not limited to: (1) traditional administration methods, such as gastrointestinal administration (such as oral administration), parenteral administration (such as 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, etc.; (2) connecting hnRNPA2B1 agonists and / or STING agonists to transferrin / poly-L-lysine complexes to enhance their biological effects; (3) encapsulating drugs with liposomes and mediating their entry into cells, which is conducive to the smooth entry of compound molecules and is protected from the hydrolysis of various extracellular enzymes; (4) using liposomes to transport active substances so that they are specifically transported to target tissues and target cells.

[0132] In addition, the product 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 for regulating anti-tumor effects are administered before, simultaneously with, or after administration of the product of the present disclosure.

[0133] In some embodiments, the compounds or products of the present application activate hnRNPA2B1 and STING to further regulate the immune system to prevent and treat tumors.

[0134] The term "subject" may 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.

[0135] 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, disease 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 tumor growth to some extent, 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 peripheral organs; (6) inhibiting, including (i) reducing, (ii) slowing, or (iii) completely preventing cancer metastasis; (7) enhancing anti-tumor immune responses, 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.

[0136] In a specific embodiment, the disease or condition to be treated is a cell proliferative disease. In certain embodiments, the cell proliferative disease 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 tumors, germ cell tumors, malignant neuroendocrine (carcinoid) tumors, 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 related to AIDS.

[0137] 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 tumors (e.g., oligotrophic astrocytoma and anaplastic follicular astrocytoma), ependymoma (e.g., mucosal papillary ependymoma and anaplastic ependymoma); medulloblastoma, primitive neuroectodermal tumor, schwannoma, meningioma, atypical meningioma, anaplastic meningioma, pituitary adenoma, brain stem glioma, cerebellar astrocytoma, cerebral aneurysm / malignant glioma, visual pathway and hypothalamic glioma and primary central nervous system lymphoma. In specific examples of these embodiments, brain cancer is selected from: glioma, multiforme glioblastoma, paraganglioma and upper tract epithelial primitive neuroectodermal tumor (sPNET).

[0138] 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 intraocular melanoma and retinoblastoma.

[0139] 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.

[0140] 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.

[0141] 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, which is 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, which is selected from squamous cell carcinoma and adenocarcinoma. In specific examples of these embodiments, the cancer is ovarian cancer, which is selected from epithelial cancer.

[0142] In specific embodiments, the cancer is selected from cancers of the gastrointestinal system. In specific embodiments, the cancer is selected from: esophageal cancer, gastric cancer (also known as gastric cancer), gastrointestinal carcinoid tumors, pancreatic cancer, gallbladder cancer, colorectal cancer, and anal cancer. In examples of these embodiments, the cancer is selected from: esophageal squamous cell carcinoma, esophageal adenocarcinoma, gastric adenocarcinoma, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors, gastric lymphoma, gastrointestinal lymphoma, solid papillary pancreatic tumors, pancreatoblastoma, islet cell tumors, pancreatic cancer (including acinar cell carcinoma and ductal adenocarcinoma), gallbladder adenocarcinoma, colorectal adenocarcinoma, and anal squamous cell carcinoma.

[0143] In specific embodiments, the cancer is selected from liver cancer and bile duct cancer. In specific embodiments, the cancer is liver cancer (also known as hepatocellular carcinoma). In specific embodiments, 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.

[0144] 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 carcinoma (transitional cell carcinoma), squamous cell carcinoma and adenocarcinoma.

[0145] In a specific embodiment, the cancer is selected from bone cancer. In a specific embodiment, the bone cancer is selected from: osteosarcoma, malignant fibrous histiocytoma of bone, Ewing's sarcoma, chordoma (bone cancer along the spine).

[0146] 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 tumors and pleuropulmonary blastoma.

[0147] 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.

[0148] 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 chondroma, fibrosarcoma, clear cell sarcoma of the tendon sheath, and Kaposi's sarcoma.

[0149] In a specific embodiment, the cancer is selected from lymphoma. In a specific embodiment, the cancer is selected from: Hodgkin's lymphoma (e.g., Reed-Stemberg cell), non-Hodgkin's lymphoma (e.g., diffuse large B-cell lymphoma, follicular lymphoma, mycosis fungoides, Sezary syndrome, primary central nervous system lymphoma), cutaneous T-cell lymphoma, primary central nervous system lymphoma.

[0150] 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.

[0151] 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.

[0152] 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.

[0153] In a specific embodiment, the cancer is selected from a cardiac tumor. In a specific embodiment, the cardiac tumor is selected from a malignant teratoma, a lymphoma, a rhabdomyosarcoma, angiosarcoma, a chondrosarcoma, an infantile fibrosarcoma, and a synovial sarcoma.

[0154] In specific embodiments, the cell proliferative disease is selected from the group consisting of benign papillomatosis, benign neoplastic disease, and gestational trophoblastic disease. In specific embodiments, the benign neoplastic disease is selected from the group consisting of skin papilloma (warts) and genital papilloma. In specific embodiments, the gestational trophoblastic disease is selected from the group consisting of hydatid cystic nevus and gestational trophoblastic neoplasms (e.g., invasive nevus, choriocarcinoma, placental site trophoblastic tumor, and epithelioid trophoblastic tumor).

[0155] In some embodiments, the compounds or products of the present application are administered as preventive drugs before tumor development to prevent tumor development or reduce the severity of subsequent tumors. In some embodiments, the compounds or products of the present application are administered as therapeutic drugs after tumor development to reduce the severity of tumor diseases including their metastasis. In some embodiments, the compounds or products of the present application are administered as both preventive drugs and therapeutic drugs, and are administered continuously or intermittently before and after tumor development.

[0156] 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 are obvious to those skilled in the art due to the disclosure of this article.

[0157] Example

[0158] The present disclosure is further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present disclosure and are not used to limit the scope of the present disclosure. Those skilled in the art may make appropriate modifications and changes to the present disclosure, and these modifications and changes are within the scope of the present disclosure.

[0159] The experimental methods for which the specific conditions are not specified in the following examples can be conventional methods in the art, for example, refer to "Molecular Cloning Laboratory Manual" (3rd edition, New York: Cold Spring Harbor Laboratory Press, 1989) or follow the conditions recommended by the supplier. The DNA sequencing method is a conventional method in the art and can also be tested by commercial companies.

[0160] Unless otherwise specified, percentages and parts are calculated by weight. Unless otherwise defined, all professional 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 applied to the disclosed methods. The preferred implementation methods and materials described herein are for demonstration purposes only.

[0161] I. Synthesis and Characterization Examples of Compounds

[0162] Example I.1: Synthesis of Compound A (FPC-1):

[0163]

[0164] *eq equivalent

[0165] 1-1. Preparation of Compound 2

[0166]

[0167] 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, the resulting suspension was stirred to allow it to naturally warm to room temperature and stirred overnight. The reaction solution was filtered, the filter cake was washed three times with ethanol, the dried filter cake was collected, and recrystallized with EA to obtain a pure product (10.1 g, off-white crystals).

[0168] 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).

[0169] 1-2. Preparation of Compound 3

[0170]

[0171] 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 N-fluorobisbenzenesulfonamide (NFSI) (9 g, 28.8 mmol) in 100 mL of anhydrous THF was slowly added, the mixture was stirred for another 60 minutes at -80 ° C. 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 with 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.

[0172] 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).

[0173] 1-3. Preparation of Compound 4

[0174]

[0175] Compound 3 (4.5g, 17.9mmol) is added in the glacial acetic acid solution (50mL) of potassium dichromate (24g, 82.4mmol).The solution is heated to reflux 16 hours, then poured into ice water.Gained precipitate is collected by filtration and washed with water.Filter cake is air-dried, added among the DCM and refluxed 30 minutes, filtered, washed with DCM, filter cake is added among the DCM and refluxed 15 minutes, filtered and washed with DCM.Merge all filtrates and vacuum concentration to obtain compound 4 (4.1g, crude product, yellow solid).

[0176] 1-4. Preparation of INT-1

[0177]

[0178] 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 under reflux for 1 hour. The reaction mixture was concentrated, and the residue was purified by column chromatography (PE / EA=5 / 1). An off-white solid (2.8 g) was obtained.

[0179] 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).

[0180] 1-5. Preparation of FPC-1-1

[0181]

[0182] 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, and 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+NH3·H2O) to obtain 240 mg of pure yellow solid.

[0183] 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).

[0184] 1-6. Preparation of FPC-1

[0185]

[0186] Compound FPC-1-1 (240 mg, 0.44 mmol) was dissolved in 11 mL DCM / MeOH (10 / 1). 5M HCl / 1,4-dioxane (5 mL) was added to the mixture and stirred at room temperature for 2 hours. The mixture was concentrated in vacuo, the residue was dissolved in a small amount of methanol, precipitated with ether, centrifuged, the precipitate was washed with ether, centrifuged and dried in vacuo to obtain 240 mg of pure product (yellow solid).

[0187] 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).

[0188] MS (ESI) m / z [M+H] + =440.3.

[0189] Example I.2: Synthesis of Compound B (FPC-2)

[0190]

[0191] 2-1. Preparation of FPC-2-1

[0192]

[0193] 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.

[0194] 1H 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).

[0195] 2-2. Preparation of FPC-2

[0196]

[0197] Compound FPC-2-1 (20 mg, mmol) was dissolved in 2 mL 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 obtain 5 mg of pure product (yellow solid).

[0198] 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).

[0199] MS (ESI) m / z [M+H] + =401.47.

[0200] Example I.3: Synthesis of Compound C (FPC-3)

[0201]

[0202] 3-1. Preparation of FPC-3-1

[0203]

[0204] 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), and bis(triphenylphosphine)palladium chloride (II) (128 mg, 0.18 mmol) was added to the mixture under argon protection, and 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+NH3·H2O) to obtain 30 mg of pure yellow solid.

[0205] 1 H NMR (400MHz, chloroform-d) δ8.63-8.52(m,2H),7.37-7.28(m,2H),4.96(s,1H),4.32(t,J=5.5Hz,2H),3.99(s,2H),3.93-3.75(m,2H),3.49(s,2H) ),3.44(d,J=6.8Hz,2H),3.36-3.16(m,2H),2.87-2.62(m,2H),2.29-2.12(m,2H),2.12-1.91(m,2H),1.28(s,9H),1.10(t,J=7.1Hz,3H).

[0206] 3-2. Preparation of FPC-3

[0207]

[0208] Compound FPC-3-1 (30 mg, 0.06 mmol) was dissolved in 4.4 mL 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, centrifuged, and the precipitate was washed with ether, centrifuged and dried in vacuo to obtain 25 mg of pure product (yellow solid).

[0209] 1H NMR (300MHz, methanol-d4) δ8.67-8.54(m,2H),7.58-7.46(m,2H),4.51-4.41(m,2H),3.87-3.77(m,2H),3.77-3.65(m ,2H),3.51(q,J=6.6Hz,4H),3.34(d,J=1.8Hz,1H),3.20-3.05(m,2H),2.20-1.92(m,5H),1.10(t,J=7.0Hz,3H).

[0210] MS (ESI) m / z [M+H] + =399.47.

[0211] Example I.4: Synthesis of Compound D (FPC-5)

[0212]

[0213] 4-1. Preparation of FPC-5-1

[0214]

[0215] 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), and bis(triphenylphosphine)palladium chloride (II) (32 mg, 0.0.045 mmol) was added to the mixture under argon protection. The reaction was carried out at 80°C for 16 hours, and the reaction solution was cooled to room temperature. Water was added to the reaction solution and stirred, and the organic phase was separated. The aqueous phase was extracted once with DCM, and the organic phases were combined and dried over anhydrous magnesium sulfate. The crude product was purified by TLC (DCM / MeOH=10 / 1+NH3·H2O) by vacuum concentration and column purification to obtain 10 mg of pure yellow solid.

[0216] 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).

[0217] 4-2. Preparation of FPC-5

[0218]

[0219] Compound FPC-5-1 (10 mg, 0.022 mmol) was dissolved in 1.1 mL 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, centrifuged, and the precipitate was washed with ether, centrifuged and dried in vacuo to obtain 9 mg of pure product (yellow solid).

[0220] 1 H NMR (400MHz, methanol-d4) δ8.61-8.52(m,1H),8.44(d,J=8.3Hz,1H),7.38(dd,J=12.5,8.5Hz,1H),7.25(d,J=8.4Hz,1H),4.44(t,J=5.1Hz ,2H),3.22(dd,J=19.7,7.1Hz,4H),3.10(d,J=3.1Hz,3H),2.16(dp,J=13.3,7.2Hz,1H),1.31(d,J=7.7Hz,1H),0.87(d,J=6.4Hz,6H).

[0221] MS (ESI) m / z [M+H] + =344.42.

[0222] Example I.5: Synthesis of Compound E (AIR-2)

[0223]

[0224] 5-1. Preparation of Compound 5

[0225]

[0226] 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 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 obtain 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 give compound 5 (15.42 g, 63.82%) as a white solid.

[0227] 1 H NMR (300MHz, Chloroform-d) δ7.16 (d, J = 7.6Hz, 2H), 7.12–7.04 (m, 2H), 3.37 (s, 4H).

[0228] 5-2. Preparation of INT-2

[0229]

[0230] 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, and PE and ultrasonic dispersion were added. The resulting precipitate was filtered and dried to obtain crude INT-2 (10.55 g) as a brown solid, which was used without further purification.

[0231] 5-3. Preparation of AIR-2-1

[0232]

[0233] 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), 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 a brown solid AIR-2-1 (1.81 g, 27.86%). MS (ESI), m / z: 305.4 [M + H] + .

[0234] 1 H NMR (300MHz, Chloroform-d) δ8.65(t,J=2.5Hz,1H),8.62(t,J=2.5Hz,1H),7.47–7.39(m,2H),4.38(t,J=6.7Hz,2H),2.89–2.81(m,2H),2.49(s,6H).

[0235] 5-4. Preparation of AIR-2

[0236]

[0237] 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 residue obtained was purified by column chromatography (DCM: MeOH = 15: 1 to 5: 1) to obtain a pure product dissolved in DCM. Then 5M HCl / 1,4-dioxane (1.5 mL) was added and stirring was continued, and precipitated with ether. The resulting precipitate was filtered, washed with ether, and dried to give a yellow solid AIR-2 (1.21 g). MS (ESI), m / z: 468.47 [M+H] + .

[0238] 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).

[0239] Example I.6: Synthesis of Compound F (AIR-3)

[0240]

[0241] 6-1. Preparation of Compound 2

[0242]

[0243] NBS (499.72 g, 2.81 mol) was added to DMF (1000 mL) and stirred until most of the solid was dissolved. The reaction system was replaced 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 allowed to warm to room temperature naturally and stirred over the weekend. The reaction solution was filtered and the filter cake was washed twice with PE / EA=1 / 1. The dried filter cake was collected and recrystallized with EA to obtain pure compound 2 (102.57 g, 28.80%) as off-white crystals.

[0244] 1 H NMR (300MHz, Chloroform-d) δ7.79 (d, J = 7.4 Hz, 2H), 7.09 (d, J = 8.8 Hz, 2H), 3.30 (s, 4H).

[0245] 6-2. Preparation of Compound 3

[0246]

[0247] 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 for at least 20 minutes at the same temperature. N-fluorobisbenzenesulfonamide (NFSI) (75.8 g, 240.38 mmol) in 250 mL anhydrous THF was added dropwise, and after stirring for 1.5 hours 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 a 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.

[0248] 1H NMR (300MHz, Chloroform-d) δ7.66 (d, J = 7.4Hz, 1H), 7.21–7.09 (m, 3H), 3.35 (s, 4H).

[0249] 6-3. Preparation of Compound 4

[0250]

[0251] 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 to 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, the filter cake was added to DCM and refluxed for 15 minutes, filtered and washed with DCM. All filtrates were concentrated to give crude compound 3 (6.72 g), which could be used without further purification.

[0252] 6-4. Preparation of AIR-3-1

[0253]

[0254] DIPEA (358.48uL, 2.17mmol) and (2-aminoethyl)(methyl)carbamic acid tert-butyl ester hydrochloride (428.46mg, 2.03mmol) were added to a solution of compound 4 (400mg, 1.36mmol) in EtOH (20mL). The reaction system was replaced with argon 5 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 a yellow solid AIR-3-1 (244mg, 39.88%).

[0255] 6-5. Preparation of AIR-3-2

[0256]

[0257] NaOt-Bu (103.92 mg, 1.08 mol), 1-(2-pyridone-ethyl)piperazine (118.92 mg, 648.81 mmol) and BINAP (101.00 mg, 162.20 mmol) were added to a toluene solution (15 mL) of AIR-3-1 (244 mg, 540.68 mmol). The reaction system was replaced with argon 5 times, and then Pd(OAc)2 (24.28 mg, 108.14 mmol) was added. The reaction system was replaced with argon three times, and the reaction mixture was stirred at 90 ° C for 12 hours under argon protection. The reaction mixture was quenched with water and extracted twice with EA. The combined organic layer was washed with water and brine, dried over Na2SO4 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%).

[0258] 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).

[0259] 6-6. Preparation of AIR-3

[0260]

[0261] A DCM / MeOH=10 / 1 solution (3 mL) of AIR-3-2 (40 mg, 72.24 mmol) 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 ether, centrifuged, and the precipitate was washed with ether, centrifuged, and dried to give a crude product of 33 mg. The crude product was further purified by Prep-TLC (DCM:MeOH=20:1, NH3·H2O was added) to give a yellow solid AIR-3 (10 mg). MS (ESI), m / z: 454.58 [M+H] +.HPLC 92.439% (220nm), HPLC 96.712% (254nm).

[0262] 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).

[0263] Example II. Activity Test

[0264] Example II.1: Activation of hnRNPA2B1 and TBK1-IRF3 pathways in tumor cells by test compounds

[0265] After stimulating A549 lung cancer cells (cell density of 1x10^5 cells per well) with 20 μM concentration of test compound E (prepared as in Example 1.5, dissolved in physiological 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.

[0266] The activation of hnRNPA2B1, TBK1, and IRF3 is shown in Figure 1 , 2 shown.

[0267] 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, the same experiment was carried out using other compounds prepared in Example I, and similar results were obtained.

[0268] The above results demonstrate that the test compound has an activating effect on the hnRNPA2B1 pathway and is a hnRNPA2B1 agonist.

[0269] Example II.2: Broad induction of type I interferon production by test compounds in various tumor types

[0270] A549 (non-small cell lung cancer), HepG2 (liver cancer), LLC (Lewis lung cancer), and B16F10 (melanoma) tumor cells were stimulated with 20 μM concentration of test compound E (cell density was 1x10^4 cells per well); cell culture supernatants were collected after 18 hours of stimulation, and IFN-β protein levels were detected by ELISA.

[0271] The test compound activates type I interferon as Figure 3 shown.

[0272] The results showed that the test compound E could significantly activate multiple cells to express type I interferon. In addition, the same experiment was performed using other compounds prepared in Example I, and similar results were obtained.

[0273] The above results prove 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 type of novel compound is defined as a hnRNPA2B1 agonist in the present disclosure.

[0274] Example II.3: hnRNPA2B1 agonist promotes STING expression in tumor cells

[0275] A549 and LLC tumor cells were stimulated with hnRNPA2B1 agonist compound E (20 μM), and the total protein of the cells was extracted at 0.5, 1, 2, 4, and 6 hours after stimulation, respectively. The protein level of STING in tumor cells was detected by Western blotting.

[0276] The protein level of STING Figure 4 shown.

[0277] The results showed that hnRNPA2B1 agonist compound E could promote the protein expression level of STING, which suggested that hnRNPA2B1 agonist may have synergistic effect with STING agonist. In addition, the same experiment was carried out using other compounds prepared in Example 1, and similar results were obtained.

[0278] Example II.4: hnRNPA2B1 agonist combined with STING agonist induces stronger type I interferon production

[0279] A549 tumor cells were stimulated with hnRNPA2B1 agonist compound E (10 μM) and STING agonist SR-717 (10 μM, purchased from Selleck, catalog number S0853). After 12 hours, total RNA of the cells was extracted and the transcription level of type I interferon (IFN-β) in tumor cells was detected by real-time fluorescence quantitative PCR.

[0280] The activation level of IFN-β Figure 5 shown.

[0281] The results showed that the hnRNPA2B1 agonist compound E combined with the STING agonist can induce stronger type I interferon production. In addition, the same experiment was performed using other compounds prepared in Example I, and similar results were obtained.

[0282] Example II.4: Synergistic anti-tumor effect of hnRNPA2B1 agonist combined with STING agonist

[0283] Mice (6-8 week old male C57BL / 6J mice) were subcutaneously inoculated with 5x10^5 LLC tumor cells. From the 10th day after inoculation, the mice were randomly divided into the following groups: control group (only tail vein injection and intraperitoneal injection, no drug administration), compound E alone administration group, SR-717 administration group, and combined administration group. Compound E was administered by tail vein injection at a dose of 15 mg / kg mouse body weight), once every four days, for a total of four times. SR-717 was administered by intraperitoneal injection at a dose of 30 mg / kg mouse body weight, once a day, for 7 consecutive times. The growth of mouse tumors was continuously observed, tumor size = length x width 2 / 2.

[0284] The results are as follows Figure 6 shown.

[0285] The results showed that the hnRNPA2B1 agonist compound E can cooperate with the STING agonist to more effectively inhibit tumor growth in vivo. In addition, the same experiment was carried out using other compounds prepared in Example 1, and similar results were obtained.

[0286] The above results suggest that hnRNPA2B1 agonists can effectively exert anti-tumor effects.

[0287] Biological evaluation

[0288] The compounds disclosed herein are defined as hnRNPA2B1 agonists by: (i) binding to the hnRNPA2B1 protein, as demonstrated by the high affinity of the compounds to the hnRNPA2B1 protein as low as 20 μM as determined by Biacore; and (ii) demonstrating by cell-based assays that the compounds can broadly induce the production of higher levels of IFN-β at concentrations as low as 20 μM.

[0289] (i) Biacore T200 detection of the binding between compounds and hnRNPA2B1

[0290] Using S series CM5 chip; amino coupling reagents (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide EDC and N-hydroxysuccinimide NHS);

[0291] Coupling buffer: 10 mM sodium acetate pH 4.0, pH 4.5;

[0292] Running buffer: 10× PBS-P+;

[0293] Analytical grade DMSO, deionized water (0.22 μm membrane filtration);

[0294] hnRNPA2B1 recombinant protein (concentration must be greater than 0.5 mg / mL, total protein amount must be at least 20 μg);

[0295] Compound: Stock solution concentration greater than 20 mM, volume greater than 30 μL, purity greater than 90%, dissolved in 100% DMSO;

[0296] Other consumables: 96-well plates and sealing films, 1.5 mL centrifuge tubes without caps, and type 2 rubber bottle caps.

[0297] Experimental steps:

[0298] 1. Place the running buffer 1× PBS and waste liquid bottle in the designated positions and place the chip.

[0299] 2. Protein coupling. Select the immobilization program in the Wizard template, check flow cell 2 or 4, select amine as the method, select specify contact time and flow rate to achieve high coupling, enter contact time 900s, flow rate 10μL / min.

[0300] 3. Prepare the required protein solution according to the instructions. Dilute the protein to the required volume with sodium acetate at pH 4.0, with a final concentration of 20 μg / mL. Place 100 μL EDC, 100 μL NHS, and 140 μL ethanolamine in the corresponding positions.

[0301] 4. The system will automatically coat the target conjugated amount of hnRNPA2B1 protein on the chip surface and automatically generate a conjugation report.

[0302] 5. Prepare the running buffer for the interaction. The running buffer for small molecule samples is 1×PBS-P+ containing 5% DMSO: 105mL 10×PBS-P+ is diluted to 1L with deionized water to make 1.05×PBS-P+.

[0303] Solvent correction stock solution containing 4.5% DMSO: 1.05×PBS-P+9.5mL+0.45mL DMSO

[0304] Solvent correction stock solution containing 5.8% DMSO: 1.05×PBS-P+9.5mL+0.58mL DMSO

[0305] Running buffer containing 5% DMSO: 1.05×PBS-P+950mL+50mL DMSO

[0306] Dilute 10mM small molecule stock solution with 1.05×PBS-P+ buffer without DMSO: 0.7μL small molecule stock solution + 6.3μL DMSO + 133μL 1.05×PBS-P+ buffer, total volume is 140μL, final concentration is 50μM. Add to 96-well plate and affix sealing film.

[0307] After the test, use Kinetics or Affinity mode to measure binding affinity.

[0308] (ii) Total cellular RNA can be extracted using TRIzol or RNA rapid extraction kit.

[0309] 24-well plate cells (about 2-3×10 5 10 cells / well) RNA rapid extraction: discard the culture medium, add 500μL RA2 lysis buffer to each well, pipet and lyse several times, transfer to the inner sleeve of the special adsorption tube, and centrifuge at 12000rpm for 1min at room temperature. Discard the filtrate, add 500μL washing buffer, and centrifuge at 12000rpm for 1min at room temperature. Repeat the washing operation once. Discard the filtrate, put the inner sleeve back, do not add Wash Buffer, and centrifuge at 12000rpm for 2min at room temperature. Transfer the inner sleeve to a new 1.5mL centrifuge tube, add 25μL Elution Buffer to the center of the membrane, let it stand at room temperature for 5min, and centrifuge at 12000rpm for 1min at room temperature. The filtrate obtained is RNA, and the RNA concentration is measured using Nanodrop One.

[0310] qRT-PCR: After the concentration of each sample was determined, 1 μg of total RNA was taken and ReverTra qRT-PCR RTMaster Mix reverse transcribes RNA into cDNA. The reverse transcription system and reaction conditions are as follows:

[0311] Reverse transcription reaction system (20 μL system)

[0312]

[0313] Reverse transcription reaction conditions (20 μL system)

[0314]

[0315] Add 60 μL of deionized water to the cDNA sample obtained by reverse transcription for dilution, mix well and centrifuge instantly. The primer sequence information used for qRT-PCR is shown in the table. SYBR Green Realtime PCR Master Mix is ​​used to perform relative quantitative detection of the mRNA level of the gene.

[0316] qRT-PCR reaction system (20 μL system)

[0317]

[0318] qRT-PCR reaction conditions

[0319]

[0320] qRT-PCR primers

[0321]

[0322] All documents mentioned in this disclosure are cited as references in this application, just as each document is cited as reference individually. In addition, it should 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 these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. 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, in, L is -(CH2)n-, n is an integer from 0 to 6; X is a halogen; R 1 For-NR 4 R 5 , where R 4 , R 5 Each independently is H or C 1-6 alkyl; R 2 and R 3 Each independently is H or C 1-6 alkyl; or R 2 and R 3 Together with the N atom to which they are attached, they form a 5-8 membered heterocycloalkyl group containing 1-3 heteroatoms selected from nitrogen, oxygen or sulfur; 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 hydroxyl, amino, halogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, hydroxyl 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 heterocycloalkylC 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.

2. The compound according to claim 1, its cis-trans isomers, its enantiomers, its diastereomers, its racemates, its solvates, its hydrates, or its pharmaceutically acceptable salts or prodrugs thereof, in, X is F or Cl; L is -(CH2)n-, wherein n is 1, 2 or 3; R 4 , R 5 Each is independently H, methyl or ethyl; R 2 and R 3 Each independently is C 1-3 Alkyl, the 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; 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 2 nitrogen atoms, wherein the 5-8 membered heterocycloalkyl group is surrounded by a 3-8 membered heterocycloalkyl group C 1-6 Alkyl or hydroxyl C 1-6 Alkyl substitution.

3. The compound according to claim 1, its cis-trans isomers, its enantiomers, its diastereomers, its racemates, its solvates, its hydrates, or its pharmaceutically acceptable salts or prodrugs thereof, in, X is F; L is -(CH2)n-, wherein n is 2; R 4 , R 5 Each is independently H, methyl or ethyl; R 2 and R 3 Each independently is C 1-3 Alkyl, the C 1-3 The alkyl group is unsubstituted or substituted with a 4-6 membered heterocycloalkyl group containing 1 nitrogen heteroatom; or 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 hydroxyl C 1-6 Alkyl substitution.

4. The compound according to claim 1, its cis-trans isomers, its enantiomers, its diastereomers, its racemates, its solvates, its hydrates, or its pharmaceutically acceptable salts or prodrugs thereof, in, X is F; L is -(CH2)n-, wherein n is 2; R 4 , R 5 Each is independently H, methyl or ethyl; R 2 and R 3 are each independently methyl, ethyl or isopropyl, the C 1-3 Alkyl is unsubstituted or unsubstituted or substituted with pyrrolidinyl; or R 2 and R 3 Together with the N atom to which they are attached they form a piperazinyl group which is substituted by a pyrrolidinylethyl group or by a hydroxyethyl group.

5. The compound according to claim 1, its cis-trans isomers, its enantiomers, its diastereomers, its racemates, its solvates, its hydrates, or its pharmaceutically acceptable salts or prodrugs thereof, wherein the compound has the following formula (II): in, R a , R b Each independently is H or C 1-6 alkyl; M is -(CH2)n-, wherein n is 1, 2 or 3; X is F or Cl; R c is hydroxyl, amino, 3-8 membered heterocycloalkyl containing 1-2 heteroatoms selected from nitrogen, oxygen or sulfur, more preferably R c is hydroxyl or a 4-6 membered heterocycloalkyl group containing 1 nitrogen atom, more preferably, R c is hydroxyl or pyrrolidinyl, most preferably, R c is hydroxy or 1-pyrrolidinyl.

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

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

8. The product of claim 7, wherein: The product is selected from: a pharmaceutical composition, a medicine, a kit or a medicine box; and / or Wherein, the product further comprises a STING agonist or is used in combination with a STING agonist, for example, the STING agonist is selected from: SR-717, MSA2, MK-1454, ADU-S100, BMS-986301, TAK-676, E7766, GSK3745417, SB11285, IMSA101, SR-8314, IACS-8803, AN3005, OX401, TTI-10001, VTX-001, IACS-8779, SOMCL-18-202, MSA-2; Wherein, the product is used to induce the production of type I interferon in a subject and / or is used to prevent and / or treat tumors in a subject.

9. Use of a compound according to any one of claims 1 to 6 in the preparation of a product for use in combination with a STING agonist to induce type I interferon production in a subject or a product for use in combination with a STING agonist to prevent and / or treat a tumor in a subject.

10. The use according to claim 9, wherein: The subject is a human or 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 deer, etc.); and / or wherein the production of type I interferon is mediated by hnRNPA2B1; and / or Wherein, the tumor prevention and / or treatment of the subject can benefit from the production of type I interferon in vivo or the increase of its level; and / or Wherein, the tumor is one or more tumors selected from the following groups: 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 blood system 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: a pharmaceutical composition, a medicine, a test kit or a medicine box; and / or Wherein, the STING agonist is selected from: SR-717, MSA2, MK-1454, ADU-S100, BMS-986301, TAK-676, E7766, GSK3745417, SB11285, IMSA101, SR-8314, IACS-8803, AN3005, OX401, TTI-10001, VTX-001, IACS-8779, SOMCL-18-202, MSA-2; and / or The product is useful for inducing the production of type I interferon in a subject and / or for preventing and / or treating a tumor in a subject.

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